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: FERROPTOTIC CARDIOMYOCYTE-DERIVED EXOSOMES PROMOTE CARDIAC MACROPHAGE M1 POLARIZATION DURING MYOCARDIAL INFARCTION
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript by Sun et.al and colleagues are exploring the role of ferroptosis cardiomyocytes derived exosomes in M1 macrophage polarization. The authors provide compelling evidence of M1 macrophage infiltration upon cardiomyocyte ferroptosis and ferroptosis derived exosomes promote M1 macrophage polarization. Overall, the data support the conclusions very well and I have only a few suggestions for improving the manuscript.
Experimental design: No comments
Validity of the findings: 1. Figure 1C, 3A, and Fig 4A: Scale bars are missing.

2. Authors quantified the ferroptosis indicator MDA and Fe+2 concentration but it looks like there is approximately a 20% difference between the Sham group and MI group. Can the author comment on what Fe+2 concentration could induce ferroptosis?

3. Although the authors used ferroptosis indicators such as MDA and Fe+2 concentration, the Authors can try ferroptosis biomarker transferrin receptor TFRC and check whether there is any difference between the shame and MI group.

4. Figure 1F: It’s difficult to evaluate the western blot data. I don’t any expression of IL-10 in the MI group and it’s hard to see any difference in NOS2 levels. Authors could have tried using Immunohistochemistry with a similar maker or better marker such as CD86 or CD206 which can differentiate M1 and M2.

5. How does the author characterize the exosomes other than the particle size quantification? Also, how to distinguish exosomes derived from the ferroptosis cardiomyocytes from other exosomes?

6. Fig 3A: Authors could have discussed a bit more about exosomes uptake by RAW 264.7 cells. Is it possible to quantify?

7. Fig 4A: How do authors identify or distinguish that these are M1 macrophages?
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: FERROPTOTIC CARDIOMYOCYTE-DERIVED EXOSOMES PROMOTE CARDIAC MACROPHAGE M1 POLARIZATION DURING MYOCARDIAL INFARCTION
Review round: 1
Reviewer: 3

Output:
Basic reporting: This work is very well designed, executed, and discussed.
Experimental design: Shengjia Sun and colleagues are trying to identify the reason behind M1 macrophage polarization in myocardial infarction and trying to establish if this is exerted by exosomes derived from ferroptotic cardiomyocytes (MI-Exo). The authors have succeeded in providing compelling evidence for the involvement of exosomes derived from ferroptotic cardiomyocytes in M1 macrophage polarization. The underlying mechanism was also identified as Wnt/ ß-catenin pathway activation. In addition, they also show that inhibition of Ferroptosis by Ferrostatin-1 results in abrogating the effect of exosomes. Overall, the data support the conclusions very well and I have only a few suggestions for improving the manuscript.

1) The authors have indicated that RAW 264.7 cells uptake DiI-labelled exosomes, but it is not clear by which pathway they are engulfed by macrophages, and inhibiting their uptake affects the resultant effect of exosomes in M1 macrophage polarization?
2) The authors have indicated the uptake of both MI-Exo & Fer-1-Exo by Raw 264.7 cells; however, it seems that uptake of Fer-1-Exo is higher than MI-Exo as shown in Fig. 3A. It would be interesting to know if this is due to the differential labeling of respective exosomes or their differential uptake. And if it is differential uptake, what is the reason behind this, and does it have any effect on subsequent experiments?
3) In figure-4A, where authors are comparing phagocytosis in the blank, MI-Exo & Fer-1-Exo, it seems there are fewer cells in Fer-1-Exo than in Blank & MI-Exo conditions. Are Fer-1-Exo toxic to macrophages?
Validity of the findings: 4)The Wnt/ß-catenin pathway involvement should be confirmed by another experiment such as Wnt signaling inhibition and its effect on macrophage polarization.
Additional comments: A few minor points.

5) the scale bar is missing in Fig.-3A & 4A.
6) What is the unit of Fe2+ concentration in Fig.1E & 2 B. This is for the readers to understand Fe2+ levels in vitro and in vivo under physiological conditions?
7)Presentation of flow data can be improved by providing staggered overlay histogram and inclusion of unstained control etc.


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

Input:
Title: FERROPTOTIC CARDIOMYOCYTE-DERIVED EXOSOMES PROMOTE CARDIAC MACROPHAGE M1 POLARIZATION DURING MYOCARDIAL INFARCTION
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have satisfactorily addressed the concerns raised by reviewers. I have no objection to recommending this manuscript for publication in PeerJ.
Experimental design: Authors have addressed concerns raised by reviewers by supplementing their work with additional experiments such as analysis of molecular markers of ferroptosis.
Validity of the findings: Please see above.
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: FERROPTOTIC CARDIOMYOCYTE-DERIVED EXOSOMES PROMOTE CARDIAC MACROPHAGE M1 POLARIZATION DURING MYOCARDIAL INFARCTION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have now answered all of my requests and suggestions.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: I am now happy for this manuscript to be published.


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

Input:
Title: FERROPTOTIC CARDIOMYOCYTE-DERIVED EXOSOMES PROMOTE CARDIAC MACROPHAGE M1 POLARIZATION DURING MYOCARDIAL INFARCTION
Review round: 2
Reviewer: 3

Output:
Basic reporting: The paper has been revised accordingly.
Experimental design: The authors have revised the manuscript with relevant experiments.
Validity of the findings: Everything looks great.
Additional comments: The flow data needs a very very minor correction. In Fig.3C, in the overlay histogram, instead of overlaying the mCherry Fluorescence channel, authors have overlaid either Forward Scatter/Side-Scatter. a very minor correction is needed.


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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: In the manuscript, the authors were aimed to investigated the prevalence of HBB and G6PD  genes polymorphisms in Senegalese populations to evaluate the risk for severe Plasmodium falciparum malaria and its specific phenotypes. The study was well designed and the results were sufficient to comment about the affect of  the studied gene polymorphisms on Malaria clinical phenotype.
Validity of the findings: the results were sufficient to comment about the affect of  the studied gene polymorphisms on Malaria clinical phenotype as well as the authors had selected suitable statistical analysis.
Additional comments: no comment


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

Input:
Title: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1.	The manuscript lay out conforms with that required for PeerJ. Whilst the English is overall very good, there are innumerable spelling and small grammatical errors throughout. In many places we have random capital letters mid sentence. There are also some strange sentences dotted throughout. For example Line 216 “The materiality threshold was set at p=005.” Could do with another very very thorough read through.
2.	Human gene abbreviations should be in italics. This has been done occasionally but not throughout.
3.	In the results section on blood indices, it is written in Table and text PNB and PNE. Can these be spelled out.
Experimental design: 1.	The study sets out two objectives, the first of which is “to  investigate  the  prevalence  of  key  malaria-protective 138 polymorphisms in G6PD and HBB genes in the global Senegalese population”.
I’m not convinced that the samples from 323 individuals can really be considered globally representative of the Senegalese population, especially, as well described in the paper, the large number of ethnic groups.
2.	And in the methods it states that “114 CTR, 170 UM and 153 SM subjects were included” which sums to over 400. Maybe best to state the number of controls in the abstract as well (ie 437 individuals not just 323)
3.	Line 219-224. This is really not very clear and surprised to see a result given in the methods section. It’s hard to understand what analysis was done but it doesn’t look correct.
“The  participant’s  average  percentage  parasitaemia  is  calculated  by taking the parasitaemia reported by all included subjects and calculating the average. From those results,  the  standard  deviation  is  determined,  and  the  results>3  standard  deviations  from  the average  are  omitted  and  a  new  mean  and  standard  deviation  calculated.  This  new average  is referred to as the mean parasitamia. We found a difference of level of parasitaemia between SM and UM groups, which were statistically significant (P < 0.01).”
4.	Line 252 “The PCR products were checked by 1.5% agarose gel electrophoresis wholly,….”
What does this mean?
5.	Line 303 2.2. Haemotological and parasitaemia parameters of the study population.
In this small paragraph there’s no mention of parasitemia. Maybe what was written in the methods should have been placed here.
It would be a good idea for the reader to define precisely what criteria are required for a classification as Uncomplicated or Severe malaria.
Validity of the findings: 1.	Age is a hugely important factor to be taken into account for the outcome of  malaria infection. I can’t see where this has been taken into account. This is also especially important when comparing areas of differing endemicity.
2.	Line 359 onwards. I’m a little concerned about the stat results as we have a significant P value whereas the 95% confidence intervals go over 1. I don’t see how this can be significant. Please see general comment below.
The HbC polymorphism  (HBB +19 G>A, rs33930165) showed a significant p value when comparing SM vs. CTR  (p value = 0.0084, OR 1.67; 95% IC 0.99–3.42) (Table 4). The comparative analysis UM vs. CTR of the other Hb polymorphisms showed significant associations with rs7946748 C>T and rs713040 C>T, respectively (p value = 0.033, OR 2.67 95% IC: 2.01–3.50) (p-value = 0.008, OR 1.67; 95% IC 363 0.99 - 3.42) (Table 4)
3.	There are lots of blood cell parameters that are associated with various SNPs but I can’t see what analyses was performed and worse still these findings are not at all discussed in the discussion.
4.	Table 2 In P value column it is written SM/MM. What is MM?
Additional comments: I am a little bewildered about the HBC result. In the abstract it states that there was no protective effect, in the results and discussion it states there was despite the Confidence intervals overlapping with 1 (see above).
As noted above the associations of snps with blood cell parameters could be interesting but we need to know more about these analyses and the co-variates used.


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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 1
Reviewer: 3

Output:
Basic reporting: Some editing required across the document. Major editing needed as the text is not in proper English. The raw data has been shared and relevant references have been used.
Line 75-76 Malaria is caused by Plasmodium species infection and affect hundred of millions of people per year
Line 77-78 Malaria disease remain a major cause of death and is still the 4th leading cause of death for infectious diseases worldwide.
Line 81-82 And  the comprehension of these mechanisms are necessary for a global malaria eradication
Line 93-94 In endemic areas, the protective effect of HBB variants (HbS and HbC) on P. falciparum malaria clinical phenotypes were reported.
Line 120 and 121. G6pd and G6pD
Line 217 The individual % parasitaemia was mesure as
Line 160 Ethnics distribution
Line 321 SNPs in Western Africa, and lefted out polymorphisms such as
Line 400 In The Gambia, a neighbor country of
Use of comma instead of decimals in numerals is not standard practice.
All abbreviations should be described. What is PNB (line 384) and PNE (line 386)
Line 266 (Ref 28-29 de Zuo) the references need to be quoted appropriately.
Experimental design: Sample size: how was the number of participants arrived at? Most studies have been done in children who are under-represented in this study, what was the rationale for the inclusion of adults?

153 SM and 170 UM cases from 2003-2015 across the regions these are very low numbers is this the actual number of cases or was there selection bias. Where the participants actively recruited across the years then followed up at admission or how exactly is this a cohort? Or where the available stored samples retrieved for the study.

Potential confounders (sex and ethnicity) are listed in the discussion, were any of them adjusted for in the analysis for the ORs?
The significance for SM vs UM rs762515 is borderline would this still hold when analyzed by sex? The severe and uncomplicated malaria cases need to be clearly defined. Is there a parasitaemia threshold?

Is the Plasmodium-positive QBC test quantitative or qualitative? What are the product details?
What is the age of the study participants?
Where were the controls recruited? if at the hospital what was the cause of admission?
Why wasn’t parasitaemia expressed as parasites/uL instead of %? Yet on Table 2 it is presented as parasites/uL
Lines 213-224 are describing statistical analysis hence should move to section 1.5
The tests should be named properly Student's test and Yates corrected X2 test
G6PD has previously been described to be X-Linked what is the relevance of analyzing the sex-specific effects underlying X chromosome loci?
Ethics section should be after section 1.5
Validity of the findings: Mean age of the participants is 25.8 years. The burden of malaria lies in children yet those <15 years of age are only 33% of the participants. Was the study done in the correct age group? What is the burden of malaria in those over the age of 15 in Senegal?

Line 300 “13.7% of mortality was observed.” What % of this mortality occurred in children? What % of the severe malaria cases died?

Table 1 - why were all the controls recruited from Dakar yet 50 %of the cases come from the other regions? The controls are also underrepresented in age <5years category.

Table 2 - what proportion of the participants had severe anemia? What proportion of the participants had bacteraemia? Parasitemia should be presented as geometric mean. Lymphocyte and neutrophil should be presented as actual counts. It’s not clear what the value of monocyte, eosinophil and basophil count is on this table.

“Global population” used in tables 3 and 4 should be replaced with “overall” is the mean that the MAF of the overall population sampled. Same change should be effected in the discussion as the word global is misleading.

Section 2.5 – the investigators are reporting results of SNPs showing protection against malaria yet the data has not been included in the document.

Lines 385-389 – what is the direction of the reported associations?

What is the role of rs7946748? Has it been described before?

Line 407- 411 “In our Senegalese population cohort, we found that G6PD-202 G>A polymorphism was not associated with protection against severe malaria at global level. However, differences were founded in the different ecological localities. The polymorphism was associated with protection against SM in Dakar Capital center, but not in the other regions (St Louis-Louga and Tambacounda-Kolda).” This text is not supported by the data presented by the authors.
Additional comments: The number of controls is missing in the abstract.

Line 79 asymptomatic malaria should be initialed as AM not UM which is normally used for uncomplicated malaria. Mild malaria should be replaced with Uncomplicated malaria UM as is the case in the rest of the paper. Line 399 states MM.

The flow of the background needs revision. Lines 75-92 should be followed by the description of the G6PD and HBB genes before describing the association with malaria endemicity. What is the prevalence of HbC in west Africa?

Consider replacing the word subjects with participants.

Line 41-43 “We investigated the prevalence of HBB (chr11) and G6PD (chrX) deficiencies polymorphisms, among Senegalese populations, the risk for severe Plasmodium falciparum malaria and its specific phenotypes, including severe and mild malaria.”
Mild malaria is a phenotype of malaria not severe malaria.

Line 140-141 We characterized polymorphisms on HBB and G6PD genes in residents from three ecological zones where the malaria endemicity was different.
Did the different ecological zones also have different ethnicity?


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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 1
Reviewer: 4

Output:
Basic reporting: The research article entitle "G6PD and HBB polymorphisms in the Senegalese population: prevalence and correlation with clinical malaria" by Fatou Thiam et al. The manuscript is  written very well and organized. However, it needs minor correction before final publication. The idea of the study of G6PD and HBB polymorphisms in the Senegalese population is interesting and will enhance knowledge for the development of malaria drugs.  I would request authors to please correct sentences and grammar so that the non-science people could easily understand your work and its importance.

The 2nd reference is a very old study. Please replace with some new article and finding and also look other references.

Introduction is very comprehensive and representing sufficient for this study. Covered all manuscript part and follow but please check references and grammar before final submission.

Additional, Authors advised to check the figures number, visibility, figures legend, Table no, and legends carefully before final submission.

Figure 1: Mark the number of samples collected from different. What is represented (0,30,60,120, 180, and 240) in this figure, and how is it important? What means outside (border number) in the figures? Is it relevant with endemic or sample collection? Would you please clarify it?

Figure 2: For better visualization and representation, I suggest plotting a lollipop plot of HBB and G6PD with all exonic and intronic regions. Please also mark all SNPs in 3D protein structure.

Table: Write the full name of MAF, HWE, SM, UM, and CTR below. It will help the reader to understand easily.
Experimental design: The overall finding of this project is a novelty in the context of G6PD and HBB polymorphisms in the Senegalese population, and this work should be publishable after minor revision.

The research questions are well defined and relevant to the scientific community, especially malaria research. Authors used a adequate number of samples to address the questions.
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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Thank you for the extensive revisions. It reads much better. It could do with another read through and a spell check as there are a number of misspelled words and some bizarre use of capitals mid-sentence. Otherwise some minor points below.
Line 51 It reads Results: Six frequent SNPs The minor allele frequency (MAF) > 3%) ….
Maybe should read: Six frequent SNPs with a minor allele frequency (MAF) > 3%
Line 138 It reads “malaria versus outcome”  Word missing?
Line 163 “Even there is an ethnic distribution from North to South regions”  word(s) missing?
Line 164/5 “For commodity”   do you mean convenience?
Line 172 “however, health conditions are different with better disease coverage..” what is better disease coverage?
Line 195 “Still, other complications such as hypoglycemia, thrombocytopenia, renal insufficiency, hepatic or even pulmonary oedema may appear alone or in combination.” This sentence seems to come from nowhere. Do you mean these conditions without malari infections?
Line 252-4 “Differences in allelic  frequencies among the three groups (SM, UM, CTR) were implemented using the logistic regression analysis method, performed elsewhere [55, 56].”
This is a strange sentence. What is meant by implemented and performed elsewhere.
Line 489-491 “It is logical that there are more adults. Because our inclusions are not oriented towards pediatric services, otherwise this will be a point to be shaved in our future inclusions.” This needs to be reworded and I’m not sure why “shaved” is used.
Line 511 “Our results showed an association between In conclusion,” There seem to be words missing before the “In conclusion…”
Table 2. Maybe include stat test used in the title.
Table 5 needs more explanation in the legend. I presume PV is P-value but the rest are? And what do the blue boxes, lines and arrows refer to?
Experimental design: Previous issues addressed. Dieureudieuf.
Validity of the findings: Previous issues addressed. Dieureudieuf.
Additional comments: Previous issues addressed. Dieureudieuf.


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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have taken into account issues raised by reviewers.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The manuscript reads a lot better than the earlier version.


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

Input:
Title: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 3
Reviewer: 1

Output:
Basic reporting: Points addressed and english corrected
Experimental design: Points addressed.
Validity of the findings: Points addressed
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: G6PD AND HBB POLYMORPHISMS IN THE SENEGALESE POPULATION: PREVALENCE, CORRELATION WITH CLINICAL MALARIA
Review round: 3
Reviewer: 2

Output:
Basic reporting: The authors have corrected all the raised comments. Thanks for the quick response. Now research article is impressive, and it could be publishable.
Experimental design: No change is required.
Validity of the findings: Not required any modification original article is fine for publication.
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: CHARACTERISTICS AND RISK FACTORS OF INFECTIONS IN PATIENTS WITH HBV-RELATED ACUTE-ON-CHRONIC LIVER FAILURE: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: This is a a retrospective analysis of 539 patients with HBV-ACLF.  The manuscript is well written but lack of innovation. The data presented do not bring new messages to the current knowledge.

Major concerns:

1- The authors used the APASL classification of ACLF that lacks data on ACLF grading.
The EASL-CLIF classification allow grading of ACLF by severity. We might except that infection rates was higher in patients admitted with ACLF 3. Please heck if you can classify the patients by ACLF grading at time of admission

2-Other scores like the Child-Pugh score, the MELD score could allow better classification of infection occurrence according to liver severity.

3-Unless I missed, it's not clear whether the patients were admitted in the ward or in ICU

4- According to above, how many patients under mechanical ventilation or underwent dialysis ?

5- Type of Bacterial infection and strains identification, resistance phenotype are not reported ?

6- Carbapenem was the most common antibiotic used. How many patients had BMR or MDRO ?

7- What was the proportion of patients who had community acquired infection and those who developed a nosocomial infection ?

8- It's not reported what incidence and type of fungal and viral infection.

9- Outcome of patients who developed infection ?
Additional comments: None


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

Input:
Title: CHARACTERISTICS AND RISK FACTORS OF INFECTIONS IN PATIENTS WITH HBV-RELATED ACUTE-ON-CHRONIC LIVER FAILURE: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors describe the prevalence and predictors of infections in patients with HBV ACLF. However, there are certain flaws in the manuscript in its present form as highlighted below
1.	English language is very poor and requires proof reading by an English expert
2.	Background shows context but needs some literature review on the topic
3.	Raw data has not been supplied
4.	Figures are well made but are a replication of what is mentioned in the text
Experimental design: 1.	A major flaw of this study is the inclusion of all patients with infection. Infection may also have been the precipitant of ACLF
2.	Ideally the authors should have only included patients who were non-infected at admission and then developed infection after 48 hours of admission to evaluate the risk and predictors of infection
3.	The authors mention cirrhosis as a severe complication of ACLF, but cirrhosis is usually a background chronic liver disease in these patients upon which they develop acute insult resulting in ACLF. Authors should clarify this
4.	Acute cholecystitis may be an infectious complication if acalculous. However, GB wall thickening may also be a manifestation of acute hepatitis
5.	Acute pancreatitis is not an infection, authors should clarify this
6.	The authors should provide reference for the HBV DNA cut offs.
7.	Were these patients HBe Ag negative and positive or HBs Ag negative and positive ?
8.	The authors should clarify, why some patients received antibiotic for more than 1 month
9.	The lines 159 – 161 in the results section should have come in the beginning of results section
10.	What was the acute precipitant of ACLF in these patients ?
11.	What was the duration of presentation in these patients
12.	How many patients had index presentation as ACLF ?
13.	As per the canionic definition, what were the ACLF grades in these patients
Validity of the findings: 1.	Discussion needs to refer more studies on infection and ACLF (Dig Liver Dis. 2018 Nov;50(11):1225-1231)
2. Relevant findings but not adequately represented
Additional comments: None


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

Input:
Title: CHARACTERISTICS AND RISK FACTORS OF INFECTIONS IN PATIENTS WITH HBV-RELATED ACUTE-ON-CHRONIC LIVER FAILURE: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Clear
Experimental design: Not applicable
Validity of the findings: Yes
Additional comments: None


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

Input:
Title: CHARACTERISTICS AND RISK FACTORS OF INFECTIONS IN PATIENTS WITH HBV-RELATED ACUTE-ON-CHRONIC LIVER FAILURE: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Fine
Experimental design: Fine
Validity of the findings: Fine
Additional comments: The paper has been revised adequately.
Please cite the following reference.
Shalimar, Rout G, Jadaun SS, Ranjan G, Kedia S, Gunjan D, Nayak B, Acharya SK, Kumar A, Kapil A. Prevalence, predictors and impact of bacterial infection in acute on chronic liver failure patients. Dig Liver Dis. 2018 Nov;50(11):1225-1231. doi: 10.1016/j.dld.2018.05.013.


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: SOIL MINERALIZED CARBON DRIVES MORE CARBON STOCK IN CONIFEROUS-BROADLEAF MIXED PLANTATIONS COMPARED TO PURE PLANTATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: - Language needs improvement (In many cases it is almost impossible what authors are trying to say).
- Some scientific terminologies are incorrectly used.
- Figures and tables are not well formatted, some tables even went outside of the page!
- Not enough background of the topic to justify how the research fits into broader filed of knowledge.
- Authors completely failed to justify why this study is important both in terms of scientific advancement and practical application.
Experimental design: - Poorly designed experiment.
- Not enough replications (in fact no actual replication, only pseudo-replication).
- Statistics used on only ~3 data points barely carry any merit to justify a conclusion.
- Soil samples are not collected in a properly designed fashion (there are missing measurements in 2017). Besides, sampling schemes are not clearly described.
- Methods are not described with enough details to be replicated by someone not part of this experiment.
Validity of the findings: As I mentioned earlier, the authors failed to justify the novelty of this research. Moreover, the poorly designed experiment makes it almost impossible to draw any data-supported conclusion.
Additional comments: I have proved some comments in the attached document.


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

Input:
Title: SOIL MINERALIZED CARBON DRIVES MORE CARBON STOCK IN CONIFEROUS-BROADLEAF MIXED PLANTATIONS COMPARED TO PURE PLANTATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: I am not a native speaker.
The context is presented correctly.
The article contains the correct structure.
Table 2, Figures 2 and 3 – please add asterix  according to the table footnote.
Experimental design: Please find suggestion in enclosed document.
Validity of the findings: Please find suggestion in enclosed document.
Additional comments: Please find suggestion in enclosed document.


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

Input:
Title: SOIL MINERALIZED CARBON DRIVES MORE CARBON STOCK IN CONIFEROUS-BROADLEAF MIXED PLANTATIONS COMPARED TO PURE PLANTATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am not a native speaker but the language and scientific terminology used still needs improvement.

Citation and references need checking – e.g. line 75 -Tian and Chang  et al., while in the reference list there is Tan and Chang…; line 90 – lack of “and” (Sanford Kucharik, 2013).

Figures need improvement - details were shown the the report.
Experimental design: The sampling protocol is still not clearly explained.
Validity of the findings: Studies in the proposed scope are not new to the contribution of forest soils to C sequestration.

The data provided needs improvement, e.g., statistical significance. In many places in the text with results it is difficult to find the data in tables or graphs.
Additional comments: no comments


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

Input:
Title: SOIL MINERALIZED CARBON DRIVES MORE CARBON STOCK IN CONIFEROUS-BROADLEAF MIXED PLANTATIONS COMPARED TO PURE PLANTATIONS
Review round: 3
Reviewer: 1

Output:
Basic reporting: I am not a native speaker but in my opinion, the article has been improved on the language side. Minor language changes were suggested. Citation – line 171 – Ma et al. instead of Ma ?; please check. Figures and Table have been improved. The article is structured correctly.
Experimental design: The sampling protocol has been improved, although there are aspects to be completed, e.g., lack of methodology to the data shown in Table 2.
Validity of the findings: Studies in the proposed scope are not new to the contribution of forest soils to C sequestration, however increase the number and area of forest soils examined. The data provided has been improved.
Additional comments: Overall the article has been improved in many ways. However, it still requires some revision before publication. In enclosed some suggestions for consideration.


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: THE ROLES OF TRAIT AND PROCESS RESILIENCE IN RELATION OF BIS/BAS AND DEPRESSIVE SYMPTOMS AMONG ADOLESCENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: See attached PDF


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

Input:
Title: THE ROLES OF TRAIT AND PROCESS RESILIENCE IN RELATION OF BIS/BAS AND DEPRESSIVE SYMPTOMS AMONG ADOLESCENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: Participants: It could be better to maximize the participants and add descriptions of relevant demographic characteristics in detail, such as gender.
Procedures: Suggest specify the date range of data/survey collection, considering the depressive symptoms would be different at different test times (month/day/year). And the study mentioned it was conducted during the COVID-19 pandemic, so it’s better to describe the pandemic where participants were.
Results: provide the results with more details, including the description data of participants in the tables or figures.
For instance: Table 1. should add the note of *p< .05, **p< .01, ***p< .001.
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: THE ROLES OF TRAIT AND PROCESS RESILIENCE IN RELATION OF BIS/BAS AND DEPRESSIVE SYMPTOMS AMONG ADOLESCENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1.The paper requires extensive language editing. Please ensure in revising your manuscript that this is attended to.
2. It is required to provide more evidence in the introduction that supports the hypotheses and to expand the findings on the role of resilience in the model.
3.Line 207: Please carefully check the symbol in this line. Currently 'ps<.001' is wrong.
Experimental design: no comment
Validity of the findings: 1.It is recommended to include the x2/df and SRMR values, in the results of the structural equations.
2.It is recommended to give more details about demographic characteristics of the study sample, for example the gender and the percentage of reported symptoms of depression.
3.Figure 2 Please carefully check all the path coefficients are standardized values in this figure. Currently 'the path coefficient from trait resilience to depression ' is greater than 1 which means it is unstandardized.
4.To test the mediation effect, the bias-corrected 95% confidence interval (CI) should be calculated with 1,000 bootstrapping samples. If the 95% CI of the indirect effect does not include 0, a significant mediation effect can be established. Please use this method to test the mediation effect in this study.
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: THE ROLES OF TRAIT AND PROCESS RESILIENCE IN RELATION OF BIS/BAS AND DEPRESSIVE SYMPTOMS AMONG ADOLESCENTS
Review round: 1
Reviewer: 4

Output:
Basic reporting: English is clear but should be revised. Punctuation sould also be revised
Literature references are updated and appropiately quoted. The background of the study is sufficiently convincing
The authors may wish to quote V. De Pascalis regarding the possible relation between BIS/BAS and optimism, which could be studied in further ressearch by the same group

The article structure,   figures and the table are OK. Raw data availability is obvious as authors analyzed an open dataset
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Please check English and punctuation across 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: HIDE AND SEEK SHARK TEETH IN RANDOM FORESTS: MACHINE LEARNING APPLIED TO SCYLIORHINUS CANICULA POPULATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: A pdf file is provided.
Experimental design: A pdf file is provided.
Validity of the findings: A pdf file is provided.
Additional comments: This MS deals with the level of tooth shape and form differentiation between the Mediterranean and Atlantic population samples of S. canicula by using two approaches. This work is well-written and structured, and the methodology applied is considered adequate. Nevertheless, this MS needs major revision prior to acceptance based on the following issues: significant bibliographic gaps have been identified, several parts need clarification, and in several cases the assumptions should be lightened. More specifically (all comments are provided analytically in the pdf file):
 It is recommended that authors include some significant references about the species (they are provided in comments in the pdf file).
 Line 25: add the important information: "from 56 individuals".
 Lines 85-86: According to a previous study length-weight relationships of S. cannicula in the eastern Mediterranean varied significantly from those of the western and central Mediterranean.
 Lines 148-150: Authors should explain why the maturity assessment was not conducted in the current study, considering that they samples the whole specimen. This should be clarified in text.
 Line 162: the ageing methodologies used by each study should be added, considering that they are different and results may vary.
 Line 167: "Based on the von Bertalanffy growth curves" based on which exactly equations? Please clarify.
 In general, this part {167-171) needs caution considering that S. canicula vertebrae are characterized by low calcification degree and annual rings cannot be easily discriminated.
 Knowing also that in most, if not all, age-studies no age validation method has been applied, solid estimations of growth parameters cannot be provided, and thus applied in meta-analysis with high confidence. As a consequence, authors should highlight that these results need further investigation based on the above "issues".
 Lines 458-461: This part needs modification. In the case of S. canicula "Higher diet diversity was observed in males than females, in immature individuals than mature ones, regardless of sex, as well as in spring in comparison to autumn and winter. Feeding intensity seemed to be influenced mainly by sex and maturity condition." meaning that sex and season can also be considered as significant factors (sexual maturity is related positively to length).
 Lines 470-471: This statement is incorrect and needs modification. It has been found that "Both mitochondrial and nuclear microsatellite DNA data revealed a strong genetic subdivision, mainly between the western and eastern Mediterranean, whereas the Levantine Basin shared haplotypes with both areas. The geographic isolation of the Mediterranean basins seems to enforce the population genetic differentiation of the species, with the deep sea acting as a strong barrier to its dispersal."
 Lines 475-476: Is there a reference to support this statement. Usually species with pelagic larvae are influenced by hydrographic conditions. Please delete or rephrase.
 Line 477: Replace with: is probably related to the genetic......
 Lines 476-478: This part should be lightened, since the samples included in this study were not examined genetically. A sentence in the end can be added: "Nevertheless, further research examining the differentiation at both molecular and tooth morphology level of the same specimens between this regions, could confirm this assumption."


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: HIDE AND SEEK SHARK TEETH IN RANDOM FORESTS: MACHINE LEARNING APPLIED TO SCYLIORHINUS CANICULA POPULATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well written, in clear and readable English.
The references are ok, and for the main scope of the article I think that the authors have provided sufficient and satisfactory background and contextualization.
The structure of the article is ok, with good figures and tables. However, for the figures that shows the morphology of teeth, as figure 3, I have one suggestion: to facilitate the reader, I suggest that the authors create two more figures. One that show the morphological comparison of the teeth of adult specimens from the Mediterranean and Atlantic populations, side by side (Atlantic male with Mediterranean male, Atlantic female with Mediterranean female), since the main morphological differences between the teeth of the two populations are found in these specimens. And a second one that shows the ontogenetic difference between neonates, juveniles and adults of each of these populations. I liked figure 3, but as a morphologist who works with fossil teeth, I think that summarizing the differences in a figure like 3 loses important visual information on the differences in the teeth of these populations which can be shown in more detail. And for the figure 5, I have one question: Does the size of the centroid have any measurement value (like centimeters or millimeters)?
The results are good, and relevant mainly because they show how RF can be a good and efficient tool for taxonomy studies that use morphometric data.
Experimental design: The article deals with new data on morphometric differences in dentition between Atlantic and Mediterranean populations of Scyliorhinus canicula, and for me the main and original point of the article was the use of RF to analyze these data. The data to be published are in agreement with the scope of the journal.
The subject addressed is well defined and the data obtained, despite being more robust when used in adult specimens of the two populations, are relevant.
There was an investigation into the differences found in the morphology of the teeth of the sampled populations of S. canicula, and it is very clear how the use of RF programs for the analysis of morphometric data (that can express these differences) was more efficient and robust than LDA. However, for the results, I felt a lack of a more complete morphological description, reporting for example the number of cusps found in hatchlings, juveniles and adults between males and females within and between the investigated populations. In my opinion, the authors were a little more concerned with showing the differences obtained between the LDA and RF analyzes used to interpret the landmarks and morphometric data of the structures, than reporting in more detail the morphological aspects of the teeth of populations analyzed.
Methodology is well explained and described.
In the abstract, I understand that stating that more than 3,000 teeth were sampled is interesting and catches the reader's attention; but I think it is more important to highlight there the number of specimens analyzed: 25 from the Mediterranean and 31 from the Atlantic. Personally I find it more relevant.
Validity of the findings: The results found are very relevant, and show mainly that morphometric data are a good source of information to be explored in structures such as teeth to help distinguish populations of the same species. Even more so if these data are added to detailed descriptions of such structures. For me, one of the highlights of the article is the demonstration of the greater efficiency of RF to analyze and interpret morphometric data (when compared to LDA) as being a great replicable methodology for studies that look for morphological differences in structures such as teeth and dermal denticles between populations of species of elasmobranchs. I see that there is great potential in using this type of analysis for morphometric data of such structures.
All underlying data are ok, however I think it is important to emphasize in the introduction that gene flow possibly occurs between the populations of S. canicula, even if at low flow, as mentioned in the final part of the discussion page 17 between lines 470 and 473, because on page 2 line 70 (introduction) the authors state there is a geographical disjunction between the populations of S. canicula, which, in my view, contradicts what was exposed in the final part of the discussion. It is clear that the analyzes of teeth sampled here for these two populations should also be carried out for other regions of the Atlantic and Mediterranean where S. canicula occurs (as the authors themselves stated) to see if there are these teeth differences between other populations. However, since there is still a genetic connection between the populations of S. canicula, the statement of a geographic disjunction can lead the reader to an interpretation of a possible speciation, which is not yet confirmed for this species.
Additional comments: Here I will expose a few corrections of small details that I found in the text:
Line 67: Eastern instead of "Western".
Line 140: I don't know if the journal allows, but usually the genus epithet is written in full at the beginning of sentences: Scyliorhinus.
Lines 445 and 465: the same as 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: HIDE AND SEEK SHARK TEETH IN RANDOM FORESTS: MACHINE LEARNING APPLIED TO SCYLIORHINUS CANICULA POPULATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: 14h March 2022
Dear Editor-in-chief,

You may find attached the revised version of the manuscript (MS) entitled “Hide and seek shark teeth in Random Forests: Machine learning applied to Scyliorhinus canicula populations”. This MS deals with the level of tooth shape and form differentiation between the Mediterranean and Atlantic population samples of S. canicula by using two approaches.

Authors have addressed several issues form previous review report. Nevertheless a major deficiency still encountered is the groupings that applied based on maturity state, as well as the calculation of growth parameters on such few measurements and by using indirect methodology.

Synoptically, the MS still needs revision mainly in terms of methodology applied: Authors should consider to delete completely the calculation of growth parameters from Methods and be specific regarding the groupings. Further minor linguistic and syntax issues are highlighted. A linguistic check is highly recommended.

A pdf file with track changes is provided for the convenience of authors.

Authors are encouraged to provide a "clean" version of the revised MS.

Sincerely,

The Reviewer
Experimental design: A pdf file is provided.
Validity of the findings: A pdf file is provided.
Additional comments: A pdf file is provided.


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: HIDE AND SEEK SHARK TEETH IN RANDOM FORESTS: MACHINE LEARNING APPLIED TO SCYLIORHINUS CANICULA POPULATIONS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors responded and accepted the points made by me satisfactorily.
Experimental design: The authors responded and accepted the points made by me satisfactorily.
Validity of the findings: The authors responded and accepted the points made by me satisfactorily.
Additional comments: The authors responded and accepted the points made by me satisfactorily.


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: HIDE AND SEEK SHARK TEETH IN RANDOM FORESTS: MACHINE LEARNING APPLIED TO SCYLIORHINUS CANICULA POPULATIONS
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: pdf with comments is provided
Validity of the findings: pdf with comments is provided
Additional comments: pdf with comments is provided


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: SEA URCHINS: AN UPDATE ON THEIR PHARMACOLOGICAL PROPERTIES
Review round: 1
Reviewer: 1

Output:
Basic reporting: I do not have any comment on the English. I would just suggest to use "an update on" instead of "an update of".
The literature overview is quite broad and updated, I consider it sufficient to cover the topic.
In the paper there are no tables and figures. I think that this is a lack of the paper. In the reviews, particularly, this kind of schemes are very useful to guide and summarize the findings.
Experimental design: The review is well organized in a systematic way.
Validity of the findings: The main failure of the paper is the lack of detailed description of the mechanisms of action of compounds showing a pharmacological / biological activity. The description is too general and lack in going deeper, but this could represent the most useful information for the reader. Most of the times the authors talk about "the extract effect" towards a specific activity, but frequently the cited papers go into more details of the specific components responsible of the activity.
More information should be added.
The conclusions should be expanded. In the present paper, they are just a general and too brief summary of the manuscript.
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: SEA URCHINS: AN UPDATE ON THEIR PHARMACOLOGICAL PROPERTIES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript #67602 by DM Moreno-García and co-authors is a synthesis of potential, medicinal properties of bioactive compounds found, in former works, in nine common species of sea urchins. Based on an extensive review of the literature, it highlights the potential of numerous, different compounds present in sea urchins with distinct potential pharmacological implications, from antifungal and anti-arasitic benefits, to anti-diabetic and cardiotoxic effects. I think this short, and very synthetic and rich review should be of interest to a wide panel of readers, from the fields of marine biology to pharmacology, and highlight the potential of these organisms to develop efficient active ingredients in the future.

The manuscript is clearly written in correct, professional English and well-structured. Only some terms refering to the anatomy of sea-urchins are not correctly used in the text and in figure 1. Please see my specific comments and suggestions on this point below.

The objectives of the review is well-presented in the introduction. However, there are significant weaknesses in the general presentation of sea urchins’ anatomy and evolution, along with imprecisions in the use of several anatomical terms and in the description of biological functions (see my comments below and in the annotated version of the ms).
Experimental design: The review method is rigorous and well described in a dedicated section (survey methodology). Sources are clearly cited and the review is logically organized following the potential parmacological effects.
Validity of the findings: While the check-list of active substances with potential pharmacological effects in sea urchins is rich, the conclusion very logically highlights that the understanding of some active principles is still needed without mentioning the need of toxicological and clinical trials before effective drugs and treatments can be developed.
Additional comments: To conclude, I find the manuscript topical and of high interest. My main concern is that the general presentation of sea urchins and their anatomy, and of their scientific value in the introduction comprises some mistakes and imprecisions.  Please find below and in an annotated version of the manuscript (comments and corrections in pdf) my detailed remarks and suggestions on the form of the manuscript.

In the abstract :

L41 « primitive » : The word "primitive" should be avoided as it may refer to a concept of final evolution that has no place in modern evolutionary sciences. It could be replaced by "ancestral", for instance. However, I don't see how the immune system of echinoids should be regarded as more ancestral than the one of other groups such as asteroids, polychetes, or sponges...?

L42 « ever-changing environmental conditions » : it is not clear why sea urchins should be impacted by ever-changing conditions compared to other marine oganisms ?

In the introduction :

L59 : no phylogenetic analsyis ever supported an ancestral position of sea-urchins at the base of echinoderms. This is very surprising.

L60-63 : this statement is not correct. There are many invertebrates that have closer phylgenetic relationships with vertebrates than echinoderms (ex : all chordates). Please amend.

L64-65 : these are general characters present in many echinoderms, they are not specific to echinoids.

L 65- 75 : this section is confusing as some of the mentioned functions are associated to the water vascular system, others are in link with the hemal system, other (ex : locomotion) are independent.  Can the authors be more specific and clearly highlight the biological and physiological roles of the hemal system in echinoids in link with the manuscript’s objectives.

L 78 :  It is not clear why benthic organisms are more frequently exposed to pathogens compared to other organisms ?

L 83 : here again, the necessity of having evolved an efficient immunitory system is not specific to echinoids but common to all organisms

Figure 1.
In the figure, some terms related to the anatomy of sea urchins are not correct. Please replace the words « spike » for « spine » and « shells » for « test ». Spine is the official word to qualify the sea urchin’s appendices, and test the official term to qualify the internal skeleton of echinoderms (while shell refers to the external skeleton present in mollusks).


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: SEA URCHINS: AN UPDATE ON THEIR PHARMACOLOGICAL PROPERTIES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The English quality is not the best and needs improvement.
The used references seem appropriate and thoroughly selected.
The article structure is adequate for a review.
I believe the review has a broad scope and has cross-disciplinary interest.
A similar review has been published in 2021 by Sibiya et al. but focused solely on the anticarcinogenic, antioxidant, anticoagulating and antibacterial properties exhibited by sea urchins derived molecules. This review is a complement focusing on antifungal, anti-parasitic, anti-inflammatory, hepatoprotective, antiviral, anti-diabetic, anti-lipidemic, gastro-protective and cardiotoxic effects.
The introduction seems appropriate but needs improvement according to the specific comments.
Experimental design: I believe this review is within the Scope and Aims of PeerJ.
The methodology used to select the papers to be included in this review are described.
The papers are adequately cited and the review is organized logically. However, some sentences are too long and recommendation are provided in the Detailed comments.
Validity of the findings: Conclusions are brief but seem adequate for a review.
Additional comments: General comments:

In this manuscript Moreno-García and colleagues provide a review on recent studies on sea urchin bioactive molecules and their potential pharmacological benefits such as antifungal, anti-parasitic, anti-inflammatory, hepatoprotective, antiviral, anti-diabetic, anti-lipidemic, gastro-protective and cardiotoxic effects. They stress that their main goal with this review is to complement a recent review from Sibiya and collaborators (2021) that focused solely on the anticarcinogenic, antioxidant, anticoagulating and antibacterial properties exhibited by sea urchins derived molecules.

The authors conclude that sea urchin derived molecules have a great potential for the development of effective drugs for many modern society human illnesses, but there is still a long way to go, since studies are lacking that elucidate the structure of the active principles, as well as toxicological, preclinical and clinical trials to demonstrate their therapeutic safety and pharmacological efficacy.

Overall, the manuscript content is relevant in view of the development of the Blue Economy. The English quality is not the best and needs improvement, has well as clarification in some minor points detailed below.

In my opinion, the current manuscript meets the quality guidelines to be published in PeerJ, after the proposed corrections (see specific comments).

Specific comments:

Line 59: replace by “…marine invertebrates of the class …”

Line 59: explain what do you mean by “the class Echinoidea at the base of the echinoderms”. The current phylogenetic tree of Echinodermata, support two sister groups – the Asterozoa (sea stars and brittle stars) and Echinozoa (sea urchins and sea cucumbers). See Reich et al. 2015. Phylogenomic Analyses of Echinodermata Support the Sister Groups of Asterozoa and Echinozoa.

Line 78: replace by “…exposed to pathogens…”

Line 83: replace by “…conditions sea urchins…”

Line 84: replace by “…innate immunitary system…”

Line 87: replace by “…animals to acquire…”

Line 91: you mention “apoptosis and autophagy” which are mainly related with echinoderm’s ability to regenerate. I think this should be clearly stated for the readers.

Line 98: replace by “…Asia-Pacific regions…”

Line 99: replace by “cardiovascular conditions…”

Line 103: replace by “…sea urchins…” and “but due to their…”

Line 120: replace by “Hepatoprotector”

Line 132: remove or add a Subheading

Lines 133-134: replace 09 by 9

Lines 138-139: in-vitro and in-vivo should be in italics

Line 152: explain the abbreviation ROS for non-specialized readers. This was done in line 213 but should be done here.

Line 158: replace “medicines” by drugs

Line 162: replace What’s more by Furthermore

Line 175: replace by “…improvement in glucose homeostasis and decrease in insulin resistance…”

Line 176-179: this should be a sentence on its own and should be better explained “Methanol extracts most likely contain saponins, which have properties similar to insulin, stimulating glucose absorption and thereby contributing to its storage in adipocytes (Soliman, 2016; Soliman, Mohamed & Marie, 2016).”

Line 185-186: replace by “…are the most effective.”

Line 191: replace by “…in vaginal, respiratory and intestinal infections”

Line 193: explain “…from the interior…”. From which part or organs were the extracts prepared?

Line 205: rewrite this sentence “which are be found in from other types of echinoderms”. Do you mean “which are known to be found…”?

Line 212-213: replace by “…leading thereby to the production of increased amounts of ROS, …”

Line 221: replace by “…attaches to the active site…”

Line 224: replace by “…that contain mainly …”

Line 229: replace by “…Dyslipidemia is having a high concentration of lipids in an organism or the blood…”

Line 237: replace by “…administered to mice which were previously provided with high fat content diets…”

Line 251: replace by “…preventive effect…”

Line 267-272: too long, divide in two sentences

Line 274: in silico must be in italic

Line 273-277: again too long, divide in two sentences

Line 279: replace “forces” by “contacts”

Line 325-327: replace by “…[catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-s-transferase (GST), glutathione reductase (GR)]”

Line 334: replace “Sea urchins” by “Sea urchin-derived molecules…”

Line 336-339: rephrase as follows “Furthermore, the reports included in this review, show that the bioactive compounds are not restricted to one body part, but instead evidence that the entire organism and even its waste products, can be used.”

Table 1: Scaphechinus mirabilis, Stomopneustes variolaris, Tripneustes depressus and T. ventricosus are not mentioned in the text only in Table 1. This should be corrected.


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: SEA URCHINS: AN UPDATE ON THEIR PHARMACOLOGICAL PROPERTIES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors met all the reviewers comments and clearly improved the menuscript. I consider it acceptable in the present form
Experimental design: The authors met all the reviewers comments and clearly improved the menuscript. I consider it acceptable in the present form
Validity of the findings: The authors met all the reviewers comments and clearly improved the menuscript. I consider it acceptable in the present form
Additional comments: The authors met all the reviewers comments and clearly improved the menuscript. I consider it acceptable in the present form


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

Input:
Title: SEA URCHINS: AN UPDATE ON THEIR PHARMACOLOGICAL PROPERTIES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have significantly improved the manuscript and have incorporated all the suggested revisions.
Experimental design: The authors have significantly improved the manuscript and have incorporated all the suggested revisions.
Validity of the findings: The authors have significantly improved the manuscript and have incorporated all the suggested revisions.
Additional comments: No additional comments


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

Input:
Title: COMPARATIVE ACTUALISTIC STUDY HINTS AT ORIGINS OF ALLEGED MIOCENE COPROLITES OF POLAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is well organized and written in a clearly understandable way. The illustrations are very good.
Experimental design: The research involves to primary aspects: a description of ferruginous extrusions, and  comparison with excrements obtained from a wide variety of extant animals, with a goal of determining what ancient creatures may have been responsible of producing fecal masses that later became replaced by iron minerals. This is a very sound experimental design.
Validity of the findings: I believe that the conclusions lack validity, a result of the authors' decision to only consider terrestrial animals for coprolites that are preserved in sediments that were likely deposited in lakebed or stream deposits. I discuss this topic in detail in an attached letter, and in annotations on my edit of the original manuscript.
Additional comments: Review Comments
2/10/22

The manuscript provides a detailed description of ferruginous objects from Miocene strata that are interpreted as being the fossilized excrement having two morphotypes. The smaller objects are inferred to have been made by tortoises, the larger by snakes.

The manuscript has many positive attributes. The writing is clear and well organized, suitable for a wide range of readers, illustrations are excellent, and numerous references are provided. Given the long-standing controversy regarding the excremental versus abiotic origin of ferruginous extrusions (which have wide geographic and chronological distribution), this paper is an important contribution. The authors provide a clear summary of the controversy, where the lack of internal structure and associated body fossils are consistent features of these occurrences. These characteristics suggest an abiotic origin. In contrast, the shapes of these objects closely resemble fecal excrements. I appreciate the fact the authors limit their interpretation to a specific locality, and do not try to generalize their findings in an attempt to present a model that is alleged to apply to other locations where ferruginous extrusions occur.

Although the manuscript has many positive characteristics, there are also some weaknesses that in my view make it unacceptable for publication in its present form. My recommendation is therefore for either acceptance with major revision, or rejection with a suggestion for resubmission. I will leave it to journal editors to make that call.

I am attaching my edit of the manuscript, which includes many comments and suggestions. These mostly involve small details, not major issues. My intent is to be helpful rather than critical, and I admit that as a reviewer I tend to be wordy. I would also like to say that although I have published some papers on the topic of ferruginous “coprolites”, I am open to accept interpretations that are different from my own. I believe that scientific research benefits from a diversity of viewpoints. As a reviewer, my goal is to evaluate manuscripts based on the quality of the presentation and the adequacy of the supporting evidence, without insisting that the conclusions match my own views, or fit traditional paradigms.

In this instance, my primary concern is related to a fundamental inconsistency between geologic evidence and biologic interpretations.  In the description of geologic setting, the ferruginous masses (“coprolites”) are reported to have been found in the Biedrzychowice Formation. This unit is reported to contain numerous paleosol  layers where plant roots and tree trunks are preserved in situ. This is consistent with the description of the paleoenvironemnt as representing “vast swamps and rush marshes”. Although paleosols are evidence of subaerial conditions, the “coprolites” occur in a clay/mudstone layer, a sediment type that is characteristic of subaqueous conditions.

Herein lies the problem. In studying 787 excrements from extant creatures, the authors chose to consider only terrestrial-dwelling forms. This selection limited the likely choices to tortoises (smaller “coprolite” morphotype, and snakes for the larger morphotype. This interpretation that has major flaws. First, it omits consideration of other creatures that might have lived in the water column overlying the clay/mud layer. This might include reptiles (crocodilians, pond turtles) or mammals (modern forms include beaver, otter, muskrat, etc.)  This ecological niche may have been occupied by ancestral equivalents during the Miocene, but it is an environment conducive to habitation. A second problem is that the clay/mudstone deposit is an unlikely habitat for tortoises and snakes. For example, the modern snake excrements all came from taxa whose habitats that include grasslands, scrubland, savannahs, woodlands, or rain forests. None of these environments are likely to produce clay beds, which require the low-energy depositional environment typical of lake beds or slow-moving streams. Tortoises dwell in dry habitats that include grasslands, savannah, and desert, where soils and sediments are typically sandy. They are not likely to be found in quiet-water environments that favor deposition of clay/mudstone.

A further issue is the authors’ utilization of known coprolite-producers based on previous reports.  Many of these occurrences are from geologic settings very different from the study area. A further complication is that for virtually all organisms, the fossil record is very incomplete, and new discoveries are being made all the time.  A similar issue occurs with the listing of known Miocene fossils from sites in Central Europe. The situation can be summed up with the axiom “Lack of evidence is not evidence of absence”.  The list of known fossil taxa from any location is likely to be very incomplete compared to the total taxonomic diversity.

In summary, the authors have provided very detailed mineralogical analysis of the specimens, using a variety of sophisticated methods.  I applaud that careful work.  The geochemical processes that could cause organic-rich fecal material to become completely replaced by siderite defy easy explanation (the roll-front replacement hypothesis cited in the text came from a paleontologist who had no credibility as a geochemist).  This is an attraction of the paper: it adds new observations to a controversial  topic. However, the contention that the objects originated as excrements from tortoises and snakes is not supported by geologic or paleocologic evidence. I see this as a fatal flaw for the acceptance of a manuscript that is otherwise an important contribution.

I welcome having my identity revealed to the authors.

George Mustoe
Research Associate
Geology Department
Western Washington University
Bellingham, WA 98225 USA.


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

Input:
Title: COMPARATIVE ACTUALISTIC STUDY HINTS AT ORIGINS OF ALLEGED MIOCENE COPROLITES OF POLAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is well written and contribute to the knowledge of coprolites, especially those that can be mistaken with concretions. However, there are important reviews to be done before it is ready for publication:

-	Some words are attached to each other and are marked in the PDF;
-	Some important papers can improve the discussion (such as Francischini et al., 2018; Milán, 2012).
Experimental design: The methods used are sound, the inclusions are described in good detail, and the figures are clear and easy to interpret. However some images can improve the paper:
- It is not clear if the inclusions of the modern excrements were analyzed, make it clear;
- Provide images of the concretions (morphologies and the internal structure) and show the diferences with coprolites;
-	The authors analyzed the excrement morphology of modern fish, amphibians, reptiles, birds and mammals. A small sample of the images taken of this variety of excrements can be added as a supplementary material.
Validity of the findings: - Usually, siderite and iron oxide masses are considered concretions, the authors say that in the Biedrzychowice Fm have numerous concretions with the same chemical composition of the coprolites. They must provide a new figure showing the differences between the concretions and coprolites;
-	The absence of inclusions must be better discussed. Crocodiles have a very effective digestive system, and are found in the Miocene deposits, while the serpents faeces described by the authors are not clear if there were bones;
-	The tunnels shown in figure 5 can be better discussed, specially the 5a. It resembles perfectly a tunnel produced by an invertebrate. Due the size, could be produced by a first stage larva of a helminth. This possibility reinforces that these structures are truly coprolites. The authors should discuss this.
Additional comments: See the PDF for more comments.


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

Input:
Title: COMPARATIVE ACTUALISTIC STUDY HINTS AT ORIGINS OF ALLEGED MIOCENE COPROLITES OF POLAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: Reviewer comments:
Thank you for asking me to review this interesting paper! It covers a very interesting subject matter that seems quite difficult to unpick and the authors have made a good (and convincing) attempt to understand these potential fossil poops!
The figures are well made and easy to interpret. The paper seems well referenced (although please note I am not a coprologist). The methods are well described and appropriate – using an interesting method to do some comparative morphological work with extant animal faeces, and I think the findings shown in this paper are sound and sensible. The paper is fairly easy to read, although I think that some minor revisions to the abstract and introduction would greatly help the reader.  I would have also liked a little more investigation into the potential diagenetic mode of preservation for faeces to become siderite, but I do understand that maybe outside of the scope of the paper.
I have one concern pertaining an aspect of the paper which I did not understand well. This maybe my fault, but I was not clear whether there are lots of siderite concretions found in this deposit which do not look like the morphotypes identified by the authors (see my comments RE: line 385). This is concerning and I would like to see how this addressed, because, at the moment it undermines the findings of the paper.
If this can be addressed then I would recommend this fun and fascinating paper for publication.
I have made a few detailed comments in my PDF attached.
All the best,
Dr Thomas Clements
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: COMPARATIVE ACTUALISTIC STUDY HINTS AT ORIGINS OF ALLEGED MIOCENE COPROLITES OF POLAND
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is well-written, with clear organization and good illustrations. Conclusions are based on abundant data.
Experimental design: The comparison of suspected coprolites with excrements from living organisms is a  well-designed strategy.
Validity of the findings: The authors do a good job of presenting evidence that supports their hypothesis. Personally, I am skeptical of the interpretation of the coexistence of snakes and tortoises as the only producers of coprolites (it's an unusual ecological assemblage), However, the origin of ferruginous "coprolite" extrusions has been a subject of much controversy, and I believe that this new study is a useful contribution.
Additional comments: The authors have paid careful attention to the comments from reviewers, and I recommend the manuscript for acceptance pending some minor changes. These are small issues that would likely be detected and remedied during final production editing. Here are some examples:
Introduction, paragraph 1: change spelling of "undegested" to "undigested".
Materals and Methods, paragraph 2:  The description of screening of sediments concludes with "Unfortunately nothing was found..,." This introduces a subjective judgement to the analysis. A better statement would be to delete "unfortunately" and just say "no fossils were found".
Mineralogy, geochemistry and structural analysis, paragraph 3. This present wording suffers from a dangling participle kind of problem caused bu the vague use of "it". : "The spectrum of the carbonate...... It can be recognized as siderite." This wording is saying that the spectrum is made of siderite.
Discussion, paragraph 4. Fix spelling for "communitaction"
Also, a  bit later, "afterall" should be "after all).
The last section of the Discussion uses the spellings "feces" and faeces" in a single sentence.

These are very minor issues that will be easy  to correct.


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

Input:
Title: CURCUMIN EFFECT ON ACANTHAMOEBA TRIANGULARIS ENCYSTATION UNDER NUTRIENT STARVATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: Article is well written. Sufficient literature is provided.
Experimental design: Study is well designed. Please provide justification for use of 100% DMSO as solvent for curcumin preparation. DMSO can interfere with study results, as reported literature indicate:
- DMSO is reported to induce apoptosis at concentrations >10% (v/v)
- DMSO stimulate prompt Acanthamoeba differentiation into a rounded cyst-like stage with a single envelope
- DMSO increases penetration of drugs into parasite cysts
- DMSO is cytotoxic
Validity of the findings: Findings are reported nicely. Please provide suitable justification to clarify that results obtained in this study are because of curcumin use only and there is no interference in study results with the use of DMSO and if DMSO interferes with study data, then please include role of DMSO also in discussion and conclusion.
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: CURCUMIN EFFECT ON ACANTHAMOEBA TRIANGULARIS ENCYSTATION UNDER NUTRIENT STARVATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript describes the effect of sub-lethal doses of curcumin on Acanthamoeba encystation, by investigating morphological parameters and expression of genes related to encystation.  In its essence, the study proposal is sound because encystation inhibition is considered an auxiliary strategy for treating infections by Acanthamoeba. The article is well structured scientifically with adequate figures, the author presented consistent supplementary material and raw data. A robust set of references are presented, but some of them are dispensable. English writing makes understanding difficult in some sections. The text is wordy and an overall revision of the English language must be done. I strongly recommend the assistance of a colleague who is proficient in English or from a professional editing service.
Experimental design: The experimental approach involves the comparison of trophozoites treated or not with sublethal doses of curcumin, in starvation or nutrient-rich conditions, regarding morphological parameters and expression of encystation-related genes. The idea is original and based on previous evidence of the curcumin effect on Acanthamoeba inactivation. An English language revision would improve the overall quality of the text to make clear the main question. Some specific issues in methodology with impact even in the title should be addressed (see specific comments)
Validity of the findings: The data supported the conclusion that curcumin inhibits Acanthamoeba encystation, but the link with Atg genes was not confirmed.  The conclusion description should be more assertive and concise in this direction.
Additional comments: Specific comments and questions are the following:
Main issues:
1.	Abstract: In the Background subsection, the term “surviving amoebas” does not make sense if one does not know they were treated. Consider including “amoebas treated with sublethal doses of curcumin”. The aim of the study is not clear and assertive, I suggest rewriting mainly the last two phrases in the Background subsection.
2.	Introduction: What do the authors consider the focus of the work? The inhibition of encystation by curcumin or the Atg genes expression? As written, the introduction section makes the central aim of the work ambiguous and does not properly connect these two topics. To my understanding, the curcumin effect on encystation is the central point, and the investigation of Atg and other genes expression will give insights into the encystation mechanisms. Therefore, the second paragraph's idea should be before the third one, and the connection of both should be improved.
3.	Material and Methods: The encystation inductor medium (PAS+5% glucose) was based on Aqeel et al (2003) as the authors said. However, Aqeel et al used PBS + 10% glucose + 50mM MgCl2. If a modification was inserted, it must be described, as well as the reason for the modification.
4.	Material & Methods:  Does the term “starvation” adequate, considering that glucose is a nutrient able to support trophozoites activity for the period used in the assays (24h)? Besides, it is clear in the work of Aqeel et al (2003) that high osmolarity is the trigger for encystation. Thus, it seems conceptually incorrect to indicate starvation as the only inducer of encystation. Several recent and old works described encystation saline (no nutrients) to induce cyst formation by starvation as the Neff's encystation medium or other. (Examples: Rolland et al 2020, doi: 10.3390/pathogens9050321; Coulon et al.   2010, doi:10.1128/JCM.00309-10; Bowers & Korn 1969, doi:10.1083/jcb.41.3.786). Why don´t you use them instead of a glucose medium? This conceptual issue demands correction in the paper and in the title (I wonder if distinct types of encystation stimuli could influence the effect of curcumin and even the Atg genes expression, an approach for future investigations.)
5.	Material & Methods: What do you mean by "irregular trophozoites"? Are they pleomorphic, presenting pseudopodia, acanthopodia, and vacuoles as active and healthy forms? If so, the term “irregular” can be misinterpreted.
6.	Discussion: This section should be shortened to address some issues more objectively.  Examples are descriptions on lines 383 to 392 and 516 to 541. Removing references and text restructuring will provide greater conciseness.
7.	Conclusions: Pieces of information previously described in Introduction and Results are unproperly repeated here. I recommend rewriting to indicate the main findings according to the aim and the significance. It must be concise.
Additional minor comments:
8.	Abstract: The methods should be described more technically.
9.	Abstract: In the Results subsection, the description of the results should follow the same sequence as the main text.
10.	Abstract: In the Results subsection, line 83: “Altogether, the data reveals that curcumin stress does not induce cysts formation…” Consider replacing by “curcumin stress inhibits cyst formation”
11.	Introduction: A revision of the English language is needed. Several phrases can be improved in the style (Example on lines 78-85). What is “cidal”?? (line 129).
12.	Material and Methods: The subsection “A. triangularis cultivation” should be the first.
13.	Results: Tables S1, S2 and S3 do not indicate the evaluated parameter (percentage viability)
14.	Results: Asterisks indicative of p-value significance are lacking in Atg 16 graphic. Include that transcriptional expression of other encystation-related genes is in response to curcumin in the caption of Figure 4.


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: CURCUMIN EFFECT ON ACANTHAMOEBA TRIANGULARIS ENCYSTATION UNDER NUTRIENT STARVATION
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: CAN HIGH-INTENSITY INTERVAL TRAINING AND SMALL-SIDED GAMES BE EFFECTIVE FOR IMPROVING PHYSICAL FITNESS AFTER DETRAINING? A PARALLEL STUDY DESIGN IN YOUTH MALE SOCCER PLAYERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Introduction clearly highlights the area but the literature sources used are limited to a handful of authors. The hypothesis for the study is clear and article structure is fine.
Experimental design: It is an interesitng question that the research design seeks to answer. As a true randomisation was carried out there are differences between the groups at baseline which limits the usefulness when looking at different training modalities. A clear description of all the tests which would allow for replication of the work.
Validity of the findings: Given that there are differences between the groups at baseline then there is a problem with appliying the findings. How can you assume one training modality is better than another when they are not starting from the same point. The authors need to address this problem as the discussion is meaningless at the moment. I would suggest either normalising the values to baseline for each group (% change or fold change are common for this) or to change the statistical analysis to an ANCOVA and using baseline test as a covariate. Until the analysis is doen correctly then the discussion is pointless.
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: CAN HIGH-INTENSITY INTERVAL TRAINING AND SMALL-SIDED GAMES BE EFFECTIVE FOR IMPROVING PHYSICAL FITNESS AFTER DETRAINING? A PARALLEL STUDY DESIGN IN YOUTH MALE SOCCER PLAYERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: -	The authors need to work with a fluent English speaker/writer to correct grammatical and punctuation errors throughout the manuscript.
-	The title is not well formulated.
-	Why you choose this comparison (SSG vs. HIIT) in the detraining period.
-	Is there a difference in results between youth players and professional players?
-	The problematic is not clear. Thanks to clear it.
-	More ideas are needed in this part.
-	L78- L81: Thanks to reformulate this paragraph
Experimental design: -	Why you choose this test battery not another ?
-	Which method used to control training intensity ?
-	Assessment procedures is not very clear.
Validity of the findings: -	I think that figure 1 and 2 are not very clear to the reader.
-	Table 5 is very complex. So, i suggest trying to simplify information in this table.
-	L430-L433: This part need to be reformulated
-	L454-L457: Thanks to revise this part
-	At the bottom of the discussion the authors should:
*Discuss limitations of the study, taking into account sources of potential bias or imprecision. Discuss both direction and magnitude of any potential bias
*Discuss the generalisability (external validity) of the study results
*Discuss the practical implications and future research
Additional comments: No additional comments


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

Input:
Title: CAN HIGH-INTENSITY INTERVAL TRAINING AND SMALL-SIDED GAMES BE EFFECTIVE FOR IMPROVING PHYSICAL FITNESS AFTER DETRAINING? A PARALLEL STUDY DESIGN IN YOUTH MALE SOCCER PLAYERS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Nice effort by the research team. The article addresses a great topic and a clear need for research within this area. The information you provided within the literature is relevant and laid out nicely leading into your purpose statement. However, there are a few edits which need to be addressed to ensure the article is sound and ready for publication. My main concern are the originality, the methodology, and the conclusions made. You may need to reformulate the title, to explain more the analysis data, and to soften the conclusions based on the number of subjects and the age of the subjects. The research team makes some pretty broad conclusions that probably need to be softened.
Experimental design: acceptable but needs a lot of explanation and clearly
Validity of the findings: nice conclusions
Additional comments: General Comments:
Nice effort by the research team. The article addresses a great topic and a clear need for research within this area. The information you provided within the literature is relevant and laid out nicely leading into your purpose statement. However, there are a few edits which need to be addressed to ensure the article is sound and ready for publication. My main concern are the originality, the methodology, and the conclusions made. You may need to reformulate the title, to explain more the analysis data, and to soften the conclusions based on the number of subjects and the age of the subjects. The research team makes some pretty broad conclusions that probably need to be softened.
Title:
Rephrase the title (change or modify the title)
Abstract:
L44: include maturity offset I the abstract.
L47: YoYo intermittent recovery test (which level 1 or 2).
Include the statistical tool used in this investigation
Include the intervention of d (small; medium or large)
Keywords:
Delete physical fitness
Change human physical condition
Introduction:
There are many interesting ideas in the introduction but they are not well organized and well classified.
What is the originality of this work ???? highlight it in the introduction
L84-85: i believe that this sentence should be placed at the beginning of the introduction
L90 : remove the parenthesis
L99: Clemente et al ?????
Check the references
Materials and Methods:
Study design:
The second assessment (after 4 weeks detraining), may be include injury in agility or sprint test????
In detraining period; what they have achieved the players during this period, how did you control that ?????? if not mentioned as a limit of this work
Participants:
Both teams training the same training (the load,….)????
More information about both teams (i.e., training weekly; friend game….)
Assessment procedures:
Include the CCI and CV% for each test (sprint, change of direction, jump, balance)
Zig-Zag test: did they control the technique of driving the ball ????? if yes, added description (inside, outside of the foot)
Anthropometry and body composition: include the maturity status of the players (The maturity status of each participant was calculated as a maturity offset (Mirwald RL, Baxter-Jones AD, Bailey DA, Beunen GP. An assessment of maturity from anthropometric measurements. Med Sci Sports Exerc 2002 Apr;34(4):689-94. PubMed PMID: 11932580. Epub 2002/04/05. eng.
):
Maturity Offset = −9.236 + 0.000278 leg length × sitting body height −0.001663 age × leg length + 0.007216 age × sitting body height + 0.02292 body weight × body height (years).
)
Training intervention:
this section needs a lot of explanation and clarification
SSG
•	number of players (1vs1; 2vs2; 3vs3; 4vs4)
•	number of the touch the ball (how many times to hit the ball: 1, 2, 3, gold games free)
•	number of set of SSG is very small……
HIIT
•	number of set
•	with or without change of direction
Results:
Summarize the result section.
The figures are well presented.
Put the first and the third table in a single table.
Add the dimentions of the piutch in the table 2.
Correct p values (p<0.05 instead of p<0.005) in the tables.
L311 and 314: more explication ??? why you used d and n2p
Discussion:
The discussion requires a lot of explanation
L441: how did you control the heart rate (i.e., 85% of maximum heart rate)
L451-452: more explication (i.e., neuromuscular strain)
Be broad in the discussion i.e., when citing a reference add some detail of the reference such as the population, the period of the intervention…..
L487-495: conclusions or limitations
Conclusions:
Reformulate conclusions

I recommend this manuscript be revised and resubmitted due to inaccurate formatting, grammar, and depth and relevancy of literature review.  In the current form, this paper does not convey an adequate message to the readership of the Peer J. Please check journal format for the Peer J and take a look at the writing tips so that each section of the paper is intact. The paper is not properly formatted for the journal. As written, this study does not meet the standards of a scholarly journal such as Peer J. The authors are encouraged to gain clarity on a) intention b) purpose c) format d) meaningful contributions to literature. I strongly encourage the authors to further pursue this area and resubmit as a new manuscript with major revisions.


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: CAN HIGH-INTENSITY INTERVAL TRAINING AND SMALL-SIDED GAMES BE EFFECTIVE FOR IMPROVING PHYSICAL FITNESS AFTER DETRAINING? A PARALLEL STUDY DESIGN IN YOUTH MALE SOCCER PLAYERS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have addressed concerns and the manuscript now reaches the desired level


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: CAN HIGH-INTENSITY INTERVAL TRAINING AND SMALL-SIDED GAMES BE EFFECTIVE FOR IMPROVING PHYSICAL FITNESS AFTER DETRAINING? A PARALLEL STUDY DESIGN IN YOUTH MALE SOCCER PLAYERS
Review round: 2
Reviewer: 2

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