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: INVERSE MODELLING FOR PREDICTING BOTH WATER AND NITRATE MOVEMENT IN A STRUCTURED-CLAY SOIL (RED FERROSOL)
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper is well written and provides interesting results on the use of inverse modelling to parameterise models for water flow and solute transport.

Detailed suggestions for consideration by the authors have been added as comment boxes throughout the pdf manuscript.
Experimental design: A critical component of this paper is the inverse modelling of soil hydraulic parameter. However, the explanation and detail of how the column data was incorporated and run in Hydrus 1-D to generate the inverse parameters is insufficient for readers to easily understand how this was undertaken.

In addition a table of the column data that was used in the inverse modelling would be useful to enable others to repeat the parameter optimisation. If this is felt to be too big to go into the paper then a much clearer table needs to be added in the supplemental files.

The paper reports on a small study, with little replication, however the excellent fit between measured and modelled data suggests that, unless the authors have been very lucky, further replication would probably have been superfluous. The controlled conditions where soil structure was accurately reproduced also added to the success of this small study.

The authors have provide a great deal of information in the supplemental files, however, more descriptive metadata would be useful if they are to be readily accessible to readers.
Validity of the findings: The findings are generally well described and supported by the data. Some extra discussion on the authors thoughts on the practical merits of having to include independent measurement for the inverse modelling, and why adding retention points might make the Rosetta results worse would be interesting.
Additional comments: I look forward to the review comments being addressed and seeing the paper published.


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

Input:
Title: INVERSE MODELLING FOR PREDICTING BOTH WATER AND NITRATE MOVEMENT IN A STRUCTURED-CLAY SOIL (RED FERROSOL)
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is a generally well written manuscript. There are some aspect where I think the authors need to explain or describe what they have done in more detail.
Experimental design: The value of l in Table 2 is quite different to the standard HYDRUS value of 0.5, yet no attention or discussion about this occurs. Having a large value of l will mean that the hydraulic conductivity decreases rapidly with water content. All the Rosetta derived values will have used 0.5, rather than values > 3 as given in Table 2. This will have a large effect on the shape of figure 3a, which is why the Rosetta derived values may have been unable to match the experimental data in figure 4. The authors show see what happens if the alter l with the other Rosetta values. Cook and Cresswell (2008) (Cook FJ and Cresswell HP (2007).  Chapter 84 Estimation of Soil Hydraulic Properties.  In Soil Sampling and Methods of Analysis. M.R. Carter and E.G. Gregorich (Eds.), Canadian Society of Soil Science, Taylor and Francis, LLC, Boca Raton, Fl, 1139-1161.) discuss how to estimate l.
Validity of the findings: Thevan Genuchten equation and Mualem equations need to be shown. The values of the parameters in this equation are shown in Table 2 but no explanation of what the parameters represent. This could be done either in the text or in the caption of Table 2. The symbols in Table 2 need correcting as they differ from Table 3 and the rest of the text and figures
Additional comments: The referencing needs to be fixed with citations missing from the reference list and references which are not cited. There are also a few minor grammatical corrections required.

Minor Corrections.
1.	Lines 48 and 49 and throughout the rest of the manuscript. Vrugy and Bouten (2002) and Wohling et al. (2008) are not in the reference list. Also for the Wohling reference et al. is not italicised
2.	Line 62. I think the reference to Mallant et al. (2007) should be Mallants et  al. (2007).
3.	Line 77. Jacques et al. (2012) is not in the reference list.
4.	Line 171. The abbreviation CI is not defined.
5.	Line 201. Replace ‘was’ with ‘were’.
6.	Line 208. I do not know what ‘equivalent’ refers to?
7.	Line 219. Replace ‘Horizontal Columns’ with ‘Wedges’.
8.	Line 255. Simunek and van Genuchten (1996) is not in the reference list.
9.	Line 323. I think Hopmans (2002) should be Hopmans et al. (2002).
10.	Line 428. Gardenas et al. is not cited.
11.	Lines 428-487. Romos et al. (2011, 2012) are not cited.
12.	Line 520. Soil Survey Staff (1992) is not cited.
13.	Table 2. Symbols used are not correct.


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: RECOGNITION OF SPECIES GROUPS OF NAUPACTUS DEJEAN (COLEOPTERA: CURCULIONIDAE) FROM ARGENTINA AND NEIGHBORING COUNTRIES
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an excellent manuscript about weevils of tribe Naupactini, written by the two leading experts in the group. It deals with the highly diverse and economically important South American genus Naupactus, focused on the species that occur in Argentina. This taxonomic contribution allows identification of the nine species groups and provides new, revised and updated information (morphological, biogeographic and on host-plants) for 37 species occurring in Argentina and neighboring countries.
The paper is well written and nicely illustrated. I only have some minor corrections and few suggestions to improve the manuscript:
Line 137. Methathoracic wing, correct to plural “wings”.
Line 171. Please check/correct  the sentence where it says “…setae of medium-length hand suberect…”. It is not understandable as it is written.
Lines 240-243. I suggest using present tense here because it is understood that these are nomenclatural changes done in the present paper. Also, placing the word “therefore” (= for that reason, consequently) is not enough because “not being a Naupactini” doesn´t provide justification for being a Tanymecini. I suggest rewording in present tense and providing a justification for your decision, something like:
“…it is not a Naupactini and meets definition of genus Eurymetopellus Emden, therefore it is herein  transferred to Tanymecini, genus Eurymetopellus, thus establishing the new combination Eurymetopellus cephalotes (Hustache).”
Line 483. Please check specific epithet of “N. lar (Germar)”.
Lines 845 and 847. There are extra words in the following pairs: “…bears resembles” (delete bears); “…is looks” (I would delete “is”).
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is an excellent manuscript about weevils of tribe Naupactini, written by the two leading experts in the group. It deals with the highly diverse and economically important South American genus Naupactus, focused on the species that occur in Argentina. This taxonomic contribution allows identification of the nine species groups and provides new, revised and updated information (morphological, biogeographic and on host-plants) for 37 species occurring in Argentina and neighboring countries.
The paper is well written and nicely illustrated. I only have some minor corrections and few suggestions to improve the manuscript:
Line 137. Methathoracic wing, correct to plural “wings”.
Line 171. Please check/correct  the sentence where it says “…setae of medium-length hand suberect…”. It is not understandable as it is written.
Lines 240-243. I suggest using present tense here because it is understood that these are nomenclatural changes done in the present paper. Also, placing the word “therefore” (= for that reason, consequently) is not enough because “not being a Naupactini” doesn´t provide justification for being a Tanymecini. I suggest rewording in present tense and providing a justification for your decision, something like:
“…it is not a Naupactini and meets definition of genus Eurymetopellus Emden, therefore it is herein  transferred to Tanymecini, genus Eurymetopellus, thus establishing the new combination Eurymetopellus cephalotes (Hustache).”
Line 483. Please check specific epithet of “N. lar (Germar)”.
Lines 845 and 847. There are extra words in the following pairs: “…bears resembles” (delete bears); “…is looks” (I would delete “is”).


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

Input:
Title: RECOGNITION OF SPECIES GROUPS OF NAUPACTUS DEJEAN (COLEOPTERA: CURCULIONIDAE) FROM ARGENTINA AND NEIGHBORING COUNTRIES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The following typographical or grammatical errors should be corrected:

Line 171 – “hand” should be “and”

328 – "withouf" should be "without"

407 – "Cyrtomon-like", with a hyphen, is used elsewhere in the text but not here

There should be consistency with having spaces after numbers and before units. e.g. line 427

738 – "oftern" should be "often"

767 – Gossypium is only spelled with one 's'

835 – "differs" should be used instead of differentiates

Other items that should be addressed:

Line 95 - the specimens borrowed from Charles O'Brien should be reported as being housed at ASUCOB, for Arizona State University Charles O'Brien collection, where the collection is now, in Tempe, Arizona, USA (http://scan-bugs.org/portal/collections/misc/collprofiles.php?collid=121)

Line 169 – there is no couplet 7 in the key! change all 8’s to 7’s (lines 138, 169, 172) and all 9’s to 8’s (lines 175, 176, 179).

Lines 315/387/432/492/571/628/723 – instead of forehead, should be frons or vertex be used? if there is some reason that forehead is preferred please specify.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This was a very well done study on a large and difficult genus of weevils and will surely be a crucial reference for anyone studying naupactines. There are only very minor errors that should be addressed.


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

Input:
Title: EFFECTS OF TABLE TENNIS MULTI-BALL TRAINING ON DYNAMIC POSTURE CONTROL
Review round: 1
Reviewer: 1

Output:
Basic reporting: Line 168: Correct section as session
Line 176: Correct a supine position to supine position
Line 339-340: Rephrase the sentence
Experimental design: Well articulated. It is stated how research fills an identified knowledge gap.
Validity of the findings: Statistically sound.
Additional comments: Interesting work.
The section on discussion would be solid if you could add more explanations citing articles in accord with your 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: EFFECTS OF TABLE TENNIS MULTI-BALL TRAINING ON DYNAMIC POSTURE CONTROL
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English language used is correct. The paper is provided a clear background of the topic. The structure of article with all figures and tables are also clearly and understandably.
Experimental design: The problem of this research is clear, and the questions are well defined. The described methods gives good information about data analysis used in paper. The paper geted ethical approval.
Validity of the findings: The results are well described and presented. The conclusions and explanation in Discussion are well stated, and all is linked to original research question and problem.
Authors presented the limitation of their study and put some directions for next studies in this research area.
Additional comments: In next studies please try to find correlation of  multi-ball  training with overall balance or postural stability. You can used some dynamic tests as Biodex balance system or some similar. In this study you provide to us info's about CoP distribution during the testing procedures.But it will be good to know the post effects of multi-ball training on the overall balance after few sessions of multi-ball training.


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: INDIVIDUAL AND FLOCK IMMUNITY RESPONSES OF NAÏVE DUCKS ON SMALLHOLDER FARMS AFTER VACCINATION WITH H5N1 AVIAN INFLUENZA VACCINE: A STUDY IN A PROVINCE OF THE MEKONG DELTA, VIETNAM
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript “#31719” by Huynh et al., entitled “Individual and flock immunity responses of naïve ducks on smallholder farms after vaccination with H5N1 Avian Influenza vaccine: a study in a province of the Mekong Delta, Vietnam” is a well-designed and well written account providing important information on humoral immune responses of ducks to the applied H5N1 vaccine under field conditions in Vietnam. This study is also essential for guiding the veterinary authorities, either locally in Vietnam or worldwide, to efficiently adopt effective booster vaccine protocol to vaccinate ducks which are commonly reared together with other avian species in hotspots areas of avian influenza viruses.
Generally, the English language of the manuscript is adequate; the quality of the figure and tables is satisfactory, the reference list cover the relevant literature adequately and in an objective manner. The topic is timely and interesting and the results are presented well.
However, the following points should be addressed:
1.	Line 36-37 and line 107: remove “at 21-day intervals”. It is anyway stated in line 37 and may be confusing with the prevaccination sampling time point.
2.	Line 57: replace “H5 HPAI viruses” with either “H5Nx HPAI viruses” or H5-type HPAI viruses” and keep it consistent throughout the manuscript (e.g. line 67, Line 102, line 123)
3.	Line 64-65: the statement “The vaccine will continue to be used in the foreseeable future because it provides a certain protective effect against HPAI to the poultry population” is scientifically not optimum. Please phrase it for the present taking into consideration the rapid evolution of these H5-type strains from clade 2.3.2→2.3.2.1a→2.3.2.1b →2.3.2.1c and possible genetic and antigenic evolution into different subclades.
4.	Line 73: keep distance between “H5N1” and “HPAI”.
5.	Line 81-83: Concerning the statement “published data on the effectiveness of vaccines against avian influenza (AI), particularly the Re-6 vaccine, in inducing antibody response in domestic ducks under field conditions remain limited”, have the authors compared their data with similar studies in different geographical localities (e.g. Kandeil et al., 2017. Avian influenza H5N1 vaccination efficacy in Egyptian backyard poultry. Vaccine; 35 (45): 6195-6201). If not please discuss.
6.	Line 93: replace “The Sample size” with Sample size.
7.	Line 111: what other factors which can lead to H5-specific antibodies other than maternal antibodies or active infections of newly born duck. If no, please remove “or other unknown factors” and rephrase the statement accordingly.
8.	Line 113: replace “which contains the HA gene of” with “which expresses the HA antigen of”.
9.	Line 181: please provide a rationale before direct stating of the results (e.g. To investigate that maternal antibodies have already depleted, prevaccination HI titers for studied ducks were evaluated”.
10.	Line 183: remove “with RAHO VI”.
11.	Line 191: replace “produced” with “showed”.
12.	Line 193: replace “H5N1 vaccine” with “Re-6 vaccine”.
13.	Line 218-219: Have the authors checked or can check the effectiveness of induced antibodies titers to protect duck after first and booster vaccination doses (Challenge infection with currently circulating H5N1 HPAIV (Clade 2.3.2.1c))?
14.	 Line 240 and Line 298: discuss other studies which are performed under field or backyard setting in different geographical localities dealing with duck and other avian species` vaccination and adopting booster vaccination protocol.


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: INDIVIDUAL AND FLOCK IMMUNITY RESPONSES OF NAÏVE DUCKS ON SMALLHOLDER FARMS AFTER VACCINATION WITH H5N1 AVIAN INFLUENZA VACCINE: A STUDY IN A PROVINCE OF THE MEKONG DELTA, VIETNAM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript by Huynh et. al. entitled “Individual and flock immunity responses of naïve ducks on smallholder farms after vaccination with H5N1 Avian Influenza vaccine: a study in a province of the Mekong Delta, Vietnam” is a straightforward to determine immunity responses against influenza H5N1 in duck smallholder farms after vaccination. vaccination has played a crucial role in the strategy for the prevention and control of H5N1 highly pathogenic avian influenza (HPAI). This study is essential for guiding the veterinary authorities.
Experimental design: The authors use well accepted methods in evaluating antibody responses of immunologically naïve domestic ducks to H5N1 avian influenza vaccine currently used in the national mass vaccination program of Vietnam.
Validity of the findings: The authors use well accepted methods in evaluating antibody responses of immunologically naïve domestic ducks to H5N1 avian influenza vaccine currently used in the national mass vaccination program of Vietnam.
However, there appears to be some careless misinterpretation of results, which needs checking and revision by the authors. Several hemagglutinin gene clades were detected in Vietnam clade 1.1.2 was predominant in 2012 and 2013 but gradually disappeared and clades 2.3.2.1a, 2.3.2.1b, and 2.3.2.1c from 2012 and recently clade 2.3.4.4 were detected. Why the authors used strain A\Ck\Scot\59 H5 as antigen in serological assay? And what the antigenic similarity between this strain and other strains circulated in Vietnam?
Additional comments: Minor Comments:
1-	line 58; “…the failure of other anti-HPAI measures.”  It is unclear the auther must explain what other measurements to control of avian influenza HP
2-	line 108 “…after the primary vaccination (21 dpv), and 21 days after the booster vaccination (21 dpbv) Can the authors use consistent abbreviation in all manuscript.
3-	In table 1 please include description for superscript letter a and b
4-	In discussion, were there any published studies in literature on the evaluated the effectiveness of the vaccine in inducing immunological responses under field conditions? These should be discussed and compared to the result presented here.


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

Input:
Title: CRISPR/CAS9-MEDIATED DELETION OF THE WISKOTT-ALDRICH SYNDROME LOCUS CAUSES ACTIN CYTOSKELETON DISORGANIZATION IN MURINE ERYTHROLEUKEMIA CELLS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is well organized and clearly written, including sufficient detail and references for the background material.
Experimental design: The manuscript describes a well defined set of experiments to study the effect of WAS gene deletion in a murine erythroleukemia cell line. The results are original and within the scope of PeerJ. Overall, the methods are described in adequate detail and the investigations are well performed. However, there is a lack of quantification and statistical evaluation of the results. The images and western blots should be quantified to enable statistical testing to rigorously support the conclusions of the study. If the experiments have been replicated enough times, this should not require additional replicates, simply quantitative analysis of the findings.
Validity of the findings: The results are convincing and the data are consistent with the conclusions, although as noted above there is a lack of quantification and statistical analysis that should be addressed. The discussion of the relevance of the observations are well stated and appropriate.
Additional comments: This manuscript is a welcome addition to the field. The one clarification that should be made is defining HMBA on line 185 of the Results section in addition to it's definition in the Methods section on line 106.


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: CRISPR/CAS9-MEDIATED DELETION OF THE WISKOTT-ALDRICH SYNDROME LOCUS CAUSES ACTIN CYTOSKELETON DISORGANIZATION IN MURINE ERYTHROLEUKEMIA CELLS
Review round: 1
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below.
Additional comments: This manuscript describes a role for Wiskott-Aldrich Syndrome protein (WASp) in regulating actin cytoskeleton organization in mouse erythrolukemia (MEL) cells. Deletion of the Was gene in locus in MEL cells using CRISPR-Cas9 genome editing technology resulted in increased G actin/F actin ratio in biochemical assays. Consistently, weak phalloidin staining for F-actin was observed in Was knockout MEL cells.  However, ectopic WASp expression in the Was knock out background successfully rescued the poorly organized actin phenotype. Furthermore, it was also shown that WASp overexpression results in Bruton’s Tyrosine kinase expression.  Although the experiments are well designed with appropriate controls, the study is not very novel as the role of WASp in actin cytoskeleton regulation in a variety of cell types has been already well established. Further experiments demonstrating the role of WASp in various aspects of erythroblast differentiation are required. The authors need to address these concerns listed below.

Major points:

1.	How does WASp deletion affect erythroblast differentiation? Does it impair enucleation or other organelle clearance from the cells?
2.	It has been shown that WASp overexpression results in Bruton’s Tyrosine kinase expression. What are the downstream targets of this kinase in the erythroblasts and how does this upregulation of Bruton’s tyrosine kinase affect erythroblast differentiation?
3.	WASp helps to regulate the assembly of Arp2/3 complex and thereby helps in dendritic branching of F-actin filaments. Are there any differences seen in the dendritic branching structure of the actin cytoskeleton in the Was knockout MEL cells? Are their differences in the rates of actin assembly and turnover?  Live cell actin dynamics assay with fluorophore tagged-Lifeact could be carried out in the WT and knock out cell lines along with rescue experiments with WASp.

Additional point:

1. The authors should provide quantification and statistical analysis of the western blots and immunofluorescence images in 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: CRISPR/CAS9-MEDIATED DELETION OF THE WISKOTT-ALDRICH SYNDROME LOCUS CAUSES ACTIN CYTOSKELETON DISORGANIZATION IN MURINE ERYTHROLEUKEMIA CELLS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors are correct to acknowledge the inherent problems with quantifying western blots that use chemiluminescence and film for detection, the absolute values aren't reliable. However, quantification gives a better view of the reproducibility between experiments than simply telling us that the presented results are representative of independent replicates, even if the takeaway message is only about trends rather than absolute values. For this reason, I strongly recommend that the quantification of the western blots be included in the primary figures. Similarly, the quantification of the image analysis should also be included in the primary figures. There is little point to include these data in the rebuttal for reviewers only, it doesn't serve the scientific community at all. We all want to know how reproducible results are, not just to see examples.


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: CRISPR/CAS9-MEDIATED DELETION OF THE WISKOTT-ALDRICH SYNDROME LOCUS CAUSES ACTIN CYTOSKELETON DISORGANIZATION IN MURINE ERYTHROLEUKEMIA CELLS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Acceptable.
Experimental design: Acceptable.
Validity of the findings: I was pleased to see that the authors had quantified their western blot and immunofliuorescence data.  However, the quantification of the western blot and immunofluorescence data has only been provided in two Supplemental figures that are labeled as for the Reviewers.  In addition, the data  in the bar graphs have not been analyzed for statistical significance, and no P values are determined. This data must be included in the main article and P values must be provided.  This data MUST be presented in the manuscript proper to allow the readers to assess the validity of the authors conclusions.

The argument that western blots are non-linear and thus it is not useful to provide quantification is not valid.  If the westerns performed by the authors are non-linear then none of that data is robust, and the authors would need to modify the protein loading of the samples and repeat the experiments under conditions where the blots are linear.  However, I will assume that they are approximately in the linear range, and that the authors can perform the quantification, as in fact they have shown in the Supplemental files.  The authors can of course discuss the issues with westerns, if they choose.
Additional comments: No 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: AWARENESS, BEHAVIOR AND ATTITUDES CONCERNING SUN EXPOSURE AMONG BEACHGOERS IN THE NORTHERN COAST OF PERU
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please see comments below.
Experimental design: Please see comments below.
Validity of the findings: Please see comments below.
Additional comments: General comments
=============
This manuscript aims to describe the awareness, behavior and attitudes concerning sun exposure among beachgoers in the northern coast of Peru. I would like to congratulate the authors for their efforts in planning the research and writing this manuscript.

Specific comments
=============

Major comments
---------------------
1. Additional detail about the methods used would benefit his paper. My understanding is that there a checklist for reporting cross sectional studies (STROBE Statement), including all the recommended details on the STROBE statement would be useful.
2. I appreciate the restrictions in terms of length of the manuscript. However, I suggest that you improve your introduction by detailing the following:
	2.1 In line 53, there is a mention to skin cancer trend. I would recommend highlighting in that first sentence what is the trend and then going to specific details by country.
	2.2 A brief summary of previous studies would benefit the description at lines 58- 61 to provide more justification for your study (specifically, you should expand upon the knowledge gap being filled)
3. Given the goal of changing sun-safety behaviour in recreational settings, I suggest providing more background for this in your introduction. Rodrigues et al (Annals of Behavioral Medicine, 2012, V45, p224; JMIR research protocols, 2017, V6)) have shown the potential of behavioural interventions to promote sun protection in recreational settings. Please consider adding these details to your introduction section.
4. The results section needs more detail. I suggest that you improve the description at lines 124- 128 to provide more detail of what these figures mean in terms of the knowledge about sun exposure.
5. The discussion section seems to be missing information on what are the implications of your findings. Additional detail would help put this study in perspective for future interventions/prevention strategies.

Minor comments
---------------------
6. The English language should be improved to ensure that an international audience can clearly understand your text. Some examples where the language could be improved include lines 38 and 198– the current phrasing makes comprehension difficult.
7. The abstract could be improved by revising the conclusions section. The first sentence is still related to results and a statement about implications of this study might be a better description.
8. The citations in text seem to be incorrect at times (line 170, 175). I would suggest checking these and your list of reference thoroughly.


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: AWARENESS, BEHAVIOR AND ATTITUDES CONCERNING SUN EXPOSURE AMONG BEACHGOERS IN THE NORTHERN COAST OF PERU
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The present study has investigated sun behavior among beachgoers. An important topic as skin cancer is a big problem worldwide. The manuscript is well-structured. The survey is based on a questionnaire.

My biggest concern with this study is the quality of this questionnaire especially the part about “knowledge about sun exposure”. These questions should deal with basic knowledge about sun exposure and skin cancer. However, I find it very difficult to answer these questions and the correct answers are not provided in the article. I think the weaknesses of these questions should be discussed in the discussion. E.g. I find the following questions difficult to answer:

-	“If I use total sun block I can sunbath without any risk”
To use “total sun block” is a very speculative situation. There is no sunscreen that blocks all radiation and no sunscreen users apply sunscreen to their all their exposed skin without missed area. The word “total sun block” is not a good word to use.

-	“Avoiding the sun at young ages (under 18 years) reduces the risk of skin cancer by 80%”
What is the right answer to this question? How do you know?

I would not recommend use of this part of the questionnaire again.

I have a few additional comments:
-	What is meant by ”negative attitudes” in the following sentence in the abstract? “Negative attitudes were more frequent in men and women presented better”.
-	Figure 1 could be omitted.
-	In Table 2 it is not clear what questions the participants had answered.
-	What year are your data collected?
-
Thanks for the opportunity to read this interesting 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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: Additional information should be added on how accurately the MAC-T and BME-UV1 cells mimic mammary gland cells assayed ex vivo from cattle.
Please include some information to place into context the suggestion that MAC-T are myoepithelial cells and BME-UV1 are luminal. Where are these cells placed in the intact mammary gland? Do they differ in lactating and non-lactating glands? How relevant are they to the process where TGFb1 may have an impact in vivo?
Validity of the findings: If no existing literature exists then a suggested question for the future would be to address how closely these mammary cell lines compare to those directly isolated or identified in the bovine mammary gland. This would provide very useful information as to whether they are useful to assess the utility of approaches such as TGFb1 inhibition for extended lactation/improved milk yield.
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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comments
Validity of the findings: no comments
Additional comments: This is a really interesting review regarding the impact of TGFb1 on two different bovine udder-derived cell line.
As this topic is probably not the most know to readers of Peerj, could the authors potentially re-structure the lay out by starting with
1) Describing phenotypic differences of Mac-T vs BME-UV1
2) Describe TGFb1 signalling in general
3) Then follow with the specifics

What this reviewer is missing a bit is the notion of whether this is exclusively specific for TGFb1, or may also be similar for TGFb2, is it important where the cell lines are coming from (as in lab sources of either lines used in the different articles cited), and whether the antibodies to CD mmarkers tested were bovine specific.


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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: - Is the review of broad and cross-disciplinary interest and within the scope of the journal?
This mini review is not of broad and cross-disciplinary interest.

-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.)?
The field has not be reviewed recently, but the manuscript presents not enough up-to-date information on the subject described.

-Does the Introduction adequately introduce the subject and make it clear who the audience is/what the motivation is?
Yes.
Experimental design: -Is the Survey Methodology consistent with a comprehensive, unbiased coverage of the subject? If not, what is missing?
Yes.

- Are sources adequately cited? Quoted or paraphrased as appropriate?
Yes.

- Is the review organized logically into coherent paragraphs/subsections?
Yes.
Validity of the findings: - Is there a well developed and supported argument that meets the goals set out in the Introduction?
No,see general comments.
 -Does the Conclusion identify unresolved questions / gaps / future directions?
No,see general comments.
Additional comments: The mini review (#29236) by Mitz Ch.A. and Viloria-Petit A.M. describes the apoptogenic and anti-proliferative effect of TGF-beta1 in two bovine mammary epithelial cell lines: MAC-T and BME-UV1, and is based mainly on data described in scientific literature in years  1995-2005, although the chapter regarding the phenotypic differences in MAC-T vs. BME-UV1 cells has been created using also more recent findings (2010-2016). The authors have undertaken this topic due to the continuous need of dairy industry to extend lactation persistency in cows,  and thus search for possible new solutions to reach this goal. Such solutions should be alternative to exogenous administration of recombinant bovine growth hormone to dairy cows. The Authors hypothesize that inhibiting TGF-beta1 signaling pathway may be helpful in extending lactation and increasing milk production, due to the fact that this cytokine exerts apoptogenic and anti-proliferative actions in bovine MECs. This hypothesis I quite risky considering the fact that no in vivo studies have been conducted to test it, and it is impossible to predict what will happen if TGF-beta1 becomes inhibited by e.g. exogenous factors, considering the fact that this cytokine plays an important regulatory role  in many different tissues.
Important issues decreasing the value of this review:
1.	The studies commented in this mini review were done at the beginning of XXI century using commercially available, immortalized bovine MECs lines. There is no data on the effect of TGF-beta1 on primary bovine MECs, isolated directly from the udder, and thus maintaining more properties of mammary epithelium within the mammary gland. This should be taken under consideration by the authors and commented in this review.
2.	Nowadays a growing body of evidence point at important interactions between  the mesenchymal and stromal tissue of the mammary gland during its development and remodeling, and TGF-beta has been shown to play an important role also in regulation of the stromal compartment in bovine mammary gland (Musters S. et al., J Dairy Sci. 2004;87(4):896-904.; Andreotti CS, et al.,Res Vet Sci. 2014 Feb;96(1):5-14). This review would be more interesting if the authors presented more comprehensive approach discussing not only the available data of in vitro studies, but also studies on animals (in vivo studies on bovine mammary gland) to show pros and cons of using TGF-beta inhibition as a potentially new method of extending the lactation in cattle.
3.	In addition, the review is lacking information about the noncanonical pathways induced by TGF-beta1, which may be of great importance. The reader of such review should be informed that aside from the classic, canonical Smad-mediated pathway, other signaling pathways, mediated by Wnt or MAP kinases may also be activated by this cytokine, and thus inhibition of TGF-beta1 activity may result in other consequences beside inhibiting MECs apoptosis.
Overall, the article is written in clear written in professional language, but in the current form it is lacking up-to-date findings regarding the role of TGF-beta1 in bovine mammary gland, thus it may be of limited interest.


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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors have addressed this reviewers concerns and 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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Revised version of previously submitted manuscript
Experimental design: Revised version of previously submitted manuscript
Validity of the findings: Revised version of previously submitted manuscript
Additional comments: The authors have answered all my comments raised in a very satisfactory manner


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: TGF-BETA SIGNALLING IN BOVINE MAMMARY GLAND INVOLUTION AND A COMPARATIVE ASSESSMENT OF MAC-T AND BME-UV1 CELLS AS IN VITRO MODELS FOR ITS STUDY
Review round: 2
Reviewer: 3

Output:
Basic reporting: With reference to remarks in my first review:
- the manuscript has been largely improved in terms of its broad and cross-disciplinary interest. It is now within the scope of the journal.
- Furthermore,  the manuscript was enriched by substantial amount of  up-to-date information on the subject described.
- Introduction has been improved.
Experimental design: -Is the Survey Methodology consistent with a comprehensive, unbiased coverage of the subject? If not, what is missing?
Yes.

- Are sources adequately cited? Quoted or paraphrased as appropriate?
Yes.

- Is the review organized logically into coherent paragraphs/subsections?
Yes.
Validity of the findings: - Is there a well developed and supported argument that meets the goals set out in the Introduction?
Yes
-Does the Conclusion identify unresolved questions / gaps / future directions?
Yes
Additional comments: The present version of the manuscript by Mitz and co-workers has been substantially improved following the suggestions of the Reviewers. In my opinion this version is well structured, informative and easy to follow, therefore I recommend the manuscript for publication in Peer J journal.


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: OPIOID DISTRIBUTION TRENDS (2006–2017) IN THE US TERRITORIES
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: not applicable
Validity of the findings: See below under general comments
Additional comments: General comments
This article is basically reporting bulk distribution of opioids in 3 US territories and 2 US states – corrected with morphine equivalents and then for population.   It doesn’t really do much more than describe these data (are these data publically available anyway?).  It isn’t able to say whether changes are due to increased/decreased doses or more/less people?  Have recent decreases due to reduced prescribing led to more illicit opioids?  Can any changes be linked to any particular events ?
The key to making this paper interesting would be to have some outcome data such as overdose/poisoning deaths in each territory.   At the moment it’s a small amount of data and a lot of speculation. If they can’t add more interesting data it would be much better as a brief report with very little introduction or discussion.  The discussion is way too long for such a thin paper.
Specific points
The exclusion of American Samoa because of low volumes makes no sense.
Buprenorphine is used as an analgesic as well as for opioid dependence.
Opioids are usually corrected to oral morphine equivalents (ie conversion rates for IV, dermal and oral preps are different) – it is unclear if that has been done in these MME.
It would make more sense to report pop adjusted OTP and analgesic opioids separately – the % of total analysis (Fig 2) is less relevant than the actual rates of each.


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: OPIOID DISTRIBUTION TRENDS (2006–2017) IN THE US TERRITORIES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The language used is mostly clear and unambiguous. A couple of recommendations are suggested below to help increase clarity to readers.
Line 181-182: “Similarly, hospitals distributed 26.6% in Guam, …”. To increase clarify it would be better to say distributed 26.6% of opioids in Guam,….
Figure 1: Caption of Figure 1 needs to be rewritten to increase clarity. It’s missing (D).
Good background provided with sufficient reference.
Line 97-98: According to table provided US Virgin Islands is populated by mainly Black/African American which is different from comparison States. This need to be highlighted as drug misuse in Black/African American population is known to be different from White populations.
The following is recommended to support/enrich discussion:
Line 227: Meperidine decreased distribution more likely to be due to change in guidelines than awareness of opioid addiction crisis. (Where has all the meperidine gone? Dobbins E, Nursing2010: January 2010 - Volume 40 - Issue 1 - p 65–66, doi: 10.1097/01.NURSE.0000365924.16631.a4).
Line 253-254: Needs to correlate low methadone use to the absence of NTP in Guam.
The tables are clear however for table 1 it would be interesting to make it easier to visualise opioids change. Maybe different colour for up and down changes (or colours in a scale). Table 2: Missing parenthesis – Hawaii oxycodone)
Figure 1A to C are well designed and easy to understand. But figure not clear what it is representing, is it mean of overtime (2006-2016) MME per person? Better to use scatter plot instead of bar graph.
Figure 2: It would be interesting to have both graphs under the same Y-axis. It would make the graphs easily comparable.
Experimental design: Good experimental design and in the scope of the journal. It clearly fills an identified knowledge gap.
When calculating MME per capita was only 2010 census used? If so, this needs to be added as a limitation to the research. Population numbers can change considerably in 10 years and using the same population to calculate MME per capita over 10 years can introduce an error to this data set.
Validity of the findings: The findings are valid and benefit to literature clearly stated.
Data is robust and raw data can be accessed online.
Line 144: Why paired t-test used to compare the MME per capita by location? It is not clear to me why using paired t-test. Although data is coming from the same database, it is about different sample groups.
Interesting conclusion highlighting answer to the research question and important points which could explain differences found.
Additional comments: This is an interesting manuscript, well written and straight to the point. More information on how some of the differences encountered could be related to specific policies and changes in the territories would have been appreciated.


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: ESTIMATING THE INCIDENCE AND DIAGNOSED PROPORTION OF HIV INFECTIONS IN JAPAN: A STATISTICAL MODELING STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: English: good
References/context: good
Structure: good
Data: publicly available (not clear if location is provided)
Figures/tables quality: good
Self-contained: yes
Experimental design: Scope of the journal: adapted
Research question: important and interesting, with real-world implications
Methods: rigorous and adapted to the research question, well described
Validity of the findings: Statistical analysis: sound and robust
Conclusions supported by analysis: yes
Additional comments: Prof. Nishiura adapts a method based on McKendrick and Morison's approach to estimate the incubation period during the 1919 flu epidemic to address a problem related to specificities of the surveillance of HIV in Japan. Indeed, surveillance of HIV in this country relies upon 1) reports of new HIV diagnosis among people who undertook voluntary testing and 2) reports of new AIDS cases among people that were not diagnosed as HIV-infected. This data does not allow the direct estimation of HIV incidence using traditional back-calculation methods, as the incidence of all AIDS cases is not available.

The proposed method method is generative and closely matches the mechanisms of HIV surveillance and evolution towards AIDS (and this could be more emphasized in the paper). It allows the estimation of past HIV incidence in a convincing way.

My only concern about this work is the handling of uncertainty. It is surprising to me that the data allows e.g. the estimation of the cumulative number of HIV infection with such a narrow confidence interval (29,059-30,167). Seeing Fig. 3, this may be related to overfitting. In that case, using a negative binomial distribution instead of a Poisson might have been preferable. The author should provide a sensitivity analysis using a negative binomial distribution, or support his use of the Poisson distribution.

+++++
Minor comments

l121: while the model is well described both in equations and in Fig. 1, the text is a bit confusing as to
which population does compartment h(t,tau) represent precisely. Line 121, it is written that it is "individuals who are either undiagnosed or in the incubation period" and it seems to me that it should be "individuals who are both undiagnosed *and* in the incubation period", as either of these events would remove an individual from the h compartment. The terms referring to HIV diagnosis and AIDS diagnosis should also be more coherent, as in the current state of the manuscript the authors sometimes use "incubating" or just "diagnosed".

l130: "rho(tau)" should be "f(tau)"

l135: I don't understand why the upper boundary of the second integral is x, shouldn't it be s? (also in following equations)

l136: It would maybe make the equations easier to read to use h(t,s) instead of the full expression in eq. 5 and  6 as h(t,s) is defined in eq. 4.

l150: there is no justification as to why a step function has to be used, is that related to data structure?

l160: very minor, it is not clear in the notation that k is an exposant

Fig2F: this plot is very confusing, the author should consider using a different coding, or maybe remove part of the information displayed.

l237: the predicted values obtained with 3 different median incubation periods are very close, although they lead to marked differences in the estimated incidence of HIV (Fig 2C). Could the author provide an explanation about this? Also, I would have liked to see confidence intervals on Fig. 3A and 3B, although I understand that it is not convenient with only one figure.

l288: the comparison of the results to Iwamoto (2017) should be a bit more detailed and discussed.


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

Input:
Title: ESTIMATING THE INCIDENCE AND DIAGNOSED PROPORTION OF HIV INFECTIONS IN JAPAN: A STATISTICAL MODELING STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: No main comments, otherwise please see general comments below.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This is a timely, useful and practical analysis of relevance for characterizing the HIV epidemic in Japan and for informing HIV policy and programming in this country. The analysis is thoughtfully and finely conducted and captures the essential features to address the research questions, and nicely using a parsimonious model. The complexity of the analytical methods is appropriate for the research questions and nature of available data.
I have few remarks in the spirit of enhancing the article’s contribution to the literature and its impact. These can be addressed through a minor revision.
1)	Introduction Line 69: I think it would be useful here to indicate the limitation of potential under-sampling in blood donor data. For example, HIV testing for blood donors in the US substantially underestimates actual HIV prevalence in the population. Accordingly, the Iwamoto et al 2017 analysis could have underestimated the prevalence of infection, and thus overestimated the fraction diagnosed.
2)	Introduction Line 86: In relation to estimation methods using HIV surveillance data, there are currently different developed methods, the use of which depends on the nature of available data. For example, the ECDC HIV Modelling Tool provides an example of available methods that are useful with availability of CD4 data (see van Sighem et al, Euro Surveill. 2017). Meanwhile, the method discussed in the present paper is useful and fitting in consideration of Japan’s data. I suggest more discussion of this issue, such as along the lines discussed here.
3)	Methods Line 121: The statement “either undiagnosed or in incubation period” seems confusing to me. I think it is just simpler to state it as “undiagnosed HIV infected persons who have not developed AIDS”, or something along this line.
4)	Results Line 235: It is interesting that there was no improvement in the frequency of diagnosis among females. This is worth discussing in the Discussion section and providing potential explanation(s). For example, could this relate to how females are being typically diagnosed, say through routine testing rather than VCT services?
5)	Results Line 264: The higher diagnosis rate among females compared to males is worth discussing/explaining in the Discussion section (actually it is paradoxical and counter-intuitive). Why is this the case, even though the focus of VCT services in Japan is to target MSM, rather than women/heterosexual populations? Could this relate to how females are being actually diagnosed (say following the diagnosis of their spouse/partner)?
6)	Discussion: A gap in the Discussion section is linking the results to the HIV epidemic among MSM in Japan, which is the core of the epidemic. Much of HIV incidence is believed to occur among MSM, but the paper hardly discusses this issue. What is and how did HIV prevalence among MSM change over time in Japan? Are there other and contextual data of the epidemic among MSM that can explain the results of this paper?
7)	Discussion: A related point, which is a limitation of the present study, is that the modeling method is on the descriptive indirect side, and does not directly model HIV transmission, such as among MSM, to generate such results. It is worth suggesting in the Discussion section that future work should build on this work to model HIV transmission in the population, with focus on the role of MSM.
8)	Discussion: Another potential work that could build on this study is to attempt to use other data, such as CD4 count data among diagnosed infections, to generate similar results and examine the differences if any. Such application can also be useful to generate inferences about HIV cascade, which is critical to inform HIV response. I suggest discussing this point.
9)	Discussion Line 304: The declines in HIV incidence is probably best explained by examining HIV transmission dynamics among MSM. I am aware of an earlier work to model the HIV epidemic among MSM in Japan (regrettably unpublished), that reached similar conclusions about the trend in HIV incidence, though through a very different approach. This is also supported by the scale-up of NGOs working with MSM in Japan. The authors may want to discuss this issue in some detail, as it provides important epidemiological context.
10)	Please check the citations and references. For example, I do not seem to find the bibliography entry for the citation of Hall et al 2008.
11)	Typo in Results Line 224: probability of diagnosis “has” monotonously improved…


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: ESTIMATING THE INCIDENCE AND DIAGNOSED PROPORTION OF HIV INFECTIONS IN JAPAN: A STATISTICAL MODELING STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Prof. Nishiura modified his paper according to the reviewers' recommendations. The new version is mostly satisfying, only one point remains questionable in my opinion.

The author compared his findings based on a Poisson distribution with an alternative approach using a negative-binomial distribution (line 249 of the new manuscript). I fail to understand how the AIC could be 847.8 using a NB distribution versus 650.7 with a Poisson distribution. If the Poisson is indeed more adapted to the question, shouldn't the overdispersion parameter of the NB distribution go to infinity so that the NB reduces to Poisson, and the AIC a bit higher because of the additional parameter? Could this very large difference between Poisson and NB be caused by a local optima found by the maximum likelihood algorithm? Adding a short clarification of this point would be enough to accept the manuscript for publication.

Other minor comments:
- citing my paper as a reference for approaches using negative binomial distributions is a bit excessive. Considering that I am reviewing this manuscript, I would be more confortable if this citation was replaced by a more general article on overdispersion for count data or simply 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: ESTIMATING THE INCIDENCE AND DIAGNOSED PROPORTION OF HIV INFECTIONS IN JAPAN: A STATISTICAL MODELING STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revision addressed my comments.
Experimental design: The revision addressed my comments.
Validity of the findings: The revision addressed my comments.
Additional comments: The revision addressed 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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 1
Reviewer: 1

Output:
Basic reporting: NO comments
Experimental design: Authors should mention the period of time selected for the references
Validity of the findings: NO comments
Additional comments: In general, the article is well written. the relevance of the references is clearly identified and the order is understandable. However, I consider that a broad section should be added and should include why most of the described adjuvants have not gone through clinical trials?, which are the biggest limitations? A deeper analysis of them with possible alternatives are necessary .

It would be interesting to include a table indicating the test phase in which each of them is located, from preclinical to clinical phases.


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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 1
Reviewer: 2

Output:
Basic reporting: This review is broad and cross-disciplinary interest. The authors present detailed review on adjuvants and vaccine strategies.
Experimental design: Survey Methodology is consistent with a comprehensive coverage of the vaccine immunology.
Validity of the findings: Conclusion addresses gaps in knowledge and future directions.1.	Please discuss protective antigen of B. anthracis which can also be used as a cytosolic delivery agent of polypeptides, nucleic acids and other noncanonical biomolecules into the cell. Please refer PMID: 28137237.
2.	Line 228 “In addition, this…. No allergic reactions to TLR receptor expression on the surface of immune cells”.  I don’t understand do you mean allergic reaction to immune cells?
3.	Since the authors covered Tuberculosis, the immune response to antigens such as ESAT-6 and Mycobacterial lipid antigens should also be mentioned Please refer PMID: 28137237, 12882829, 17351619

4.	Please look for the formatting and English mistakes. I will cite few example.
a.	Line 68-70: This sentence is wrong. Thus, an…….adaptive immune response”
b.	Line 89-92 “These epitopes ….their surface”. Epitopes composed of B- cell ? Please reframe sentence.
c.	Line 150.. “David et al…” Reference is wrong.
d.	Line 205 “chemokines(Fu et……”  and Line 244 “activity (Deng…”Please check formatting.
e.	Line 222 “production of INF-…..” It should be IFN
f.	Font title and all Capital letters at some place.
g.	Line 374 Heat Labile toxin is referred as LT. LT is often used for lethal toxin of B. anthracis. Please change the abbreviation.
h.	Line 446 “The immunogenicity of adjuvanted and non-adjuvanted”. I am not sure adjuvanted is correct term here. Please restructure sentence.
i.	Line 278-279 “HBcAg can……epitopes” Sentence is not clear.
j.	Figure 2 legends : mediates the production of type I IFNs, such as IFN a/b/g.
k.	Table 1: “Described above” Please describe properly. Also add a column for references.

5.	Please include chemical structure/formula of different Pam lipopeptides in table.

6.	Table 3. Please include citation of original references for MALP2(PMID:
9166424). Please check other places too.
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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article is well written and grammatically acceptable. However the authors have failed to add many recent references in the area which will be very beneficial to the readers. Further, the manuscript lacks the details about the mechanistic action of various adjuvants. The reference styling is good and the tables and figures are drawn right. However, there is no labelling and captioning of the diagrams. The authors shall also cite the sources where from the figures have been taken and wherever required, the necessary permissions must be sought.
Experimental design: The authors need to study further the left over classes of adjuvants and to add those with proper citations.
Validity of the findings: The work is relevant and meaningful addition to already existing literature.
Additional comments: The paper should be revised thoroughly revised with the above considerations in mind.


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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 1
Reviewer: 4

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Lei et al., review the recent literature related to adjuvants for peptide vaccines focusing in 4 general groups: Pater Recognition Receptors, Virus Like Particles platforms, Bacterial Toxins and other organic and inorganic nanoparticles.  The review is of general interest for life sciences scientist and academics (and in particular, to immunologist and vaccinologist), to introduce and broadly inform on this particular area of the adjuvants field. I have not found similar reviews at least in the past 2 years; I therefore consider this article will be a useful update in the field. The article is clearly written, figures and tables are relevant to the content of the article. Most of relevant prior literature is cited, however, I consider review will importantly benefit from including other important adjuvants that have been important to develop vaccines against several virus and bacterial induced diseases. For the section of PRRs agonists, the inclusion of adjuvants based on Outer Membrane Proteins from Gram-negative bacteria (i.e. Neisseria, Salmonella) will be of interest. For the VLP section, the inclusion of plant virus VLP platforms (i.e. Tobacco and Papaya Mosaic Virus) will enrich the review. In addition, the inclusion of outer membrane vesicles and Generalized Modules for Membrane Antigens (GMMA) will also benefit the text. Finally, I recommend the inclusion of chitosan as adjuvant in the section of organic and inorganic nanoparticles.
Please check references of Azmi F., on line 569 and 571 seems repeated.


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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The article is written in clear and unambiguous, professional English used throughout. Literature is well referenced.
Experimental design: Sources are adequately cited.
Validity of the findings: Conclusion are well stated and provide future directions.
Additional comments: I will like to congratulate authors on doing commendable job in revising manuscript. I am overall satisfied with the current version.


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: APPLICATION OF BUILT-IN ADJUVANTS FOR EPITOPE-BASED VACCINES
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Authors have properly responded to almost all my previous suggestions, the manuscript is importantly improved,  however, is still missing the brief description of Papaya Mosaic Virus vaccine adjuvant platform and the description of Salmonella porins as adjuvants/immunomodulators as requested previously. (in particular the description of Salmonella porins used as adjuvant will strength the message for figure 6). In addition,  the repeated references in the manuscript have not been corrected:
771 Azmi, F., Ahmad, F.A., Skwarczynski, M., and Toth, I. 2014a. Recent progress in adjuvant discovery for peptide-
772 based subunit vaccines. Hum Vaccin Immunother 10:778-796.
773 Azmi, F., Ahmad, F.A., Skwarczynski, M., and Toth, I. 2014b. Recent progress in adjuvant discovery for peptide-
774 based subunit vaccines. Hum Vaccin Immunother 10:778-796.

I consider these will help to further improve this review and extend its broad and cross-disciplinary interest.


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: COMBINING LEGACY DATA WITH NEW DRONE AND DGPS MAPPING TO IDENTIFY THE PROVENANCE OF PLIO-PLEISTOCENE FOSSILS FROM BOLT’S FARM, CRADLE OF HUMANKIND (SOUTH AFRICA)
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article follows a recent trend in which paleoanthropologists and vertebrate paleontologists are increasingly using sophisticated tools, methods, and datasets from the Geographic Information Sciences (GISciences) in their explorations for and analyses of fossil-bearing deposits around the world.  It would be appropriate for the authors to include citations to some of this recent work in paleoanthropology and vertebrate paleontology (published in journals like JHE, JVP, AJPA, Evol Anthro and others), and to thereby place their work within this historical context and evolving tradition within our disciplines. (Archaeologists have been using sophisticated geospatial tools for much longer than paleoanthropologists).  The writing is clear and unambiguous, although many small suggestions and edits have been included on the uploaded pdf, and these will improve the tone, clarity, and the sense of the manuscript.   The Figures and Tables are relevant and high quality and appropriately labeled for the most part.  Figure 1 depicts what looks like a DEM of South Africa, yet the color ramp is not defined.  Figure 1 also includes a topographic map yet doesn’t label the contour interval.  Please include the map projection or UTM zone for Figure 2 below the scale bar.
Experimental design: The design of the research is clear, its aims are well within the scope of PeerJ, and its methods are rigorous, utilizing state of the art geospatial techniques to clarify long-standing confusion as to the location of fossil sites at Bolt’s Farm. The Methods are described in sufficient detail and the Supplementary Data are relevant and provide added value to this work (esp. the detailed faunal lists and notes concerning biochronology).
Validity of the findings: The findings are valid and help to clarify long-standing confusion as to the identity and location of fossil-0bearing sites at Bolt’s Farm, resulting from a long and episodic history of work there by different teams separated by long temporal gaps. The authors have done the field a service by restudying the collections from Bolt’s Farm at different institutions on different continents. The biochronological conclusions are necessarily limited in their scope and finality, but this is mostly a result of limitations in the data themselves rather than a problem with the current research design or execution.
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: COMBINING LEGACY DATA WITH NEW DRONE AND DGPS MAPPING TO IDENTIFY THE PROVENANCE OF PLIO-PLEISTOCENE FOSSILS FROM BOLT’S FARM, CRADLE OF HUMANKIND (SOUTH AFRICA)
Review round: 1
Reviewer: 2

Output:
Basic reporting: Some minor edits are suggested on the pdf itself in terms of grammar and wording. It is a well written paper with relevant information and high quality figures that will certainly provide guidance for future scientists that are interested in field work at this site or the fauna that has been recovered from there.
Experimental design: All of the previous maps of the site, as well as any potential sources of information (like field journals) were accessed for this study, giving the sense that this truly is an exhaustive description of the site. Methods for how various maps were combined with current aerial photography were clear.
Validity of the findings: The recommendation of using the original names of the pits when possible is useful and as a great example of how research at several Cradle sites could move forward with more information that is better translated across research groups. Errors in naming are likely seen at many sites that have had different research projects over the years, and this paper serves as a good example of potential “best practices” in moving forward with rectifying some of the errors. It’s possible other researchers who have worked at the site or will in the future may not agree with the recommendation, but at minimum this paper illustrates which pits are named or renamed by various projects. The data used for the biochronology estimates seem robust and will be interesting in light of new absolute dates coming out of the Cradle.
Additional comments: This paper, “Combining legacy data with new drone and DGPS mapping to identify the provenance of Plio-Pleistocene fossils from Bolt's Farm, Cradle of Humankind (South Africa),” revises the map and names of Bolt’s Farm assemblages, as well as provides maximum and minimum dates via biochronology for those assemblages when possible. The background provided about the excavation history of Bolt’s Farm is valuable, and the tables of site names and related information through time is an extremely important contribution to the field.

Though it is mentioned in the paper that this study did not have a goal of comprehensive faunal lists for each site within Bolt’s Farm, such a contribution would certainly be valuable as a follow-up in the future.


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

Input:
Title: USING LIDAR TO ASSESS THE DEVELOPMENT OF STRUCTURAL DIVERSITY IN FORESTS UNDERGOING PASSIVE REWILDING IN TEMPERATE NORTHERN EUROPE
Review round: 1
Reviewer: 1

Output:
Basic reporting: Clear and unambiguous, professional English used throughout.
Experimental design: Here is where it comes my main concern to the methods side of this manuscript. Are you using all 2 grid cells data (10 x 10-meter n=7319 and 30 x 30-meter n=1002) be used in Principal Component Analysis (PCA)? You have a data set where you have to build the model now. In the future, you will get new samples that you have to predict using that prebuilt model. PCA is an unsupervised method although you may split data into the Training set and Test set to be sure it will generalize. It was no considered in MS.
Validity of the findings: The conclusions should be appropriately stated, should be connected to the original question investigated, and should be limited to those supported by the results.
Additional comments: This manuscript titled “Using lidar to assess the development of structural diversity in forests undergoing passive rewilding in temperate Northern Europe” combined LiDAR and Principal Component Analysis (PCA) to address structural diversity of forests. I find the topic of the paper very interesting and potentially useful by different users. Forest management is increasingly important for many purposes such as climate adaptation, modify stand structure and maintains biodiversity etc.

In this sense, providing a monitoring and interpreting structural development  method for forest structural homogeneity and heterogeneity is very much welcome. This study is generally well written. However, I believe that the MS needs some clarifications and improvements before being considered for publication.

The main points to be addressed are:
Page 15- Lines 248-249. Stensballe area is no mention stand age range. In your manuscript, Stensballe was classed both into Mid-aged and Old stand (Table 1). In Figure 3., Stensballe is not consistent with your groups. The PCA outcomes did not indicated a good classifying for the Stensballe. You divided Vorsø into seven areas according to the history of the island but the development of structural diversity may be not consistent with your groups. In this sense my question is which is near the Stensballe real structure status ?? (management group or PCA outcome?)

Page 16- Lines 278-286. Here is where it comes my main concern to the methods side of this manuscript. Are you using all 2 grid cells data (10 x 10-meter n=7319 and 30 x 30-meter n=1002) be used in Principal Component Analysis (PCA)? You have a data set where you have to build the model now. In the future, you will get new samples that you have to predict using that prebuilt model. PCA is an unsupervised method although you may split data into the Training set and Test set to be sure it will generalize. It was no considered in MS.

Page 26- Lines 512-516. The eigenvectors of the PC3 of PCA_10m (Table S5.) shows the Percentile05_ExclGround and Percentile10_ExclGround loading greater than CanopyCover. It may not say PC3 of PCA_10m meaning that it has the most open canopy layer.

Table S5. The eigenvectors of the first five axes of PCA_10m show the PC1 have the highest proportion of variance (about 0.654). The PC2-PC5 are relative vary low proportion of variance (about 0.088-0.032). For assessing the development of structural diversity, the outcome may not be sufficiently.

Maybe you can try count pixels value (proportion) at different PCA axis and plot map. To show structural diversity consist of different PCA axes values distribution in forests.

Additionally I’ve found some minor concerns:
Figure 1, 4,5 is can't easy to read. The site location labels should be consistent with Figure 1.

Figure 3. The x-axes labels should be consistent with Figure 1.


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

Input:
Title: USING LIDAR TO ASSESS THE DEVELOPMENT OF STRUCTURAL DIVERSITY IN FORESTS UNDERGOING PASSIVE REWILDING IN TEMPERATE NORTHERN EUROPE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors used air-borne lidar point cloud-derived metrics treated by PCA (principal component analysis) to analyze a unique Danish rewilding site, and try to find the relationship between the data and the existing vegetation development. The method is very straightforward but the manuscript is hard to read and understand.
Experimental design: The field experimental design is fine.

It’s difficult to understand the methods based on the description. I suggest the authors provide a figure to visualize the grid sampling method from line 254 to 268.

From line 261 to 264, it said "The 25 metrics were found in existing lidar literature and selected on a criteria of being used in comparison to manual ground surveys in previous studies of for being alike such metrics (Fig. S1)." However, I didn’t find what the "25 metrics" are in Figure S1. I didn’t find what the “25 metrics” in other part of this manuscript, neither.
Validity of the findings: It’s very hard to see which field site is young or old in the figures and tables. For example, for young stands 1/2/3, the abbreviation Y1/2/3 can be easier for readers to identify in figures and table.
Additional comments: I suggest the authors to resubmit the manuscript after major revision.


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

Input:
Title: USING LIDAR TO ASSESS THE DEVELOPMENT OF STRUCTURAL DIVERSITY IN FORESTS UNDERGOING PASSIVE REWILDING IN TEMPERATE NORTHERN EUROPE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Clear and unambiguous, professional English used throughout.
Experimental design: The submission should clearly define the research question, which must be relevant and meaningful.
Validity of the findings: The data analysis should be robust, statistically sound, and controlled.
Additional comments: 1.I thank you for your responses, as you say “this study is interdisciplinary, meaning that it concerns forest ecology and remote sensing aspects at the same time”. Your results are compelling, the effects of different management strategy should be discussed the details in the manuscript. ie. The managed Sten_M/O forest stage is not the same as others due to tree harvest.

2.It’s difficult to understand the explains for relative low proportion of PCA axis (like as PC2-PC5) . The explanation is pretty thin. The Values of PCA_10m axis 5, which is a gradient from bare ground to low and sparse vegetation, subscribing intermediate values to all kinds of taller vegetation. LiDAR data are just in this way. For assessing the development of structural diversity, the outcome may not be sufficiently.

3.The manuscript is written in standard, unambiguous language. If there is a weakness, it is in the statistical analysis (as I have noted above) which should be improved upon before Acceptance.


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: METHOD FOR THE QUANTITATIVE EVALUATION OF ECOSYSTEM SERVICES IN COASTAL REGIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: English was mostly very good – have indicated a few spelling errors in the attached PDF.

The manuscript develops the indicator/assessment literature relatively well. However, the manuscript would greatly benefit by better introducing tidal flats as these are the subject of the paper. Explaining their characteristics and the difference between man-made and natural tidal flats would help those who are not experts with this ecosystem. The Shiosai Nagisa tidal flat pictured in Figure 4, looks more like a concrete structure than what I would think of as an artificial tidal flat. Explanation of the characteristics is needed up front.

The structure of the article works generally and figures are of sufficient resolution. However, there is a significant portion of data that is not presented and hinders adequate peer review. For instance, from the 12 sub-goals of the index the authors propose, only food provision is presented in terms of the data used, and how scores were calculated. This can be presented in a more refined way such as a table or tables, but equations and descriptions should also really be provided. These can be placed in supplementary materials, but should be available.
Experimental design: The research question could be defined more clearly – it is not clear whether the purpose of the index is to compare between man-made flats and natural flats or just assess all different flats.

While some methods are very well developed, others are vague. It would benefit hugely by more detail on each sub-goal, how the scores were aggregated and how they were weighted.

The authors refer to the theory behind setting reference points but then apply the same reference point (highest score across all areas) to every sub-goal. The reference point will be the main driver behind the index and therefore I would recommend development of a clearer methodology for choosing reference point for each goal – about what should be an ‘optimum’ reference point. The conceptual model approach that the authors’ used for investigating resilience and pressures is an excellent way to think about these ideas.
Validity of the findings: It is difficult to truly understand the validity of the results given there is a lack of information presented for each of the goal models. It is necessary, either in the text or in the supplementary, to provide a model for each sub-index. It is not sufficient to present just one sub-index (food-provision).

Food Provision – reference point is highest catch across all years and all sites. This is not necessarily a desirable reference point – this could be a year subject to intense overfishing. Reference point should be related more to sustainability and can differ between sites.

Water Front Use – Scoring SN as 0 because it is a closed facility is potentially unfair. Might be better to leave this out of the water front use assessment?

Discussion would benefit by being less fractured and a more holistic view of the results. What are the most interesting points? In order to validate the index, results should not simply be accepted. Validating indices is extremely difficult but a more nuanced discussion can help to interpret results better; also, composite indicators can always be iteratively improved over time by refining different areas or utilising better data. Some discussion of this would be welcomed (see ‘Navigating uncertainty in environmental composite indicators’). Interpreting your results and things that look odd – i.e. some scores are 0 for some tidal flats and 100 for others – will likely want refinement. As this type of indicator is overly sensitive (all or nothing) and could have a large impact on the overall index.

Given this it is not entirely clear how the index can be used for management. This ties back to the goal of the index and what it is to be used for.
Additional comments: Although this paper is described as being modeled on the Ocean Health Index - it is significantly different. The Ocean Health Index theoretically assesses sustainability of large ocean areas, with multiple habitats and a large range of benefits. Adapting the OHI for this scale is a large change and therefore requires justification for many of the decisions taken. The OHI has strong theoretical backing for many of its goals and therefore by trying to adapt the OHI, it means having to justify clearly all your sub-indices (see other regional OHI papers). It may be wiser to pull-back on the comparisons to the Ocean Health Index given how different this is.


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

Input:
Title: METHOD FOR THE QUANTITATIVE EVALUATION OF ECOSYSTEM SERVICES IN COASTAL REGIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall, the paper is well written and figures and tables are well presented and support the article.  In particular, the introduction Is very well written, thoroughly explained, and flows together quite nicely. There are times where the text can be redundant and a little lengthy which, while I understand the authors want the text to be thorough, can be quite distracting for the reader.  I have included some suggestions below for the introduction as well as an annotated pdf, which is attached.  Some areas of the methodology are unclear and lack appropriate references and this should certainly be addressed.

Suggested chnages to grammar are below for the Introduction with added comments for the remainder of the paper in the annotated pdf....

81 Halpern et al. (2012) proposed the Ocean Health Index (OHI) as a method for
82 comprehensively quantitating ocean health. The OHI defines healthy oceans as those that
83 sustainably deliver a range of benefits to people now and in the future and enables the
84 comprehensive evaluation of not only benefits gained from ecosystems but also social and
85 economic benefits (Halpern et al., 2014). The OHI scores the state of each ocean against its
86 reference point (Samhouri et al., 2012) which enables the
87 degree of achievement of each goal to be evaluated. The OHI also enables sustainability and
88 changes in policy or regulatory systems to be taken into account and can be adapted flexibly to
89 match the quantity and quality of data, as well as changes in geographic scale
90 (Elfes et al., 2014; Halpern et al., 2015; Lowndes et al. 2015; Seling et al., 2015). One key
91 advantage of the OHI over other approaches is that it enables the scoring and quantification of
92 services that are difficult to quantify in economic terms.

93, 94 Remove this sentence since it doesn’t quite flow with the rest of the paragraph…..”Meanwhile, the biodiversity of coastal areas has been severely reduced due to the habitat degradation of numerous organisms resulting from various human activities.”
96:  Remove “As such”
99 Remove “In addition”
103:  In
104: addition, since the effect of the environmental improvement project strongly depends on the
105 ecosystem states, it is necessary to carefully consider the states for appropriate
106 evaluation of the project (Zhao et al., 2016). Moreover, because the ecosystem states after
107 creating artificial tidal flat (or wetland) change over time (Palmer, Ambrose & Poff, 1997;
108 Kuwae, 2005), the effect of the environmental improvement project should be evaluated
for several years following completion.  Therefore, development of an evaluation method that considers the states and changes of ecosystems is also a challenge in evaluating the benefits of the environmental improvement project.
.
Experimental design: The research question seems to be well defined and relevant to environmental issues outlined in the introduction.  While many of the goal and sub-goal criteria are well presented, some are not well referenced or explained at all (please see comments of thresholds for DO, research papers, rare species ranking criteria, etc in annotated pdf).  In particular, there was very little information on water front use for research, use for education, use for recreation, and weighting of rare species.  How were these sampled, what was the sample size, etc?  It seems like none of this is reported in the paper.  This makes it difficult to access how rigorous or robust the methodology actually is and the lack of detail needs to be addressed before the paper is considered publishable.  I have considerable concerns on the sample design, results, and interpretability of this study if the sample design is not presented.

Another major concern of mine is the impact of not standardizing the wet weight by unit area or unit effort (number of fishing days) for each tidal site.  How does area affect other metrics as well considering the two artificial tidal areas are the smallest in area and have the least amount of coastline.

On a positive note, the example the authors provide for calculating scores is nicely written and helped clarify the process of calculating the index for the goals and sub-goals that were well defined.
Validity of the findings: I feel like this paper is missing a major synthesis of the index that is being proposed.  What are the strengths of this index?  What are the weaknesses?  How did area and length of coastline affect metrics, was standardization important?  The discussion and conclusion need to be expanded in order to bring the paper back to the original focus of the paper as outlined through the introduction.
Additional comments: I think the article is interesting but is lacking some key information regarding how the authors calculated their goal and sub goal criteria.  Please see my comments regarding the experimental design.  The discussion and conclusion sections need additional detail as well in order to give support to the original research question.  While the introduction was well written, the conclusion doesn't quite address the original research question and simply focuses on the findings of each site rather than also synthesizing the pros, cons, and caveats of using this index.


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: METHOD FOR THE QUANTITATIVE EVALUATION OF ECOSYSTEM SERVICES IN COASTAL REGIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have developed the overall paper very well since the previous revision. Development of the introduction has clarified many aspects of the study and the supplementary materials have been vastly improved to give much needed background to the paper.

I feel that the structure of the paper could be reworked to make it easier to follow and generally improve the reading of the paper:

Section 2.7 :Evaluated tidal flats and evaluation year. Bring these descriptions of the tidal flats further up in the paper - either describe them briefly in the introduction or right at the start of the methods.

Lines 132-155: Move these further down the methods. First talk about the overall construction of the index and bring in the sub-indices afterwards.

Section 2.5: This mixes together overall index construction and specific sub-goal examples and that is confusing. Pick one example and keep it concise.

Section 2.6: Move directly below where you talk about likely future state. You move on to pressures and resilience then back to talking about aspects related to LFS.

I think the above restructuring will vastly help the flow of this section, which is crucial in setting up the paper.
Experimental design: I think the experimental design has been improved and the authors have provided adequate justification for their methods.
Validity of the findings: The authors have significantly expanded their discussion to help validate their findings as well as provide discussion and quantification of some uncertainties. They also recognise where improvements might be made.
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: COMPARATIVE GENOMICS OF 16 MICROBACTERIUM SPP. THAT TOLERATE MULTIPLE HEAVY METALS AND ANTIBIOTICS
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Due to their high genetic diversity and adaptation ability, bacteria play an vital role in bioremediation of heavy metal pollution. This manuscript reported the tolerance of 16 strains of Microbacterium spp. to heavy metal and antibiotics, and many genes related to Cr(VI) reduction, heavy metal tolerance and antibiotic resistance were identified. In general, this manuscript merits to be published in Peer J.
1. Line 26, Since "Sixteen different strains....." was mentioned at the beginning of Abstract, ANI among the sixteen isolates was suggested to provide.
2. Line 89, what was criterion to select the 16 isolates? Were they selected randomly?
3. Line 124-125, Since 16S rRNA gene is duplicate gene, was it identical in one strain? which one was selected to generate phylogenetic tree?
4. A phylogenetic tree on the basis of chrR gene is suggested to provided.
5. "sp." should not be italic in table1-table3.
6. It is interesting that two concentrations (0.5 mM and 1 mM) were adopted to the ability of Cr reduction. Why?