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: PROTEOMIC ANALYSIS OF DIFFERENTIAL ANTHER DEVELOPMENT FROM STERILE/FERTILE LINES IN CAPSICUM ANNUUM L.
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this manuscript by Pei et al., the authors perform proteomic analysis of male sterility and fertile pepper lines. The data presented could be potentially valuable. There are some issues which need to be clarified:
1, Abstract in Page 4 and Page 7 is different, please check it.
2, I have noticed that some abbreviations were introduced at the beginning of this manuscript, while it was appeared the full name again at the end, such as the DAP, KEGG etc.
3, Keywords: “Proteasome” should change to “Proteomic”.
4, The English must be deeply edited，especially some long sentence, which makes reader difficult to understand. As it is presented, the manuscript is far from the standards of the scientific (biochemical) language. Besides that, the format in this manuscript also needs to be carefully modified, such as line 74, line 86 ,line 93 and so on.
5, The literature on proteomics of male sterility in pepper is not fully described in the background part, for example Cheng et al 2019 ijms.
6, Figure 1, It is necessary to indicate what period the letters in the figure represent.
7, Are buds or anthers taken for proteomic analysis? From Figure 2, it seems that anthers are taken.
8, The format of references should be uniform, for example, Latin names, gene name and mutant name need to be italicized (line 537, 500, 546 etc.); the first letter of the quoted title should accordance, most of them are lower case, while line470, 510 and so on are capital; line 470 have no volume or page number. Line 568: reference title is wrong,please check it. The name of the reference journal need to be unified, whether to use abbreviation or full name.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: PROTEOMIC ANALYSIS OF DIFFERENTIAL ANTHER DEVELOPMENT FROM STERILE/FERTILE LINES IN CAPSICUM ANNUUM L.
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper needs to be proof read for grammar and spellings. I found several instances and few of them are mentioned below:

1) Line 34: "diffenrent"; Line 39: "Encyclopaedia"; Line 52-53: "This study largely improves our understanding of key fators through proteome analyses and revealed components of metabolic pathway impacted the anther and pollen development in the male sterility". This line does not make sense grammatically.

2) Line 66: "The sporogenous cells were surrounded by" this switches the paragraph from present to past. Please check this.

3) Line 44: The sentence starts with this "cytoplasmic male". It should be "Cytoplasmic male...".
Experimental design: All bioinformatics methods seemed to be thoroughly performed and documented.

Just one minor question: Why was a fold change of 1.3 used to detect differentially abundant proteins? I have seen people using a cut-off of 0 or 1 or 1.5 but never seen anybody use 1.3 before. Was there a rational behind using such an arbitrary cut-off?
Validity of the findings: The authors seemed to have applied appropriate bioinformatics tools and techniques to determine the genes and pathways responsible for male sterility in Capsicum annuum L. The results seem to be compelling and statistically reliable.
Additional comments: Please double check all the figures for clarity, a few figures are not clear and appear to be pixelated.


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: PROTEOMIC ANALYSIS OF DIFFERENTIAL ANTHER DEVELOPMENT FROM STERILE/FERTILE LINES IN CAPSICUM ANNUUM L.
Review round: 2
Reviewer: 1

Output:
Basic reporting: All issues seem to have been addressed well. Thanks!
Experimental design: N/A
Validity of the findings: N/A
Additional comments: N/A


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: INFLUENCE OF DNA EXTRACTION KITS ON FRESHWATER FUNGAL DNA METABARCODING
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is interesting, they evaluated the effect of DNA extraction kits on the fungal DNA metabarcoding for freshwater samples. They have evaluated 3 DNA extraction methods with few sample sites in Japan. The manuscript is clear and generally well written. They mentioned that limited information are available in studies of fungi using metabarcoding in freshwater, but believe they can find more references and some externally than Japanese publications. Ex Coble et al, 2019 P1060 indicated some publications in it https://www.fs.fed.us/pnw/pubs/journals/pnw_2019_coble001.pdf . The structure of the manuscript is well done and they indicated all elements in the supplement files. I just find a bit disappointing in the analysis with all genus, species and taxonomy determination of fungal group that nothing is stated in the manuscript about organisms or group of organisms Fungal group family in the analysis. You have the info in Supple. Table S2 but nothing about it in the manuscript. The manuscript will benefits to have some abundance figure about the finding and limitation as well on the identification to the Phylum leve. genus, species....?  They did great statistical analysis and well used the diversity analysis but they can I believe can go more on the analysis.
Experimental design: The disigned seem good, they have a limited number of samples and they indicated the limitation in this. However due to limited studies in the field this well presented. They present a good bioinformatics pipeline and statistics, but I think they can go more far in the analysis with the Abundance taxa figures and indication of key species. They can also add a FunGUILD analysis to their data to separate the fungal ID in different group by niches. Do you have more saprotroph, pathogen...  It seem that only evaluating the overall diversity seem limited in it. Do you have any important group of fungi that could have been missed?  The inclusion of Internal control with qPCR is great.
Validity of the findings: The validity of the finding seems correct. They dont have a clear answer about the cut filter why more low, could be interesting to highlight what is detected specifically there?  The stats seems robust.
Additional comments: The manuscript by itself is well done, clear and they present a good statistical method to reach their conclusion. However I think more analysis in taxonomic side can be accomplish with the abundance to get a better description and evaluation of the findings. Maybe some group of organisms can show a different pattern?
Specific comments:
Abstract-Methods, first time: need a whitespace between "extractmicrobial" extract microbial
L102, should put a short description of the DNA extraction, not just reference, also when reference can be confusing to get 2 references about a protocol, Ex. L103 and 111 why 2 reference seems self citing?
L118, any URL link?
L133, why do you say no rarefaction curve were performed, the figure S1 is a rarefaction curve, not sure I follow here why not? Also seem you can indicated at what number of sequences you are getting saturated, maybe 2000, 5000 sequences?
L236, will be interested to have info on Phyllum difference before to state this sentence. Any diff in niches?


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: INFLUENCE OF DNA EXTRACTION KITS ON FRESHWATER FUNGAL DNA METABARCODING
Review round: 1
Reviewer: 2

Output:
Basic reporting: Background literature should acknowledge that the technique currently referred to as 'DNA metabarcoding' is not as new as the authors imply.  In the fields of molecular ecology and microbial/fungal ecology, this was referred to as 'cultivation independent', 'marker gene sequencing', 'amplicon sequencing' approaches.  The first published references to similar work occurred in the 1980's (analysis of microbes from sea water) and it became more routine in the 2000's when soil DNA extraction kits were commercialized.  See additional comments for details.
Experimental design: Experimental design seems to be optimized to detect differences among sites not DNA extraction protocols (only one replicate per a site).  I don't think this should prevent publication of this paper, but it should be acknowledged by the authors.  The current experimental design seems underpowered to detect protocol differences.  A power analysis could provide some insight.  See additional comments for details.
Validity of the findings: These findings show that similar OTU richness and beta diversity patterns are detected in freshwater fungi using 2 different DNA extraction kits when following the manufacturer's protocol.  The authors should be cautious about extrapolating these results more generally, as their experiment seems underpowered for detecting biodiversity differences due to DNA extraction kit (no replicates within sites).  See additional comments for details.
Additional comments: PeerJ 66935

Summary

This paper describes the use of three different DNA extraction protocols using two different kits on freshwater fungal metabarcoding results.  No significant differences in richness or beta diversity were found between the two kits when using the manufacturer’s protocol.  The paper is straightforward and clearly written.

Major comments

The paper is very brief and there is plenty of room for the authors to include full methods to describe their molecular biology approach, move from Appendix to main text, and bioinformatic methods by including program parameters/settings.  The paper would also benefit from the additional description of the fungal groups that were detected from these sites and from each kit such as stacked bar charts or heatmap--this information is already provided in the supplement but it would be nice to summarize this as a figure in the main text.  Unfortunately, it seems that there is only one biological replicate per site-protocol combination.  The authors could set up future work by calling for increased replication to enable statistical comparisons among protocols with sufficient power to detect differences in DNA extraction protocol (if they exist).  Currently, the variation due to site differences seems to be greater than any protocol differences, making it difficult to detect variation due to protocol and this should be acknowledged.

Minor comments

Title - The word ‘DNA’ is repeated three times in the title, consider changing to something like ‘Influence of DNA extraction kits on freshwater fungal DNA metabarcoding.’  The sequencing of environmental DNA is implied, could also add to your list of keywords.

L17 - ‘rapidly expanding technique’. This technique has been around since the 80s and has been ‘routine’ since the 2000’s.  Consider rephrasing to ‘environmental DNA metabarcoding is a common technique for ...’

L39-41 - Sequencing signature regions of DNA from environmental samples is not as new as is repeatedly stated in the abstract and here.  Acknowledging this in your introduction is recommended.  It would also be a good idea to cite the paper that first coined the catchy term ‘metabarcoding’ Taberlet et al. 2012.

L68 - ‘ there are no examples of ... ‘ consider adding ‘that we are aware of’.

L70 - ‘experimental processing after DNA extraction is inhibited by PCR’.  Consider rephrasing to ‘inhibitors carried over from DNA extraction have been shown to inhibit PCR and limit the number of amplified template that can be successfully sequenced and analyzed in downstream biodiversity analyses’.  Also consider listing what some common PCR inhibitors are, what they do (ex. impair Taq polymerase activity in PCR), and how common these inhibitors are in freshwater (compared to soil where this has been studied more intensively).  This paragraph mentions humic acids twice, is this a common inhibitor in freshwater studies?  Some common issues when extracting fungi are physically breaking down chitinous cell walls and co-extracting polysaccharides, consider mentioning these issues.  Other common environmental DNA extraction issues includes DNA extraction efficiency, ug of DNA extracted and how much of it is high molecular weight (not degraded).  Consider at least mentioning these other DNA extraction concerns as for some people this could affect choice of DNA extraction kit.  Also consider mentioning some of the traditional ways researchers have handled PCR inhibitors, ex. dilution of DNA extract prior to PCR, addition of BSA, betaine, beta-mercaptoethanol, DMSO, gel extraction of extracted DNA, flocculation, etc.  This would give more context to why you are focusing on DNA extraction ‘kits’, to avoid using potentially hazardous chemicals, as well as improving reproducibility and standardization.  There is plenty of room to expand on these topics in the intro/discussion to give more context to the importance of your study.

L94 - Please add one line to explain how benzalkonium chloride reduces DNA degradation in addition to your citation.

L111 - There is plenty of space here to provide the PCR cocktail and thermal cycler settings in addition to your citations so that readers don’t have to look up another paper to understand what you have done.

L121 - forward and reverse reads were ‘paired’ not ‘concatenated’.

L122 - I was not aware that CD-HIT-OTU could address sequence errors, does it now include a denoising step or remove low quality bases/sequences?  Please include the settings for that here.  For each program used in your bioinformatic processing, include the parameters that were set (in addition to the papers you have cited so that readers don’t have to look up another paper to understand how data was processed in this study).  Were the conserved SSU and 5.8S gene regions trimmed from the ITS1 spacer region?  If it wasn’t done don’t worry about it now but do mention in the methods as this will affect how your OTUs clustered (not as sensitive, could probably detect slightly more OTUs if the conserved ends were removed).

Q - Did you quantify how much DNA that was extracted from each DNA extraction protocol? Please provide this info in the main text, it would give readers an idea of the type of yield they could expect from these protocols/kits.  Did you take any gel pictures to see if it was high molecular weight (high quality) DNA?  Did you notice any significant autofluorescence?  DNA shearing? Did you attempt to assess DNA purity using any other complementary methods, ex. nanodrop to check for any obvious co-extracted proteins/carbohydrates, etc.  If so, please mention in the text and consider including pictures in the supplement.

L156 - be sure to add a citation for each major R package that you used, ex. lme4

L167 - 169 - I’m not clear about the benefit of including the Raup-Crick index.  Can you include another line to the text explaining why not being sensitive to differences in number of OTUs is a benefit for readers like myself who are not familiar with this index?

L183 - 15-372 OTUs per sample is a huge amount of variation that looks like it’s related to site differences as opposed to protocol differences.  It would probably be a good idea to run a power analysis, I suspect this experimental design is under-powered to detect protocol differences (it looks fine to detect among-site differences).

L191 - consider changing OTU composition to ‘community composition’ here and on the next line.

L198 - because you don’t detect any significant differences among the DNA extraction protocols, consider adding a power analysis to see if you have enough data to detect a protocol difference if there was one.  It could be that with the samples you have, you would only have the power to detect a very large effect.  The current experimental design seems geared towards detecting among-site differences.  If you include a power analysis, you could then set up your next paper by calling for the need to include additional replicates (more replicates from one site for each kit).

L215 - blood and tissue kits are not commonly used with fungi as fungi tend to have plant-like DNA extraction problems (thick cell walls, polysaccharides) whereas extracting DNA from blood is easy which probably explains why this kit is not often used on fungi.  Not sure that previous use of blood and tissue kits to detect animals is relevant here.

-Please provide any additional information on data accessibility - you have already provided a DDJB accession but I was unable to find the record, can you double check that the data is public and accessible?  Consider depositing your OTU sequences and OTU x sample table and R code to an online repository (Dryad, GitHub, etc.)

-This paper would benefit from further description of the fungal taxa that were detected by each kit and how they compare.  This is present in the supplementary material, but the results could be summarized in a figure to present in the main text so that readers can which fungal groups were detected from water overall and by each kit, ex. heat map, network, or stacked bar plots.


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: INFLUENCE OF DNA EXTRACTION KITS ON FRESHWATER FUNGAL DNA METABARCODING
Review round: 1
Reviewer: 3

Output:
Basic reporting: The objective of the work developed in this manuscript was to evaluate the effect of DNA extraction kits on the results of fungal DNA metabarcoding for freshwater samples, namely of rivers and lakes/ponds. Overall, the manuscript is well written.

The introduction section cites mostly works using eDNA to target fish (e.g., L41). Although these examples are valid, I suggest author also include examples of other works focusing on other organisms like metazoan, eukaryotic communities (e.g., Leduc et al 2019; Antich et al 2021). These are just 2 examples; author may find other articles more suitable.
Also, reference support should be given at L48-50, when authors state that “there are fewer such methodological studies in fungi”; and L129-130 “which corresponds to 2–9 reads, a typically used criterion in eDNA metabarcoding studies”.

L64-65. This sentence should be rephrased. Please see paper by Hermans et al 2018, where they tested DNA extraction methods to simultaneously extract bacterial, fungal, plant, animal and fish DNA from soil, leaf litter, stream water, stream sediment, stream biofilm and kick-net samples, as well as from mock communities. This paper might be useful also to the discussion section, to compare with results obtained here.

References
Leduc, N, Lacoursière-Roussel, A, Howland, KL, et al. Comparing eDNA metabarcoding and species collection for documenting Arctic metazoan biodiversity. Environmental DNA. 2019; 1: 342– 358. https://doi.org/10.1002/edn3.35
Antich, A, Palacín, C, Cebrian, E, Golo, R, Wangensteen, OS, Turon, X. Marine biomonitoring with eDNA: Can metabarcoding of water samples cut it as a tool for surveying benthic communities?. Mol Ecol 2021; 30: 3175– 3188. https://doi.org/10.1111/mec.15641
Hermans, SM, Buckley, HL, Lear, G. Optimal extraction methods for the simultaneous analysis of DNA from diverse organisms and sample types. Mol Ecol Resour. 2018; 18: 557– 569. https://doi.org/10.1111/1755-0998.12762
Experimental design: I think it is important that authors clearly state their hypotheses in the last paragraph of the introduction. Did you expect one of the kits to perform better? Or you expected both to perform equally?  Did you expect differences between rivers and lakes/ponds?

Regarding the experimental design, the most important question I see is why you tested for filter cutting or not, using the Blood and Tissues kit, but did not do the same test for the Powersoil?  It is really important for the readers to understand your reasoning. Would authors also expect differences in performance?

Regarding statistics, author presented only the main effects. It could be interesting to look at the interactive effects “dna extraction x sampling site” for both number of OTUs and community composition. Also, why authors decided to test for latitude and longitude (L172) effect on OTU composition? This is not clear in the text and was not stated and an objective of the work previously. Finally, on L174. authors explain the indicator taxa analysis and they state that it “was performed to determine which OTUs had significantly different frequencies between DNA extraction kits”; but below you say that you used the two matrices, i) relative number of reads and ii) presence/absence. This seems a bit confusing since presence/absence matrix will not provide information of OUT frequencies. Please clarify this question.

Others small details below to increase clarity of methods:
L92. Do authors measured any physical-chemical parameters of the water (Table S1)? If not from this study, do author have these data from previous studies? Having information of content of organic matter for instances, would be important to support your results and discussion.
L95. How long between samples collection and filtration? Please add this information.
L96-97. Why authors selected the GF/F filter (0.7um pore size) and not Sterivex type filter (0.2um pore size)? Would you expect differences in the performance? See paper Spens et al 2017. I think it is important to explain this choice and maybe add some points to discussion.
L104. Can author give an estimate of the size of filter pieces after cut? This can be important for other researcher to replicate your study.
L127. Why did authors used the NCBI database and not other options like UNITE? Maybe add a small sentence to justify your choice.
L149. Please correct: ΔCt of ≥3 cycles and is usually considered > ΔCt of ≥3 cycles is usually considered.

References
Spens, J., Evans, A.R., Halfmaerten, D., Knudsen, S.W., Sengupta, M.E., Mak, S.S.T., Sigsgaard, E.E. and Hellström, M. (2017), Comparison of capture and storage methods for aqueous macrobial eDNA using an optimized extraction protocol: advantage of enclosed filter. Methods Ecol Evol, 8: 635-645. https://doi.org/10.1111/2041-210X.12683
Validity of the findings: I think the section results and discussion can be improved to increase the value of the manuscript. Once author include their hypotheses in the introduction, it will make the discussion clearer already.

L197-202 Authors state that “variation within the same study site (…) was smaller”. But, it is interesting to notice that overall variation among pond/lake sites was also much smaller in the Raup-Crick; but rivers did not follow that pattern. Do authors have and explanation for that?

I think the paragraphs L208 and L224 should be rephrased and discussion better integrated. With the current structure, and by the end of paragraph L208-223, it seems like Blood and Tissue kit is the best option. Just in the paragraph L224-233 author refer that water samples with high content of organic matter (for instance) might result in PCR inhibition, and so using a kit that removes more inhibitors, like the Powersoil, could be a better option. I understand the separation to show first the results for OTUs and after for PCR inhibition, but maybe integrating and discussing together can improve clarity. And, adding information on the physical-chemical characteristics of your sampling sites would definitely give support to your results.
Additional comments: Minor typos on Reference list:
L295. Systematicreview > Systematic review
L347. Ofnon-native > of non-native
L356. Year here is 2017, but on L.95 is 2016. Please confirm and 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: INFLUENCE OF DNA EXTRACTION KITS ON FRESHWATER FUNGAL DNA METABARCODING
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Peerj-66935

Summary

This paper is greatly improved over the original submission.  I can see now that the study was performed in a much more stringent manner than I had originally thought.  The newly added background in the introduction and details added to the molecular biology and bioinformatic steps greatly improves the paper.

Major comments

None

Minor comments

L24 - change to ....of DNA extraction...  remove the word ‘the’

L29 - remove square brackets, just do ... DNA metabarcoding results; i.e., operational ...

L125 - change physicochemical to physical-chemical

L129 - change ‘Molecular experiments’ to ‘Molecular biology’

L148 - please clarify, was the second PCR to add Illumina sample indexes?  If so, indicate the index kit that you used, ex. Nextera?

L286 - just a comment, there are numerous early diverging fungal lineages that have not been identified to the species level that are commonly found from freshwater samples, search Rozellomycota/Cryptomycota but all early diverging fungal lineages are difficult to classify as so many are currently unnamed (Blastocladiomycota, Chytridiomycota and everything in between).    It would be interesting to know if you detected any of this highly diverse group (Cryptomycota) from your own samples as this could explain why you were unable to identify so many OTUs.  In GenBank they are often only labelled as ‘uncultured fungal clone’ or ‘environmental sample’.  During LCA processing, these types of vague identifications really hinder taxonomic assignment.  They are best identified using phylogenetic methods with reference sequences used to represent this group (try BLASTing a few of these by hand and looking at the distance tree results provided by the NCBI BLAST method).  It would actually be nice to add a few sentences about this common freshwater group to your discussion to highlight how common and diverse they are and how challenging they can be to properly identify because none of these samples actually have species names associated with them (except for the well-known chytrid parasite Rozella allomycis).

L286 - also, it would be good to add one sentence (perhaps in the methods section) to address how specific the primers are that you used in this study, are they expected to amplify non-fungal taxa such as macroinvertebrates?

Fig 1-legend.  Add “Upper-left box shows Japan and lower right box shows sampling sites.” or something similar.


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: INFLUENCE OF DNA EXTRACTION KITS ON FRESHWATER FUNGAL DNA METABARCODING
Review round: 2
Reviewer: 2

Output:
Basic reporting: I want to acknowledge the very extensive work authors have performed in the MS. I am glad that all my questions and suggestions (and other reviewers) have been addressed. I think the MS has improved in clarity and general interest to the readers.
Authors have now included in the introduction section the questions addressed in this MS. This really helps the readers to understand the objectives. However, I think adding hypotheses (if possible) would benefit the MS. For instance, you state that you will address the following questions: “(1) whether the degree of PCR inhibition and (2) DNA metabarcoding results (i.e., species number and composition) differed depending on the DNA extraction kit used and (3) whether these effects vary by habitat type”. I understand that authors have not measured physical-chemical parameters of the water and so this data is not available. But if we think these are environmental samples (and unless these are super oligotrophic streams and lakes), we would expect some organic matter and even humic acids to be present, for instances. As so, I would hypothesize that the kit Powersoil would be more efficient preventing PCR inhibition compared to the blood and tissue. The same exercise can be made for the sites: based on your knowledge on the streams and lakes/ponds, can you hypothesize that one of these could have more inhibitors and so in one of these type of sites one of the kit would provide better results? Or characteristics are similar and you expected them to perform the same?
Experimental design: The section on methods is much improved now. One thing I am missing is description in this section of how you performed the analysis of functional traits with FUNGuild. Another thing that is missing is the description of effect of the Latitude and longitude. You say in the methods you will test this, but nothing in mentioned in the Results and discussion section. Only in the legend of Fig.4 some information was added. Please also describe it in the results section.
Validity of the findings: No comment
Additional comments: Some minor comments/suggestions:
L42. Please consider changing “as well as” to “including”.
L68. Please remove repeat word “fungi”.
L71. eDNA metabarcoding has been also applied in wetlands (e.g., https://doi.org/10.3996/042014-JFWM-034). Please provide references for other examples stated.
L81. Please provide a reference.
L122. Please correct “beforehand to allow”.
139. Consider changing “duplicated” to “replicated”
L252. If the community composition is similar, shouldn’t the P-value be >0.05? in the text you have P<0.0001. Please clarify.
L470. Please add to the legend the functional assignment of species.


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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The title is not very clear; I would suggest "Sexual dimorphism in shell size of the land snail Leptopoma perlucidum (Caenogastropoda: Cyclophoridae)" (The fact that this study was carried out in Sabah is probably not sufficiently relevant to be in the title; only differences in size were found, not in shape; and it seems more logical to mention the shell size immediately after "sexual dimorphism" and not only after the species name.)

In the Abstract, "islands in Borneo" should be changed to "islands off Borneo"

I noticed numerous stylistic and linguistic details in the text that could be improved during the editing process.

The Introduction should be supplemented with a brief paragraph in which the distribution of gonochorism and hermaphroditism in the Mollusca is outlined, because this limits the taxa in which research on sexual dimorphism is possible.

In the Introduction (L. 82), it says: "To our knowledge, sexual dimorphism in terrestrial caenogastropod shells has only been studied in four genera: Cochlostoma, Obscurella, Barnaia, and Streptaulus." However, we also studied it in the genus Plectostoma:

>>for colour (Sexual dimorphism in shell coloration of Plectostoma (Caenogastropoda: Diplommatinidae) is caused by polyenes; M Schilthuizen, I Sipman, H Zwaan - Journal of Molluscan Studies, 2018)

>>for shape (Schilthuizen, M., Rosmaineh B. Rosli, Abdul Muji B. Mohd. Ali, M. Salverda, H. van Oosten, H. Bernard, M. Ancrenaz & I. Lackman-Ancrenaz, 2003. The ecology and demography of Opisthostoma (Plectostoma) concinnum s.l. (Gastropoda: Diplommatinidae) on limestone outcrops along the Kinabatangan River. In: Kinabatangan Scientific Expedition (Maryati Mohamed, B. Goossens, M. Ancrenaz & M. Andau, eds.). UMS, Kota Kinabalu, Malaysia.

I don't understand how sexual dimorphism can result from "selection pressures among different distant geographical populations" (L. 69-70).
Experimental design: In L. 124 it says, "the ANOVA is considered a robust test against the normality assumption (Zar, 1999)". I presume what is meant is that the ANOVA is considered robust against violations of the normality assumption.

In the description of the geometric morphometrics, the package 'plotTangentSapce' is said to have been used (L. 140). Wouldn't this be 'plotTangentSpace'?

In L. 96-97, it might be good to add that the sexual anatomy of L. perlucidum has been described by Jonges (1980; Bijdr. Dierk.)
Validity of the findings: In L. 184, it says, 'This study of L. perlucidum sexual dimorphism in shell form is the first investigation of land snails in the family Cyclophoridae'. This is somewhat misleading since to the uninitiated reader it may suggest that no studies in Cyclophoridae have ever been conducted. I would suggest instead: 'This is the first investigation into sexual dimorphism in shell size and shape in the Cyclophoridae.'

The last few sentences of the discussion do not seem very appropriate, since this study is mainly about sexual dimorphism, not about geographic variation. I would suggest to end on a positive note, namely that this study shows that, in L. perlucidum at least, no shape differences (which would further complicatie a taxonomic revision of the genus) are found between males and females.
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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. Basic reporting
Being non-native English, I nevertheless commented on some sentences (see 4 below). This is a mixture of linguistics and biology.
The context is clearly indicated.
Experimental design: 2. Experimental design
Unfortunately, my apologies, I do not know that scope sufficiently well.
The research question is well defined. To some extent there is a knowledge gap, next to many similar gaps for other taxa.
The methods are clearly described.
Validity of the findings: 3. Validity of the findings
There is no novelty for gastropods, but only for a particular genus. The findings are not conspicuously different from what we know for other genera that have been investigated. The impact will be meagre.
I feel not competent to judge the statistics. I fully rely on the corresponding author.
The conclusions are clear, correct, and linked to the research question.
Additional comments: 4. Comments, questions of the referee
42 The "effect" of a location is something dubious. Drift is possible equally well.
44 "the between"??
46 Geographical variation is a very well known phenomenon for every taxonomist.
67 Adapt wording
69 Is sexual dimorphism a selection pressure?
72 Adapt wording since there is no geographical dimorphism
83 "Both"?
92 Explain why "reflected lips" is relevant and adapt the wording since without such a lip the snail will probably be immature.
221-223 This applies to all gastropods with potential sexual dimorphism, not only Leptopoma. This is well known among taxonomists.


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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: There are many studies of sexual dimorphism using mollusks as subjects, dating back to the 19th century.

A quick Google Scholar search revealed papers on the sexual dimorphism in Plectostoma (Schilthuizen et al. 2017, cited in the paper), Arinia (Maassen 2008), and Chondrina (Baur et al. 1993) as well. Obviously a more thorough search is needed.
Experimental design: The sample sizes from Gaya Island are not large enough to support the significant findings reported in the paper. Using a power analysis (80%), at least 13 individuals from each sex needed to be used to achieve significance at p<0.05.

There a better and more thorough ways to analyze shell outlines that with a set of nine landmarks, including the addition of more landmarks, curve capture, and Fourier analysis. A more robust analysis of shell shape is warranted, especially when looking for potentially subtle differences between sexes.

In addition to shell shape, geometric morphometrics can assess shell size through analysis of centroid size if the landmarks are scaled appropriately to start.
Validity of the findings: The sample sizes from Gaya Island are not large enough to support the significant findings reported in the paper (see power analysis comment under Experimental design).

Given the small number of landmarks, it is unclear whether sexual shape dimorphism is actually present or not.
Additional comments: The discussion says very little other than that females may be larger than males in certain populations of this species. No explanation nor proposed reason for this difference is provided. The discussion regarding the taxonomy of the genus and the number of nominal species lacks context – the reader doesn’t know how many species are based on size alone, what the overlaps of the species are, etc.


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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revision has addressed all the points raised by myself and other reviewers in a satisfactory manner. I have no further comments.
Experimental design: The revision has addressed all the points raised by myself and other reviewers in a satisfactory manner. I have no further comments.
Validity of the findings: The revision has addressed all the points raised by myself and other reviewers in a satisfactory manner. I have no further comments.
Additional comments: The revision has addressed all the points raised by myself and other reviewers in a satisfactory manner. I have 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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 2
Reviewer: 2

Output:
Basic reporting: Sexual dimorphism in shell size of the land snail Leptopoma perlucidum (Caenogastropoda: Cyclophoridae)
The authors have taken the comments of the reviewers seriously and have considerably adapted the manuscript at several relevant points.
I see no way for additional improvement.
Edmund Gittenberger
Experimental design: --
Validity of the findings: --
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: SEXUAL DIMORPHISM IN SHELL SIZE OF THE LAND SNAIL LEPTOPOMA PERLUCIDUM (CAENOGASTROPODA: CYCLOPHORIDAE)
Review round: 2
Reviewer: 3

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The revisions that the authors have made in this version have improved the manuscript significantly. I have included some minor editorial changes in the text.


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

Input:
Title: PREDICTING HABITAT SUITABILITY AND RANGE SHIFTS UNDER PROJECTED CLIMATE CHANGE FOR TWO OCTOCORALS IN THE NORTH-EAST ATLANTIC
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: Your models use environmental data from a lat/long grid. Given the latitudinal range of your study the pixels in the south have >50% larger area than the pixels in the north. We know that variables like slope (which you are using) are sensitive to the area used in their measurement and that this affects their use in SDMs (e.g. Dolan et al https://doi.org/10.1016/j.dsr.2008.06.010). This should at least be mentioned in the discussion, but better be rectified by using equal area grids for your modelling.

Background sampling - there are several issues with the background sampling that I'd like to see addressed. You provide depth limits for the target species in the intro (50m), but your background sampling doesn't filter areas by depth. However there is a de-facto depth limit introduced by filtering to your substrate layer, which you should acknowledge. Your study area includes many areas much deeper than 50m, and table 1 reports both species seen below 100m. Including these area as background, which are wildly different from your samples, will artificially inflate your evaluation stats (see the modelling standard papers you cite in the intro).  While substrate restriction is on the surface a perfectly reasonable assumption to make, it doesn't seem to have restricted your background data beyond the shelf. I'd like to see this tested by your data. What % of your presences are in the least rocky area? I'd guess there are a lot of Alcyonium that come out as being on a different substrate. If the data do not support this assumption (i.e. a significant % of observations are outside your rocky ground layer) then it seems inappropriate to use this as a background filter.

Also the rock filter does nothing to address the inherent sampling bias in your data, which is skewed to shallower, near-shore sites.  There are some pretty standard methods that could adress this, e.g. selecting more background data closer to your presence points (e.g. https://onlinelibrary.wiley.com/doi/full/10.1111/ecog.04503).  Given the spread of data a simple distance filter for the background data would probably cover all bases here.
Validity of the findings: Prediction maps

There are a cluster of presence points in the central north sea which the model fails to pick up. I'm guessing this is because you have chosen a 0.5 cutoff for your display. I would rather see a wider range of suitability indices displayed. There is no justification for the 0.5 threshold and I think its more useful to see the lower value predictions. Similarly for the future predictions this 0.5 threshold selection is restrictive in allowing the reader to interpret the results. There are a variety of methods for selecting a cut-off threshold (i.e. maximum sensitivity and specificity or based on percentile of presence points predicted - see maxent manual), any of these options would be better than the arbitrary selection of 0.5 which seems to be mistakenly intrepretted as a probability of presence.  This is really important for assessment of future predictions.
Additional comments: Intro

lines 76-77 unnecessary repeat of distribution of E. verrucosa
lines 150-2 - Does this mean all other variables were kept at present-day values? If so please state this in the methods.

Results

line 253 - your suitability index is *not* a probability of presence and should not be called this. Reference to this should be removed everywhere in the manuscript.
line 238 - its not surprising that depth is the biggest predictor given that the majority of your samples are inshore & shallow, and most of your background points are from much deeper water.

Discussion

line 302-7 - I think you should speak about the observations of A. digitatum on low % rocky areas - it looks like there are lots of obs in very low % areas.

The critical factor for calcite (and aragonite) is the saturation state (omega). The calcite saturation horizon in the N. Atlantic is several thousand metres (e.g. feely et al https://www.pmel.noaa.gov/pubs/outstand/feel2633/feel2633.shtml). No predictions of future saturation state (that i'm aware of) have suggested that NE Atlantic will undersaturated at 50m (or anywhere close) in the near future. Also, its quite difficult to talk about future impact when your future scenarios don't include any change in saturation state (despite these layers being available and used in other, similar studies).

Can you show the thermoclines on your figures? You could easily generate these from your environmental layers. I think this would support your point that northern limit of E. verrucosa is linked with thermocline.

line 349 - I'm not convinced that a single minimum observation is a robust temperature minimum value. One outlier observation could skew this number. I think its better to present a 99th percentile value. Also this is the minimum observed annual mean temperature not the minimum bottom temperature experienced by the corals.  This is an important distinction.

Figures

I find the blue dots difficult to distinguish, particularly in the scandanavian fjords, please consider changing the colour or make the country border-lines more faint. Also the scale is incorrect for the majority of latitudes on display, please reproject to an equal area grid or remove the scale bar.

Fig 4. I'm not sure what you mean by "the model was then re-run to generate future predictions". Surely the model trained on the original data was projected into your future climate layers (all but 1 of which was identical to the original layers).

Table 1 - While the min/max values observed are interesting, it would be good to show the spread of this data (e.g. include a median)


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: PREDICTING HABITAT SUITABILITY AND RANGE SHIFTS UNDER PROJECTED CLIMATE CHANGE FOR TWO OCTOCORALS IN THE NORTH-EAST ATLANTIC
Review round: 1
Reviewer: 2

Output:
Basic reporting: Jenkins and Stevens report on a regional species distribution modeling approach for two species of subtidal temperate octocoral species. For the vast majority of the document, the writing is clear and the figures are well drawn (but could be slightly revised to improve engagement). The analyses appear well done and well reported (in general) but there are some areas for clarification.
Experimental design: There are some queries that I have regarding the experimental design, which are embedded as comments on the attached PDF. Generally, these are not major issues, but need some consideration.
Validity of the findings: Appear valid and insightful.
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: PREDICTING HABITAT SUITABILITY AND RANGE SHIFTS UNDER PROJECTED CLIMATE CHANGE FOR TWO OCTOCORALS IN THE NORTH-EAST ATLANTIC
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have taken on board the majority of comments made by both reviewers. The methodology is now much more robust.  This has resulted in the re-running of the models using significantly different datasets and unsurprisingly the results have changed too.
Experimental design: no comment
Validity of the findings: The only question that remains for me is the future projection for A. digitatum. The model suggests that NONE of the current range of A. dig will be suitable. This appears to be driven by the dramatic difference in Calcite saturation state exhibited by the underlying environmental layer. I understand this result is driven by the underlying data that the authors cannot control, but I think its a bit of a problem for the credibility of this study, and will very likely be over-interpreted by some readers (I could easily see a headline off of this saying beloved UK coral doomed). I'd recommend the authors do a little bit more to interpret these results and make sure this isn't the takeaway.

I would be inclined to present a model without the calcite variable to examine the relative influence of that variable on these predictions. I'd also discuss whether the real limitation is the threshold of positive or negative saturation state rather than the absolute value (you could treat the calcite layer as a binary under/oversaturated grid). It appears that the state goes from ~+4 everywhere on the shelf to ~+2. Clearly the predicted state is outside the observed range, and this is why it gets low suitability in the future projection, but omega >1 still represents an oversaturated state that is likely within Alcyonium tolerance.
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: TRAJECTORY PATTERNS AND FACTORS INFLUENCING PERINATAL FATIGUE AMONG CHINESE WOMEN FROM LATE PREGNANCY TO 6 MONTHS AFTER DELIVERY
Review round: 1
Reviewer: 1

Output:
Basic reporting: First, please check the English language of this paper and make necessary editing because I have found several problematic sentences and terms.
Second, the title seems not consistent with the work done by the authors because the authors classified subjects into two groups with different changing patterns of fatigue. The title means an overall changing patterns of fatigue, which should be provided in this paper.
Third, please consider whether logistic regression is appropriate to answer factors associated with the trajectory, I do not think these identified fixed factors can be associated a changing pattern.
Fourth, please consider latent variable growth model because there are many external indictors of the latent variable, fatigue.
Fifth, in the introduction, please clearly indicate the clinical significance of this research topic, i.e., whether the identified changing patterns could guide clinical intervention.
Sixth, the measure of fatigue is a foreign language version, I do not think it is valid for assessing fatigue in Chinese pregnant women.
Seventh, the psychological factor of anxiety is inadequate. Please provide your considerations, for example, why not included depression.
Eighth, the authors should provide a theoretical model of perinatal fatigue to inform the clinical research design of this study.
Experimental design: See above
Validity of the findings: See above
Additional comments: See above


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

Input:
Title: TRAJECTORY PATTERNS AND FACTORS INFLUENCING PERINATAL FATIGUE AMONG CHINESE WOMEN FROM LATE PREGNANCY TO 6 MONTHS AFTER DELIVERY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Pugh’s framework were used as the theoretical framework to explore the related factors that influence perinatal fatigue. However, authors did not provide sufficient information to demonstrate how this framework work. Examples of the four types of factor  - situational, physiologic, psychologic and performance - can be illustrated in the form of figure.
Experimental design: The methods and results sections of this paper leave out a lot of critical information, such as when the study was conducted, the flowchart of the participants which include information about drop-out reasons  at each time point and a table comparing basic information about the included and non-included subjects. A checklist from the appropriate reporting guideline should be used to guide the author in writing.
Another issue is about the scale used to measure fatigue- Postnatal Accumulated Fatigue Scale (PAFS). The authors do not provide information on whether this scale has been translated and validated in the Chinese population.
Validity of the findings: one major  problem with this article is that many of the conclusions in the discussion were not directly related to the results. For example，in line 235, the  conclusion "fatigue may be strongly associated with individual internal substances. Interleukin-6 (IL-6) ..." seems very arbitrary.
Additional comments: no comment


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

Input:
Title: TRAJECTORY PATTERNS AND FACTORS INFLUENCING PERINATAL FATIGUE AMONG CHINESE WOMEN FROM LATE PREGNANCY TO 6 MONTHS AFTER DELIVERY
Review round: 2
Reviewer: 1

Output:
Basic reporting: None.
Experimental design: None.
Validity of the findings: None.
Additional comments: None.


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: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The author studied generating a reliability scale for clinical brain activity test scores based on medical scans and demographic data. The idea of proposing a decision support system is not new as some papers have already been published by using different methods; the only novelty is the use of an unsupervised missing value imputation methodology. The presentation is not organized logically. Also, the quality of written English is poor, I would suggest the authors have their manuscripts checked by a proficient English language speaker.

Some suggestions for further improvement:

The summary abstract is not concise. I do recommend the re-writing of the abstract and grammar check through the document.

The Introduction section is well-written, and I propose to schematize all the discussion in several paragraphs (to facilitate the reading): (1) motivations, (2) the overall approach, (3) main contributions.
However, the main challenges to the field and the needs and benefits of this study are missed in the introduction.

Many important and recent papers are ignored by the authors. This research area is not new and there are many other papers in the literature.

A "Related Work" section is completely missing (much could be extracted from the introduction), which highlights the most important techniques (at least for the chosen problem), and above all a comparison with the technique proposed in the paper. A final table summarizing the technical aspects of the various methods would be very useful.

A general discussion of the limitations and expectations should be inserted.
Experimental design: The 'unsupervised' approach used in the comparisons is not explained or discussed. There is a brief mention that it can be intuitively deduced in a referenced work, but at least a basic description should also be present in this work. Also, and more importantly, there is no state-of-the-art for generating a reliability scale test scores method considered in comparisons. If there is not something specific to clinical brain activity data, I believe there are several general domains with good performance that could be considered.

The authors need to compare their results with other existing methods and add a table of comparison to the article. In addition, a statistical test should be made.
Validity of the findings: The authors didn't have performed a sufficient analysis of the uncertainties in their results, for example, the numbers of significant figures and tables are not justified. The need for statistical analysis is required to make statements of the significance of differences in results. The authors have to explicitly study the uncertainties of the experimental variables that represent the key to the measurements. The performance metric plots results are not clear. Please explain Figure 4 in the text.
Additional comments: Line 221 - 240: The paragraph is really too long. It makes the meaning difficult to understand. It can be divided at least into 3 sentences.


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: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article validated an efficient method of the imputation of missing clinical test score values i.e. unsupervised DUCF method. This can enhance the application of machine learning in the field of medical science. The article is well structured with clear and unambiguous professional English. I have some minor queries.
1. Provide the equation to calculate the Absolute error percentage. The information provided in the manuscript is a bit confusing.
Experimental design: 1. As DUCF consists of two stages, one is calculating user-based similarities and the other is predicting the missing values, Although the authors have mentioned the approaches for the first step in detail but did not provide enough information regarding the methodology used to predict the missing values based on the similarities.
Validity of the findings: 1. As mentioned in line 165, the number of independent parameters is 603 while the number of samples is 816. As the number of parameters is very high so it can cause a Curse of dimensionality, any comments on this.
2. In line 235, the authors have reported that 1-norm distance i.e. MAN performs better. Provide the explanation behind this observation.
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: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 1
Reviewer: 3

Output:
Basic reporting: This paper proposed a new method to impute the missing clinical test scores using unsupervised dementia-related user-based collaborative filtering, and validate the reliability of predicted scores using MRI analysis. The study itself is innovative, and is within the interest of medical research field. However, the aim of this study should be justified at first, that is, why the MRI analysis can be used to validate the predicted clinical scores. Some details about the prediction method should be specified.
Experimental design: 1.	There are two aims in this paper, 1) impute clinical test scores, and 2) validate the reliability of imputed scores using MRI and demographics analysis. However, the title “Generating a reliability scale for clinical brain activity test scores based on medical scans and demographic data” only mentions the first aim, which is actually described less than the second aim. I suggest the author to change the title by including both aims.
2. Line 198, Page 6. It is unclear to me how the missing score is predicted. Please add more details about this.
3.	Line 135, Page3. “…, the ratio of the absolute difference between two to the test score range gives the error percentage.”  It is unclear that which ratio was used, ratio between absolute difference and “two to the test score range”, or the ratio between two and the test score range? What does “two” mean in this sentence?
Validity of the findings: A fundamental question of this study is the rationale of verifying the imputed/predicted missing clinical test scores with morphometrics and demographics. The author should firstly justify why the MRI analysis can be used to evaluate the reliability of the imputed clinical scores. In other words, how are the MRI analysis metrics correlated with full clinical test scores (without removing data)?
Additional comments: 1.	There are too many abbreviations throughout the paper, which are not commonly used. Please consider to eliminate the unnecessary abbreviations to make the paper easy to read. For example, “recommender systems” (RS), collaborative filtering (CF).
2.	Line 165, Page 5. “it is yielded” should be “it yields”.
3.	Line 174, Page 5. “is required” should be “requires”.


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: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors addressed all the comments. Therefore, the manuscript is acceptable in its present form.
Experimental design: The authors addressed all the comments. Therefore, the manuscript is acceptable in its present form.
Validity of the findings: The authors addressed all the comments. Therefore, the manuscript is acceptable in its present form.
Additional comments: The authors addressed all the comments. Therefore, the manuscript is acceptable in its present form.


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

Input:
Title: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 2
Reviewer: 2

Output:
Basic reporting: All the mentioned changes have been incorporated into the revised manuscript. It presents clean and professional English throughout with sufficient field background literature survey.
Experimental design: The research is within the Aims and scope of the journal. The manuscript has provided a new methodology i.e., Dementia-related user-based collaborative filtering for filling the missing values. This work would have a high impact to improve the accuracy of the models.
Validity of the findings: The underlying data provided is statistically sound and robust. The conclusions are well stated.
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: DEMENTIA-RELATED USER-BASED COLLABORATIVE FILTERING FOR IMPUTING MISSING DATA AND GENERATING A RELIABILITY SCALE ON CLINICAL TEST SCORES
Review round: 2
Reviewer: 3

Output:
Basic reporting: I thank the reviewer for addressing my comments. I am satisfied and have no further comments.
Experimental design: No comments.
Validity of the findings: No comments.
Additional comments: No comments.


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

Input:
Title: APPLICATION OF HYDROPHILIC POLYMERS FOR THE PREPARATION OF TADALAFIL SOLID DISPERSIONS: MICROMERITICS PROPERTIES, RELEASE AND ERECTILE DYSFUNCTION STUDIES IN MALE RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 'no comment'
Experimental design: 'no comment'
Validity of the findings: 'no comment'
Additional comments: The manuscript "Application of hydrophilic polymers for the preparation of Tadalafil solid dispersions:
Micromeritics properties, release and erectile dysfunction studies in male rats (#68972)" is very well written with strong scientific pieces of evidence. I recommend acceptance for publication as 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: APPLICATION OF HYDROPHILIC POLYMERS FOR THE PREPARATION OF TADALAFIL SOLID DISPERSIONS: MICROMERITICS PROPERTIES, RELEASE AND ERECTILE DYSFUNCTION STUDIES IN MALE RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: In all the manuscript, english grammar and sentences must be reviewed.  For example:
Page 7, line 59. Please correct “Entity (NCE) entities...”
Page 7. Please correct sentences from line 64 to line 68.

Page 7. Indeed, choice PDE5 or PDE5I

In line 75, please write empirical formula correctly, with the numbers adequately.
Experimental design: In line 93, please indicate the tadalafil aqueous solubility.

In section 2.1. Please indicate the weight (in mg) of drug and polymer for each preparation. Insert a sample table with the preparation conditions, if necessary.

In section 2.3 Why the solution needs to be filtered? Are you sure that all the weighed drug is solubilized? Contrary, it could be generated a concentration error!

In section 2.6 Please indicate correctly the heating flow. And, it was only one scan?

In 2.10.1. The 5mg/Kg of TFL, it was also oral administered? Please specify.
Validity of the findings: In lines 115-117. Please justify the advantages of using their preparation method compared with the enlisted.

According to the described in lines 118 to 135, please justify very well the novelty of this study.

In all results and discussion sections, please indicate specifically the results related with each study. In section 3.1, please indicate the figure or table number.

The described in lines 385-387: It is not desirable the sticky SDS in the wall of equipment-glassware. In order to solve that, the glass surface must to be modified or another surfactant agent must to be added.  This problem affects the stability and polydispersity of systems.

Please indicate the possible administration route of the prepared systems and justify why the 2-5 micrometer of particle size, irregularities and polydispersity could not affect.

For Figure 2, from section 3.3 must be improved. Please indicate the characteristics bands in all the FTIR spectra. Indeed, add the FTIR spectra off all polymers without TFDL.

In figure 3, endo and exo legends are too small.

In section lines 465-468, please indicates the following: The CMC (concentration micellar critical) for the Kolliphor p188. Is this CMC rebased under the preparation conditions?  If micelles are formed, why the irregular morphology obtained? Please explain how the polymer and TFL could be distributed in the prepared systems.
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 HYDROPHILIC POLYMERS FOR THE PREPARATION OF TADALAFIL SOLID DISPERSIONS: MICROMERITICS PROPERTIES, RELEASE AND ERECTILE DYSFUNCTION STUDIES IN MALE RATS
Review round: 1
Reviewer: 3

Output:
Basic reporting: All the related comments are in the additional comments box
Experimental design: All the related comments are in the additional comments box
Validity of the findings: All the related comments are in the additional comments box
Additional comments: 1.	Authors have not mentioned the ratio of all the polymers in the abstract (Koliphore 188, Kollidon® VA64, and Kollidon® 30 polymers in a 1:1 ratio). There are three polymers but the ratio is mentioned for only two polymers (unknown).
2.	Authors needed to perform HPLC for the estimation of Tadalafil from the formulation. The results from the UV spectrometer are not authentic most of the times because of interactions.
3.	How was the EE calculated? Was it a direct or indirect method? Please mention.
4.	USFDA has approved Tadalafil for oral use only, how could authors compare drug efficacy when given parenterally?
5.	Authors have not done optimization for the formulation to find the appropriate ration of drug and polymers.
6.	Authors need to add more relevant references e.g. doi: https://dx.doi.org/10.1016/j.ijpharm.2014.07.022, https://doi.org/10.1016/j.molliq.2018.02.091, doi: https://dx.doi.org/10.1016/j.mehy.2015.03.003, DOI: https://dx.doi.org/10.3109/10717544.2015.1105323; https://doi.org/10.1016/j.molliq.2017.11.081; DOI: https://dx.doi.org/10.26717/BJSTR.2020.31.005129


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 HYDROPHILIC POLYMERS FOR THE PREPARATION OF TADALAFIL SOLID DISPERSIONS: MICROMERITICS PROPERTIES, RELEASE AND ERECTILE DYSFUNCTION STUDIES IN MALE RATS
Review round: 2
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
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 HYDROPHILIC POLYMERS FOR THE PREPARATION OF TADALAFIL SOLID DISPERSIONS: MICROMERITICS PROPERTIES, RELEASE AND ERECTILE DYSFUNCTION STUDIES IN MALE RATS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Meets expectations
Experimental design: Meets expectations
Validity of the findings: Meets expectations
Additional comments: The authors have answered all the queries satisfactorily.
I recommend the manuscript to be accepted in this 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: HEALTHCARE APPLICATIONS OF SINGLE CAMERA MARKERLESS MOTION CAPTURE: A SCOPING REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall the paper is well written and easy to follow, with a clear message. Figures are also very clear. A couple of comments:

It would be helpful to note early on that “single camera motion capture systems” refers to both hardware-based solutions (e.g., Kinect, Leap Motion), as well as video-based offline solutions (e.g., OpenPose, DeepLabCut).

Line 245 states, regarding neurological injuries in adults: “in all cases voluntary movement is affected.” Is this a general statement about neurological injury? Or about the studies being included?
Experimental design: Was there a reason for not including psychiatric patient groups (e.g., autism, schizophrenia)? Or were they included under one of the other headings?
Validity of the findings: The authors comment on single camera motion capture being a much more accessible solution to kinematic analysis than more complex, expensive systems. While I very much agree, I wonder if the authors can comment on how ‘accessible’ the different single camera systems may be in different circumstances. For example, Kinect V2 requires a computer set up to run the Kinect, making this at least somewhat more cumbersome than video-based methods that only require a camera. On the other hand, video-based methods may require more expertise and/or computing power to actually process the data. This is particularly true of DeepLabCut, and to a less extent also OpenPose. Newer methods, such as MediaPipe doesn’t have this restriction as much, but have other limitations in its processing possibilities.

It would be helpful to indicate which system of motion capture was used in each study. For example, “The ability to detect bradykinesia, slowness of movement associated 296 with Parkinson’s, had strong agreement with clinical scores using measurements of wrist 297 supination and pronation, and hand opening and closing movements; however, they found that 298 finger tapping contributed little predictive power to the model due to inaccuracies with tracking 299 (Lee, Sinclair & Jones, 2019)”.  Knowing which systems succeeded in which tasks and populations (which is indicated in some paragraphs, but not others) would be quite useful information to include throughout the results, in my opinion.

As an extension of one of my previous questions, it may be useful to comment in the discussion on which method(s) may be most promising (e.g., based on what they can do, but also hardware requirements, etc), perhaps for different patient groups, or in different data acquisition contexts. If this is possible with this type of review.
Additional comments: No comment


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

Input:
Title: HEALTHCARE APPLICATIONS OF SINGLE CAMERA MARKERLESS MOTION CAPTURE: A SCOPING REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English is clear and professional and absolutely suitable to be read by an international audience.
The literature references show a detailed and in-depth knowledge of the clinical applications of markerless systems.
The structure of the article is consistent with PeerJ standards. In particular, there is an abstract divided into four sections: Background, Survey methodology, Results and Conclusions. The introduction describes correctly the field and its importance by clarifying which is the aim of this review (e.g. from lines 107 to 109 “this review provides insight into the healthcare application of SCMoCap to inform clinicians…” or from lines 111 to 114 “a scoping review was therefore conducted with the aim of investigations: which SCMoCap systems …”). In my opinion, the introduction should cite other previous reviews in the field of markerless analysis. The survey methodology is well-written since inclusion and exclusion criteria are reported.
To the best of my knowledge, the field of markerless analysis has not been reviewed recently and for this reason I can affirm that it is necessary a review that summarizes and clarifies all the information provided in the literature in this period.
Experimental design: In my opinion, the investigation is sufficiently rigorous and reflects the aims and scope of the journal. The methods are described with sufficient details. In particular, the search strategy, the inclusion and exclusion criteria are reported and there is also a figure which represents the flow chart of the review with which another investigator can replicate the same investigation.
The unbiased coverage of the subject is guaranteed by the presence of two investigators who have selected and read all the articles.
The sources of the selected articles are adequately cited.
The best point of this review is the organization of the paragraphs because the field of applicability of the markerless systems is very wide because it could include neurological injuries in Children and Adults, Neuromuscular Disorders and Frailty. The presence of these paragraphs makes the reading of the results clearer and more fluent. In addition, each paragraph contains a brief introduction about the selected disease for the readers who are not confident with it.
Validity of the findings: The conclusions are well written since they are connected with the research questions reported in the introduction.
In particular, both advantages and disadvantages of markerless systems are reported. I agree with the sentence reported from lines 499 to 501 since in my opinion markerless systems are suitable especially for screening purposes.
In the conclusions, unresolved questions and future directions are identified. In particular from the lines 558 to 563 the article reported the improving of tracking accuracy in presence of occlusions or of movements out of plane relative to the camera orientation as a future research.
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: PREVALENCE OF FOOD ADDICTION USING THE YALE-C SCALE IN MEXICAN CHILDREN WITH OVERWEIGHT AND OBESITY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors have some interesting data here that contribute to the growing literature on food addiction in children. However, there were some major concerns that need to be address before this manuscript is reading to be published:

-The authors present no hypotheses and do no outline there analyses on the association between food addiction in children and demographic variables anywhere in the introduction. These analyses are also not included in the abstract.

-There was a major meta-analysis on food addiction in children and adolescents recently published. This should be discussed in the intro and discussion.

https://onlinelibrary.wiley.com/doi/full/10.1111/obr.13183

- Given this meta-analysis, the comments about the lack of literature on this topic should be removed or lessened. However, the authors can and should highlight more the lack of research in Mexico (only one other study) in children where obesity rates are so high.

-There is no need for Table 2. This isn't a meta-analysis or structured review.
Experimental design: -The authors spend too much time focusing  on the endorsement of specific symptoms and how it differs by demographics. The YFAS-C is a scale that is meant to be either summed up or used as a diagnosis. Each symptom is not meant to stand on their own. Considering most to least endorsed in a table is reasonable, but the authors focus on this too much.

-The authors appear to interpret results that are non-significant. Non-significance should be clearly stated.

-The authors sometimes make statements about food addiction starting in childhood and continuing through adolescence. Given that this study is cross-section, the data can't support this statement. The cross sectional nature of the data should be a limitation and the need for longitudinal research highlighted.

- Information was missing from the methods, including what participants were told the study was about and whether they were compensated. Was a Spanish version of the YFAS-C used? Was it validated? Was it internally consistent?
Validity of the findings: -Through out this paper and in the abstract, the authors need to highlight that this is a sample of only children/adolescents with overweight/obesity who are seeking treatment. This likely inflates the prevalence and reduces the ability to detect differences by BMI and alters the symptoms endorsed. This should be clearly indicated in all interpretation of the findings.

-The discussion begins to feel very repetitive. A discussion of next steps for this literature (specially in Mexican children and adolescents is of importance).

-The lack of gender differences is notable and contrasts with typical eating disorders and should be discussed.

-The findings overall should be discussed in context of the prior meta-analysis.
Additional comments: The authors refer to substance abuse when it should be substance dependence when describing the YFAS-C in the methods

Gearhardt is referred to with a male pronoun, but is a female researcher.


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: PREVALENCE OF FOOD ADDICTION USING THE YALE-C SCALE IN MEXICAN CHILDREN WITH OVERWEIGHT AND OBESITY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Comments to the Authors:
In this study, the authors aimed to assess Prevalence of food addiction using the Yale-C scale in
overweight and obese children in Mexico. I have a number of minor and major comments.
1-	Please mention to the global prevalence of food addiction in the introduction section before the prevalence of food addiction in Mexico.
2-	Please mention "Food Addiction" with the abbreviation "FA" once in the text of the article. Then in the following times, the abbreviated form should be used in the text of the article. Also for the term YFAS in the same way in the text.
3-	The word “significant” does not make sense in the discussion section of the article. Please rewrite this sentence.
“The coincidence of our results may be justified by the fact that we included a significant number of adolescents (67%)”.
4-	In dissection section, line 219-222, grammatical errors seem to have occurred in this section, and the pronoun his needs to be corrected.
“In his initial study using the modified version of YFAS for a pediatric population, Gearhardt reported a 7.2% prevalence of FA. In contrast, in another study by Tompkins and collaborators (24) on overweight and obese adolescents, the reported prevalence was 30.7%.”
5-	The English language should be improved to ensure that an international audience can clearly understand your text. Please review and revise the manuscript for grammatical mistakes.
Experimental design: -
Validity of the findings: -
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: PREVALENCE OF FOOD ADDICTION USING THE YALE-C SCALE IN MEXICAN CHILDREN WITH OVERWEIGHT AND OBESITY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is clear and appropriately reviews the literature. The analytic approaches are appropriate.
Experimental design: The approach is clear and appropriate.
Validity of the findings: The findings are valid and data are appropriate.
Additional comments: The authors have addressed all my concerns


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

Input:
Title: AGEING- AND AAA-ASSOCIATED DIFFERENTIALLY EXPRESSED PROTEINS IDENTIFIED BY PROTEOMIC ANALYSIS IN MICE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript entitled “Ageing- and AAA-associated differentially expressed proteins identified by Proteomic Analysis” by Jinrui Ren et al. investigated the proteomic profiling of abdominal aortic aneurysm (AAA) in ApoE-/- male mice of different ages (10 or 24 weeks). They identified age- and AAA-related proteins were associated with the response to oxidative stress, coagulation and platelet activation, and complement and inflammation pathways.
Experimental design: None.
Validity of the findings: Authors identified AAA-related protein profile in ApoE-/- male mice model. How similar is the profile to human AAA patients?
Additional comments: 1.	Quality of Figure 2, 3, 4 and 5 could be improved. Do authors have images with higher resolution?
2.	Figures should be referred in the same order as presented. Figure 2A should be referred before Figure 2B. Figure 3B and 3C should be referred before Figure 3D.
3.	Table 1 showed GO and KEGG pathways terms with top 5 count number of DEPs, rather than 92 DEPs as stated in text line 221-224.
4.	Were Figure 4 and Figure 5 switched? Figure 4 showed DEPs co-expression network, hub proteins cluster, and transcription factors predict and Figure 5 showed the significantly enriched GO, KEGG pathways, and Mesh words, not as stated in the manuscript.
5.	Authors need to cite literature for roles of Tpi1, Eno1,and Prdx1 in text line 257-261.


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: AGEING- AND AAA-ASSOCIATED DIFFERENTIALLY EXPRESSED PROTEINS IDENTIFIED BY PROTEOMIC ANALYSIS IN MICE
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript by Jinrui Ren et al. contains some interesting data, and breaks some new ground. The concept that aging related proteins in circulatory may contribute to the development of AAA and this could have high physiological significance. To the authors credit, the data are in general supportive of the major conclusions. However, there appear to be some mistakes and flaws in the illustration and interpretations of some of the data and there is limited evidence for the overall concepts provided at this stage in the study.
Experimental design: The experiments design is very interesting to compare the different ages on the development of AAA.
Validity of the findings: 1.	1B shows Ang II causes significant increases in Aorta diameter in both ages, while the combination of age and Ang II have profound effects. However, do authors know how many the higher expressed proteins from AAA groups were secreted proteins? Did any published data indicate where these proteins coming from?

2.	In table 2, mistake in Fgb protein Rank number, should be number 10. Please correct it. In addition, Figure 1B showed the AngII woks in AAA youth mice, but authors did not show Hub proteins in AAA youth lab. Did authors compared the AAA youth VS Normal youth alone in Hub Proteins? Are there interesting proteins?

3.	Rows 310-312, the authors wrote ”dysregulation of glucose metabolism contributes to…….various disease, such as cancer….”. This is not an accurate description. Especially in cancer, the tumor growth require large amount of ATP and nutrients, causes the switch to the fast ATP-generating process, that is Glycolysis. So, it is not surprising, if the aortic wall expansion, leading to the changes in glycolysis. But it should be discussed more in the text. Maybe this is the reason why restricting glycolysis improve AAA development.
Additional comments: Mistakes: Figure 4 and Figure 5 figures were not consistent with the text in the manuscript. Figure5 refers GO, KEGG analysis. Figure 4 were String analysis.  Please correct it.


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

Input:
Title: AGEING- AND AAA-ASSOCIATED DIFFERENTIALLY EXPRESSED PROTEINS IDENTIFIED BY PROTEOMIC ANALYSIS IN MICE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Authors have successfully responded to my comments.
Experimental design: None.
Validity of the findings: None.
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: AGEING- AND AAA-ASSOCIATED DIFFERENTIALLY EXPRESSED PROTEINS IDENTIFIED BY PROTEOMIC ANALYSIS IN MICE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors answered my questions very carefully by supplying with more information about the secreted protein analysis in aged-related AAA formation. And authors had corrected their mistakes for the order of the figures.
Experimental design: None
Validity of the findings: Thanks to authors for offering more interesting data on secreted proteins. It is very interesting data from Table 2 and Figure 1B, indicating the age alone causes dramatic increases in several HuB proteins of DEPs, while four gens of them are secreted proteins. In mice, Vwf and Prdx2 may come from liver, which suggests the inter-organ communication from the other organs to regulate the AAA formation in old mice. Interesting data in terms of  physiological significance.
Additional comments: Please make sure to upload good quality of figures.


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: EXPLORING THE PERCEPTION AND READINESS OF PHARMACISTS TOWARDS TELEPHARMACY IMPLEMENTATION; A CROSS SECTIONAL ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The introduction part of the study is well explained. but I would like to suggest you to improve the description in the very first paragraph, in order to avoid the content repetition.
Experimental design: even though it had some limitations on the no. of participants, the methodological part can be well accepted.
Validity of the findings: your study primarily aimed to assess the perception of pharmacist towards telecommunications and the findings that obtained through the statistical analysis are similiar to the same type of study conducted in other countries as well. statistical analysis can be well accepted.
Additional comments: As a reader i would like to know that whether tele pharmacy is adapted in any part of the country, as this is as an innovative study in Pakistan for assessing the perception of pharmacist towarrds implementation of telepharmacy.


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: EXPLORING THE PERCEPTION AND READINESS OF PHARMACISTS TOWARDS TELEPHARMACY IMPLEMENTATION; A CROSS SECTIONAL ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Thu authors conducted a study on exploring the perception and readiness of Pakistani
pharmacists towards telepharmacy implementation. This is a timely research. I highly appreciated the authors effort to conduct such an important study. This is a good piece of work. The manuscript has a good potential and can be accepted after further revision. I have some observation which I think could help the author to improve the manuscript. My ocomments are below:

1.	The authors have done interesting work. Congratulation on your work on this topical issue.

2.	The abstract needs to be improved. I would suggest the authors follow the order of: What is the problem? Why is the problem important? What have others done to solve the problem? What are you doing to solve the problem? In addition, expand a bit of the method part in the abstract. Please remove the software name from the abstract section.

3.	The authors provided a narrative description of telepharmacy and its importance in healthcare service, especially for pharmacists. They also add a short literature review from lines 92-101. However, it was difficult to get a depth idea about the topics with such a short literature review. Include a literature review section where you present the results of earlier studies from the field.

4. The English language should be improved to ensure that an international audience can clearly understand your text. In particular, the Abstract, Introduction, Method, and Result section need major revision of English language editing. I suggest you have a colleague who is proficient in English and familiar with the subject matter review your manuscript.
Experimental design: 5. The article is original and within the scope of this journal.

6.	The author nicely articulated the evolution of telemedicine and its importance in healthcare service worldwide in the introduction section. They also explained telepharmacy which is the main topic of this current study. However, the authors did not indicate in the introduction section if this was the first study in Pakistan. Then, what was the novelty of this study? The author explained very little about the justification of the study which is not sufficient to understand the motivation to conduct this study. Need a clear explanation of the knowledge gap and addressing approach of this study. Therefore, I recommend adding a separate paragraph on the justification of the study by mentioning the knowledge gap and how to address them in this study. Please also explicitly mention the hypothesis or research question/objective if any. I suggest authors organize the introduction section from a more general to a more specific character.

7. The ethical standard is ok.

8.	The study designs and methodology are well explained. In the method section, the authors should clearly specify the sampling technique used in this study so that it is clear to the reader. For the online study, many authors used convenience sampling while others use the snowball sampling technique. However, both are associated with section bias. Thus, can you please explain how did you avoid the selection bias in your study?

9.	In method, the study instrument (2nd, 3rd, 4th, and 5th sections) used in this study was globally validated? If yes, please use a valid reference (otherwise it does not make sense). If not, how did you validate the survey items? If the survey item was author-produced, please estimate the internal consistency of the items.

10.	Why the authors did not consider other sociodemographic factors such as urbanicity, education, monthly income, area of work, job experience as a pharmacist which could have a major influence on the telepharmacy implementation?

11.	Please expand the statistical analysis section a bit (perhaps you can add why you used the chi-square test).
Validity of the findings: 12.	In the results section, the data presented in the tables are easily understandable and represent the tests performed. Highly Appreciated.

13.	In Table 1, to present “age”, you mentioned both percentages and mean (sd). Please use only one of them.

14.	In the discussion section, the author provided a narrative interpretation of the results but lack an explicit discussion on comparing present study findings with previous studies. Therefore, I suggest to expand the discussion accordingly (Answer your objective/hypotheses and link your results to the results of earlier studies in the field).

15.	Add a separate section on implication and policy recommendation.

16.	Add some strengths to your study.

17.	The limitation section must be improved. All limitations such as sampling biases, response bias, cross-sectional study consequences, other factors consideration, etc should be mentioned. Also, write about the future line of research on this topic.
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: EXPLORING THE PERCEPTION AND READINESS OF PHARMACISTS TOWARDS TELEPHARMACY IMPLEMENTATION; A CROSS SECTIONAL ANALYSIS
Review round: 2
Reviewer: 1

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


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

Input:
Title: EXPLORING THE PERCEPTION AND READINESS OF PHARMACISTS TOWARDS TELEPHARMACY IMPLEMENTATION; A CROSS SECTIONAL ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have substantially improved this section.
Experimental design: The authors have substantially improved this section.
Validity of the findings: The findings have well-validated.
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: OVEREXPRESSION OF AMCBF1 ENHANCES DROUGHT AND COLD STRESS TOLERANCE, AND IMPROVES PHOTOSYNTHESIS IN TRANSGENIC COTTON
Review round: 1
Reviewer: 1

Output:
Basic reporting: This work needs more experimental work to validate the transgenic materials generated, before doing the phenotyping analyses. Therefore, I have to reject it in its current form.
Experimental design: This work needs more experimental work to validate the transgenic materials generated, before doing the phenotyping analyses. Therefore, I have to reject it in its current form.
Validity of the findings: This work needs more experimental work to validate the transgenic materials generated, before doing the phenotyping analyses. Therefore, I have to reject it in its current form.
Additional comments: No additional comments