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: REVIEWS ON COMMON OBJECTIVES AND EVALUATION INDICATORS FOR RISK COMMUNICATION ACTIVITIES FROM 2011 TO 2017
Review round: 1
Reviewer: 2

Output:
Basic reporting: Clear and unambiguous, professional English used throughout. The article must be written in English and must use clear, unambiguous, technically correct text. The article must conform to professional standards of courtesy and expression.
The paper is well written and does not need any further improvement of the language. Sentences are easy to read and enable me to fully understand what the authors intended.

Literature references, sufficient field background/context provided. The article should include sufficient introduction and background to demonstrate how the work fits into the broader field of knowledge. Relevant prior literature should be appropriately referenced.
In the Introduction section, the authors address the definition of “risk communication” and its relevance for difference research and policy fields. Therefore, it is a challenging task to give an exhaustive definition in order to represent this variety (lines 62-63). The authors made a good job but I would suggest to add some other references, e.g. Lundgren & McMakin (2009) and, more generally, the work done by professionals, consultants, … who tired to translated definitions into practices. Through the years, risk communication was born as a discipline, to be defined and addressed, and followed by research and tentative interventions to make it works and be effective in understanding and properly managing risks, no matter the field. So, in this sense, I would stress that risk communication definition and aims are also given by this kind of literature. The authors will refer to some examples of these sources further ahead.

Professional article structure, figures, tables. Raw data shared. The structure of the article should conform to an acceptable format of ‘standard sections’ (see our Instructions for Authors for our suggested format). Significant departures in structure should be made only if they significantly improve clarity or conform to a discipline-specific custom.
The article structure is well organised and it fits with the suggested format for PEERJ Review articles.

Figures should be relevant to the content of the article, of sufficient resolution, and appropriately described and labeled.
Ok

All appropriate raw data have been made available in accordance with our Data Sharing policy.
Yes

Is the review of broad and cross-disciplinary interest and within the scope of the journal?
Yes, it is confirmed both by the search strategy and the results themselves.

Has the field been reviewed recently? If so, is there a good reason for this review (different point of view, accessible to a different audience, etc.)?
To my knowledge and brief online research, no recent review addressed this field. Recent works preferably reviewed risk communication in specific field, e.g. food safety, cancer, human health in general.
Does the Introduction adequately introduce the subject and make it clear who the audience is/what the motivation is?
Yes, see above for other comments to improve this section, even if it is already well organised.
Experimental design: Article content is within the Aims and Scope of the journal.
In my opinion, this paper falls within this following point:
PeerJ does not publish in the Physical Sciences,the Mathematical Sciences, the Social Sciences, or the Humanities (except where articles in those areas have clear applicability to the core areas of Biological, Environmental, Medical or Health sciences).
Rigorous investigation performed to a high technical & ethical standard. The investigation must have been conducted rigorously and to a high technical standard. The research must have been conducted in conformity with the prevailing ethical standards in the field.
Confirmed
Methods described with sufficient detail & information to replicate. Methods should be described with sufficient information to be reproducible by another investigator.
Overall, the reader understands the process made by the authors to search for the relevant articles and define the final simple. However, some questions about the Survey Methodology section arose and need major revision at some points. I would also express my concern about the choice to insert in the search procedures articles published in Japanese. How did this focus on the Japanese language serve and improve your analysis and results? I don’t want to prefer a language out of another one, I just want to say that including papers from Japanese literature could possibly bias e.g. against the fields of intervention, for example Japanese literature could be more focused on nuclear risk communication, whereas the Dutch one could be more focussed on flood risk communication. Your aim was to conduct an international review, so many other languages could have been considered and added to the search and results, according to pre-defined criteria of inclusions.
-	Line 116: why did you choose the 2011-2017 period? I can understand 2017, as you have begun the analysis in 2018, but I wonder why you went back to 2011 precisely
-	Line 117-118: I would not have included this kind of work, as they did not evaluate interventions. How many articles fall into this category? Please consider not including them in the study
-	Please identify better the beginning and the end of the description of both the first and the second rounds of eligibility assessment. E.g. the first round goes from line 126 to 157 and refers only to title and abstract, then the second…
-	Line 140: there is no need of the statement in brackets
-	Lines 143: they were divided into six groups: why? Which were these groups? What were their purpose?
-	Lines 143-146: why and how did you decide these three criteria? Just because of the study aims, other? Why did you assign the retrieved articles to one of these tree categories (as described in lines 142-154) already in the initial screening process? Would it had been enough just to assign a YES/NO to the articles according to the fact that they evaluated risk communication? Basically, I don’t understand why you performed this classification twice (lines 166-167), as in my opinion it was unnecessary in the initial phase. In addition, what about participatory approach in evaluating risk communication practices? Did you find anything about?
-	Line 172, in line with the phases in the disaster management cycle: please provide a reference for that
-	Lines 178-180: non-profit organisations: I would re-allocate this group, because these figures cannot be considered as “citizens”. In addition, it is reductive to put together in the same category “other” figures that have different roles in risk communication and intervene at different levels. Please, consider revising this classification and definition of target groups. Accordingly, I see the -same problem in lines 181-182, I refer to “(2) other (e.g. mass communication)”, where “other” could have been substituted by specific communication types and formats. As I can see from Table 1, “Other communication type” (N=214) doubles “Individual/small group communication”, making me supposing that it is not a residual category.
-	Lines 166-189: in these lines you report all categories assigned to the selected articles: which coding approach did you adopt to code? How and when did you identify, define and name the categories?
-	Lines 183-189: this is a great concern for me, as what you named “indicators used for evaluation, including desired or intended impacts on target audience” are not to be intended as indicators, because they clearly represent intended impacts of risk communication interventions. I mean, categories from 1 to 8 are example of aims to be achieved by e.g. a risk communication campaign, and they are not indicators to be used to measure whether an aim has been reached or not (to this extent, you could for example use the expression “evaluation instruments” used in line 268). You already address this difference in lines 263-267. Please, fix this point. Consider revising in Table 1 too and all concerned sections (e.g. lines 272-275, 315-356, …).
-	Lines 213-217: why did you searched these documents for definitions? You had better use these sources to identify a set of evaluation criteria upon which to base and guide your analysis, for instance to fix the point I mentioned above. Finally, consider the presence of these documents and guidance in your Introduction.
Is the Survey Methodology consistent with a comprehensive, unbiased coverage of the subject? If not, what is missing?
The subject under investigation – i.e. risk communication has been extensively searched in the main academic search engines.
Are sources adequately cited? Quoted or paraphrased as appropriate?
Yes
Is the review organized logically into coherent paragraphs/subsections?
Yes
Validity of the findings: Impact and novelty not assessed. Negative/inconclusive results accepted. Meaningful replication encouraged where rationale & benefit to literature is clearly stated.
-	Lines 251-252: why did you mentioned nuclear/radiological disaster and climate change here? I would expect stressing these points in the discussion section, together with an explanation of why there is a need to highlight this result
-	Lines 277-290: I am sorry but I did not catch the meaning of these lines and their usefulness in this part of the manuscript
-	Table 2: did you consider the possibility to indicate the research techniques used to evaluate risk communication? The “Instrument” column could be filled in with the technique, e.g. online survey, focus group, usability study, … the methodological approach (qualitative, quantitative, both…) and the theoretical framework beneath.
-	Lines 320-323: I am not sure that this is as higher as intended, for example in this study field the aim “change in risk perception” has a higher percentage.
-	Lines 358-385: this part would be better moved to the Introduction section, because it does not show results. Moreover, going ahead with the reading, and I am more and more persuaded that the documents you refer to in these lines can be considered as those who guided your selection of the criteria for the analysis. In this sense, this could shed more light to your results, in terms of what coincide and differ, as you already mentioned (e.g. lines 400-402).
-	Lines 414-420: I take your point in pointing out this difference among definitions but this sentence seems to be out of the initial premises of the study, as it is not devoted to compare risk communication definition. So please consider where and how to collocate this part.
Conclusions are well stated, linked to original research question & limited to supporting results. The conclusions should be appropriately stated, should be connected to the original question investigated, and should be limited to those supported by the results. In particular, claims of a causative relationship should be supported by a well-controlled experimental intervention. Correlation is not causation. Speculation is welcome, but should be identified as such.

Is there a well developed and supported argument that meets the goals set out in the Introduction?
-	Lines 425-427: where did you discuss these results? This could be an additional frame to classify and discuss results.
Does the Conclusion identify unresolved questions / gaps / future directions?
Overall, the Discussion section tries to interpret results in order to explain the differences in risk communication research and practices that emerged from the retrieved and analysed papers. The authors offer interesting interpretations and comments, that I definitely agree with, but that one could have expected. Some suggestions for future directions are given in the Conclusion but I try to ask you whether more intriguing insights could have been gained to better guide for example future research on risk communication, both in terms of applicability and evaluation? Any other suggestions with particular reference to crisis situations? Crisis situation is one of the three main categories risk communication is divided into, and what’s happening today due to the new SARS-Cov-2 pandemic shows that risk communication needs to be re-thought and this can be made only after a thorough evaluation of interventions made during the emergency.
Additional comments: Please make any general comments not covered by the 3 areas above. This text is sent to the author.
In this study, the authors tried to measure how much risk communication has been done and how it was evaluated, whenever evaluated, spacing throughout different fields of investigation and application of this discipline. To this extent, the authors reviewed recent English and Japanese literature to (lines 104-105)
-	Select papers within these premises
-	Appraise evaluation processes
-	Identify indicators used to evaluate risk communication activities across sectors.
I really appreciated the possibility to read and review this work, as I fully agree that there is need to re-think what risk communication is and the extent to which it is able to help people facing risks. One of the major challenges of risk communication, as it is within this study’s aims and as the new pandemic is showing us, is that risk communication practices lack effectiveness, especially during crisis and emergency situation. All principles defined and applied so far seem to fail and this utterly calls for a deeper understanding of what is risk communication and what mines its credibility and applicability. In this sense, lines 106-108 stand out as guidance for future work to fix this point.


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: REVIEWS ON COMMON OBJECTIVES AND EVALUATION INDICATORS FOR RISK COMMUNICATION ACTIVITIES FROM 2011 TO 2017
Review round: 2
Reviewer: 1

Output:
Basic reporting: I appreciate the opportunity to review this study for a second time.  The authors have addressed my concerns in this section.
Experimental design: The authors have addressed my concerns around study design, and I do not have any new concerns for them to address.
Validity of the findings: The authors have addressed my concerns that impact the validity of the findings.
Additional comments: I do not have any additional comments for the authors at this time.


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

Input:
Title: REVIEWS ON COMMON OBJECTIVES AND EVALUATION INDICATORS FOR RISK COMMUNICATION ACTIVITIES FROM 2011 TO 2017
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comments, these sections have been definitively improved according to the reviewers' suggestions.
Experimental design: the method section is more and more clear and easy to follow; data collection has been better organised into coherent paragraphs.
Validity of the findings: Results have been more deeply discussed in terms of uncovering what risk communication research and practice need to effectively help risk governance both in peace and crisis times.
Additional comments: I would suggest to changing the title into:
Evaluating risk communication interventions: a review of research and practices across fields


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: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper by Huandg el al. describes the reproductive modes of the solanum mealybug. I found the paper very interesting and adding new important data to our knowledge about developmental and evolutionary biology of insects. Solanum mealybug is a relatively new species in China and can cause important damaged to the crops.
The paper is written well, generally in proper English and is accompanied by nice figures that nicely complement the text.

There is one main point of the criticism, which I want Authors to address:
According to the recent views, there are TWO main reproductive strategies: viviparity and ovoparity. Ovoviviparity is in fact a specific type of the viviparity where developing eggs are retained within the body of the mother. Based on nutritional relationships between maternal and embryonic organisms two modes of viviparity are recognized: matrotrophic and lecidotrophic one. Please check the recent literature by Blackburn (1999) and Ostrovsky 2013 and Ostrovsky et al. 2016 on the proper terminology. Also the recent papers by Bilinski and his colleagues (Bilinski et al., 2017, 2018, 2019, Jaglarz et al., 2018, 2019; Tworzydlo et al., 2013, 2019) describing various aspects of viviparity in earwigs could be beneficial. Please correct and rewrite the first paragraph of the Introduction.
In line 75 Authors write that viviparity has been reported in Arixeniaesau and cite the paper by Benoit t al (2015). First of all, there should be Arixenia esau, secondly, the paper of Benoid focuses on the tsetse fly, and most importantly there are much more numerous examples of viviparous insects, even within earwigs. Please check carefully and correct that fragment.

Line 78 – please correct the sentence: “(…) young are produced by means of eggs (…)” is quite confusing. It should rather read: “The offspring is nourished by the reserve materials accumulated in the eggs during oogenesis (…).

Line 86: change “scale species” into “”scale insect species”.
I also think that a comprehensive English polishing and language editing will improve the manuscript.
Experimental design: The data presented in the paper meet all standards.
The Methods are presented properly with all sufficient details.
Validity of the findings: I found the findings interesting and well presented.
Additional comments: Taking into account all the aforementioned data, I strongly recommend acceptance of the paper after some minor corrections.


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: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is overall well-written, but there are several areas where the language needs to be clarified or corrected. These are especially concentrated in the figures and figure legends. Specific suggestions are listed below.

The authors have listed several of the key citations relevant to the field, including Blackburn 1999 and the publications by Gavrilov-Zimin. However, as described below, their findings are not consistent with the definitions of ovoviviparity provided in these references. In addition, as suggested in the language changes below, it is not clear which of the previous publications they have cited have already studied the same species, P. solani.

The authors have done a very nice job of providing the raw data, videos, and sequence information that will make this publication useful and relevant for future research. One note: the authors describe the process of assembling a de novo transcriptome for this species. They provide the raw data via SRA, but the de novo transcriptome should also made available. This is a substantial contribution the authors provide, and should be described more fully, unless this specific P. solani assembly has already been described elsewhere, in which case that needs to be referenced.

Suggested language changes:

- the word egg should be clarified throughout the manuscript. In insects, the word egg can refer to the structure that starts as a developing oocyte within the mothers body, as well as the egg stage of an insect lifecycle. The authors use the term inconsistently throughout the manuscript, for example, they state that in viviparous species "egg development occurs inside the mother's body" (line 72), but that in ovoviviparous insects "young are produced by means of eggs, not the body of the mother" (line 78). In order to clarify their meaning, they should restate the latter to say "offspring exit the mother's body during the egg stage, rather than during the larval stage".

- change "ovoviviparous / viviparous ones " to "ovoviviparity / viviparity" (line 94)

- clarify the meaning of the statement "but the females produced wax" (line 227). Do you mean that, while oocysts were not observed, there was maternally produced wax?

- clarify the meaning of the sentence "so the researchers thought the first nymph was produced by scale insects such as P. solani and another invasive mealybug..." (lines 230-232). Do you mean to say that you wondered if there was some cross contamination from another species? How was this ruled out?

- clarify if "Vennila et al considered the scale insect to be able to reproduce in both ways" (line 233) refers to the same species, P. solani, as this study, or does this refer to a different species?

- change "eggs are under stress" to "eggs under stress" (line 259)

- clarify why the research on yeast-like symbiotes is relevant to the study in question (line 264). Are the authors suggesting that the presence / absence of symbiotes might explain the presence of non-developing eggs?

- clarify the meaning of "no longer resemble an egg on a microscopic level" (line 299). Do they still have an eggshell? Do you mean that the embryo within the chorion is fully developed and close to hatching?

- use more specific language to describe the question at line 319: "Although the reproductive mode of P. solani has been previously described, the actual process may not be simple" (lines 318-319). What remains unknown, prior to the current study?

- the language in the conclusion needs to be reworked to resolve ambiguity. Specifically the phrases: "one thing needs to be made clear" (line 325), "ovoviviparity is defined as the mother giving live birth, basically to larvae" (line 328, and this sentence requires a reference), and "we conclude that the reproductive 'product' of P. solani is an egg" (line 329). As currently written, it is not easy to understand what feature of P. solani reproductive biology makes this species distinct from other ovoviviparous species, and according to which definition of ovoviviparity this claim is being made.

- The legend of figure 1 contains several sentence fragments, which should be corrected, e.g. change "finished observation device, and contained two separate adult females" to "The final observation device contained two separate adult female".

- change "The first 5 minutes" to "After the first 5 minutes", and clarify the meaning of "the hatching process was basically completed" (figure 3 legend).

- typo at "analyze d" (figure 4 legend)

- change "and the vitelline membrance had been detached" to "at the point at which the vitelline membrane detached"; "Eggs hatch under the mother's body" to "Eggs raised under the mother's body"; and change "(F)" to "(E)". (figure 5). Additionally, is that the vitelline membrane or the chorion? Can the authors please clarify how this structure was defined / described?
Experimental design: The manuscript addresses the question of whether the solanum mealybug, like other scale insects, demonstrates ovoviviparity. In addition, the authors describe and investigate a phenomenon of two egg types, one developing toward hatching and one not.

The authors investigate the first question through classic observational description. The descriptions and experimental manipulations they provide are sufficient to answer the question.

The second question is addressed using a variety of techniques, including electron microscopy and RNAseq.

The exact purpose and findings of the electron microscopy and RNAseq experiments should be described in further detail. From my understanding of their findings, the mealybug lays two kinds of eggs: one that develops toward hatching, and one that does not. The authors show that these can be unambiguously identified by the presence of eyespots at the moment of oviposition.

The authors mention a few factors that may explain the presence of these non-developing eggs, including that development may have arrested around the time of blastokinesis or that these eggs never underwent any form of development, and may instead be a nutritive propagule for other offspring to consume. However, their description of the results of the electron microscopy and RNAseq experiments do not help to distinguish between these scenarios.

The authors should seek to answer the following questions using the data they already have:

    - From the microscopy data for eggs without-ES, is there any evidence that any development had taken place? Are there any remaining embryonic structures? Could the authors perhaps identify an embryonic stage at which development arrested for the eggs without-ES?

    - From the RNAseq data, what genes are active in the eggs without-ES? Do these provide any suggestion of the function of these eggs as nutritive propagules, or as embryos arrested at some stage of development?

    - Many insects lay non-viable eggs. Do the authors have any evidence that the scale insect is laying a larger proportion of non-viable eggs than other scale insects? than other parthenogenic species? Is there anything to suggest that these eggs without-ES are a unique structure beyond a non-viable egg?

The authors have gathered a very rich set of data with the electron microscopy and RNAseq experiments, but without more context, these data are essentially not utilized in this manuscript.
Validity of the findings: The authors state that the mealybug does not qualify as ovoviviparous, and suggest an alternative term to describe their findings: semi-ovoviviparous. However their results are not consistent with this finding. I believe this to be due to a misunderstanding of the definitions of these reproductive categories. From the entomological literature, the definition is that "ovoviviparous species oviposit eggs at an advanced stage of embryological development, and the larva exits the egg shell during or immediately following oviposition" (Meier et al 1999, Ovoviviparity and viviparity in the Diptera, Biol. Rev.). This is precisely what the authors describe here, as they state at lines 228-229: "After egg laying, the yolk membrane was detached from the egg (eggshell), followed by the appearance of 1st instar nymphs quickly crawling away from the lower part of the mother.  It took a short time (< 30 minutes) to complete this process...". The creation of the category 'semi-ovoviviparity' is therefore unnecessary and would further confuse the situation in describing reproductive categories.

This subject has been discussed thoroughly in the literature (Blackburn 1994, Meier 1999, Weber 1953 vs. Sellier 1955), and my interpretation of this discussion is that there is a continuum between full viviparity (live birth) and full oviparity (eggs laid immediately following fertilization), and that ovoviviparity has been used to variably describe any number of states in between those extremes and along that continuum. Given this, it is most important to describe the actual details of the timing of oviposition, hatching, and development, which the authors have already done nicely. Beyond that, defining the category is important in linking these new descriptions to similar descriptions from the literature, which is where this manuscript currently falls short.

In order to rectify this, I believe that the authors should restate their conclusions to state that they find this species to exhibit ovoviviparity, consistent with other scale insects, or else provide additional and much stronger evidence that this species indeed exhibits a reproductive mode distinct from any ovoviviparous mode previously described. This would require changing the "Discussion" section of the abstract, as well as most of the text of the conclusion.
Additional comments: I appreciated reading this article, and especially enjoyed the excellent use of classical description methods to answer a straightforward question: does P. solani give live birth? I also appreciated the transparency with which the data was shared.


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: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript of Huang et al. provides new information about the reproductive biology of one of the scale insect species. This information is rather simple and rather usual for parthenogenetic species. All the novelty can be expressed in the phrase “Females lay two sorts of eggs, viable and non-viable, but we do not know why is it so”. Such kind of reports are usually published as “Short communications” and I would advise to reduce the present 23-pages to several pages only, deleting all common information, which the authors abundantly compiled from the well-known literature.
Experimental design: To my mind, methods, which the authors used for study of the material, are not suitable. Main morphological structures (like legs, antennae or eyespots) of the developing embryo of scale insects are clearly visible in the drop of acetic acid or ethanol under the usual stereoscopic microscope; it is not need to use complicated electron microscopy for this purpose. As for the RNA-sequencing I am absolutely unable to understand for what it was need in this case. It is clear without any studies that developing embryos and non-viable eggs have different biochemical composition, but the authors do not really study of this composition and only say that there is the difference in the data of RNA-sequencing. It gives nothing for the understanding of the reason, why two sorts of eggs are laid.  The only way to clarify such reason is detail cytogenetic studies of the parthenogenesis, in particular female meiosis in Phenacoccus solani. However, the authors totally avoid any cytogenetic investigations in their work.
Validity of the findings: no comment
Additional comments: In addition, I would advise to be more careful with a terminology. For example, “oocyct” is a resistant, thick-walled spore containing the zygote in Protozoa (see, for example, https://www.biologyonline.com/dictionary/oocyst or any other dictuonary). I am even unable to imagine what the authors mean, applying this term to the scale-insects. Also, the term “semi-ovoviviparity” seems to be rather strange and unclear. Previously the term “incomplete ovoviviparity” was used in the literature, cited by the authors, for the situations when the only part of embryogenesis occurs inside of the mother’s body….


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: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am satisfied with all corrections incorporated into the manuscript. I will be be happy to see the final published version of the paper!
Experimental design: n/a
Validity of the findings: n/a
Additional comments: I strongly suggest acceptance of the paper.


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

Input:
Title: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript is considerably improved, especially in the areas of defining the reproductive mode in relation to past literature, and in providing context for the RNAseq results.

The typo "analyze d" is still present in two places in the legend of Figure 5.
Experimental design: No comment.
Validity of the findings: The statement "These eggs did not hatch, just as a chick cannot hatch without a hen" seems to be in contradiction with the finding "Therefore, we suggest that the mother’s body has no effect on the success of egg hatching". Their results show that the reason that eggs without eyespots did not hatch is not related to the presence / absence of the mother. I suggest that the former statement, "just as a chick cannot hatch without a hen" should be removed.
Additional comments: In my original review I stated "This subject has been discussed thoroughly in the literature (Blackburn 1994, Meier 1999, Weber 1953 vs. Sellier 1955), and my interpretation of this discussion is that there is a continuum between full viviparity (live birth) and full oviparity (eggs laid immediately following fertilization), and that ovoviviparity has been used to variably describe any number of states in between those extremes and along that continuum."

The authors have used the second part of my statement, starting at "there is a continuum...", in the abstract and conclusion of their paper. As stated above, this was intended as my interpretation of the situation, and was not meant to be reproduced as the authoritative word on ovoviviparity. I ask that the authors replace these sentences (lines 49 - 52, and 362 - 365) defining ovoviviparity with one based on the published literature, for example, the publications listed above

Especially, I do not view this statement as a finding from the paper, and so I do not think it is appropriate for the authors to claim "We determine that there is a continuum..." (line 362). This sentence should be removed.


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: REPRODUCTIVE PATTERN IN THE SOLANUM MEALYBUG, PHENACOCCUS SOLANI: A NEW PERSPECTIVE
Review round: 2
Reviewer: 3

Output:
Basic reporting: I have no additional comments in comparison with my previous review. The authors provided a row of changes and additions in the text and I think that the paper may be published in the present form. I can not agree with some approaches and some terms, but the authors have a right to express their own views, of course.
Experimental design: no comments
Validity of the findings: no comments
Additional comments: no comments


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

Input:
Title: MITOCHONDRIAL DESTINY IN TYPE 2 DIABETES: THE EFFECTS OF OXIDATIVE STRESS ON THE DYNAMICS AND BIOGENESIS OF MITOCHONDRIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript is generally well-written and clear throughout – with occasional specific sections that have been identified in General Comments. The background to the subject is clear and literature well referenced and relevant. The structure is sensible, although the manuscript would benefit from work to increase brevity. The review topic is cross-disciplinary and of excellent potential interest to a broad audience; there are multiple reviews published on various aspects of the manuscript, however very little that draws them together with the focus of the current manuscript. The Introduction adequately introduces the subject (although the description of mitochondrial OXPHOS lines 43-53 seems unnecessary – is textbook – and could be replaced by a brief introduction to insulin resistance/adipocytes).
Experimental design: I am not sure this manuscript is really a systematic review rather than a regular review with search terms included in methodology. The search does not seem reproducible by externals – 20 keywords with unspecified combinations and subjective eligibility criteria. page 7, lines 68-74. I don’t think that this undermines the manuscript – but rather just suggests a different label from “systematic review” to “review”. Literature included does appear to be unbiased and comprehensive, and is cited appropriately. Organisation is logical.
Validity of the findings: Argument is well developed. Conclusions are well stated and indentify future direction – although some specific queries are highlighted in “General comments” below.
Additional comments: The manuscript is fairly long and would be improved by careful condensation and brevity.
Major points:
•	p8, line 93: “Signalling molecules can interact with effectors on the surface of mitochondria” is very vague and does not appear to be explained subsequently – other than an indication that oxidative/nitrosative stress can cause mitochondrial dysfunction and further ROS production. The sentence prior to line 93 discusses transcription factors activated during inflammation, and alterations in insulin signalling molecules. If the authors are implication mitochondrial involvement purely through ROS/RNS then this section should be made less ambiguous.
•	p9, line 126 on: the stated dogma of fusion alleviating stress and fission contributing to quality control/mitophagy is now somewhat more nuanced, recent evidence has also implicated fusion (or at least Mfn2) in mitophagy, mito transport and interactions with ER (reviewed in Dorn 2019, https://doi.org/10.1146/annurev-physiol-020518-114358, and also see the very recent: doi: 10.3389/fcell.2020.00221 and doi: 10.1080/15548627.2020.1749490).
•	p9, lines 139-150: these paragraphs more appropriate under sub-heading 1.2 as they don’t mention mito fission/fusion at all? Or maybe adapt this section by adding in discussion of literature that implicates mito dynamics/cristae morphology – and OPA1 in particular – in maintenance of mtDNA?
•	p8, line 110 and p10, line 184: by “soft separation / disconnection of the ETC”, do you mean mild uncoupling of ETC that decreases correlation of metabolic input with ATP/ROS production?
•	Figures 2 and 3 could be clearer by making arrows more distinct and re-arranging the diagrams where-ever possible to minimise arrow cross-over. Do make sure that the figure legends support the whole of the figure.
•	Discussion of ref 59 on p13-14 could be made both briefer and clearer (in particular “135 proteins were different in type 2 diabetes compared to those of the control (without type 2 diabetes), and 200 proteins were less active” = “135 proteins were upregulated in type 2 diabetes compared to those of the control (without type 2 diabetes), and 200 proteins were downregulated”  – I don’t think that the paper measured activities?)
•	The majority of section 4 is quite basic knowledge – could the essential aspects be incorporated into a short intro paragraph of section 5 instead? (ie lines 350-351 and 366-8, with very concise supporting text?)
•	The discussion of the potential benefits of uncouplers such as DNP in T2D (p17) does not include any mention of the well-known dangers of this drug (numerous deaths due to overheating and energy failure that accompany uncontrolled uncoupling when taken illicitly). Equally, description of Mdivi-1 should potentially be tempered by mention of this drug’s ability to induce apoptosis.
Minor points
•	p8, line 81 “a result of IR or whether insulin resistance itself” – use abbreviation throughout once introduced
•	p8, line 84 “hypertrophic increase” = “hypertrophy”?
•	p25 Figure 1:
o	legend should make reference to A and B to direct reader around figure;
o	also explain DAMPS and PAMPS; also “violate insulin signalling” seems somewhat dramatic and uninformative – “block/prevent/impinge on/downregulate insulin signalling leading to a decrease in the intracellular pathways that stimulate GLUT4 translocation to the plasma membrane”?;
o	mitofusins 1 and 2 are show in the figure as “Mtf-1” and “Mtf-2” however are referred to via the standard “Mfn-1/2” within the main text.
•	p8, line 95: “which are is associated”
•	p8, line 113-4: “… based on the effects of certain drugs used to treat type 2 diabetes. The drug mechanism of action is aimed at improving insulin sensitivity …” grammar not great, and meaning somewhat unclear; should the second sentence read “Among the mechanisms of action of these drugs that lead to improved insulin sensitivity, is the suppression of ATP generation …”?
•	p9, line 137: “Two other members of this family are mitofusin 1 (Mfn1), and optic atrophy type-1 (OPA1). Mfn1 and Mfn2 control…” surely better as “Two other members of this family are mitofusins (Mfn1 and Mfn2), and optic atrophy type-1 …”?
•	p11, line 230: “to date, its properties have been extensively studied in humans” – should this be “have not been”?
•	p12, line 239: “correlation between the HOMA-IR index and the Matsuda index” clarify that this means correlation of MOTS-c with EACH, and also insert some explanation of Matsuda index – even just “the alternative insulin-sensitivity index Matsuda index”.
•	p12, line 250: this statement is attributed to ref 46 – which is actually a commentary on the primary research paper Qin et al Int J Cardiol. 2018 Mar 1;254:23-27. doi: 10.1016/j.ijcard.2017.12.001, but this paper is not cited. Is there a reason for this? (I cannot easily access ref #46 just now)
•	p13, line 296-7: redundant as covered in paragraph above.
•	p13, line 304 & 306: “importation” = import?
•	p13, line 318 does not make grammatical sense
•	p16, line 402: the sentence starting “Mitochondrial fusion….” is unnecessary as it just repeats information from earlier in the review. (indeed, this whole paragraph could be much tighter)
•	p16, line 421: “Drug transporters aimed at mitochondria” is ambiguous – drugs that are transported in to target mitochondria? or mechanisms for transporting drugs in? or just drugs targeted at mitos?
•	p16, line 434: “proton force (Δp)” – meant to be proton motive force?
•	p18, line 487: “To targeting molecules involved…” this sentence does not make grammatical sense.


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: MITOCHONDRIAL DESTINY IN TYPE 2 DIABETES: THE EFFECTS OF OXIDATIVE STRESS ON THE DYNAMICS AND BIOGENESIS OF MITOCHONDRIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: This review covers a wide range of issues linking mitochondrial function and type 2 diabetes. The authors have summarised a great deal of work and provided an interesting review article. The style and language is clear (although some of the content is presented in a fragmented manner see comments below). The background and literature is covered sufficiently, but sometimes lacks depth. The work is provided in good context. Figures are clear and well integrated with the text. There are several reviews on mitochondria biogenesis and assembly, but not many on the relevance of mitochondria function to T2D. I find the title a little vague (mitochondrial 'variations' may mean different thing to dfferent people and the scope of the review later on is heavy on mtDNA aspects.
Experimental design: Overall logical presentation of material albeit a little disjointed.
Specific points:
-lines 206-208: not clear the relationship between mtDNA and nDNA. A balance in the expression of the two genomes for mitochondrial protein is needed, but this is not discussed in any great detail.
-line 209: they authors mention 'signal molecules' and then go on to say 'one of those proteins': signal molecules generated by mitochondria can be distinct ROS, metabolites of the TCA cycle, proteins etc. It is not clear whether the authors will refer to all of those or not.
- lines 222-224: the generalisation is too far fetched, as the cases of mitochondrial derived peptides are only two mentioned here.
- the section 5 on chemerin is introduced too abruptly in the text and seems out of place.
- line 421: 'drug transporters'??? Reading the rest of this section I understood that the authors mean modulations or drug transport vehicles like the TPP-moiety etc but not transporter proteins. This is confusing.
- line 472: the statement that proteins smaller than 10 kDa can freely diffuse across the OM is not correct. Small molecules less than 5 kDa are freely diffusible through the VDAC channels but not proteins smaller than 10 kDa. In fact there are several examples of proteins around 10 kDa (eg many substrates of the MIA pathway) that do not freely diffuse but through the TOM import gate
- line 475: the inner membrane 'consists of cardiolipin' is inaccurate: cardiolipin is in high concentrations in the IMM, but it is also present in the plasma membrane, and the IMM also contains other lipids.
- lines 475-488: it should be clearer that this whole discussion here concerns the targeting not to mitochondria in general but to the specific matrix  sub-compartment. In fact none of the technologies mentioned can target any molecule to the intermembrane space or indeed to the OMM or the IMM.
- reference 91: historically, the capacity for DNA to be delivered to the mitochondrial matrix when fused to a matrix protein was shown already in 1989 (Vestweber and Schatz, 1989, Nature 338:170-2
Validity of the findings: Arguments are mostly well developed. Conclusions are valid, and relevant literature referenced.
Additional comments: This is a nice effort to bring together several aspects of mitochondrial biology and their relevance to T2D. I find the review interesting. It is hard to follow in parts because it tries to cover different aspects  (mitochondrially derived peptides, dynamics, mito proteomics, autophagy). All of these could be in fact separate reviews in their own right. However, scanning all of these aspects has some value for people working in T2D as it highlights the multitude of ways mitochondria can be linked to this disease. To streamline the review I would suggest taking out the part on chemerin as it does not really fit so well, and takes the reader to yet another line of research (mitochondria and inflammation).


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: MITOCHONDRIAL DESTINY IN TYPE 2 DIABETES: THE EFFECTS OF OXIDATIVE STRESS ON THE DYNAMICS AND BIOGENESIS OF MITOCHONDRIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Good, no further comment specifically on basic reporting, see below for general comments.
Experimental design: Good, now has appropriate designation as review rather than systematic review.
Validity of the findings: Good, no further comment specifically on basic reporting, see below for general comments.
Additional comments: I appreciate the changes that the authors have made in line with reviewers’ comments. I think that the text is now more focussed and the supporting figures are much clearer for the reader.

General points:
- could do with overall check of grammar and flow of wording throughout, following authors' Track Changes, particularly focussed on newly-written or adapted sections - eg p2, line 43 "Process oxidative phosphorylation (OXPHOS) enables ATP generation in mitochondria. Membrane electrochemical gradient stimulates ...  " should be "The process of oxidative phosphorylation (OXPHOS) enables ATP generation in mitochondria. A trans-membrane electrochemical gradient stimulates ...."

Minor points noticed:
-p4 line 156 (with “All Markup” selected) – mitofusins (Mfn and Mfn2) = mitofusins (Mfn1 and Mfn2)
-p5 line 172 – “Mitofusin may play a role” = “Mitofusins may play a role” or “Mitofusin-2 may play a role” or “Mitochondrial fusion may play a role”?
-p6 paragraph starting line 200 – check where underlining is intended
-p7 line 258 – “Perhaps that was the number of human mtDNA in blood cells is an important indicator of various metabolic disorders” does not make grammatical sense – “perhaps the number of mtDNA molecules in human blood ….”?
-p8 line 284 – not clear what this means: “Therefore, mitochondrial dysfunction with fission upregulation prevents the elimination of damaged mitochondria producing ROS and RNS that exceed the antioxidant activity is likely initiating factors in inflammation, aging, and age-related diseases [48].” Should it be something like “Therefore a combination of mitochondrial dysfunction plus upregulation of fission of mitochondria that produce ROS/RNS to an extent that exceeds antioxidant capacity, is likely to be an initiating factor in ….”?


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: MITOCHONDRIAL DESTINY IN TYPE 2 DIABETES: THE EFFECTS OF OXIDATIVE STRESS ON THE DYNAMICS AND BIOGENESIS OF MITOCHONDRIA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revised version has taken into consideration hte majority of the points raised. Shortening the review also helps in focus.Broad interest and good reviewing of the jcurrent knowledge in the foeld sufficiently covered.
Experimental design: Good citing of literautyre, clealry improved in the revision.
Validity of the findings: Clear logic, good flow of the text
Additional comments: I am pleased to see the revision and the points that have been addressed by the authors. This is a much improved version which I am happy to recommend for publication.


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

Input:
Title: ENGAGING INEXPENSIVE HANDS-ON ACTIVITIES USING CHLAMYDOMONAS REINHARDTII (A GREEN MICRO-ALGA) BEADS TO TEACH THE INTERPLAY OF PHOTOSYNTHESIS AND CELLULAR RESPIRATION TO K4–K16 BIOLOGY STUDENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: I find the manuscript mostly clear and unambiguous. I note some areas that could use clarity in that they may be missing a word, need greater or less explanation.  It may be a bit more editorial than necessary.  Additionally, I list a few questions regarding the introduction below.  References and background seem sufficient, unless noted below.

Questions regarding the introduction:
Could the authors explain the advantage of using a motile strain of green algae vs. non motile?  I’d like to know if it is the novelty of being able to expand the species of unicellular algae that can be studied, using a well-known model organism, etc. – I see that this is in the discussion, but I think it is helpful to have a brief reference in the introduction as well.
Is there evidence for the available kits not working well? (ie. did your group test them? Survey K16 teachers?)

Writing clarity:
Lines 23-25:  awkward wording, suggestions in red: These kits are expensive, can be applied to only non-motile algae, and provide protocols that lack detailed specifics for trouble shooting and does not provide customization guidelines for different grade levels.
Lines 47-49: awkward wording, suggestions in red:  Our comparative studies of photosynthesis and cellular respiration in the10E35 and in 4A+ strains showed that 10E35 relatively has a higher respiration rate and a lower photosynthetic rate than 4A+.
 Line 90: maybe add “fun and engaging way” to sound more technical?
Line 93: “conduct their own independent experiments” – this shows the activity is not just a ‘cookie cutter’ lab.
Line 95: remove “in” before “in dark”
Lines 98-100:  It seems redundant to have both “functionally uncharacterized” and “unknown function” in this sentence.
Lines 105-107:  Add a statement that acknowledges that cellular respiration is still going on when the cells are in the light, but the net production of O2/use of CO2 from photosynthesis vastly outweighs the O2 consumption/CO2 release during cellular respiration (a very common misconception is that cellular respiration doesn’t occur in the light)
Lines 113-116:  this is just a very long sentence. Maybe according to grade level, availability of resources, and allow better time management?
Experimental design: I appreciate the level of detail that went into validating methods for this new teaching tool. I have a few questions below and some suggestions for clarity for the materials/methods provided for those using this for their own teaching.

For clarification – when referring to biological replicate – you are meaning 3 separate replications of the experiment, each with 3 internal replicates?  Apologies, I got lost in the large amount of data presented in the supplemental data files.  If this is not the case, I would suggest having more replicates to strengthen your findings.

dx.doi.org/10.17504/protocols.io.bgpyjvpw/ Are there copyright issues with having the protocol previously published?

Text_S1
Item 21: aluminum foil
Item 30: it seems that calling it a hemocytometer or counting chamber might be more flexible to users?

Text_S2
In order to have the greatest possible flexibility to educators likely working on a tight budget or who may already have some of these supplies, it would be helpful to have a disclaimer with the supply list that there may be other vendors for the general supplies that are cheaper or that they already work with.
The easiest way to do this is adding a column to the left that has the generic term for the item (ie. 1.5 ml centrifuge tubes, yarn, disposable inoculating loops and needles, hemocytometer, etc.) as they may already have these items/be able to find cheaper. For example, the Tygon E3603 laboratory tubing is $211 (I realize you get a ton of tubing, but maybe a school would only want a small amount and look to pick it up at a hardware store) and it could be broadly represented by providing the dimensions.

It may be beneficial to provide a basic protocol on how to use a hemocytometer in a teaching setting? Again, this could be a supplemental text, similar to S1 and S2.
Validity of the findings: Analysis of data appears robust (provided the replicates are as stated in section 2) and statistically sound. A few specific comments:

Lines 280-282:  I don’t typically see p values referred to as percentages in scientific papers?  Additionally, some of this information regarding the type of tailed test could be removed to condense the results and included in the figure legends (and kept in the data analysis section of the methods) to condense the text in the results.

Thinking of this article in terms of describing teaching activities, it would be helpful to have a figure/supplemental text for specifically trouble shooting common problems.

I think in terms of audience that teachers using this activity would benefit from the separation of technical details from the learning activities, pre-class assignments/background, assessments aligned to learning goals. Perhaps this could be combined with trouble shooting and supplies into a single supplemental text that is information on teaching?

Some of the items in the discussion, such as customization by grade level, could be in their own figure to highlight and provide more options (as it sounds like you have them with 14 other activities!) - this could be similar to Table 9.
Additional comments: This manuscript describes a holistic picture – using basic scientific research to create engaging activities for K4-16 students.  I very much appreciate that this work was generated in collaboration with high school and undergraduate students.

This manuscript has a huge amount of data!  For me, there is important information that has two broad audiences and you will not be able to reach them in a single manuscript.  Review instructions state that decisions are not made based on any subjective determination of niche audience, so this may be out of line, but the work seems like 2 different manuscripts that differ on scope and audience:  1) a methods paper providing all data (potentially include data from verification of the other developed activities?) with optimizing this measure of photosynthesis and cellular respiration - color change, pH, time course experiments, and statistical analysis – geared towards the broader scientific community and fitting in the PeerJ journal, and 2) a teaching module designed for K4-16 educators that has the protocols, and much less technical information (suggestions include International Journal of STEM Education, Frontiers in Education: STEM Education, American Biology Teacher).

If I am incorrect with regards to my assessment of the scope of PeerJ, I would advise a greater separation between teaching-specific information (simplicity and clarity) and basic data.   If PeerJ looks for this type of comprehensive work, I applaud having a platform that is open access and can appeal to multiple, historically separated, audiences.


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: ENGAGING INEXPENSIVE HANDS-ON ACTIVITIES USING CHLAMYDOMONAS REINHARDTII (A GREEN MICRO-ALGA) BEADS TO TEACH THE INTERPLAY OF PHOTOSYNTHESIS AND CELLULAR RESPIRATION TO K4–K16 BIOLOGY STUDENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Here, the authors designed a useful and inexpensive protocol as a tool to teach in a fun and easy way the basic concepts of photosynthesis. Work is of great importance at the educational/academic level. However, some points can be improved, such as the ones I point out below:
-I suggest deepening in the basic concepts of the photosynthesis process and highlight its biological importance. This could help you to strengthen your work justification.
-I suggest a change in the order of information in the introduction section. Line 87 to 101 is about your results, so, I think that this information should appear after line 110.
-There is information that doesn't have references, please include the original works from which that information was taken. i.e., lines 103, 107, 334, 541.
-The link that you provide for “Carolina QuickTips Making Algae Beads” reference doesn't work, please check it.
-It's highly recommended that the figures show all the necessary data to can understand them even without seeing the figure legend. In this regard, I suggest including some missed information in the figures. For example, in Fig. 1, please indicates which vials correspond to DI or tap water; Fig 2, please indicates the concentrations of the beads, etc. For more details please check the attached pdf file.
-Tables 1 to 7 are repeated or complementary information of figures, thus, I suggest sending them to supplementary material.
Experimental design: The information obtained in this investigation has very educational and academic significance, which will be helpful for many educators and institutions.
The research question was well defined and I think that the methods used were appropriate. Also, these were described in detail. I only have a few suggestions:
-Considering the nature of your investigation and intending to facilitate the reading of your results, I suggest adding one figure to show the color-scale expected when you use the pH indicators as the bicarbonate indicator and the phenol red solution.
-I suggest describing that phenol red solution is used as a pH indicator too, as a bicarbonate indicator (line 236).
-For the evaluation of the effect of cell density in line 234, I suggest writing the two concentrations of beads used.

About the data analysis, I appreciate that you provide all the statistical analysis, however, I found them difficult to read. I suggest founding a better way to describe them. For instance, you can consider the way you describe it on lines 378-379. In this example, you summarize the information and put them in parentheses. Also, consider that it's not mandatory to describe all the numeric data obtained in your statistical analysis. Because you provide all your analysis in detail in supplementary material; you can always refer to that.
Validity of the findings: I consider that all the experiments here described were well performed, and the results are supported by statistical analysis. The conclusions are well stated.
Additional comments: Please refer to the attached file for more details.


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: ENGAGING INEXPENSIVE HANDS-ON ACTIVITIES USING CHLAMYDOMONAS REINHARDTII (A GREEN MICRO-ALGA) BEADS TO TEACH THE INTERPLAY OF PHOTOSYNTHESIS AND CELLULAR RESPIRATION TO K4–K16 BIOLOGY STUDENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall, the paper has a logical flow of ideas and is self-contained.  There are some grammatical issues in the paper, in particular those around use of tense.  The Background section of the Abstract would benefit from an additional sentence connecting the ideas from sentence1 to 2 (lines 20-23).  The third sentence in the Abstract Background has grammatical problems and would benefit from the use of colons and semicolons (lines 23-25). It is unclear to the rationale for including the last sentence of the Abstract Background (lines 28-31). The article is missing in the second sentence of the Methods portion of the Abstract (lines 24-36).  There are grammatical issues with the last sentence of the Abstract Methods (lines 36-39).  Tense and verb-subject agreement issues in the second sentence of the Results section of the Abstract (lines 41-44).  The fourth and last sentences of the Abstract Results section are a different tense from the rest of the section (lines 45-47, 50-52).  The first sentence of the Introduction needs commas separating the coordinating adjectives and the sentence ends with a phrase that is not grammatically correct (lines 55-56).  The second sentence of the Introduction is a run on and would benefit from being split into two (lines 56-60). These are some of the initial edits that should be made.

In the Introduction section, BIO-RADs “Photosynthesis and Cellular Respiration Kit for General Biology” should be added to the list of currently available commercial kits, as it offers the most comprehensive activity guidelines and includes detailed troubleshooting prompts (lines 60-64, 70-72).  It is unclear what the advantage would be of using motile versus non-motile algae for photosynthesis and respiration assays (lines 67-70).  Authors should articulate the rationale for use of Chlamydomonas over Chlorella, as the commercial kits use the latter.  Related, the authors should explain the advantage of making the beads over purchasing pre-made beads, such as the ones available from BIO-RAD.  Given the number of additional reagents and time needed for making the beads, it is unclear if this protocol is more cost effective in comparison to the BIO-RAD kit, which one can purchase for $181 in the US.  The author’s should include a detailed cost comparison to support this claim.  The authors should read Canbay, Kose, and Oncel (2018) for a published protocol for making algae beads using Chlamydomonas (lines 67-68).

The figures are extensive and well organized.  For the sake of reducing confusion in your target audience (K4-12 teachers), it is recommended that the hand-written labels on the glass vials be cropped out.  Given the importance of subtle color changes, the images should have a consistent white-balance applied.  The authors are encouraged to look at Figure 2(A), specifically.
Experimental design: It is unclear if the focus of this article is the modification of existing protocols for use with Chlamydomonas or the use of this technique as part of a teaching module.  If the former, then I would encourage the author’s to read the paper by Canbay, Kose, and Oncel (2018) that describes production and analysis of immobilizing Chlamydomonas in beads.  The author’s should describe how their protocol differs and/or is superior to the protocol described in the 2018 paper.  If the latter, then it is unclear why the author’s would choose to include a mutant strain that is not available from the Chlamy Stock Center.  The authors do include a list of strains that are available, but results from this activity using those strains was not provided (lines 544-547).  For a K6-12 teacher, it will be difficult for them to know what expected results are.  The authors can provide a table or figure describing expected outcomes using exemplar strains available from the Chlamy Stock Center.

In the “Preparation of Chlamydmomonas…” subsection, the URLs do not work because of the inclusion of the forward slash at the end of the link (lines 160 and 166).  The chemical analysis of the “tap water” should be included to define “tap water” for this study and confirm that chemicals that would modify photosynthesis/respiration, such as Acetate or Sulfur-containing compounds, are not present.
Validity of the findings: This paper modifies a published protocol and applies it to existing activities with a new twist, which is the plastic tubing “bracelets”.  The tubing allows for better visualization of the pH changes due to the activity of organisms in individual beads, in comparison to glass vials or microcentrifuge tubes.  The “bracelet” modification is meant to capture the attention of students, and this should be effective in doing so, however, data was not provided.  The existing, commercially-available beads can also be repackaged into this “bracelet” form.  The authors sufficiently addressed the caveats of the plastic tubing.
 Given the number of institutions this protocol has been used at, the inclusion of student and teacher opinions would have strengthened the argument that the learning module is engaging.  The authors also claim that this protocol is most cost-effectives than existing protocols or kits.  The authors should include a detailed list of all reagents and equipment required, with their respective cost of use, to support this claim.
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: ENGAGING INEXPENSIVE HANDS-ON ACTIVITIES USING CHLAMYDOMONAS REINHARDTII (A GREEN MICRO-ALGA) BEADS TO TEACH THE INTERPLAY OF PHOTOSYNTHESIS AND CELLULAR RESPIRATION TO K4–K16 BIOLOGY STUDENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors followed the recommendations made to improve their work.
The introduction became more complete and clear with the changes made. Also, the figures were improved and now is easier to see what they want to show.
I appreciate that you move the tables to supplemental material. In this way, it is easier to read the manuscript.
Experimental design: The changes made to the descriptions of statistical analyses improve considerably your manuscript. However, I think that it is very important to mention which was the p-value used in your analyses. You can mention it in the "Imaging and Data analyses" section or, you can point it out in the results section, by enclosing the p-value in parentheses when necessary. Nevertheless, it is important to note that the p-values usually are presented as decimals, as already mentioned by another reviewer. So, I suggest writing in these ways in the manuscript, and if you want to conserve the percentages data in the Data files, you can do it, too.
Validity of the findings: I consider that this work will have an important impact on the educational and academic level.
I think that the experiments were properly developed and the conclusions are well stated. Also, the changes made to the "Materials and methods" section improve considerably the manuscript.
Additional comments: I appreciate the changes made. I think you improved the manuscript a lot. I consider that it is almost ready for publication.


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

Input:
Title: A NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comments here, see my attachment
Experimental design: no comments here, see my attachment
Validity of the findings: no comments here, see my attachment
Additional comments: no comments here, see my attachment


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 NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Good English throughout in general. One or two corrections are made in the attached pdf.

Introduction could be expanded a little with greater explanation of why now. See general comments for more detail.

Structure and plates in general OK but the conclusions need to be revised as contain much discussion. There is inadequate discussion of the materials in the M&M section.
Experimental design: Research is original.

Questions well defined and meaningful
Validity of the findings: Conclusions are in the main good. I however think that significantly broader discussion is required around the proposed epizoic nature of new species.
Additional comments: Review comments
This paper erects a new species to accommodate material previously assigned to an inappropriate existing taxon, and thereby highlighting important diversity. Whilst much of this work has already be hinted at, or noted in various other publications, the current work is important as it formerly pulls together these notes and thoroughly addresses the implications. I would like to congratulate the authors for this fascinating paper and let them know that I greatly enjoyed reading it.
That said, this paper needs to be revised and restructured in many places. I have made numerous suggestions within the attached PDF where I feel changes or greater elaboration is needed to help the reader more clearly understand what is being attempted. Below I have listed some of the more important points which are based on comments made within the text of the attached pdf. Of the these, the most important to address are; that most of the “Conclusions” are, in fact, mostly discussion points; the introduction needs to be expanded to better contextualise the study, that there is content in the “History of Study” section that is better placed in the material and methods and; that they need better evidence to support their interpretation of this ophiuroid as an epizoic organism.
As such, I am pleased to recommend this paper for publication but only after major revision.

1.	The Conclusions are, in the main, not conclusions but a general and very useful discussion about the broader relevance of the findings of this work. As such, the paper needs to be re-structured accordingly.
2.	Please expand the Introduction slightly to say specifically which aspects of these specimens are providing the impetus for the study. Something along the lines of it has not been adequately described/accurately assigned and this has broader implications but it would help to state this explicitly and why it is being done now.
3.	I think the evidence for O. floorae being epizoic on Bourgueticrinids is poor. This association is as likely, if not more likely, to be the result of both organisms being washed together during the obrution event that formed the associated Lagerstatten from which these fossils were collected from. This is because only one arm is draped around the stem, not 4, or all 5 arms closely clinging to the stem or cup, as seen in multiple examples from the Carboniferous. That they lived in the same environment, there is no doubt, but I do not think the current evidence of “general resemblance” to known modern epizoic ophiocanthids is strong enough to interpret the fossil co-occurrence as an epizoic relationship. Please also, see further comments made on lines 224-225. As such, significantly more evidence of epizoic behaviour in O. floorae is needed than is currently provided or the various statements regarding its epizoic habit need to be revised throughout the paper (in the “Conclusions”, Abstract and, Introduction).
4.	Whilst I can see that images of the association between the crinoid and ophiuroid have been published previously (Jagt 2000a) an image of it must be included in this paper if the authors want to maintain that the relationship is epizoic or discuss the palaeoecological implications. As such, in Figure 5 (or a separate figure) please show the association between the ophiuroid and the bourgueticrinid stem.
5.	In the log (figure 3), I am assuming that the black globular shapes are a representations of flint bands. There are also other shapes on here which may indicate bioturbation. Please can a key be included? The log would be further improved if chronological dates are added (these are mentioned in the text so it should not be too hard to do) and that the columns are labelled (particularly for the column with IVa-1 etc. as I am unsure specifically what these refer to). Finally, can they please show where the specimen was located within the log. In the text it is fairly explicit as to where it is from but the arrow on the log instead points to “one of the more spectacular storm levels”. Whilst interesting, storm levels have little apparent relevance to the rest of the paper.
6.	I am confused about the status of the other specimens assigned to Ophiocantha? danica by Jagt 2000a but not mentioned at all in the current paper. I assume that they are not part of the new species but I think it would be valuable to add some discussion about the status of it as there is so little material currently included in the new species and yet there is published reference to more material from the same site that was formerly placed within the same taxa by one of the co-authors.
7.	See pdf for further 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: A NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The English is excellent.
Experimental design: No experimental design per se in a paleontological taxonomy study. The methods and examinations follow standard protocol for this type of study.
Validity of the findings: I agree that the fossil is an ophiacanthid and due to the fact that it was epizoic on a crinoid, it is more likely to be an Ophiomitrella than an Ophiacantha. However, it would be helpful for readers if you would explain why you exclude other possible genera. The LAP is very similar not only to Ophiomitrella but also to Ophiacantha. Without knowing the radial shield and possible granules on the disc, why can't it be an Ophiacantha?

You state (row 199) that Ophiomitrella is paraphyletic, which would mean that all Ophiomitrella species share a common ancestor but one or more groups that also share this ancestor are excluded. On the phylogenies by O'Hara (latest version in Christodoulou et al. 2019), Ophiomitrella includes species with different ancestors, which means it is polyphyletic (as is stated in O'Hara et al. 2018 not 2017). As long as we don't know in which group the type species falls, we cannot decide, which of these are to be considered Ophiomitrella, but there are 3 clades. Can you really decide that floorae is closest to the clade that contains clavigera and conferta? Why not the clade with globulifera or the one with stellata? Perhaps you can comment a little more on this. Considering that Ophiacantha is likewise polyphyletic and intermingled with Ophiomitrella on the tree, this is difficult and should be discussed a little more broadly.
Additional comments: Another paleontological mystery has been cleared up, very nice.
Just out of curiosity, I do not have the original paper (Jagt 200) at hand and wonder how this animal looked when it was wrapped around the crinoid stalk. Was it removed or are we suppose to see this in the images? The ophiuroids wrapped around crinoid arms that I have seen were orientated with the mouth towards the crinoid. Here it was only one arm that attached to the stalk, but was it broken off or how?


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 NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I have already wrote before that this is very interestging MS that should be published in PeerJ. I see that in its current form, all corrections and suggestions proposed by me and other reviewers have been made. Basically I would suggest publishing the article as it is, but I have found some editorial mistakes that need to be corrected (Am attaching pdf).


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

Input:
Title: A NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 2
Reviewer: 2

Output:
Basic reporting: All fine
Experimental design: Fine
Validity of the findings: Fine
Additional comments: Whilst, I remain unconvinced of the geological evidence for an epizoic relationship between this individual ophiuroid and the crinoid, I can see that it resembles some modern epizoic ophiuroids. The amendments made in the paper addressing this point are fine. As far as I am concerned, all the other changes suggested by the reviewers have been thoroughly addressed and it is a pleasure to recommend publication. My congratulations to the authors for such a nice 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: A NEW OPHIACANTHID BRITTLE STAR (ECHINODERMATA, OPHIUROIDEA) FROM SUBLITTORAL CRINOID AND SEAGRASS COMMUNITIES OF LATE MAASTRICHTIAN AGE IN THE SOUTHEAST NETHERLANDS
Review round: 2
Reviewer: 3

Output:
Basic reporting: The English is excellent
Experimental design: There is no experiment as such but the methods are appropriate
Validity of the findings: Valuable study.
Additional comments: The authors have revised the manuscript diligently and I have only a few minor corrections to suggest.

rows 51-52: 'attempt at', I think, the verb attempt does not go with 'at'
row 111: change danica to small d
row 225: the historical samples are not rare (but very valuable), the species seems to be rare in collections
Fig 5F-G seems to be the previous caption in the pdf but is changed in the Word document, and please attribute these to S. Stöhr.


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: MINING OF PROGNOSIS-RELATED GENES IN CERVICAL SQUAMOUS CELL CARCINOMA IMMUNE MICROENVIRONMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: A little more background on the significance of microenvironment related diagnostic and prognostic markers would be better to provide.
Experimental design: Methods should describe in more detail why specific scores and databases mentioned are used/selected.
The reason to choose the mentioned data analysis methods to search for the three immune-related scores should be described.
Validity of the findings: How different/similar was external data set for validation of the correlations of 18 immune microenvironment related genes?
Authors should talk about the significance of their findings in more detail.
Additional comments: Figure 8: All acrony’s full form should be mentioned in the description of the figure


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: MINING OF PROGNOSIS-RELATED GENES IN CERVICAL SQUAMOUS CELL CARCINOMA IMMUNE MICROENVIRONMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: PeerJ: #45770v1
Title: Mining of prognosis-related genes in cervical squamous cell carcinoma immune microenvironment

This work reports on the applicability of the immune-related scoring approach to screen immune-related prognostic markers in cervical squamous cell carcinoma immune microenvironment. Two levels of screening were applied step by step using ESTIMATE algorithm and WGCNA and network analysis to identify prognostic genes. Considering the biological heterogeneity of cervical squamous cell carcinoma it is worth carrying out bioinformatic studies similar to this present study to contribute to the current efforts to identifying prognostic biomarkers. This work is within the scope of the journal. The authors used the English language professionally throughout the manuscript and the manuscript was appropriately structured following PeerJ standard. Figures are of publication quality and appropriately annotated. Raw data is available as supporting information files. I have pointed out minor points.

1. Cervical squamous cell carcinoma should be abbreviated as CSCC throughout the manuscript

2. Although the context of this study well-reported with sufficient citations, to aid the reader additionally, the biological heterogeneity associated with cervical squamous cell carcinoma should be discussed in details (with appropriate citations) in the Introduction section, as because the International Federation of Gynecology and Obstetrics (FIGO) staging lacks heterogeneity of CSCC

3. As this study focuses on the applicability of the immune-related scoring approach, the authors should introduce a comparative discussion (with appropriate citations) of different type of scoring approaches in the Introduction section

4. Supporting information materials should be appropriately cited throughout the  manuscript (For example, Line 80, “shown in ImmuneScore.genes.ids.txt” should be written as “shown in ImmuneScore.genes.ids.txt, Supplemental File”)

5. In-text citations and references are not in PeerJ style

6. Figure 3 caption, “he relationships between levels” should be “The relationships between levels”

7. In general, all the abbreviation used should be annotated in its first appearance
Experimental design: The objective of the study is well defined and computational experiments are well structured and seem to be performed accurately following standard strategies. Authors should address the following minor issues.

1. A flowchart representing the workflow of the computational experiments performed in this work should be added in the Materials and Methods section to make the method section more comprehensive

2. A brief description of each of the ImmuneScore, StromalScore and ESTIMATEScore which is associated with the ESTIMATE algorithm should be added at appropriate place in the manuscript (maybe under “Computational methods of multiple immune scores and result determination” in the method section)

3. Line 123, appropriate citation(s) should be added for “Cytoscape software”
Validity of the findings: The results are presented stepwise in a logical manner and also in details. Statistical analyses have been performed and reported. The results are well discussed and concluded appropriately answering the original research question. There are only minor points to be taken care of.

1. Line 136, The sentence “The first three scoring systems with most obvious correlation with others including ImmuneScore (R=0.59), Co_inhibition(R=0.59) and LCK(R=0.62), indicating that the consistency among the immune scores calculated by different algorithms to a certain extent.” should be rephrased

2. Line 157, appropriate citation(s) should be added for Kaplan-Meier method

3. Line 182-183, “As shown Fig.5A,” should be “As shown in Fig.5A,”

4. Line 186, appropriate citation(s) should be added for “dynamic shear method”

5. Line 198, The sentence “The detailed enrichment results were shown in yellow enrich.txt” should be written as “The detailed enrichment results were shown as supporting information file (yellow_enrich.txt)”

6. Line 220, The sentence “Moreover, a total of 26 genes (Table 2 and lst.genes.txt)” should be written as “Moreover, a total of 26 genes (Table 2 and lst.genes.txt as supplemental file)”

7. Line 229, “summarized in lst enrich.txt” should be “summarized in lst.enrich.txt (supplemental file)”

8. Line 252, “contrib1-GPL14951.txt” file should be added as supplemental file

9. Line 258, SIRPG gene should be reported for weak association compared to other genes

10. Any future application of this work flow and applicability of this approach in mining prognosis related genes in other biologically heterogeneous carcinoma should be discussed in the discussion section
Additional comments: No additional comments


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

Input:
Title: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is interesting and understandable, has good spots for reflection. The structure, figures and tables are clear. And the result is valuable and it will be of interest to some researchers. However, it needs major revision before being accepted.
Experimental design: In my opinion, the number of meteorological stations is not enough. Please explain why you select these four meteorological stations of Pakistan?
Validity of the findings: No Comments
Additional comments: It is an interesting piece of work, but there still exists some problems in the manuscript. The specific suggestions are shown as follow:
(1) line 18: “Mann Kendall” should be “Mann-Kendall”. Please revise this expression throughout the manuscript.
(2) The English language should be improved. For example, line 19 “It is concluded MMHS method indicated that all stations are suffering……”.
(3) line 48: The reference doesn’t conform to PeerJ standards.
(4) Some words are incorrect in this PDF version, e.g. lines 50, 55. Please check and revise throughout the manuscript.
(5) There are many unreasonable citations in the manuscript, e.g., lines 58, 64, 70.
(6) line 65: Please use the full name of “NECT region”.
(7) line 71: “Songnen plain” should be “Songnen Plain”.
(8) line 80: “indicates” should be “indicated”.
(9) line 85: Please add “and” before “its”. Or rewrite this sentence.
(10) line 90: Please explain why you choose only four stations? Or please explain why you select four meteorological stations of Pakistan? In my opinion, the number of meteorological stations is not enough.
(11) A map of the locations of the study area and the meteorological stations should be added in this manuscript.
(12) line 126: The reference should be at the end of this sentence.
(13) lines 128, 144, 160, 167, 184, 188: “Where” should be “where”.
(14) line 176: I think the reference “Yang et al, 2019” should be deleted in this sentence.
(15) line 227: The superscript of “ith” is unreasonable.
(16) In the discussion part, the limitations of this study should be added.
(17) In the conclusion, authors should supply your future 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: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This manuscript entitled “Evaluation of wet and dry event’s instability and trend based on meteorological drought index” by Muhammad H Shaukat et al. mainly discusses the temporal variation of wet/dry events in Pakistan using SPI calculated by  observed data of four stations(Multan, Bahawalpur, Barkhan and Khanpur). The results of EEMD and MRA show prominent non-linear trend which is consistent with the Mann Kendall (MK) and Spearman’s Rho (SR) tests. And the Mean Marginal Hilbert Spectrum (MMHS) and Continuous Wavelet Power Spectrum (CWPS) are applied to detect the instabilities of wet/dry events. Similar results also are captured by Hilbert Huang Transformation (HHT). Furthermore, the differences of EEMD and MRA are also presented in this study, finding that EEMD has better performance in reconstructing SPI series. And SR test is better than MK test in describing the trend of dry/wet events. The manuscript is in general well organized and scientifically interesting for the community. Therefore, I recommend publishing the manuscript after addressed the following comments.

General Comments:
1. In Section 3(Line 246-281), the authors use the residuals of EEMD and MRA to describe the trend of dry/wet condition (Figure1, Figure2). However, the long-term trend exhibited in residual of EEMD is quite different from the one from MRA. Why?
2. In Table 2, all instantaneous frequency indexes of IMF2 and IMF3 of Bahawalpur is the same, which is very unusual. I think the authors should check their results make sure the results are right.
3. Figure 4c&4d: the location of the capture does not agree with the title in the figures, and the corresponding analysis (Section 3, Line 316-320) is confusing, too.
4.Table 3: I don’t understand what exactly the period means in table 3. For instance, which period 1990s refers to? And how is the test statistic in period 1990s calculated?
5. Section3 (Line342-346): The authors try to prove whether the trend obtained by MRA residuals (Figure 2c-d) can be verified by the MK and SR trend test (table3) in Barkhan and Khanpur. However, the analysis here is not consistent with what the table and figures show. For example, the residual shows a decrease of SPI in Barkhan in 1998 (Figure 2c) while the MK and SR tests shows a increasing trend (Table 3). It’s recommended that authors check it out and make corrections or explain it.

Minor Comments：
1. Section 3, Line 334-335: “It turns out that the SR test indicates the consistency with residuals of EEMD in mostly periods as compared to MRA.” “compared to MRA” should be “compared to MK”.
2. Authors should also pay attention to the citation format. For example, in Section 1, Line 59, “(Bin Li et al., 2013) utilize the data of daily precipitation (1960-2015) of 35 meteorological stations of the lancing river basin in China” It should be changed to “Bin Li et al. (2013) utilize the data of daily precipitation (1960-2015) of 35 meteorological stations of the lancing river basin in China”.
3. Reference (Line484-486 & Line503-504): The DOI is missing.


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

Input:
Title: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Comment on the “Evaluation of wet and dry event’s instability and trend based on meteorological drought index”

In this study, the authors aim to evaluate variability and trend of the wet/dry event of the four stations in Pakistan by using several methods. The differences between the results based on different method are also contrasted. Through careful reviewing, the reviewer think that the present manuscript is badly written and lacks of clear logicality. And importantly, the authors paid more attention to the method rather than the results or the merit of this study. Therefore, I recommend the article a rejection at least a resubmission after a major revision.
Other comments:
1.	In the abstract, the authors seem to evaluate the applicability of the method in detecting the variability of the wet/dry event, because more than 2/3 part is used to the introduce the methods.
2.	The reviewer can not obtain clear goals of the present work, as well as the present understanding and main issues in the literature.
3.	I suggest the authors focus on one or two methods and highlight the new findings.


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

Input:
Title: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 2
Reviewer: 1

Output:
Basic reporting: No Comments
Experimental design: No Comments
Validity of the findings: No Comments
Additional comments: The authors have addressed all 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: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors modified the manuscript according to my comments/suggestions. I therefore recommend an acceptance of publication.


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

Input:
Title: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors revised the manuscript mainly by polishing the manuscript. However, the weak analyses and weak contrast or merit are still major shortcomings of the manuscript, which has been pointed out in the previous review cycle. The revised manuscript still does not convince me enough. Therefore, I would suggest rejecting the manuscript.
Experimental design: no  comment
Validity of the findings: no  comment
Additional comments: The authors revised the manuscript mainly by polishing the manuscript. However, the weak analyses and weak contrast or merit are still major shortcomings of the manuscript, which has been pointed out in the previous review cycle. The revised manuscript still does not convince me enough. Therefore, I would suggest rejecting the manuscript.


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

Input:
Title: EVALUATION OF WET AND DRY EVENT’S TREND AND INSTABILITY BASED ON THE METEOROLOGICAL DROUGHT INDEX
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I have no further comments on the manuscript.