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

Input:
Title: A COMPARATIVE STUDY OF ROOT CANAL SHAPING USING PROTAPER UNIVERSAL AND PROTAPER NEXT ROTARY FILES IN PRECLINICAL DENTAL EDUCATION
Review round: 1
Reviewer: 3

Output:
Basic reporting: The author should mention the importance of using NiTi rotary instrumentation by the undergraduate dental student with more details, the author can refer to this study:
Comparative study of root-canal shaping with stainless steel and rotary NiTi files performed by preclinical dental students
Technology and Health Care 23 (3), 257-265
Experimental design: Was the participation of students in this study mandatory or voluntary?
is there any acceptance form for participation in this study?
is there any relation between students grades in this course and participating in this study
All these points should be mentioned in the materials and method section
Validity of the findings: The findings are valid
Additional comments: The manuscript provides a good idea but needs several modifications to be more valuable work where I have the following comments;
Introduction section:
1. The author should mention the importance of using NiTi rotary instrumentation by the undergraduate dental student with more details and the author can refer to this study:
Comparative study of root-canal shaping with stainless steel and rotary NiTi files performed by preclinical dental students
Technology and Health Care 23 (3), 257-265
Methods section
 Was the participation of students in this study mandatory or voluntary?
is there any acceptance form for participation in this study?
is there any relation between students grades in this course and participating in this study
All these points should be mentioned in the materials and method section

The students had applied root canal treatment to a total of 12 extracted human teeth in the phantom laboratory
what is the instrumentation technique do the student used  stainless steel file with step back technique

Canal straightening and the incidence of ledges were assessed on digital images with AutoCAD 2007.
This part needs more expansion

Discussion section
Narayanaraopeta & AlShwaimi (2015) announced that five of the six schools employed
 NiTi rotary instruments. Please this statement needs more details and clarification
Muñoz et al. (2014) showed that students also engaged in satisfactory shaping via reciprocal movement with a single file. but in your study, you used rotary systems. and you could improve your study by comparing a rotary system with a reciprocating single file system

These researchers assessed the amount of straightening of the channels prepared by the third-year students, who had no experience with the rotary instruments, in the extracted molar teeth.
Is there no effect of the difference between dentin in the root canal  and resin in simulated root canals on the apical transportation you can refer to the following article:
Assessment of apical transportation caused by nickel-titanium rotary systems with full rotation and reciprocating movements using extracted teeth and resin blocks with simulated root canals: A comparative study
In severely curved canals, the master apical size should be smaller than #30; Where is the reference?
In our study, the incidence rates of deformation and fracture were 17% for PTU and 9% for PTN. Can you explain this difference in fracture rate between PTU and PTN

Ledges occurred in 19% of the canals prepared with PTU, whereas ledges formed
in 4% of PTN cases. Does swaggering motion in PTN has any effects in reducing ledges

Within the limitations of this study, What are the limitations of your study?
and what is your suggestion to improve teaching and using NiTi systems by undergraduate dental students


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

Input:
Title: A COMPARATIVE STUDY OF ROOT CANAL SHAPING USING PROTAPER UNIVERSAL AND PROTAPER NEXT ROTARY FILES IN PRECLINICAL DENTAL EDUCATION
Review round: 2
Reviewer: 1

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


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

Input:
Title: A COMPARATIVE STUDY OF ROOT CANAL SHAPING USING PROTAPER UNIVERSAL AND PROTAPER NEXT ROTARY FILES IN PRECLINICAL DENTAL EDUCATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript is written in clear English in professional method, with sufficient background and referenced appropriately.
The article structured in  an acceptable format .
Figures  relevant to the content of the article, of sufficient resolution, and appropriately described and labeled.
The manuscript is self-contained with relevant results to hypotheses.
Experimental design: The manuscript has  a  clear define  research question, with  high technical standard.
Methods  described with sufficient information and can  be reproducible by another investigator.
Validity of the findings: The findings of this manuscript are rationale & benefit to literature .
The conclusions are well stated, linked to original research question and  supported by  results.
Additional comments: this manuscript could be considered suitable for publication now and provide  valuiabl information in field of teaching endodontic


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: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The aim of the study was analyses the effect of fallow treatment on nutrient dynamic and bacterial composition in the subsoil plots in China. The paper fit the scope of Peer J. and it had a good data, however the actual version of Ms had several problems. The English grammar should be improved to ensure that an international audience can clearly understand. I suggest to the authors that the paper must be revised by a native English-spoken before submitting the paper. Additionally, the actual version of the introduction is very general and without a clear context. The study had no a clear general question. What are the novel results presented? What is the conceptual basis for test the hypotheses of the present papers? The authors did not show clearly the results of previous studies related with the factors of fallow treatments and vegetation types used in the present paper. The experimental design must be explained clearly. The presentation of result must be improved. The actual version has no a clear narrative history. Which are the hypotheses that the authors want to test? The sections of results seem with no clear link among them. Therefore, the discussion is very speculative, because the narrative history of this study is not clear. For this reason, this paper requires improve significative before accepted for its publication in PEER J.
Experimental design: Methods
The English language should be improved to ensure that an international audience can clearly understand your text. The current phrasing makes comprehension difficult. Below I show examples where the language must be improved. Additionally, experimental design must be explained clearly. A specific comment of methods used are below:

L107-121: Improve English language of this paragraph.
L107-109: I can not understand the experimental design. Each treatment has three plots (replicates)? The authors mentioned that “There were four parallel plots for each treatment” (L107-108) and below they mentioned: “and the plots were randomly distributed” (L109). Are the plots of each treatment together? Please improve the explanation of experimental design.
L109-114: move these sentences to introduction and explain clearly how this plant species affects soil fertility and soil microbial community.
L114-130: but the authors mentioned before: “We have set five treatments in the study area” (L107).
L156-157: Was the inorganic C concentration measured of the extraction for microbial C analysis? If the soil has carbonates, a correction is needed for SOC and Microbial C.
L200-202: Move these sentences after ANOVAs explanations.
L202-203: The time factors must be analyzed with RMANOVA.
Validity of the findings: Results
The presentation of result must be improved. The actual version has no a clear narrative history. Which are the hypotheses that the authors want to test? The sections of results seem with no clear link among them. I recommend doing an integration analyses of physicochemical variables and nutrients within microbial biomass as principal components analyses or another multivariate analyses. Why did the authors only show the Spearman correlation with 3 specific bacterial group? They must explain, mainly if the MDS analyses are no different when used all sampling dates.

L211-221: This paragraph is a narration of table 1, and the readers can not see the effect of fallow treatment on physicochemical variables and nutrient concentration within microbial biomass. I recommend moving as supplementary material the Table 1 and include in the text the Fig S2. Therefore, this paragraph must rewrote showing the principal results from Fig. S2.
Fig. S2: this figure must be following the two-way ANOVA. When the interaction is significant they can not compare de main factors (vegetation and manure). For example, TN of August sampled presented how if the factorial was not significant, while TN of October samples presented how if the factorial was significant. However, the interaction was significant for TN of August sample, but it was not for October samples (Table1). The authors must improve the presentation of these results.
L224-231: However, when the analyses did with all sampling data, the value of stress is 0.136, meaning no differences related to treatments (Fig. 1). Therefore, the authors must review their conclusion. Mainly if gamma diversity was also no significant.
Table 2: I suggest move table 2 to supplementary material.
L233-239: Improve English grammar of these sentences.
Additional comments: 1. Introduction
The English language should be improved to ensure that an international audience can clearly understand your text. The current phrasing makes comprehension difficult. Below I show examples where the language must be improved. Additionally, the actual version of the introduction is very general and without a clear context. The study had no a clear general question. What is the novel results presented? What is the conceptual basis for test the hypotheses of the present papers? The authors did not show clearly the results of previous studies related with the effect of fallow treatments used in the present paper on the nutrient dynamic and composition of microbial community.

L38-40: rewrote sentences as: Nevertheless, studies have also pointed out that spontaneous vegetation for fallow without fertilization is risky, because nutrients input is still necessary (Reference?).
L43-47: Improve these sentences, because they are not clear.
L42-49: The structure of these sentences must be improved, because their context is not clear.
L58-61: Improve these sentences. I recommend that the authors show the main results of the previous study (which nutrient availability increased with fallow).
L61-66: I recommend that these sentences shoe in a different paragraph. Additionally, the authors must show the results of previous papers of effect of fallow in nutrient and bacterial composition in subsoil.
L66-67: What is the context of this sentence?
L67-68: Context?
L73-76: Context?
L50-76: The structure of this paragraph must be improved. The actual version is a several sentences with no clear context and link among them.
L78-79: Which do changes of microbial communities reported the cited authors?
L82-84: I can not understand if the 5 years field experiment finish or why the authors reported only the first year. Why do authors report the results at the end of 5 years field experiment?
L81-86: The authors must show the objective clearly and the related hypotheses. What is the new knowledge of the present paper?

4. Discussion.
 The discussion is very limited and very speculative. The authors must improve the narrative history of the present paper.

L246-247: this is not true, microbial C biomass was not significant among fallow treatments.


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: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 1
Reviewer: 2

Output:
Basic reporting: No Comment
Experimental design: The experiment is well design
Validity of the findings: The findings in the present study is well presented.
Additional comments: This article provide basic information for improving fallow management that enhance soil fertility and increase its productivity. I suggest for publication in after incorporating few comments below:
Please add the value or percentage of the results obtains in abstract.
Line 24: Instead of more nitrogen and more ammonia.. Mention the percentage by which it is more..
Line 28-29: How Blastopirellula connected to MBN but not to MBC.
Line 308: Mention specific soil  characteristics.
Few typographical errors should be taken care.


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: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The aim of the study was analyses the effect of fallow treatment on nutrient dynamic and bacterial composition in the subsoil plots in China. The paper fit the scope of Peer J. and it had a good data. The new version improved some sections, but the paper requires more work. In particular, the conceptual basis must be improved in the Introduction and the hypothesis still remains very general. Therefore, the discussion is very speculative. For this reason, this paper still requires improve before accepted for its publication in PEER J.
Experimental design: The material and methods section were improved by the authors. I suggest that the time factors must be analyzed with RMANOVA.
Validity of the findings: 3. Results
The results section was improved by the authors. However, some sentences must be improved, as well as Figure 2 (see below).

L237-238: Improve English grammar of this sentence.
L241-249: This paragraph must be improved, because require a clear narrative history. The current version does not explain which treatments affects the studied variables. The authors must present the main results obtained, without to describe in detail the figure 2. I suggest that the authors must define which are the main narrative argument for improve the redaction of these sentences.
Fig. 2: This figure must be improved, because it is not clear. The current version assume that all variables were affected by interaction. In the case that interaction was significant, I suggest that the differences among means use letters (Upper- and Lowercase for within manure treatments and within vegetation cover). In the case that the interaction factor was not significant, I suggest that the authors show the principal factor with statistical differences.
L251: I suggest: “…significant differences; therefore, we focused…”
L253: Changed “which indicates a decrease in differences” by “which indicates that differences decreased”
L253-256: Improve English grammar.
Additional comments: The aim of the study was analyses the effect of fallow treatment on nutrient dynamic and bacterial composition in the subsoil plots in China. The paper fit the scope of Peer J. and it had a good data. The new version improved some sections, but the paper requires more work. In particular, the conceptual basis must be improved in the Introduction and the hypothesis still remains very general. Therefore, the discussion is very speculative. For this reason, this paper still requires improve before accepted for its publication in PEER J.

In detail:
1. Introduction
The conceptual basis must be improved in the Introduction. The authors hypothesize that mixed plants and farmyard manure may accelerate changes in subsoil nutrients and bacterial communities under short-term fallow management. However, they did not explain why they expect these results. There a several studies related with how the chemical composition of organic matter affect soil microbial composition and its activity, and therefore soil nutrients transformation. I recommend that the authors explain why the mixed plants improve soil microbial activity.  The hypothesis still remains very general, it must be improved.
L40-56: I recommend reducing this paragraph with the most important sentences.
L57-75: I recommend delete this paragraph, because this version looks like a paper list related with fallow.
L93-104: These sentences must be improved. I suggest that the authors report the main results of how the fallow affect soil nutrients transformation. It is very important that the authors explain soil processes reported in previous papers.
L105-107: What is the context of these sentences?

2. Materials and Methods.
The material and methods section were improved by the authors. I suggest that the time factors must be analyzed with RMANOVA.

3. Results
The results section was improved by the authors. However, some sentences must be improved, as well as Figure 2 (see below).
L237-238: Improve English grammar of this sentence.
L241-249: This paragraph must be improved, because require a clear narrative history. The current version does not explain which treatments affects the studied variables. The authors must present the main results obtained, without to describe in detail the figure 2. I suggest that the authors must define which are the main narrative argument for improve the redaction of these sentences.
Fig. 2: This figure must be improved, because it is not clear. The current version assume that all variables were affected by interaction. In the case that interaction was significant, I suggest that the differences among means use letters (Upper- and Lowercase for within manure treatments and within vegetation cover). In the case that the interaction factor was not significant, I suggest that the authors show the principal factor with statistical differences.
L251: I suggest: “…significant differences; therefore, we focused…”
L253: Changed “which indicates a decrease in differences” by “which indicates that differences decreased”
L253-256: Improve English grammar.

4. Discussion.
The discussion is still very speculative. The authors must improve the narrative history of the current version and this section must be strongly linked with the other sections, mainly with introduction and results.


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

Input:
Title: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 2
Reviewer: 2

Output:
Basic reporting: See attached PDF
Experimental design: See attached PDF
Validity of the findings: See attached PDF
Additional comments: The responses were satisfactory. However, minor changes are made in the present review.


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

Input:
Title: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 3
Reviewer: 1

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: The ms may be accepted as it is and suggested for publication.


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

Input:
Title: IMPROVEMENT OF SUBSOIL PHYSICOCHEMICAL AND MICROBIAL PROPERTIES BY SHORT-TERM FALLOW PRACTICES
Review round: 3
Reviewer: 2

Output:
Basic reporting: The aim of the article is very interesting but unfortunately, major corrections are necessary.

Principally, I think that the methods described are inadequate and need be improved but if the authors consider that was correct please improve the description and response to other reviewers. For these reasons, the discussed findings and conclusions aren’t clear in light of the methods and results presented.

Introduction
L62 I suggest delete “Therefore, hybrid vegetation was used in our study.”
L89-92 Please move the hypothesis to the final paragraph
L93-107 These paragraphs are outside in reference to the development of introduction, please review and improve.
L107-110 Here appears another hypothesis that will be redundant and/or oppose (partially) with the former. Please review and improving and put the (one or two) hypothesis (ses) after the description of the way of this study in L110
Materials & methods
L132-L134 Please homogenizes all the units (preferent to mg g-1) for better comprehension by non-expert readers.
L223- You include time as a factor (three dates) in the experimental design but don’t use in statistical models. I see that previous reviewer suggest RMANOVA and I agree with him. Your response was that included time as a factor in fig 4 and table S5 but these are unsatisfactory. Completely analyses were performed not was ANOVA per date but as RMANOVA (Between subjects: fallowing treatments; Within subjects: Sampling date and interaction (fallowing X sampling date))
L223- Please describe how performed the PCA, the number and type of variables are important for interpretation…
L233- Why Spearman’s rank test? Other analyses are parametric, please expand the arguments for statistical selection
Results
L235-239 The effect by treatment in PCA is not clear, the distribution in two axes simultaneously is difficult to describe in the sense that not a pattern is consistently in all dates. I suggest not focuses on the results in PCA before describing individual behavior of nutrients in RMANOVA.
L238 Please improve the description of these 3 groups that are important, an if this is very important supplementary data is not the correct site. The actual form is very confusing.
L240 If you performed a multivariate analysis using SOC and C/N ratio (constructed with SOC) you are autocorrelating and this is a basic error that influences a bad interpretation.
L239- You do not describe that two-way anova was used to identify the effect of fertilization and vegetation! This a necessary for understanding the analyses since methods, in current form the paper surprises continuously with results derived from analyses not described.
Discussion
L282 ammonia (NH3+) is incorrect, in methods you refer that quantify NH4+ this is ammonium! After that you correct the term, I suggest using only ammonium.
Experimental design: No comment
Validity of the findings: The discussed findings and conclusions aren’t clear in light of the methods and results presented.
Additional comments: Need to improve and clarify the strongest the material and methods for validating the presentation of the results and validate the findings. The actual response to former reviewers is not satisfactory by improvement methods.


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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper didn't show enough details of the methodology. For example, the authors mentioned a model algorithm was established in Line 142 and the visual interpretation method was used in Line 149. However, no details of these methods were given in the paper.
Experimental design: The authors should improve the visualization of the experiment results, e.g. explicitly show the changing areas in Figure 2 and Figure 4.
Validity of the findings: The article fails to show the accuracies of land cover classification and change detection.
Additional comments: The authors should present the main contributions of this paper compared with existing studies in the section of Introduction.


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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article deals with the Monitoring and Analysis of a Small Port Expansion Based on Multi-source and Multi-temporal High-resolution Imagery. This is an interesting topic. However, the current title is not appropriate, I recommend the authors replace the Multi-temporal High-resolution Imagery with Remote sensing data. The writing is clear. Overall, the manuscript is publishable, but, needs a major revision first. There are many grammatical and structural mistakes in the manuscript and I am not able to highlight/revise all of them. So, please revise the manuscript and improve it considerably in terms of grammatical and structural mistakes.
The literature is suitable for the purpose of the article.
Experimental design: 1) Within Scope of the PeerJ journal.
2) Research question well defined, relevant & meaningful. It is stated how the research fills an identified knowledge gap.
3) Structure conforms to technical & ethical standard.
4) Although the experimentation is appropriate, the paper would gain in interest if it would provide a more in-depth discussion about the selected topic.
Validity of the findings: 1) Data is robust, statistically sound, & controlled.
2) Conclusions are well stated, linked to original research question & limited to supporting results.
3) Speculation is welcome.
Additional comments: 1.	The authors should thoroughly address the novelty of this study, because so many works have been done in this point.
2.	The section of introduction did not well review and cite existed works.
3.	The section of study area is unclear and redundant, there are many unrelated contents of your study.
4.	Line 109 This paper studies the expansion process for the port of Ajmr, which has been under construction since 2002. However, nearly no expansion occurred from 2002 to 2009, so only images from 2009 to 2018 were selected for preprocessing. How can the authors prove this?
5.	The spatial resolution of these selected remote sensing data are definitely different. The authors should address problem properly.
6.	A sketch map should be added in the revision to illustrate the detailed processing.
7.	The section of Survey methodology, I want to see some substantial details about the field validation.
8.	The detailed symbols of different Land use should be added and well illustrated.
9.	Line 158 the visual interpretation method was used to analyze the spatio-temporal dynamic changes in the land… What’s the accuracy of the detection? The accuracy assessment is essential to this study.
10.	The equations should be listed in order.
11.	I want to know why the 5% and 95% are regarded as the thresholds?
12.	Currently, the results and analysis are separated. They should be an integration.
13.	The discussion is missing and unsatisfied, the author should make the best effort to modify this section. I want to see a considerable improvement in this regard.
14.	The conclusion I not well stated and is written more as an abstract. A clear conclusion needs to be written.
15.	All tables/figures: Caption needs to completely explain the table, including what the abbreviations mean. Work on text clarification and formatting, improve figures, list of the abbreviations would be useful in this case to easily understand 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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 1
Reviewer: 3

Output:
Basic reporting: This manuscript failed to show why it is important to map small port expansion and did a poor job in demonstrating why this work is necessary. To improve, please focus on why mapping small port expansions are important,  what are the current state-of-art methods to do so, what are the knowledge gaps, and how this work can contribute to fill in the gaps.
Experimental design: Research questions were no clear in this manuscript, and therefore I cannot evaluate whether the methods taken here is novel or helpful.
Plus, when mapping port expansion (Figure 2), only a very small area are used, but for vegetation cover change, the authors decided to use a much larger region. Are you assuming that a small port will have affect vegetation in a much larger region? If so, what is your basis for this assumption and why was this area in Figure 3 chosen?

Please clearly state your research questions, and also why your approach can answer these questions. And properly show your assumptions for port expansion and vegetation change.
Validity of the findings: No comments.
Additional comments: This manuscript might be OK for a course project, but not for a research journal. To work towards a manuscript worth publishing, I recommend the authors 1) clearly identify the importance of the research subject; 2) review the latest developments in your research area; 3) identify the current knowledge gap in this field (your research question); 4) state your proposed solutions and the outcomes.


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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The article has met the standards.
Experimental design: The article has met the standards.
Validity of the findings: The article has met the standards.
Additional comments: The authors have made suitable revision to my comments. Here are some minor points that should be revised before publication.
1. Please add a legend to Figure 6.
2. Please move the content of discussion to the Conclusion Section.
3. the accuracy is 1 m -> the resolution is 1 m.


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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 2
Reviewer: 2

Output:
Basic reporting: As I can see from the previous review, you have corrected most of the reviwers comments. Especially for the research questions, and also why your approach can answer these questions. I mean the scientific problems.
Experimental design: no
Validity of the findings: no
Additional comments: no


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: MONITORING AND ANALYSIS OF THE EXPANSION OF THE AJMR PORT, DAVAO CITY, PHILIPPINES USING MULTI-SOURCE REMOTE SENSING DATA
Review round: 2
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors did not address my previous comments and I have serious concerns about the quality of the paper. In addition to the previous comments, here are a few more.

1.	The current discussion reads like a conclusion. Please do a research about what a decent discussion looks like and improve it.
2.	L411 - I do not think the authors can conclude that the construction of the port did not have a significant impact on the ecology of the area. How do you define 'ecology'? Did you do any test?
3.	L423-425 – these statements are not supported by your data. I am shocked to see these conclusions.


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: DIFFERENTIAL EXPRESSION OF ATWAKL10 IN RESPONSE TO NITRIC OXIDE SUGGESTS A PUTATIVE ROLE IN BIOTIC AND ABIOTIC STRESS RESPONSES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript need to be carefully checked. I could find may typo in the current version.
Suffience background and refs were provided.
The article structure, figure, table and raw data are well prepared.
Experimental design: The authors provided novel findings to provide the role of WAKL10 in biotic and abiotic stress resistance. Experiments are well designed and performed. However, experimental details of how many biological and experimental replicates of experiments were performed need to be provided.
Validity of the findings: The finding of manuscript is novel and important for the understanding of NO responsive gene WAKL10 in biotic and abiotic stress resistance. Although statistical analysis was performed, but some labels are missing the figures. Some of conclusion are not well stated.
Additional comments: In this manuscript, the authors investigated the role of Arabidopsis WAKL10 in biotic and abiotic stress responses. The T-DNA insertion mutant of WAKL10 is insensitive to GySNO treatment, and more susceptible to Pst DC3000, suggesting the dual role of WAKL10 in NO signaling and pathogen resistance. Sufficient background and refs were provided in the manuscript, and the experiments were well performed.
However, the manuscript still need to be improved for publishing in PeerJ.
1. First of all, the manuscript need to be carefully written. I could find quite a lot of typo in the manuscript.
For instance, I was confused with the name of ‘WAKL10’. In the figures, only ‘atwalk’ appears. So, please carefully check and give the full name of WAKL10/WALK10 in the beginning of the manuscript.
Line 139, concentration of ‘5x 105 10 CFU mL-1’, please check.
Pst DC3000 (avrB) should be Pst DC3000 avrB.
‘(Student’s t-test with 95 (*) and 99 % (**) confidence levels. A ‘)’ was lost in the end of the sentence.
… …

2. The authors also investigated the role of atgsnor1-3 for chemical treatment and pathogen treatment. However, why the authors include gsnor1 in this study is unclear, and no further discussion was found for the role of GSNOR in biotic and abiotic stresses.

3. line 216-217. Although PR1 is one of the marker for SA mediated immunity in Arabidopsis. However, the conclusion that ‘due to down regulation of SA-dependent PR genes’ with only PR1 expression may not sufficient.

4. The major point of this manuscript is the role of WAKL10 in ETI. It is reported that HR phenotype, ROS accumulation and iron leakage are accompanied with ETI. The atwakl10 mutant exhibits high level of HR, ROS accumulation and iron leakage, indicating WAKL10 negatively contributes to AvrB triggered ETI in Arabidopsis. This is inconsistent with the bacterial growth results, which makes the authors conclude that WAKL10 is positive to ETI. How would the authors explain this results?

5. Detail information of biological and experimental replicates performed in each experiments need to be described.

6. In the figure legends. The authors mentioned that ‘(Student’s t-test with 95 (*) and 99 % (**) confidence levels). However, some of the ‘*’ was lost in the figures. For instance, Fig. 1C, 2C, 3B, 5B

7. Primer ‘R’ for genotyping need to be provided in table 1.
In addition to this point (but not necessary for this revision), the authors showed that several WRKY and DREB binding site are appeared in the promoter region of WAKL10. Confirmation of WAKL10 expression in wrky and/or dreb mutants under Pst infection or salt treated conditions may provide new evidences to understand how WAKL10 was regulated under biotic and abiotic conditions.


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: DIFFERENTIAL EXPRESSION OF ATWAKL10 IN RESPONSE TO NITRIC OXIDE SUGGESTS A PUTATIVE ROLE IN BIOTIC AND ABIOTIC STRESS RESPONSES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Nitric oxide offers a very important regulatory role in plant response to environmental stresses while NO production is an important feature of plants under biotic and abiotic stresses. In this study an ARABIDOPSIS WALL ASSOCIATED KINASE-LIKE 10 (AtWAKL10) loss-of-function mutant atwakl10 was used to investigate for disease phenotype after Pst infections using both virulent and avirulent strains. Moreover, mutant line was also tested for tolerance against two NO-donors, drought and salinity stress responses regarding germination/survival rates. The gene expression profiling by qPCR also gave interesting data also. However, there are some improvements suggested as below which may be incorporated before the acceptance of this paper.

Throughout the text there is an inconsistent use of WALK10 and WAKL10. Please correct as appropriate. Add some information on atgsnor1-3 and its susceptibility to Pst, since, this mutant has been used in almost all experiments as a positive control with high pathogen growth or HR. There are some languages issues found throughout the text which can be improved with some effort by the authors.

The discussion section is well elaborated. I would suggest to add on a the possible relationship of WALK10 with NO tolerance, and that, why mutant plants showed mortality by adding NO donor (CysNO) in MS media compared to wild-type.
Experimental design: Add in M&M section, what statistics was used. If one-way ANOVA, please mention about F statistics along with number of repeats.
Validity of the findings: The study overall was well conceived and planned. The gene expression profiling, ion-leakage and staining experiments are convincing. The data obtained is of significance in the field of stress biology where a novel AtWAKL10 gene function has been associated with disease resistance and abiotic stress tolerance / susceptibility.
Additional comments: Line 48: Please rephrase. Omit "one of the smallest". "known" not “knows”
Line 51: Please omit miraculous
Line 52: Change citation order throughout the text. Write old ref. first in ascending order. Cite Liu et al., Cell (2004). Essential roles of S-nitrosothiols in vascular homeostasis and endotoxic shock.
Line 53 and 58: add Delledonne et al, 1998 (Nature).
Line 59: please rephrase.
Line 60: Omit atmospheric.
Line 61: replace “may give significant damage” with “cause significant damage”
Line 64: Rephrase “is a key feature in response to”
Line 74: Also cite Jones and Dangl, Nature 2006. The plant immune system.
Rephrase “pathogen recognition obstructs pathogen progression”
Line 84: rephrase “ little is known about its catalytic enzyme GCs”
Line 97: Replace both references with primary references for NO addition to proteins. Add Stamler’s work e.g. PNAS 1992 “S-Nitrosylation of proteins with nitric oxide”
Line 101: Replace “CySNO” with CysNO
Line 102: rephrase “most major”
Line 117 and 130: replace ±23°C with 23°C ± ??
Line 134: Add primary reference.
Line 139: 5 x 105 10 CFU mL looks like a typo. Please mention the inoculum dose for Pst vir and avr, separately.
Line 152: replace “then let to cool down” with” allowed to cool-down.
Line 156: Omit reference. The method described is sufficient. Add OD or cfu at which leaves were infiltrated.
Section 1.6: Mention experimental controls and number of repeats.
Line 170: Omit reference
Line 184: rephrase “Arabidopsis/rice was selected as conserved region”
Line 194 -196: Omit methodology. It is already there in M & M. what is MV?
Line 198: write cell death frequency (CDF) and use abbreviation onward.

Fig. 1(A): Mention in the fig legend, what is F+R and B+R. Omit ladder labelling but bar for the size of the gene band.
Fig. 2(B): Considering almost 2 log difference in pathogen growth for atwakl10 plants compared to wild-type at 2 and 4 dpi depicted by a single asterisk * saying the difference is significant at 95% needs to be rechecked for any possible errors. in my view it could be highly significant at 99% (**).
Fig. 1(D): this figure is not used in the text. Looks like showing morphological attributes of mutant lines compared to Col-0 (wild-type)?

Fig. 2(C): Seems like fold-change in PR relative expression not just relative expression. A high relative PR gene expression at 0 hpi (control) is surprising, since PR gene expression is triggered only after pathogen interaction.

Line 221: replace “within” with “in”
Line 225: Please add significance level (P≤0.01) or (P≤0.05).
Line 231: “posited” ??

Fig. 3(B): A control at day 0 is missing for comparison.

Line 246: Delete reference.
Line 249: Table S1 is missing for commentary.
Line 267: Replace “played” with “play”
Line 269: Italicize DREB2 and 19
Line 271: Please rephrase “the atwakl10 lines were deprived of water for two weeks”
Line 297: Please rephrase “CDF percentage about 80%.”
Line 343: replace “slat” with “salt”
Line 354: delete reference
Line 359: please rephrase “We hope that this article will add to our knowledge about NO biology during plant defense and environmental insults”


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: DIFFERENTIAL EXPRESSION OF ATWAKL10 IN RESPONSE TO NITRIC OXIDE SUGGESTS A PUTATIVE ROLE IN BIOTIC AND ABIOTIC STRESS RESPONSES
Review round: 2
Reviewer: 1

Output:
Basic reporting: Professional language checking need to be applied to ensure that international audiences could clearly understand.
References are improved and sufficient information is provided.
The structure of article, figures and tables are clear.
Experimental design: The authors provided novel findings to illustrate the role of AtAWLK10 in plant development, biotic and abiotic stress responses. Sufficient details of methods are described.
Validity of the findings: Comparing with previous version, number of replicate in each experiment and statistical information is provided.
Additional comments: The authors provided interesting finding for the role of AtWALK10 in plant development, biotic and abiotic stress responses, indicating AtWALK10 is important for NO signaling which plays viral role in differential each processes.
However, the major issue of this study is the bacterial growth assay in Figure2 and Figure3. The authors showed in Figure 2 that the Pst population and PR1 expression almost not changed at different time point in Col-0. Similarly, in Figure 3, Pst AvrB growth at 2 and 4 dpi are similar (or even reduced). Those are inconsistent with previous reports. (This may due to unsuccessful infiltration or other factors. However, the authors applied at least three times for each experiments.) Therefore, the conclusion addressed from those experiments might misleading.


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: DIFFERENTIAL EXPRESSION OF ATWAKL10 IN RESPONSE TO NITRIC OXIDE SUGGESTS A PUTATIVE ROLE IN BIOTIC AND ABIOTIC STRESS RESPONSES
Review round: 2
Reviewer: 2

Output:
Basic reporting: This is a revised submission and authors have incorporated almost all the suggestions and managed to clarify many ambiguities.
A few necessary comments have now been incorporated in the main file using Track changes option.
Experimental design: OK
Validity of the findings: OK
Additional comments: A few minor suggestions be addressed.


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

Input:
Title: TOWARDS A BARNACLE TREE OF LIFE: INTEGRATING DIVERSE PHYLOGENETIC EFFORTS INTO A COMPREHENSIVE HYPOTHESIS OF THECOSTRACAN EVOLUTION
Review round: 1
Reviewer: 1

Output:
Basic reporting: English fine, but a number of complex terms are used, of which the meaning is not clear to me, anyway.

References fine

structure fine.

not sure what this means precisely.
Experimental design: NA
Validity of the findings: Negative results (i.e. phylogenetically unresolved thoracican taxa) needs discussion and explanation
Additional comments: Reading this MS, I first wondered that any success has been achieved in unraveling the barnacle tree of life! However, the reverse seems to be true, insofar as the sessile barnacles and some stalked groups is concerned, for which molecular phylogenies largely congruent with morphological data have been published. I think the paper should emphasise these positive and important findings, and also clearly  explain why a mass of thoracican taxa included in fig. 3 (scalpellids, balanomorphs etc) have unresolved relationships.

I am not entirely sure what “little integration across taxonomic levels” actually means, or  “lack of significant overlap in taxonomic entities” or  “lack of overlap of characters” or indeed “overlap in represented taxa”. I suspect that some other readers are also unsure. Please explain clearly.

I can also see that there are difficulties in getting consistent results with basal relationships of groups, and that this needs much further study.

Fig. 3 clearly incorporates a lot of data, and indicates a number of significant relationships but is almost impossible to use, because if one zooms in to see names, the relationships are no longer visible. Why not also use colour for individual thoracican clades (e.g. balanomorphs) and add names down to family or superfamily level, as appropriate? This would then emphasise what we do know. The other question is one of support for clades, which is pretty essential really. An alternative would be to add a standard cladogram with support for individual nodes.

OK, so there’s a vast arc (NNE-SWS) of unresolved thoracicans, including scalpellids, balanomorphs etc. amongst which Ibliformes reside. Is there not some difference in the data available (quality, amount?) for these taxa? It seems bizarre, that their close relatives should yield phylogenetically important information, yet they do not. This needs explanation, surely.

A list of what we do know about barnacle evolution from molecules would help. This might include:

  - Basal position of Ibla

  - Monophyletic Sessilia, divided into clades of balanomorphs and verrucomorphs

-	a highly resolved tree for the balanomorphs (Perez-Losada et al. 2014), with considerable detail

-	a detailed tree for the Scalpellidae, which is congruent with morphology (e.g. Lin et al. 2015)

and other points as appropriate.

Andy Gale
-


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: TOWARDS A BARNACLE TREE OF LIFE: INTEGRATING DIVERSE PHYLOGENETIC EFFORTS INTO A COMPREHENSIVE HYPOTHESIS OF THECOSTRACAN EVOLUTION
Review round: 1
Reviewer: 2

Output:
Basic reporting: The language throughout is clear and the manuscript easy to read. The data publication is insufficient. The phylogeny is presented only as an image in a pdf file (despite the authors noting in the Methods the difficulties caused when phylogenies are not deposited in electronic repositories). The only link in the data section is the list of trees included as input to synthesis. Other ata I would expect to see:
* the genebank accessions used to reconstruct the additional tree
* the full output of the propinquity pipeline (including the synthesis tree)
* the occurrence data from GBIF
Experimental design: In general, there is not nearly enough detail in the methods section to understand the results being presented, and it would be impossible to reproduce the results.

The methods sections refers to the combination of the tree alignment graph method of Smith et al 2013 with the propinquity pipeline of Redelings and Holder. I am not sure this is accurate. The OpenTree project switched from the Smith et al method to the Redelings and Holder method in 2016 (see this blog post; https://blog.opentreeoflife.org/2016/04/08/new-version-of-tree-apis/). I am not aware of any workflows that combine the two synthesis methods, so the manuscript definitely needs clarification here.

If the synthesis was done using the propinquity method, then including the output from the propinquity pipeline would provide enough information to understand and reproduce the results (including the version of software packages, the exact inputs, and parameters used for synthesis). If propinquity was not the method used, the steps used to synthesize the trees needs to be much more clearly described.

The authors also describe using molecular data to “reconstruct an additional tree following the approach of Owen et al. (2015)”. Again, there is not enough detail here to understand the inference of this tree. The Owen paper is itself a synthesis phylogeny. Is the additional tree inferred directly from sequences and added to the synthesis method, or is the additional tree a synthesis tree? Is this one of the 27 trees included in the collection at https://tree.opentreeoflife.org/curator/collections/kcranston/barnacles?
Validity of the findings: In general, the lack of detail in the methods make it difficult to assess the findings.
Additional comments: No additional comments


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

Input:
Title: TOWARDS A BARNACLE TREE OF LIFE: INTEGRATING DIVERSE PHYLOGENETIC EFFORTS INTO A COMPREHENSIVE HYPOTHESIS OF THECOSTRACAN EVOLUTION
Review round: 2
Reviewer: 1

Output:
Basic reporting: The paper is much improved, but contains one glaring error. Lines 340-342, states that balanomorph taxa listed are basal and close to the ibliformes. this is not true - they are highly derived.

Other than this, I recommend publication.
Experimental design: N/A
Validity of the findings: Fine
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: TOWARDS A BARNACLE TREE OF LIFE: INTEGRATING DIVERSE PHYLOGENETIC EFFORTS INTO A COMPREHENSIVE HYPOTHESIS OF THECOSTRACAN EVOLUTION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The new version addresses most of my concerns with the first submission, including providing the tree files, removing the section on additional molecular-based analyses, and including GBIF and GenBank data.

I would prefer to see enough information in the methods sections or the supplemental materials to fully describe and / or reproduce the synthesis results. The manuscript simply says 'trees were assembled into a synthesis tree using the propinquity pipeline' and the files provided in peerj-33472-Barnacle_synthesis_original_files.zip are the output files from the last step of the pipeline. What about the parameters used for synthesis, and version of software packages? This is simple to do - propinquity provides all of the output needed. For example, see the 'All pipeline outputs' link under Downloads on https://tree.opentreeoflife.org/about/synthesis-release/v10.4.
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: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is a very well written and interesting article. The language was clear and the literature cited was adequate. The authors share the raw data. I have only noticed two minor spelling mistakes:
-line 239: "re-run analyses" should be "re-ran analyses"
-line 314: "shows" should be "show"

Also, the panels in Figure 2 are not labeled. They should be labeled with respect to the caption.
Experimental design: This article aims to test the relationship between paranoia and threat perceptions and whether or not this relationship was moderated by perceptions of group cohesiveness. While they found evidence that paranoia increased sensitivity to threats, their results did not support the prediction that paranoia was related to perceptions of group cohesiveness. Overall the research question was well defined and meaningful. I have only a few comments/suggestions to clarify the writing and experimental design:

1- Somewhere in the introduction it would be good to provide some statistics on how common paranoia is in the general population and what the distribution looks like.
2- In section 2.1, under Participants, the authors indicate that they had data on pre-existing paranoia. They could explain this a little more. When was this data collected, for what purpose? Was the GPTS instrument given again to the participants or is this from this previous data?
3- Under section 2.2.Design, they introduce a new concept: fairness of responder decision. There is not mention of this concept in the theory up until now. This is a bit confusing. I believe the authors should make the theoretical connections between fairness and paranoia and threat more explicit early on.
4- The justification for the Information-theoretic approach is not clear. Why is this method used? What is the main advantage? And how exactly does this method address issues related to multiple hypothesis testing?
5- On page 229, the authors mention they have included an incomprehension variable but it is not clear how this variable was measured. It needs to be specified.
Validity of the findings: I believe the analyses and the overall results are clear and robust. I had one main question and perhaps a suggestion. I wondered if the authors had enough variation on their paranoia variable and whether or not the mean score was very low. Perhaps they could speak to this in the paper. If the think there is enough variation and there are enough people with high scores, perhaps they could do some additional analyses focusing only on the lowest and the highest paranoia people to see if it makes a difference in terms of perceptions of group cohesion.
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: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Review “Paranoia and conspiracy: group cohesion increases harmful intent attribution in the Trust Game”

The manuscript is well-written and reports a well-executed experiment. I have several comments about aspects that I found unclear or limitations that might be addressed.

1. Lines 118-122: Cohesiveness of the responder pair was operationalized as whether the responder pair did or did not communicate. Please discuss whether this is a valid operationalization. What are the strengths of this operationalization? What are the limitations?

2. Lines 172-181: Responder decisions were pre-collected in the lab and MTurk participants were aware of this. Please discuss whether this has threatened the internal validity. This kind of online game is very different from regular social interaction in which one party does something and the other party responds. Now the participants did something and then were linked to a response that had already happened in the past. (Right?) Does that matter? I mean, is it possible that highly paranoid participants perceived this situation differently than regular participants, which influenced the behavior or the highly paranoid participants in this task? In other words, please discuss whether the experimental situation may have been perceived differently by highly paranoid participants and whether this has introduced confounds. (I mean, a trust game is a strange game, even when played face-to-face (e.g., why does the experimenter multiply the transfers?), I imagine that someone who is slightly paranoid, could have multiple things to be paranoid about in such a game.)

3. Line 258: Tables 1 and 2 report results from an ordered logistic model. I do not understand this. The outcome variables (attribution of harmful intent, attribution of self-interest) were measured on a continuous scale from 1 to 100 (lines 195-198). So why not do a standard OLS regression to test the hypotheses? I appreciate that the authors did effort to report a sophisticated analysis, but in my view, the analysis should not be more complicated than is needed. (Related to this, I do not fully understand the reported regression coefficients that were averaged over multiple models. This is a bit unusual, so just for communicating the results to readers who may be a bit skeptical toward unfamiliar things, please consider reporting the regular non-averaged regression coefficients.)

4. Lines 279-280: Why would we expect there to be a QUALITATIVELY different response? Does P5 not suggest that the strongly paranoid individuals are more sensitive to group cohesion (e.g., at the slightest sign of cohesion, they attribute harmful intent). This would be a QUANTITATIVE difference, right?

5. Lines 285-286: The conclusion drawn here is “the scaling of the response to changes in social threat is similar in all individuals” (high vs. low paranoia individuals). Please discuss whether the study had sufficient power to observe this. As the authors probably know, power for such interaction effects (or moderation) is complicated and strongly dependent on the population differences in the slopes (e.g., Stone-Romero et al., 1994, Journal of Management, 20, 167-178). Please consider this issue and appropriately qualify your conclusions if needed. For example, maybe the data support the conclusion that there are no large differences in scaling of responses. In my view, that would be a fine conclusion.

6. Related to the point above: What was the distribution of individual differences in paranoia (GPTS scores)? Does the sample distribution or the small number of people with high scores reduce the power to find that highly paranoid individuals respond differently?


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: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is a well-written article, that does present an interesting piece of research that is written in professional English throughout.

There is  (on the most part) sufficient literature/context provided on the background – however, as the focus is linked to conspiracy theory beliefs, I would like to draw the author (s) attention to a relevant paper by Imhoff and Lamberty (2018, https://doi.org/10.1002/ejsp.2494).  In this paper, they propose that paranoia and belief in conspiracy theories are distinct (albeit correlated) constructs.  The author (s) may wish to visit this paper, as I feel it would be useful to include this within their rationale and may also help support their conclusions (where it has been shown that paranoia did not interact).

In addition, the paper finds that harmful intentions were attributed to a cohesive group (vs non-cohesive).  This reminds me of another paper that may be relevant to the author (s).  Grzesiak-Feldman and Suszek (2008 https://doi.org/10.2466/pr0.102.3.755-758) simply found that conspiracy theories are seen to be more plausible if they are a tight formed group.  I believe your research supports this, where you have extended this by showcasing the effects in a live interaction.

The article is structured well, where the Tables and Figures are clear.  There is also clear raw data linked within the article.

The hypotheses are tested within the Results, and it is clear to see the links between each hypothesis.
Experimental design: The research is original, with a well-defined research question.  There is no research to date that has examined conspiracy thinking in live interaction, so this offers novelty to the research.

The research is based on a high-powered sample, that was pre-registered.  This brings in a strong sense of rigor to the research.

In the design, it would have been good if the author (s) included that they are also examining paranoia as part of their analyses.  I do think this should feature in the design section.
Validity of the findings: The data is based on a high-powered sample, so I am confident that the findings are robust and statistically sound.

Conclusions are strong, where the limitations are discussed.  I wonder if it could be considered a limitation that belief in conspiracy theories (or mindset) was not measured?  I would predict that someone who is more likely to endorse conspiracy theories, may attribute a higher degree of harmful intent.  We know from Imhoff and Lamberty that conspiracy theories and paranoia are different so I would predict that the conspiracy theory measure would have interacted.

I also wondered about a power imbalance – where from the literature, those who are feeling powerless, are more likely to believe in conspiracy theories.  With the set-up of the game (2 vs 1), could this have, as an outcome of the design, bred feelings of powerlessness, leading to the harmful intent?
Additional comments: This is an interesting paper, where the novelty of examining a live interaction, within a high-powered sample sparked by interest.  The paper is well-written, where some of the limitations were already addressed within the Discussion itself.   I have included a couple of points above that the author (s) may wish to consider.


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: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 2
Reviewer: 1

Output:
Basic reporting: The writing is clear and the revised manuscript is very satisfactory.
Experimental design: The revisions are clear and sufficient.
Validity of the findings: Again, revisions are satisfactory.
Additional comments: Thank you for carefully addressing all the comments brought up by the reviewers.


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: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 2
Reviewer: 2

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


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

Input:
Title: PARANOIA AND CONSPIRACY: GROUP COHESION INCREASES HARMFUL INTENT ATTRIBUTION IN THE TRUST GAME
Review round: 2
Reviewer: 3

Output:
Basic reporting: No further comments.
Experimental design: No further comments.
Validity of the findings: No further comments.
Additional comments: Thank you for addressing the points of clarification that I raised in my review.  I am happy with all the responses and congratulate you on an interesting piece of research.


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: RANDOM ALLOGENEIC BLOOD TRANSFUSION IN PIGS: CHARACTERISATION OF A NOVEL EXPERIMENTAL MODEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. In general, the clear, unambiguous, professional English language was used throughout; however:
a.	It could at times be more professional/scientific. Examples include lines 72-73 (“… and therefore hardly represents a relevant blood group antigen”; line 155 “… no hints for systemic inflammatory response were found…”; Lines 194 – 196: “Hints for … are missing.”;
b. Confusing terminology was sometimes used. e.g. “Whole blood” should be used rather than “full blood” in several instances.
c. Numbers at the start of sentences should be written rather than as numerals. E.g. line 125: “Sixteen…” rather than “16…”
d. The sentence across lines 84 to 86 is confusing as it implies that the guidelines apply to the dates 12/2014 till 3/2015, whereas the authors presumably mean that the experiments that they conducted were performed between these dates.
e. The sentence across lines 170 and 171 “The tissue wet to dry ratio as surrogate of edema formation was not increased following transfusion” is confusing as wet to dry weights were not performed pre and post-transfusion, rather they were performed on post-mortem tissue across different groups of pigs.
f. What is the context for the word “single” in line 179 within the sentence “We found that single high-volume allogeneic blood transfusion in healthy pigs…”?
g. While PeerJ does not specify it in their author instructions, I would suggest that an in-text reference at the end of a sentence should be placed before rather than after the full stop. This better links the reference to the statement that it applies to.

2. Other studies have reported on porcine transfusion models (e.g. Patel et al. 2011. Am J Physiol Renal Physiol; Biagini et al. 2018. Braz J Med Biol Res; Wozniak et al. 2018. Anesthesiology; and Masuda et al. 2018. Vox Sang.), albeit not in the context of organ cross-talk. These previous models should be described in the introduction.

3. The structure of the manuscript was logical and facilitated comprehension of the study and its findings.

4. In general the figures and table were well-designed and relevant; however, figure legends were insufficient for the reader to interpret the figures without reference to the manuscript text. Please address this limitation. Please also see points below for where clarifications and improvements are suggested:
a. Figure 2: Is this missing a 0h time-point? Are the decreases in haemoglobin and haematocrit in the control group significant? Do the asterisks represent P<0.05 vs. baseline or vs. control group?
b. Figure 3: Is this missing a 0h time-point? Do the asterisks represent P<0.05 vs. baseline or vs. control group?
c. Figure 4: should the y-axis title say “relative impedance” rather than “realtiv impedance”? Would it be more appropriate to have a graph for each lung area so that transfusion vs. control can be compared?
d. Figure 5 and Figure 6: There is a lot or unnecessary repetition of labels in these figures. For example in Figure 5: “lower lung” and “upper lung” could just be stated once above the appropriate column and no need to state the analyte for each graph title; “IL-6”, “TNF-alpha” and “iNOS” could just be stated once per row, and again no need to have this as a graph title; “control” and “transfusion” could also just be stated once per column rather than on every graph.
e. Table 1: This is a very large table. It may be an idea to split into two tables – maybe one for haemodynamic data and one for respiratory data.
f. Table 2 and Figure 2 appear to present the same data, just that in the table it is the actual results, while in the figure it is transformed to % baseline. Is this correct? If so are both necessary?

5. Raw data is included as supplements. Some additional labelling would facilitate interpretation of these data.
Experimental design: 1. The manuscript meets the scope of PeerJ.

2. The research question ultimately posed by the authors is whether blood-borne factors can precipitate organ cross-talk. More specifically, the research question posed by the authors in the manuscript is whether a porcine model of whole blood transfusion is a suitable model in which to investigate the mechanisms of organ cross-talk.

3. In general the study described by Ziebart et al. meets acceptable technical and ethical standards; however, I do have some concerns that need clarification:
a. It appears that the transfusion was of leucodepleted whole blood. Yet inflammatory cytokines are hypothesised to be responsible for initiating organ cross-talk via the blood stream. Did the authors consider processing the whole blood into plasma which could be frozen for future experiments? This approach would have allowed increased flexibility in the time intervals between whole blood collection and the transfusion experiments. And the inflammatory cytokines would still be present in the plasma. Furthermore, this approach would have avoided the increased in Hb and Hct evident following whole blood transfusion.
b. Other animal models of blood transfusion utilise blood collection from conscious donor animals without the need for euthanasia, and this seems to be a more ethical approach. Why were the donor pigs euthanised following blood collection? Was this due to the volume of blood collected, and if so what percentage of estimated circulating blood volume did this represent? If too high, could this have been avoided by using older pigs as the donors (I understand that adult German Landrace pigs reach 200 – 300kg in weight). For example, in Australia and the UK, the minimum weight requirement for blood donors is 50 kg. Assuming a circulating blood volume of 70 mL/kg, then collection of a 450 mL unit of whole blood represents 13% of the circulating blood volume. Furthermore, if the experiment used plasma and not whole blood (see earlier point), then units could be collected over time for subsequent experiments, reducing the number of donor pigs required and possibly making scheduling experiments more flexible.
c. In the discussion (lines 238 – 242) the authors state that they did not perform “simultaneous blood withdrawal” to avoid risk of circulatory overload as this would also have removed factors within the transfused blood, and because of the haemodynamic changes and complexity of the procedure. Would withdrawal of a smaller volume of blood prior to transfusion been an alternative to reduce risk of circulatory overload while at the same time minimising these other risks?

4. In general an appropriate level of detail in the Materials and Methods section was provided; however, some aspects do require clarification in the manuscript.
a. Blood collection: should state the average (+/- SD) volume of blood collected. What percentage of estimated circulating blood volume does this represent? If not a multiple of 450 mL then how did this impact the ratio of whole blood blood to anti-coagulant? Is it scientifically valid to transfuse whole blood from ‘underweight’ collections?
b. It is stated that whole blood was collected into Composelect blood packs, but these come in two formats – one in which the whole blood is leucofiltered prior to processing, and one in which the whole blood is separated and the red cells are leucofiltered. I assume that the former were sued; however, the manuscript should clearly state which ones were used. If the former, was the whole blood leucofiltered prior to storage at 4C? If the latter, then how was the whole blood separated? Furthermore, it should be stated clearly in the manuscript that these blood packs are CPD anticoagulated.
c. How long was the donor blood stored prior to transfusion? Were cytokine levels measured in the blood to be transfused?
d. How was the transfusion performed? Was it warmed prior to transfusion? Was it administered via a transfusion giving set (with a filter to remove aggregates)? What was the average volume (+/- SD) transfused?
e. How long was the baseline period? Was this sufficient time for the animals to recover from the instrumentation?
f. What type of blood samples were collected throughout the experiment? E.g. were they EDTA, citrate or heparin?
g. The reference gene used in the real-time PCT experiments should be specified in the Materials and Methods section and not just in the Figure 6 legend
Validity of the findings: 1. Ziebert et al. present a novel and interesting study describing the esablishement of a porcine model in which they plan to investigate the hyposthesis that blood-borne factors can mediate organ cross-talk. Presumably they will do so by transfusing whole blood from an injured pig into a healthy pig and observing the transfused pig for the development of injury. In the data presented, transfusion of whole blood between healthy pigs proceeded without evidence of acute transfusion reactions or haemodynamic, inflammatory or respiratory complications.

2. In general the data presented were robust, statistically sound and controlled. However the following points require clarification.
a. The methodology states that FiO2 was set to 0.4 (line 100) yet in table 1 of results FiO2 is reported throughout to be 0.39. Why the discrepancy? Furthermore, what was there any protocol to alter ventilation settings in line with respiratory function?
b. Was physiological data collected more frequently than the four time-points for which results are presented (baseline, t=0, t=4 and t=8)? The sentence in the discussion (lines 190 – 191) suggests that in addition to the increase in mPAP, transient increases in MAP and CVP were observed; however, these are not evident in the data presented in Table 1, nor in the raw data included as supplement 1. I would suggest that if more frequent data (e.g. hourly) are available that these be included in the manuscript.
c. As per the previous point, it appears that arterial blood gas samples were collected and analysed more frequently than presented in Table 1. Again, I would suggest that including these data rather than presumably averaging out measurements over a number of arterial blood gas samples would be preferable, even if only as supplementary data.

3. Where the authors have extrapolated from the findings of their study, the speculations they have made appear for the most part to be reasonable. However, some statements supporting their extrapolations require clarification.
a. The authors state that “Appearance of TRALI depends on the gender of the blood donor, and is particularly relevant in multiparous women.” However, TRALI may be either antibody-mediated, in which case the statement is true, or it may be mediated by biological response modifiers related to blood component storage duration. Given the latter mechanism, it is important for the methodology to be clear on the length of tiem that the blood was stored at 4C prior to transfusion. Furthermore, measurement of cytokine levels in the blood to be transfused would have further minimised the risk of TRALI in the porcine model.
b. The authors state that “Animal care reasons and the need for additional blood donor animal required a strict limitation of the animal numbers…” however, I would argue that it is unethical to limit the number of subjects per group if it means that a study is under-powered to answer its primary question. As per the earlier comment, I think that the blood donor part of the study could have been better designed from both an ethical and a technical perspective.

4. The conclusions of the study are reasonably well stated; however, the following
a. Statements such as “high volume… blood transfusion” are difficult to make in the context of the manuscript not stating either the normal circulating blood volume of German Landrace pigs or the percentage of blood transfused.
Additional comments: The manuscript submitted by Ziebart at al. describes the design of a porcine model suitable for the subsequent investigation of organ cross-talk. Presumably they will do so by transfusing whole blood from an injured pig into a healthy pig and observing the transfused pig for the development of injury. In the present study, transfusion between healthy pigs proceeds without evidence of unexpected haemodynamic, inflammatory or respiratory complications, with the caveat that a transient pulmonary arterial hypertension was observed.

Some general points to address:
1. Typographical errors: line 72: “gene” not “gen”; line 74: “barely” not “barley”; line 86: “Landrace” not “land race”; presumably the word “blood” is missing in the sentence from lines 238 – 239;
2. Numbers at start of a sentence should be written: line 86: “Twenty” not “20”;
3. Line 86: “Sus scrofa domestica” should be italicised
4. Whether “granulocyte clots” form as part of the pathogenesis of TRALI is not clear. Please amend the sentence from line 202 – 204.


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: RANDOM ALLOGENEIC BLOOD TRANSFUSION IN PIGS: CHARACTERISATION OF A NOVEL EXPERIMENTAL MODEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: •	Clear and unambiguous, professional English is used throughout the article
•	Literature references are sufficient; sufficient information about the field background.
•	The article includes an  introduction and background to demonstrate how the work fits into the broader field of organ crosstalk. The relevant prior literature is appropriately referenced.
•	From the reviewers point of view the phenomenon of brain-lung interaction in men and pigs needs to be introduced a more detail, including potential mechanisms and risk factors. Some typical clinical presentations in the ICU setting would probably be helpful.
•	The hypothesis and results are discussed as a “plus theory”, based on the notion that transfusion represents the systemic exposure to an “injectable” active component responsible for the cross-talk. However, a “minus theory” is possible, where transfusion impair immunocompetence and confers immunosuppression responsible for various observations during organ cross-talk. Even if transfusion is accompanied with proinflammatory signs in humans, the examination of the activity of specialized  T cells could be discussed.
•	The way the main hypothesis is described is not sound and remains unclear. One pathophysiological mechanisms of organ cross-talk includes the release of mediators from an injured organ into the blood stream. However, it remains unclear why allogenic blood transfusion would mimic the mechanisms of organ cross-talk because blood transfusion does not necessarily imply that active compounds (mediators) are being transfused. Even if transfusion was associated with systemic infusion of active compounds, it is not clear whether these compounds represent the mediators that are released by organ A to induce remote organ injury in organs B, C, or D. Therefore, the main objective of the present study is limited to the characterization of the baseline immunogenic effect of allogeneic blood transfusion in pigs (line 75 ff.) It is a transfusion study. Not an organ cross-talk study, and therefore, the statement (line 177) that this study established a novel model for the characterization of remote organ injuries is not valid and should be removed. Furthermore, it is suprising and not substantiated in the introduction why the authors hypothesize that transfusion “neither inhibits inflammatory response nor impairs the pulmonary function” (line 77ff). The scientific rationale why transfusion would impair pulmonary function needs to be mentioned in the introduction.
•	Professional article structure, figs, tables. Raw data shared is given.
•	The structure of the article is in agreement with the format of ‘standard sections’
Figures are relevant to the content of the article, of sufficient resolution, and appropriately described and labeled.
All appropriate raw data have been made available in accordance with the Data Sharing policy.
•	The submission is ‘self-contained,’ and represents an appropriate ‘unit of publication’, and includes the results relevant to the hypothesis.
•	Line 54: “well-know” and “not fully understood” are potentially contradictory
•	Line 233: again, the interpretation that this is a study about organ interaction should be removed because the design of the study does not include organ interactions
•	Line 249: I agree with the authors that blood transfusion “may represent a (novel?) model to characterize remote organ injury”, unfortunately, this study does not help answer this hypothesis.  Future studies are needed to address this hypothesis, but the aspect or organ cross-talk should be removed from this manuscript.
•	Figure 2: absolute leukocyte counts and hemoglobin level should be given
•	Figures 5 and 6: The normalized mRNA expression should be displayed in relation to controls (=1), otherwise it is very hard to interprete differente scaling of the various graphs. The differentiation between lower and upper lung lobes and cortex vs. hippocampus are not discussed. Why is it important to make that differentiation and how can the results be interpreted?
Experimental design: •	Original primary research within Aims and Scope of the journal.
•	The submission has  defined the research question, the relevance remains slightly unclear because the notion of organ cross-talk is not defined specifically
•	The conclusions include the hypothesis that remains unanswered
•	The reviewer feels that the study time of 8h between transfusion and end of study needs to be explained in the context of the brain Lung crosstalk and the cited lterature. Why did the authors choose this interval? In the cited paper: Systemic Pao2 Oscillations Cause Mild Brain Injury in a Pig Model by Klaus U. Klein et al.  the authors found only a mild cerebral inflammation after a 20 h protocol of pigs treated with ECMO, wich itself is a device with a relevant inflammatory impact. What is the translational relevance of the present hypothesis and study design?

•	The  investigation is performed to a high technical & ethical standard.
•	The research  has been conducted in conformity with the prevailing ethical standards in the field.
•	Methods described with sufficient detail & information to replicate.
•	Only the procedure of drawing donor blood needs smoe additional details. The authors used  a leucocyte depleting collector bag and stored the bags at 4°C.  Hence the blood transfusion is a composite of red cells and platelets. The authors should render this more precisely and add the duration of blood storing or the timeline. Did they start on day 1 -4 with blood donation followed by day 5 – 10 with transfusion or did they perform blood donation and transfusion on the same day. This information will help to reproduce the study model by another investigator.
•	Why did the authors before “top-load” blood transfusion rather than transfusion after withdrawal of an equivalent amount of blood. Top-load transfusion may dilute the effect of potential mediators that are being transfused and may cause inflammatory organ injuries. Top-load infusion of electrolyte solution creates lesser dilution of baseline circulating mediators as compared to electrolyte infusion after withdrawal of an equivalent amount of blood volume. The study design of transfusion and electrolyte infusions is therefore reducing the probability of significant differenced between both groups with respect to circulation inflammatory parameters and functional assessment of remote organs (lung, brain).
Validity of the findings: See above re hypothesis and rationale, and limited validity of last sentence of conclusions.
Additional comments: See above.
I suggest to rewrite the article, giving it a different perspective and stressing the assessment of immonogeneic effects of transfusion, NOT of organ cross-talk, because the latter is misleading and not supported by the design and the results of the study.


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

Input:
Title: RANDOM ALLOGENEIC BLOOD TRANSFUSION IN PIGS: CHARACTERISATION OF A NOVEL EXPERIMENTAL MODEL
Review round: 2
Reviewer: 1

Output:
Basic reporting: 1. In general, the clear, unambiguous, professional English language was used throughout; however:
a. It could at times be more professional/scientific. Examples Lines 40 – 41: “No hints for systemic inflammatory response....”
b. When data values are written, ‘3.5’ rather than ‘3,5’ should be used to describe 3 and a half as this is the convention in English.
c. Some examples of odd wording. Example line 125 “piped”

2. The Introduction places the study into the context of organ cross-talk. Literature was well referenced & relevant.

3. The structure of the manuscript was logical and facilitated comprehension of the study and its findings.

4. In general the figures and tables were well-designed and relevant; however, figure legends were insufficient for the reader to interpret the figures without reference to the manuscript text. Please address this limitation. Please also see points below for where specific clarifications and improvements are suggested:
a. Figure 1: The figure legend requires some clarifications. The duration of the baseline period should be noted in the figure (from the manuscript this appears to be 20 minutes). Abbreviations (e.g. EIT) should be defined in each figure even if they have already been defined in the manuscript. In the sentence “The hemodynamic and spirometric parameters were recorded permanently.” is ‘permanently’ the correct word? Perhaps the authors mean ‘continuously’? The sentence “Lung and brain were analysed histopathological and biochemical after the end of the experiment” appears incomplete. Furthermore, what is meant by “biochemical” as no reference is made to biochemical analyses in the manuscript’s Materials and Methods section.
b. Figure 2: If blood samples were collected every 2 hours and underwent hematological analysis, then all of these data should be shown on the graphs. In particular, the 0h time-point that occurs immediately following transfusion should be included. An immediate increase in hemoglobin, hematocrit and erythrocytes would be expected with the whole blood transfusion. The description of what the asterisks represent should come at the end of the figure legend rather than at the start.
c. Figure 3: If blood samples were collected every 2 hours and underwent ELISA analysis, then all of these data should be shown on the graphs. The description of what the asterisks represent should come at the end of the figure legend rather than at the start. Do the asterisks represent P<0.05 vs. baseline or vs. control group, presumably the latter? The figure legend states that “Parameters are displayed as percent of baseline” however the data in the graphs are presented as pg/mL. It is unclear which group the asterisks in the TNF-alpha graph refer to: the control group or the transfusion group? Are the symbols in the IL-6 graph correct as it appears that black circles are used for one of the data-sets rather than grey circles as indicated in the figure legend.
d. Figure 4: If the comparison is being made between control and transfusion groups then there should be a graph for each lung area so that transfusion vs. control can be on the same graph.
e. Figure 5: The second and third sentences of the figure legend are repetitive of each other. The figure legend should specify that these analyses were based on cryopreserved lung samples, as well as very briefly describe the procedures used to generate these data.
f. Figure 6: As per Figure 5.
g. Table 1: Please refer to earlier comments on the presentation of values such as “3,6” and the inclusion of only selected time-points. The table legend states that “No significant intra- and intergroup changes of hemodynamic parameters were observed after blood transfusion or fluid infusion, apart from temporary increase of the pulmonary arterial pressure after transfusion.” however, CVP was also increased after transfusion compared to baseline but  without being increased compared to the control group.
h. Table 2: Please refer to earlier comments about the presentation of values such as “6,2” and the inclusion of only selected time-points.
i. Table 3:  Please refer to earlier comments about the presentation of values such as “7,5” and the inclusion of only selected time-points. The two sentences of the figure legend are contradictory. Furthermore, while lactate levels decrease, AaDO2 levels actually increase. Values for pH should be provided in more detail, i.e. 7.456 rather than 7.5.

5. Raw data is included as supplements. Some additional labelling is still required to facilitate interpretation of these data. For example, in DataSetS1 presumably each worksheet represents a different experiment; however it is unclear to what the numbers “1 – 4” represent. Presumably dependent periphery, dependent central, non-dependent central, and non-dependent periphery; however which is which? What do the four different values represent – four different fields of view? The summary worksheet should specify which experiment the values come from. Grouping should be also be displayed. TableS1, S2 and S3 are datasets rather than tables?
Experimental design: 1. The manuscript meets the scope of PeerJ.

2. The research question ultimately posed by the authors is whether blood-borne factors can precipitate organ cross-talk. More specifically, the research question posed by the authors in the manuscript is whether a porcine model of whole blood transfusion is a suitable model in which to investigate the mechanisms of organ cross-talk.

3. In general the study described by Ziebart et al. meets acceptable technical and ethical standards; however, I do have some concerns that need clarification:
a. Data were collected more frequently than presented in the figures/tables. For example, arterial blood gases were collected hourly (as per TableS1) however, only Baseline, 0h, 4h and 8h data were included in Table 3, meaning that 6 data points were excluded. What is the rationale for collecting these data but not presenting or analysing them? Would including these data alter the findings of the study?
b. Based upon total number of pigs (20), number of donor pigs (8), and number of control pigs infused with fluid (4) it can be inferred that there were eight pigs transfused with whole blood. This isn’t clearly stated anywhere in the manuscript. Reviewing the supplementary data and tables it appears that lung sections were scored for 12 pigs (presumably 4 controls plus 8 transfused), but tables S1, S2 and S3 don’t have data for experiment 3. This leaves the transfusion group with only 7 pigs. Please clarify these numbers in the manuscript, and if data from experiment 3 was not included in the manuscript then the rationale for its exclusion should be provided, and data from this experiment should also be removed from the histological results.

4. In general an appropriate level of detail in the Materials and Methods section was provided; however, some aspects do require clarification in the manuscript.
a. Blood withdrawal: Isn’t the CPD anticoagulant present in the primary pack (into which the blood is collected) and the SAGM additive solution in the satellite pack (into which the leucoreduced whole blood is passed)? Furthermore, should the leucocyte reduction filter have been pre-wet by passing the SAGM through into the primary pack before passing the whole blood/CPD/SAGM mixture back through the filter into the satellite pack?
b. Experimental protocol: Information presented in this section may be better placed in the Anesthesia and Instrumentation section. For example the information from lines 140 – 151 regarding monitoring and background Sterofundin infusion.
c. Post mortem analysis: Was it serum or plasma? Both are stated in line 157.
Validity of the findings: 1. Ziebert et al. present a novel and interesting study describing the establishment of a porcine model in which they plan to investigate the hypothesis that blood-borne factors can mediate organ cross-talk. Presumably they will do so by transfusing whole blood from an injured pig into a healthy pig and observing the transfused pig for the development of injury. In the data presented, transfusion of whole blood between healthy pigs proceeded without evidence of acute transfusion reactions or haemodynamic, inflammatory or respiratory complications. However, according to raw data presented in TableS1, two of the transfused animals developed hypoxaemia (PaO2/FiO2 < 300) following transfusion, although these were the only two animals that had a baseline PaO2/FiO2 of <400. This should be presented in the results and discussed in the discussion. Did either of these two animals also display evidence of pulmonary edema (by histology or wet/dry weight anlaysis)? If so then transfusion-related acute lung injury would be indicated.

2. In general the data presented were robust, statistically sound and controlled. However the following points require clarification.
a. In my experience all of the time-points for which data are collected are presented in a manuscript. However, in the figures and tables only data from specific time-points are presented in this manuscript. While these additional data points may not increase the power of the study nor provide new findings, these data are important to provide a more complete picture of the experiments and for the reader to be satisfied that the conclusions made by the authors are valid.
b. The sentence in the Discussion (lines 218 – 220) suggests that in addition to the increase in mPAP and CVP following transfusion, an increase in MAP was also observed; however, this is not evident in the data presented in Table 1.

3. Where the authors have extrapolated from the findings of their study, the speculations they have made appear for the most part to be reasonable. However, some statements supporting their extrapolations require clarification.
a. Further to the earlier point, what are the implications in that 2 out of the 7 or 8 pigs in the transfusion arm developed hypoxaemia?
b. The authors state that “Appearance of TRALI depends on the gender of the blood donor, and is particularly relevant in multiparous women.” However, TRALI may be either antibody-mediated, in which case the statement is true, or it may be mediated by biological response modifiers related to blood component storage duration in which case no differences between female and male donors have been reported.
c. What are the implications of the transient increase in mean pulmonary arterial pressure? This has previously been associated with the transfusion of soluble factors present in stored but not fresh red cell concentrates (Tung et al. Critical Care Medicine 2012; Baron et al. Anesthesiology 2012; Solomon et al. Blood 2013; Fung et al. Vox Sanguinis 2013) and yet the whole blood transfused into the pigs was fresh.

4. The conclusions of the study are reasonably well stated; however, the implications of the hypoxaemia development need to be considered prior to being able to make statements such as “high volume random allogeneic blood transfusion in pigs neither impaired pulmonary integrity…”.
Additional comments: The manuscript submitted by Ziebart at al. describes the design of a porcine model suitable for the subsequent investigation of organ cross-talk. Presumably they will do so by transfusing whole blood from an injured pig into a healthy pig and observing the transfused pig for the development of injury. In the present study, transfusion between healthy pigs was reported to proceed without evidence of unexpected haemodynamic, inflammatory or respiratory complications; however, a transient pulmonary arterial hypertension was observed and two of the transfused pigs developed hypoxemia (PaO2/FiO2 < 300) post-transfusion.