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 EARLY PLIOCENE EXTINCTION OF THE MEGA-TOOTHED SHARK OTODUS MEGALODON: A VIEW FROM THE EASTERN NORTH PACIFIC
Review round: 1
Reviewer: 3

Output:
Basic reporting: -This paper claims that the cosmopolitan Megalodon became extinct in the early Pliocene based on the last records of the species in California, without using any of the appropriate analytical methods. Accordingly, the authors base their conclusions on the assumption that the age of the last record of a cosmopolitan species in a particular region provides a reliable estimate for the date of its global extinction. Extensive literature across disciplines proves that this assumption is erroneous (i.e. species can go extinct after they were last recorded) and that quantitative methods that take into consideration the declining probability of detection are necessary to make robust extinction time estimates.
-Based on their flawed logic, the authors strongly criticize an earlier estimate of the timing of Megalodon’s extinction that does make use of global sampling and modern statistical methods. Their criticism, however, is unprofessional and ambiguous as they do not explicitly say where the other authors are wrong in their age assignments (and only present percentages of data they consider need revision). See general comments to authors for details.
-The authors do not *use* of the most recent references on the species, although they cite them (see general comments below).
-This paper is also overly long and includes many unnecessary tables and figures that add nothing to the story. It is also poorly written in parts.

I strongly recommend against publication of this article as it lacks of scientific merit.
Experimental design: -This article is not original as this question has been studied previously based on more robust analyses.
-The authors claim that there is a knowledge gap on the extinction of Megalodon, ignoring the most recent works that have been done (there are at least 5 papers published on this). While it is true that there is still a lot to be said about the extinction of this species, the authors don't recognize previous efforts and substantial findings, and they don't offer anything new or relevant.
-This work is also impossible to replicate as it is based on a qualitative and subjective observations.
Validity of the findings: - The conclusions are not linked to original research.
-Speculation is presented as main interpretation.
Additional comments: -This paper suffers from many issues that makes it unsuitable for publication. These include sloppy writing (species names not in italic, parentheses missing, incomplete sentences, etc.), exaggerated statements (e.g. that the extinction of Megalodon is understudied), outdated references (e.g. for the maximum body size of Megalodon), wrong assumptions (see basic reporting), lack of replicability (see experimental design) of their results, among others.
-This paper stands on the basis of a criticism to an early work, which is fair enough, but their criticisms are unfounded. It is also very clear that the authors fail to understand the analytical methods used in Pimiento and Clements (see comment on that paper below).
-The authors further over-interpret their qualitative work, using it to draw all kinds of conclusions about the causes of the global extinction of Megalodon, but only on the basis of their fieldwork in California and disregarding previous research. Why not simply write a paper about the last records of Megalodon in California?

On Pimento and Clements:
When thinking about the extinction of Megalodon, the first obvious step is to look at the work of Pimiento and Clements. Because this is an open access work that include the R code and data, I was willing to re run their analyses using the “correct” dates from Boessenecker et al. to see if a whole new paper with a revised date of extinction was that necessary. However, this was not possible because all criticisms to the dating of Pimiento and Clements (2014) are superficial and ambiguous. For instance, the authors main criticisms are that 1) “rather than use numerical dates from the literature, much of their dataset (88% of data consists of dates artificially stretched to fit stage ‘bins’”, that 2) “Many Piacenzian stage occurrences in New Zealand, Australia, and Europe are based on outdated stratigraphic determinations, and 3) “in many other cases (n=15, 34% of the dataset), poorly dated specimens dated to “Pliocene” are given an age of 5.3-2.6 Ma despite lacking concrete minimum dates, perhaps artificially inflating the number of true Piacenzian-age occurrences”.
First, which dates exactly are wrong (providing percentages does not help)? How do they know the dates have been stretched (I re-read the paper in question, including the supplement that provides an extensive explanation on the ages, and I see no evidence of this)? And even if they have, how is this a problem? Note that Pimento and Clements  bootstrap the timing of each record from a uniform distribution between its upper and lower age. So, if the authors are aware of a new date for one of these records, if it fits between the range assigned, it still is analytically correct, except that it will reduce the uncertainty.

If these authors feel so strongly to revise the time of extinction of Megalodon, and to show that it was actually early Pliocene and not the late Pliocene, I suggest that they:
1. Make a better effort to understand the analytical approach presented in the earlier study.
2. Read the literature on estimating extinction times in the fossil record so that they understand the difference between that and LADs.
3. Re run the analyses presented in the earlier study with the new, revised dates.
They could perhaps submit a response to the PlosOne paper showing their new results.

Alternatively, since the paper does have some valuable components (e.g. the examination of the Californian sequences, valuable stratigraphic evaluation, description of specimens, etc.), the authors could write a manuscript on the last records of Megalodon in California and make inferences on local extinctions. Perhaps their analysis could elucidate a local extinction in California in the early Pliocene, before the final, global extinction in the late Pliocene... Such a study would be of great interest and could be used to refine the time of the global extinction of Megalodon, if necessary.


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 EARLY PLIOCENE EXTINCTION OF THE MEGA-TOOTHED SHARK OTODUS MEGALODON: A VIEW FROM THE EASTERN NORTH PACIFIC
Review round: 2
Reviewer: 1

Output:
Basic reporting: Writing: There are still some errors.
Literature references and background: The authors still ignore the work on one particular author in the introduction.
Professional article structure, figs, tables: There is no figure of the analytical output.
Experimental design: Original primary research: The core of this work is not original, as the questions addressed here have been studied somewhere else before.
Research fills an identified knowledge gap: It does provide an adjustment of the extinction time, however, it was not possible to assess the analyses as the methodology of statistical analyses is not explained, if anything, their data treatment seems to be inappropriate.
Methods described with sufficient detail & information to replicate: No.
Validity of the findings: Conclusion are well stated, linked to original research question & limited to supporting results: No for the section on the causes of extinction.
Additional comments: Despite the new analyses, in which the authors adjust the time of extinction of Megalodon to between 4.14 and 3.5, as opposed to 3.5 and 2.6Ma, the authors continue 1) disregarding previous works on this species, particularly those of one author [I cannot think of a reason to do such a thing, unless they have personal, non-scientific reasons, which I think is rather unprofessional]; 2) implicitly assuming that last occurrences and time of extinction can be used interchangeably; and 3) misinterpreting the work of Pimiento and Clements (e.g. by stating that their ages were stretched to fit a time bin, which is not the case, and concluding that such practice artificially expands age ranges, which is not the case either).

There is no way to evaluate the analytical methods of this paper because they are not explained. However, given the information provided, the new analyses were not performed correctly, as the authors seem to continue not understanding the methods used in Pimiento and Clements. For instance, they excluded a number of records without subepochal age control, which is unnecessary given that all ranges are/should be re-sampled 10,000 times, bootstrapping the timing of each record from a uniform distribution between its upper and lower age to account for uncertainty. It is not possible to see if they preformed any re-sampling, as no details on the methods is provided. Most striking, they don’t include a plot, or figure on the results of their analyses.

Abstract
I suggest to be rewritten.
-It is good scientific practice to acknowledge previous works on the subject, especially if the explicitly address the question you aim to answer. “The timing of the extinction of O. megalodon is thought to be Pliocene” should be replaced with “The timing of the extinction of O. megalodon *has been proposed to be between 3.5 and 2.6Ma*”.
-Remove “although reports of Pleistocene teeth fuel speculation that O. megalodon may still be extant” because it is misleading: 1) Previous works have already assessed the timing of extinction of this species, and there is no longer any speculation. 2) The Pleistocene records are still fossil and by themselves, don’t fuel any speculation on the existence of the species today. 3) The only speculation that exists is on the media.
-Why well-sampled and well-dated upper Pliocene strata is needed? Don’t assume the reader knows that the last records are important to assess time of extinction, explain.
-L36, no space after ;
-L36-37: “youngest reliable records of O. megalodon are early Pliocene, suggesting an extinction at the early-late Pliocene boundary (~3.6 Ma)”. The authors insist in regarding last occurrence as extinction time, which is not the case.
-In general, the abstract does not have a logic flow: When rewriting, consider: Start with a few sentences on a basic introduction to the field, then provide a more detailed background, then state the problem being addressed here, then provide an overview of your results, then explain what the main results means compared with previous studies and how it adds to previous knowledge, finally provide some sentences on a the general context of your results and the broader perspective.

Introduction
Again, it is good scientific practice to acknowledge previous works on the subject, especially if the explicitly address the question you aim to answer.
-L50-52: Pimiento et al. 2016 specifically addresses the geographic distribution of megalodon.
-L58-59: Pimiento and Balk 2015 estimated a max TL of 18m. If you have reasons to not believe this work, you should still acknowledge it (e.g. Otodus megalodon is estimated to have attained a body length of 16 m (Gottfried et al., 1996, *BUT SEE PIMIENTO AND BALK 2015)
-L62-68: When providing what has been done to understand this species, the authors insist on ignoring the work of Pimiento (body size: Pimiento and Balk 2015; geographic distribution: Pimiento et al. 2016). I am now convinced that the authors have reasons beyond science to exclude the work of this particular author.

Material and Methods
-L114: predating the… (?)
-L126: “instead relying (?) upon the ages cited within the paleontological papers reporting the fossil occurrences”. Misleading. See Text S1 from Pimiento and Clements and it is clear that they revised the ages: Here some examples:
--PaleoBioDB# 18548: Early Pliocene age clearly explained in text. However, when studying specimens in the collections of the Florida Museum of Natural History (FLMNH), a Hemphillian age was regarded for this locality…
--PaleoBioDB# 18577: Early Pliocene age clearly explained in text and confirmed when
studying specimens in the collections of the FLMNH.
-L127: “Readers are instructed to consult the Paleobiology Database (Pimiento and Clements, 2014:2). Misleading. Justifications are explicitly provided in text S1.
-L138-139: Poor writing
-L149: Explain what OLE does. The authors don’t seem to be concerned with this paper being so long, therefore, they should also spend some lines explaining the analytical methods. They don’t provide any detail on how analyses were performed, and therefore, this reviewer cannot evaluate it (e.g. It is not possible to assess if the authors indeed resampled their data (or how many times) or if they performed any simulations.
-There is absolutely no reason to exclude records with lack of subepoch control when using OLE and the methods of Pimiento and Clements (i.e. because uncertainty is taken into account by re-sampling the fossil data 10,000 times, bootstrapping the timing of each record from a uniform distribution between its upper and lower age).

Results
The word “corrections” should be replaced with “adjustments” throughout.
-L 559-560: The data from Pimiento and Clements is not artificially stretched to fit stage ‘bins”. It is clear from reading their paper and looking a Tables S1-2 that they used min and max ages as reported in the literature. Therefore, the bins used correspond to those ages and were not stretched to fit any stage bin. Hence, it is incorrect to state that this artificially expanded the age range. It is true that when finer resolution is available it should be used instead, but the way the authors are phrasing this here is misleading. Rephrase or remove.
-L575-580: There is no reason to exclude records from Bone Valley Formation as Morgan, 1994 confirms its age.
-587-589: Again, there is no reason to exclude records with broad ranges as the methods used in Pimiento and Clements account for time uncertainties.
-The authors say in the abstract that they used the median and in the results that they used the modal value to estimate extinction time. Which one is it? Cannot evaluate this as no details on the analyses are provided.
-L604-607: Why should we believe that those are the results you got? Where is the plot? Fig 10 does not represent a plot of the output from OLE, as it only consist on a line drawn artificially. How can the modal value 3.51 and the youngest possible extinction 3.17 Ma. These two values should be the same. There is something wrong with these analyses but impossible to judge.
-The OLE needs some simulations as presented in Pimiento and Clements. Did you perform any? How many of the simulations fall in the range proposed?
-L720-723: The authors insist to use last records and time of extinction interchangeably. That is simply wrong and there is plenty of literature explaining why.

Discussion
The tone of the critiques to Pimiento and Clements continue to be unfair. As in Wang and Marshall (2015), the lack of resolution of fossil data is problem of paleontology, and authors using such data should not be blamed for it. After all, they do what they can with the data available, and use the best methods available to cope with the problems of the fossil record. Consequently, poor resolution in the results of Pimiento and Clements should not be interpreted as bad practice, as the authors here implicitly state.
-L825-829: It is not true that this study has a better resolution than that of Pimiento and Clements, their estimated time of extinction is 3.5-2.6, whereas this one is 4.1-3.5. So the vetting used by the authors did not improve the resolution at all.
-The section on possible causes for the extinction of Otodus megalodon seems to be a repetition of what has been published elsewhere and seems out of place here. At least, there is no correspondence between this and the data they used.
-L842-844: The most recent papers on the timing of the closure of the central American seaway are not considered here (Farris et al. 2011; Montes et al. 2012a; Montes et al. 2012b; Bacon et al. 2015a, b; Montes et al. 2015; Jaramillo et al. 2017).


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

Input:
Title: APPLICATION OF RELAY PUNCTURE TECHNIQUE IN TREATING PATIENTS WITH COMPLICATED LOWER EXTREMITY ARTERIAL DISEASES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Mostly clear and professional language but multiple small errors, including grammar and spelling mistakes (for example, branchial frequently used instead of brachial), occasional errors in syntax. Some sentences too long and unclear. The paper should be reviewed/edited by a native English speaker prior to submission to an English language journal.
More context and background should be provided. In particular the authors should defend further why they think the relay technique is superior to a hybrid approach.

I think reference 9 is not relevant as a justification for the relay approach as this paper deals with aortoiliac occlusive disease, ie comparing aortofem bypass to an endovascular approach. Reference 8 is similar. I think the authors should quote evidence that a hybrid approach to the CFA (ie CFA endarterectomy and endovasculat treatment distally) is inferior. There are multiple papers in the literature showing that a hybrid approach is safe and feasible

The article was professionally structured. Figures 1 and 2 are poorly reproduced and difficult to interpret. Raw data provided although clinical data would be more relevant in addition to ABI
Experimental design: Design - a retrospective cohort study with small numbers. Inclusion  criteria not well-defined. The degree of disease should be classified, ie TASC criteria. RCC provided. How many patients in total in the institution were treated during the study period? What was their outcome? What was the experience of the treating radiologists (i.e. how many previous cases etc)
Aim of paper – research question
 The authors define what they see as the research question, but I am not convinced it is relevant. The authors criticise hybrid procedures as they require vascular surgeons. The paper is written by interventional radiologists. In many countries, open and endovascular procedures are carried out by vascular surgeons not radiologists , and usually either in an operating theatre or a hybrid room, therefore performing hybrid procedures is straightforward and avoids puncturing the brachial artery unnecessarily. Even in the UK for example, where interventional radiologists perform endovascular procedures, it is common to perform hybrid cases where the vascular team treat the common femoral artery via an open approach and the interventional radiologists treat disease distally with access established. The relay puncture technique therefore would have less relevance.

Methods
The authors mention that in seven cases, patients had stents implanted in the femoral artery which could then not be punctured. Do they mean the common femoral (CFA)? It is unusual to stent the CFA.
Methods otherwise well described
Can they elaborate on the SAFARI technique.
It is unclear which lower extremity artery was punctured for access. In the discussion section they refer to accessing via the SFA. This is not common practice. Can this be clarified.
Validity of the findings: The relay puncture technique was found to be feasible and safe. the authors describe good technical success rates and low complication rates. The authors report that 87 stents were used in 21 patients. This seems high given that they were not treating iliac lesions. Did primary angioplasty fail in all the patients? And can the authors specify which arterial segments were stented? Do they have a policy of primary stenting?
The 1 year follow-up is quite short. Did the 6 patients with occluded stents at 1 year require treatment? And which stents occluded?
The authors used a closure device for the brachial artery. Most surgeons would not favour this approach.
Additional comments: Overall a small retrospective study with good results for this technique albeit only short term results quoted. My main issues are with applicability of this technique as I don't feel it is superior to current (i.e. hybrid) approaches although may have a role in some cases


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

Input:
Title: APPLICATION OF RELAY PUNCTURE TECHNIQUE IN TREATING PATIENTS WITH COMPLICATED LOWER EXTREMITY ARTERIAL DISEASES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article must conform to professional standards of courtesy and expression.

Literature references, sufficient field background/context  provided.

Professional article structure, figs, tables. Raw data  shared.

Self-contained with relevant results to hypotheses.
Experimental design: Original primary research within Aims and Scope of the journal.
Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.
Rigorous investigation performed to a high technical & ethical standard.
Methods described with sufficient detail & information to replicate.
Validity of the findings: Impact and novelty not assessed. Negative/inconclusive results accepted. Meaningful replication encouraged where rationale & benefit to literature is clearly stated.
Data is robust, statistically sound, & controlled.
Conclusion are well stated, linked to original research question & limited to supporting results.
Speculation is welcome.
Additional comments: The problems statement, design, and methodology are clear and well written. The results section is well-written, focused. The writing needs to be improved.


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

Input:
Title: APPLICATION OF RELAY PUNCTURE TECHNIQUE IN TREATING PATIENTS WITH COMPLICATED LOWER EXTREMITY ARTERIAL DISEASES
Review round: 1
Reviewer: 3

Output:
Basic reporting: Just note two errors line 115 and line224: Branchial instead of brachial.

Introduction, figures:OK
Experimental design: Aims of the study are: to report efficacy and safety of a new endovascular technique and the METHODOLOGY seems crucial.

In which population??
This is a RETROSPECTIVE study, and we need to know how the patients have been selected: are they consecutive patients? How many procedures have been made by the unit during the study period?

The functionnal status is described but it should be precised whereas the patients present a critical chronic ischemia or acute ischemia.

The arterial lesions should be more precisely described: topography, length, degree of obstruction, thrombosis... The TASK classification may be useful.

More precisions about the inclusion in the procedure: was surgery ever contreindicated or impossible? More generally, does the procedure always fits with the scientific recommendations?

Choosing a technique because of a "cost problem" may be questionable in terms of ethical concerns. (line 129 130)
Validity of the findings: The retrospective nature of the study and the small size of the population do not allow any extrapollation of the results in order to change our practice.
I appreciate your paragraph about the limitations of the study. Don't you think it should be take place BEFORE the conclusions?
Additional comments: The technique described by the authors seems interesting but its value (safety) and its indications have to be re evaluated after multicentric and comparative studies.
By now  the present study only allows to say it is a feasible technique in the hands  of an experienced and skilled equipe.


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

Input:
Title: APPLICATION OF RELAY PUNCTURE TECHNIQUE IN TREATING PATIENTS WITH COMPLICATED LOWER EXTREMITY ARTERIAL DISEASES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The problems statement, design, and methodology are clear and well written. The results section is well-written, focused. Authors' responses are clear and point-to-point.


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

Input:
Title: APPLICATION OF RELAY PUNCTURE TECHNIQUE IN TREATING PATIENTS WITH COMPLICATED LOWER EXTREMITY ARTERIAL DISEASES
Review round: 2
Reviewer: 2

Output:
Basic reporting: I appreciate the corrections done by the authors.
English language remains somehow crtical and I made some suggestions in the pdf attached file
Experimental design: Poulation description and  methodology have been corrected
Validity of the findings: It is still impossible to extrapollate this data to our daily practice.
The small size of the population ant the retrospective character still reperesent the weakness of this study.
I made some suggestions to the authors , specially in the conclusions
Additional comments: in my point of view, the interest of the paper is to describe this"new"technique, to defend its feasability on the basis of this preliminary data in order to arouse future larger, multicentric and comparative trials.


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 ORIGIN OF OXALIS CORNICULATA L.
Review round: 1
Reviewer: 1

Output:
Basic reporting: The ms. by Groom et al. describes the occurrences of Oxalis corniculata in space and time.
I regard this as a very interesting contribution to the question on the origin of this taxon. I agree with the authors that the question cannot be finally solved (as theoretically all scientific questions). Overall, a refreshing study, relevant for the evaluation of neophytes, in this case worldwide.
The evidence they provide is relatively strong. I like the combination of different approaches and the old illustrations. Still, the phylogenetic data by Vaio where the American natives O. dillenii and stricta also group together with O. corniculata may be a counter-argument. By just looking at the tree an American origin seems more likely.  Of course, long-distance dispersal may also have happened before the mrca of Oxalis corniculata from America to Asia. Perhaps the authors should discuss the available phylogenetic data in more detail. Besides the infraspecific problems of O.c., the inclusion of all putative related species in the phylogenetic study by Vaio should be discussed.

A different approach is worth to be mentioned:  that a population genetic approach may reveal the origin based on the assumption that the highest genetic diversity represents source areas. Especially for neophytes this makes sense. It has been nicely shown for Oxalis pes-caprae in the Mediterranean with a South African origin.

Discuss the use of seed morphology to distinguish O. species (corniculata from relatives). Best illustrate it with photos. This would strengthen the arguments.

I found some more (secondary) literature:
-	Bauhin cites O.c. for Heidelberg around 1600 (publication?).
-	Graebner 1914 for Basel (Synop. Mitteleur. Flora)
-	Gmelin 1806 (flora badensis) also mentioned it.

After these minor revisions the ms. should be fine for PeerJ.

Minor points:
l. 94, 123, 266, 304 O. => write out Oxalis (at begin of sent.)
l. 95 use => used
l. 137 Earlier? Than…
l. 139 one taxon
l. 145 flowers of this illustration are in bud and are of little diagnostic use. Another herbal BOOK? from circa
Experimental design: see Basic reporting
Validity of the findings: see Basic reporting
Additional comments: see Basic reporting


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 ORIGIN OF OXALIS CORNICULATA L.
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment, except for a suggestion:

There are, in addition to taro, several plant species that currently have wide (cosmopolitan or pantropical) distributions and it may be interesting to refer to some of them (and especially to literature on their historical biogeography, if anything is available). These include Lagenaria siceraria (African origin, but earliest archaeological evidence in Mexico?; China?); Zea mais, Capsicum frutescens,  Capsicum chinensis, Sesamum indicum, Pteridium aquilinum, Momordica charantia, Dysphania ambrosioides and Carthamus tinctorius. It appears to be a general problem to determine the origins of plants that are associated with agriculture and human migrations? How did others try to solve the problem?
Experimental design: No comment, except for a suggestion:

Genetic diversity is barely mentioned, yet this may provide an important additional avenue of research. If, for example, all European, African and American collections of Oxalis corniculatus are genetically similar or identical, while the Asian ones are genetically diverse, then the main findings of this paper can be further verified and supported.
Validity of the findings: No comment, except for the suggestion of the potential value of genetic studies as a future direction.
Additional comments: The paper is well conceived and interesting to read, with no obvious errors of fact and/or logic.

There are some very minor errors that can be corrected, perhaps during copy-editing (lines 93, 137, 167, 281, 302, 379 and 441).


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

Input:
Title: A LARGE LATE MIOCENE CETOTHERIID (CETACEA, MYSTICETI) FROM THE NETHERLANDS CLARIFIES THE STATUS OF TRANATOCETIDAE
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript by Marx et al. " A large Late Miocene cetotheriid (Cetacea, Mysticeti) from the Netherlands clarifies the status of Tranatocetidae" represents interesting additions to our knowledge of cetacean evolution. The authors described a new species of Cetotheriidae and they tested the idea that the Cetotheriidae and enigmatic taxon Tranatocetidae indeed form separate clades. Based on the several morphological characters indicated by authors and the result of phylogenetic analysis, the authors clearly indicated that Tranatocetidae and Cetotheiirade should be integrated into single family, Cetotheriidae.

The description of the specimen is detailed and the discussion is clear and solid. I think this paper will be widely received by people interested in the evolution of marine mammals. I have a few comments and suggestions that I think it will improve the paper, all of them are minor.

Lines 37-44: three distinct morphotypes of Cetotheriidae
The authors mentioned that at least three distinct morphotype is found in the Cetotheriidae. However, the authors just mentioned on body size and fossil occurrence of each morphotypes and not mentioned on the morphological characteristics for each morphotypes. Please indicate the morphological characteristics for each morphotypes.

Line 124
The words "the level of the parietal" should be replaced by "the level of the anterior edge of the parietal"?

Lines 130-131: sharp cranial rim
The authors described as "a sharp cranial rim surrounding the proximal opening of the facial canal", but it is not shown in the figure. Please indicate it in the figures.

Lines 173-180: diagnosis
This part of diagnosis is on the diagnostic characters for genus Tranatocetus, so, perhaps, this part should be included in the section of the emended diagnosis of the genus.

Line 180: diagnosis
The authors mentioned here that "Differs from T. argillarius in being slightly larger". I think this might include ambiguity; it is difficult to clearly suggest that the slight body size difference is not due to the individual variation, but due to the phylogenetic distinction.

Line 271
The words "paroccipital cavity" should be replaced by "paroccipital concavity"?

Lines 493-499: niche partitioning, extinction of the Cetotheriidae
The eschrichtiids are also known as benthic filter feeder, and several early members of the Eschrichtiidae have been recovered from the Pliocene deposits. Perhaps, they might have occupied similar ecological niche (small, benthic suction feeder). Do they have also an impact on the extinction of the Cetotheriidae?

Toshiyuki Kimura
Gunma Museum of Natural History


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

Input:
Title: A LARGE LATE MIOCENE CETOTHERIID (CETACEA, MYSTICETI) FROM THE NETHERLANDS CLARIFIES THE STATUS OF TRANATOCETIDAE
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comments
Validity of the findings: No comment
Additional comments: I find the text well written and well structured, concise and to the point. Particularly, I find the description well written and the figures are supporting the text very nicely. The authors argue well for their opinions on both the taxonomic position of the new material and the overall phylogeny of the groups investigated and have used standardly accepted methods to reach their conclusions. I find only minor suggestions, mainly to improve readability.


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

Input:
Title: A LARGE LATE MIOCENE CETOTHERIID (CETACEA, MYSTICETI) FROM THE NETHERLANDS CLARIFIES THE STATUS OF TRANATOCETIDAE
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors carefully and appropriately revised their manuscript and implemented all of my previous suggestions. On the author's response to my previous comment on the diagnosis, I agreed the author's response. I do not find any problem, and now I think this manuscript is ready for publication.

Toshiyuki Kimura
Gunma Museum of Natural History


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: EVERYTHING YOU ALWAYS WANTED TO KNOW ABOUT GENE FLOW IN TROPICAL LANDSCAPES (BUT WERE AFRAID TO ASK)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The study provides a comprehensive review of how landscape effect, particularly, anthropogenic disturbances influence the gene flow in tropical ecosystems. By analysing the studies across 157 tropical species, the authors conclude that the use of individual-based approaches rather than a community-level investigation is appropriate to understand the Landscape Effect on gene flow. Furthermore, the authors review the methodologies often used in the concerned field of investigation and suggest that next-generation sequencing and maximum likelihood-based analysis is appropriate to generate comparable results. In their conclusion, the authors highlight the lack of information for tropical species and emphasize that the future investigations must include tropical species across geography.

The topic of the investigation is definitely interesting, however, I feel that the paper does not introduce the subject adequately. As a result, the core question of the present investigation appears blurred. I have mentioned a few points below, which, I hope, would help to improve the manuscript and bring out the motivation for the investigation.

1. Authors need to expand on the Landscape Effect, with and without anthropogenic disturbances
2. Broadly discuss how the landscape effect influence gene flow using
3. Describe and link functional connectivity to the above discussions
4. Expand Isolation by Resistance, its mechanism and outcome in connection to the Landscape Effect
5. Optionally, you can introduce a flow chart describing the link across all that will be helpful in visualizing the importance of the present investigation
Experimental design: The methods for literature survey and analysis seem fine to me. However, I have concerns regarding the study design and in formulating the objectives of interest. Please see the comments in "Basic reporting"
Validity of the findings: The goals set out in the Introduction needs improvement for clarity
Similarly, bring a comprehensive outlook to the subject in the Discussion section

Moreover, the work, in its present form, seems incomplete and requires clarity on a conceptual platform to connect the ideas appropriately
Additional comments: The title seems quite uninformative. I suggest the authors modify the title to justify the objective of the study
Line 51	You may remove the 'comma' before ‘found’
Line 55-68	The phrases ‘habitat degradation’ and ‘environmental change’ have been used in association with the biodiversity response. Do these phrases convey the same message or they underlie different concepts. Add or modify the sentence for clarity
Line 76-77	In the earlier paragraph, authors have described community- and population-level studies for biodiversity response to habitat degradation. However, here, they have mentioned population and individual level studies. Please clarify or modify!


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: EVERYTHING YOU ALWAYS WANTED TO KNOW ABOUT GENE FLOW IN TROPICAL LANDSCAPES (BUT WERE AFRAID TO ASK)
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall, this manuscript deals with a very interesting and timely topic that is cross-disciplinary and definitely within the scope of the journal. This specific topic has also not been reviewed before. The intro nicely outlines the motivation for the study. Indeed, the entire manuscript is very well written.
Experimental design: I found no major flaws in the analysis, but I think more insights could be gained from the analyses.
Validity of the findings: I cannot recommend to accept the manuscript in its current form, because it is not correct to talk about “gene flow” when the reviewed studies actually analyze genetic structure, and also because I found the discussion to be too general and not specifically related to the findings of the shown analysis. I provide details comment on these issues under point 4 of this review form and encourage the authors to resubmit the manuscript after revisions.
Additional comments: Major comments
1) I do not like how the manuscript talks about “gene flow”, but actually compares measures of genetic differentiation and structure. Especially the population-based metrics (e.g., Jost D, FST etc.) are not only affected by gene flow, but also by drift, which is essentially a function of (effective) population size. This is well-known from basic population genetic theory, but the importance of this for landscape genetic inferences was recently highlighted by Prunier et al . - Methods in Ecology and Evolution “Contribution of spatial heterogeneity in effective population sizes to the variance in pairwise measures of genetic differentiation”). Similarly, individual-based metrics are not only influenced by gene flow, but also by mating schemes, space-use behavior etc., especially across small spatial scales. You need to explain this in the discussion of your manuscript, so that readers are not misinterpreting your results. I recommend to not talk about “gene flow” at all, but instead about “genetic connectivity” in a broader sense or even about “genetic structure”, and then explain what exactly this entails (i.e., gene flow and other processes).

2) One general concern I have is the combination of data/results from plant and animal species. I realize that it is currently difficult to draw cross-species conclusions, but I don’t think it is correct to combine results from flying and non-flying insects, from plants and animals, and from terrestrial and aquatic systems without at least discussing the importance of different species traits for landscape genetic patterns. Also, you mention biases in previous studies, but you also have a strong bias towards insects in your quantitative analysis (Fig. 7 and 8, Data S2 and S3). You need to discuss this in much more detail. There is an interesting paper by Lino et al. that was recently published in Mammalian Biology (“A meta-analysis of the effects of habitat loss and fragmentation on genetic diversity in mammals“). It focused in genetic diversity and mammals, but you still might want to check out that paper and potentially include it as one of your references.

3) In addition to considering species traits, some more details on the studies are warranted. For example, in lines 163-164: I think in order to make sense of such counter-intuitive results, readers would appreciate more details on a) the study design employed by each study, especially concerning the spatial scale of analysis and the sampling scheme, and b) the study species (see above). For example, a small scale analysis on flying insects is much less likely to find an effect of habitat loss than a large scale analysis of ground beetles. You explain this for one of the studies (on army ants) later in the discussion, but it is important for all the studies you use.

4) Finally, I found the discussion to be too general. It is basically a list of things that should happen in landscape genetics (use of improved methods, use of NGS approaches, considering time-lags…). The need to consider all of this has been highlighted before and it is not clear exactly how tropical systems are different or how your study added to this existing knowledge. Thinking some more about the implications of species traits and study design might help you to get more info out of the analyses, but I already think there is a bit more in your data than you report. Isn’t it interesting that the bias towards mammals is strongest outside of the Americas? Isn’t it surprising that most amphibian studies use haplotype data? You could also discuss that several studies have used habitat suitability to estimate resistances, but suitability is often a poor descriptor of functional connectivity, so you could advise against this for future tropical studies. I found it surprising that no study has looked at road effects in reptiles, and no amphibian study assessed dispersal via waterways. I also think that a more detailed overview of study aims and outcomes would be nice. You mention these briefly in lines 224-229, but it the info is missing from the results. Overall, I suggest to derive more novel insights from the results you have already obtained and can additionally obtain by considering species traits and study design more explicitly.

Minor comments:
57: In the sentence “While these metrics underpin ecosystem function, they are not always be…” – either delete the “be”, or change to “While these metrics underpin ecosystem function, they MAY not always be…”

119: Not sure why you are citing the Balkenhol et al. 2016 book here. Perhaps a better citation would be chapter 3 in that book (i.e., Waits & Storfer “Basics of population genetics: quantifying neutral and adaptive genetic variation for landscape genetic studies”. (Also, the Balkenhol et al book is listed twice in the reference list (a + b)…)

123: Fig. 1 does not show which studies are individual- vs. population-based.

139: Please explain why you did not perform a more formal meta-analysis.

145: But looking at Figure 2 and Data S1, it really seems like the bias is mostly towards amphibians, while mammals, reptiles and insects are all rather similarly represented. I’d change your conclusions accordingly. Also see my comment on Fig. 1 concerning a mammal bias outside of the Americas.

146-148: I don’t understand what this sentence means. Please reword to make it clearer.

165-166: It would be important to know whether studies tested for IBD separately, or after accounting for the effects of landscape variables. After all, straight-line and effective distances are often highly correlated and when testing for IBD separately, it will be supported, even if the actual driver is the landscape matrix.

170: Is “isolation by deforestation” the same as detecting a positive effect of forest cover on gene flow? If so, I would reword to “isolation by non-forest”, because non-forest is a pattern (just like elevation), while deforestation is a process.

176: Most studies have focused on amphibians.

182: Citing Dyer 2015 here is not appropriate, because he actually argued that thus far, landscape genetics has not evolved into a truly distinct and independent discipline. Aside, I suggest to change “interdisciplinary discipline“ to „interdisciplinary field“.

188: Amphibians yes, but mammals not too much more than insects or reptiles.

194-208: While I agree that the future will see many more landscape genetic studies using NGS approaches and resulting SNPs, this whole paragraph deals with a very general aspect of landscape genetic s and is only marginally related to your own study. I suggest to shorten this and move it to the very end of the discussion.

211: Might be good to mention some of these improved methods, especially some that were not yet mentioned by Richardson et al. (e.g., commonality analysis – Prunier et al.,Molecular Ecology (2015) 24, 263–283; extended causal modeling - Fourtune et al. Am Nat. (2018) 191:491-508). This is again something quite general.

213-214: This is incorrect. More than one variable can be partialled out in a partial Mantel test before assessing the effect of the focal variable. See e.g., the implementation of partial Mantel tests in r package ‘ecodist’.

215: But see Franckowiak et al. 2017 (PLoS ONE 12(4): e0175194) for a severe critique of model selection with pairwise data.

216: Add “models” after “population effects (MLPE)”

223: Again, I don’t think the Balkenhol et al. 2016 book is a good reference for this statement.

240: No, species with limited dispersal are not generally expected to be most impacted by habitat loss and fragmentation – it all depends on the scale of habitat loss, and species with high dispersal capability will also be highly susceptible to habitat loss. See also Evers and Didham (Biol. Rev. (2006), 81: 117–142 ; Fig. 2B) and Lino et al. (Mammalian Biology, A meta-analysis of the effects of habitat loss and fragmentation on genetic diversity in mammals).

263: I wonder whether it is desirable / realistic to have one modelling framework for all landscape genetic studies. I feel that landscape genetic studies are too diverse for this in terms of spatial and temporal sampling scheme, scale of sampling, landscape and species characteristics, etc.

Fig. 1: It looks as if the vast majority of studies outside of the Americas has focused on mammals. This is another bias you might want to report.

Fig. 3: Need to mention that 2018 was not yet complete by the time you did this analysis

Fig. 4: A pie chart would be more appropriate for this figure

Fig. 5: Need to explain the abbreviations of the different methods

Fig. 7: It is not clear to me whether the significances shown by the different symbols are for the effects size (as shown in S3), or for the statistical tests in the original studies.

Fig. 8: Not sure that the ordering of this is very informative. Would it be better to order by species names, or even to move this mix of taxa to the supplement and here focus on a few species for which multiple landscape variables were tested? In the text, you use Fig. 7 and 8 to compare effects sizes for individual- vs. population-based studies, but for that is would be better to show individual- and pop-based results on the same panel…

Data S1: The column “N samples” sometimes lists just a number, sometimes it states “individuals” and sometimes “specimen”. Please explain these differences. Also, it would be important to know what level of analysis the studies used (pops or individuals), how many analytical units they had (a study with 600 individuals conducting population-based analyses with 3 populations has an effective N of 3) and whether the study was included in your quantitative analysis (because the citations are not listed in S3…). Some of this info is presented in S3, but it would be easier for readers to have it all in one table. Finally, some of the studies have been conducted in aquatic environments, but this is never mentioned in the main text.

Data S2: Please explain how “deforestation” is different from “% forest cover”

Data S3: Need to explain the abbreviations for the genetic distance metrics. What does “Pop_ind” mean? The s.e. are not shown in the Figures, but I think they should be used to calculate confidence intervals.


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: EVERYTHING YOU ALWAYS WANTED TO KNOW ABOUT GENE FLOW IN TROPICAL LANDSCAPES (BUT WERE AFRAID TO ASK)
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am happy with the new version of the manuscript. I have gone through the authors’ response to the reviewers and happy with the modifications incorporated. I accept the manuscript in its present form with a few minor suggestions mentioned below, which can be incorporated during proof-reading.

P75-79: I suggest simplifying the sentence.
P89: Please check “specie’s”
P185: I suggest explaining the term “Electrical resistance” in landscape genetics which is used as an analogy to resistance in a network. This will bring clarification to the readers of the manuscript.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No comment


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

Input:
Title: EVERYTHING YOU ALWAYS WANTED TO KNOW ABOUT GENE FLOW IN TROPICAL LANDSCAPES (BUT WERE AFRAID TO ASK)
Review round: 2
Reviewer: 2

Output:
Basic reporting: Basic reporting is fine.
Experimental design: Study design is fine.
Validity of the findings: See my general comments below.
Additional comments: The new version incorporates most of the previous reviewer comments, but not all. The manuscript generally has improved, however, I still think that one main issue has not adequately been addressed, i.e., the inappropriate use of the term “gene flow”. In your reply to that comment, you basically say that many previous studies have used the term incorrectly, so it’s okay to continue to do so. That is obviously a horrible way to argue. I also don’t see why the definition of landscape genetics by Balkenhol et al justifies your terminology. After all, that definition describes a broad field (in which one process of interest is gene flow), while you are conducting a certain analysis (involving various genetic metrics that are influenced by gene flow, among many other factors) from which you then try to make very specific inferences (concerning gene flow). I am fine with leaving the term in the title, but I still think you should be much more careful in your use of the term throughout the manuscript.  You must also incorporate a very clear statement where you explain that even though you are interested in gene flow, the studies you reviewed use various genetic metrics of genetic differentiation and structure; that these are often used as surrogates of actual gene flow estimates; and that genetic differentiation/structure is also influenced by many other processes. This statement should be either in the beginning of the discussion, or in the methods. In your reply, you actually acknowledge that equating genetic structure with gene flow is not straightforward by stating “Genetic structure, on the other hand, may not necessarily comprise gene flow, as it can result from drift acting on isolated populations, adaptive processes or both” – so then why do you think it is okay to act in your manuscript like gene flow and measures of genetic structure are the same?

Additional /new comments:
“Interestingly, several studies found an effect of habitat suitability on gene flow, suggesting that habitat suitability models are useful when proposing ecological corridors or forecasting the impact of future climate on gene flow (Franklin & Miller, 2009)” – But you should state that this is generally not the case, as many species will still move through habitat that is not suitable to live in. See Scharf et al. (2018, doi: 10.1186/s40462-018-0136-2), Keeley et al (2017, doi: 10.1016/j.landurbplan.2017.01.007), Abrahms et al. (2016; doi: doi: 10.1111/1365-2664.12714), Mateo-Sanchez et al (2015; doi: 10.1007/s10980-015-0194-4), …

“…a limit of two predictor variables that can be simultaneously analyzed (in partial Mantel tests)…” – this is still incorrect. A partial Mantel test can be used to partial out as many covariates as you want, but you can only assess the impact of one variable that you do not partial out. Please correct this statement.


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 VALUE OF DIFFUSION TENSOR TRACTOGRAPHY DELINEATING CORTICOSPINAL TRACT IN GLIOMA IN RAT: VALIDATION VIA CORRELATION HISTOLOGY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Authors do not discuss other difusion MRI methods besides DTI.

No discussion of the effects of edema. Was edema evaluated either by imaging or by histologic examination?

There are no behavioral tests to establish that the “disruption of tracts” seen by imaging and histology had expected effect on the motor function of the rats.

Methods do not clearly describe how CST was selsected. (line 220-221) In the DTT, the initial point was the motor cortex area, and the cerebral peduncle 221 was the seed point.  How was motor cortex defined? Where in the cerebral peduncle

How were the sites of histologic evaluation selected? How were the registered to the sites of DTI measurements? How can the authors know that these measurements are representative of the same region as the DTI measurements? It seems as if only 5 HPF were examined per rat. How were these selected?

The histologic examination does not investigate tract integrity per se.  There are no myelin stains or other efforts to examine whether the tracts are in continuity or not.

Given that the motor function of the animals was not assessed, the histologic integrity of the tracts themselves were not addressed, and the colocalization o the imaging and histologic findings were not described, the conclusion is not fully supported by the findings.
In conclusion, our results demonstrate that DTI imaging may quantitatively evaluate the injury degrees in fiber tracts involving brain gliomas in rats. Our data suggest that these quantitative parameters may be used to enhance the efficiency of delineating the 284 relationship between fiber tracts and tumors.

Minor points:

Line 85: degree of damage rather than damage degree

Line 219 “right” hemisphere is not clear from methods whether this is the injected or non-injhected hemisphere.

Line 239 should be affected not effected

Line 260 unclear awkward  phrasing “were classified and predictive of total resection or partial resection”

Error in line 266: charactering

Unclear line 224
“the corresponding fiber tracts were disrupted by the tumors using 3D reconstruction DTT imaging”

Results section is very dense and hence difficult to read and follow.  This section could benefit from introductory and concluding sentences .
Experimental design: Authors do not discuss other difusion MRI methods besides DTI.

No discussion of the effects of edema. Was edema evaluated either by imaging or by histologic examination?

There are no behavioral tests to establish that the “disruption of tracts” seen by imaging and histology had expected effect on the motor function of the rats.
Validity of the findings: Given that the motor function of the animals was not assessed, the histologic integrity of the tracts themselves were not addressed, and the colocalization o the imaging and histologic findings were not described, the conclusion is not fully supported by the findings.
Additional comments: No additional comments


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

Input:
Title: THE VALUE OF DIFFUSION TENSOR TRACTOGRAPHY DELINEATING CORTICOSPINAL TRACT IN GLIOMA IN RAT: VALIDATION VIA CORRELATION HISTOLOGY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Language: I am not a native Speaker. For that reason, I am not able to proof the used language. I did not note any serious error.

Background / context: The used literature helped to follow the results and supported sufficiently the discussion. The publication from Berlin should be discussed because there were found similar finding like in this study in brain Tumor patients (Specific DTI seeding and diffusivity-analysis improve the quality and prognostic value of TMS-based deterministic DTI of the pyramidal tract).

Structure: article is well structured. Some Images of the figures seem to be very small and might be enlarged.
Experimental design: Visualization of all 3 grades of CST injury should be visualized (not only 2 grades). Additionally, there is a bias to compare the CST injury grade (which bases on the relative FA-value) with the rFA-value (which is calculated by the ipsilesional FA-value and the contralateral FA-value) - Fig 2.

Some data are repeated in the text, the tables and the figures. I also recommend to add p-values in the tables.

I am not able to proof the histological analysis.
Validity of the findings: To my Knowledge, this is the first study analyzing the diffusivity parameters in rats where Tumor cells were implemented. The shown results go in line with data published in patients.
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 VALUE OF DIFFUSION TENSOR TRACTOGRAPHY DELINEATING CORTICOSPINAL TRACT IN GLIOMA IN RAT: VALIDATION VIA CORRELATION HISTOLOGY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The report is acceptable for publication.
Experimental design: acceptable
Validity of the findings: acceptable
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: MORPHOLOGICAL IDENTIFICATION AND MOLECULAR CONFIRMATION OF THE DEEP-SEA BLUE AND RED SHRIMP ARISTEUS ANTENNATUS LARVAE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is very well written and fills an important knowledge gap in decapod morphology and ecology.   The identification of larval stages across decapods is still lacking for most species, so these types of papers that use BOTH morphological and molecular methods for larvae-adult matching are very important.   I would suggest the authors add the reference below to line 88-89.

Bracken-Grissom, H.D., Felder, D.L., Volmer, N., Martin, J. and K.A. Crandall. 2012. Phylogenetics Links Monster Larva to Deep-Sea Shrimp. Ecology and Evolution. 2(10): 2367-2373.
Experimental design: The experimental design was well-designed and executed.  I would like to see a few additions before this paper is accepted for publications:
1.	Why did the authors use different genes (COI vs. 16S) for the protozoea I pool and all known larval stages?  Both are sufficient for DNA barcoding, however I find it strange to switch from one marker to the next.  An explanation should be provided.
2.	As I am sure the authors are familiar with the presence of pseudogenes, especially within COI, so I would insist there is mention to how the authors examined the sequences for the presence and removal of pseudogenes in the COI data.
3.	Lastly, and most importantly, the larval-adult matching studies only work if a reliable ADULT specimen is available for larval matching.  GenBank is absolutely rampant with identification errors.  I would like to see the authors provide an expanded and clearly written explanation as to why they are 100% confident in the adult identifications used to match the larvae.  I noticed that a co-author is cited at the end of line 195 (also on the present paper) but an expanded justification about the morphological confirmation of adults is needed.  Also, several of the GenBank numbers provided in the line 194-195 are either 1) unpublished or 2) not released.  This also makes me feel very uneasy, without expansion on this.  For example, JX403858 is NOT from Roldan et al. so the appropriate papers needs to also be cited. I do warn caution from my own experience.
Validity of the findings: Once an expanded justification on the adults identification is expanded the findings are valid.
Additional comments: Overall, I think this is a very valuable paper that can advance the field of fisheries and decapod taxonomy.


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: MORPHOLOGICAL IDENTIFICATION AND MOLECULAR CONFIRMATION OF THE DEEP-SEA BLUE AND RED SHRIMP ARISTEUS ANTENNATUS LARVAE
Review round: 1
Reviewer: 2

Output:
Basic reporting: I was interested in reading the manuscript PeerJ-29455 regarding the morphological and molecular confirmarion of the larval stage PZI of Aristeus antennatus.  The topic is interesting and the approach, based on mprphological and molecular studies offer valuable offer to the literature.

The low number of the individuals studied may consist a controversy among the reviewers and other researchers, but - according me - this number reflects the general population's
situation.

Some references are too old and I suggest to authors to replace them with more recent ones.
Experimental design: The experimental design is efficient and according the more recent experimental design. The unique gap is the depth of each sampling station (please insert a coloumn in Table 1 indicating the depth of each station). Did the plankton samples of A. antennatus taken from 0,5-1 m?
An other point is the missing of Gennadas spp samples in summer. For comparative reasons the coexistence of both species caught in the same period could improve the results of the manuscript.
Validity of the findings: In general, it is an interesting manuscript providing novel, detailed information on the larval stages of both species (A. antennatus and G. elegans) and improves the knowledge to the taxinomists and marine biologists. The Discussion is well documented and well stractured.
Additional comments: The paper, however, is unclear in some lines and needs to be clarified before publication. For example, these lines are reported to the depth stratum of the sampling stations, on the absence of Gennadas spp samplings from the summer surveys, some changes in the Introduction.

Replace some references which are old


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 RUNNING KINEMATICS OF FREE-ROAMING GIRAFFES, MEASURED USING A LOW COST UNMANNED AERIAL VEHICLE (UAV)
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1) Clear, unambiguous, professional English language used throughout.

Excellent. No comments.

2) Intro & background to show context. Literature well referenced & relevant.

The introduction and background are very good. However, I would suggest the authors to consider expanding the part on gait dynamics, from line 74 to 89, including the following information:
According to Hildebrand (1976) and Cartmill (2002), the lateral walking gait of camels, giraffes and other few species differs from the lateral walking gait of others cursorial species by one parameter, which is diagonality (i.e. “% of stride interval that footfall of the forefoot follows hind on the same side” – Hildebrand, 1976). Depending on diagonality, the lateral walking sequence can follow a “Horse rule”, which will develop in a trot at running speed, or a “Camel rule”, which will develop in a pace at running speed (Cartmill et al., 2002). The lateral walk of giraffes follows the “Camel rule” (Cartmill, 2002, reporting observations of Hildebrand).
The diagonality (sensu Hildebrand and Cartmill) is equivalent to the phase difference explained by Alexander and Jayes in the figure 2 of their cited work of 1983 (Although slightly different in the calculation). In Alexander and Jayes (1983) only camels lies on the line “lateral walk to pace”, giraffes were not included in their sample.

I would suggest changing the sentence that begins at line 81. According to Hildebrand and Cartmill (cited above), and confirmed by a robot model simulation (Suzuki et al., 2016), the intermediate speed gait of giraffes should be pace.

-	Cartmill, M., Lemelin, P., & Schmitt, D. (2002). Support polygons and symmetrical gaits in mammals. Zoological Journal of the Linnean Society, 136(3), 401-420.
-	 Suzuki, S., Owaki, D., Fukuhara, A., & Ishiguro, A. (2016, July). Quadruped gait transition from walk to pace to rotary gallop by exploiting head movement. In Conference on Biomimetic and Biohybrid Systems (pp. 532-539). Springer, Cham.

Literature is relevant and well referenced. I have found only a little discrepancy.
Line 73: The cited work is missing in the reference list: Biancardi and Minetti, 2012
Biancardi, C. M., & Minetti, A. E. (2012). Biomechanical determinants of transverse and rotary gallop in cursorial mammals. Journal of Experimental Biology, jeb-073031.

3) The structure of the submitted article should conform to one of the templates.

No comments.

4) Figures should be relevant to the content of the article.

The figures are good and relevant.

5) The submission should be 'self-contained,' should represent an appropriate 'unit of publication,' and should include all results relevant to the hypothesis.

No comments.

6) All appropriate raw data has been made available in accordance with our Data Sharing policy.

No comments. Thank you for providing raw and supplementary data.
Experimental design: 1) Original primary research.

No comments.

2) Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.

Excellent. Besides the methodological relevant question of kinematic data collection in field, the problem of the giraffe locomotion is interesting, with different knowledge gaps to fill.

3) The investigation must have been conducted rigorously and to a high technical standard.

Excellent

4) Methods described with sufficient detail & information to replicate.

Excellent. The methodology is extensively explained. Calibration, data processing and validation of the methodology are very useful.

5) The research must have been conducted in conformity with the prevailing ethical standards in the field.

No comments.
Validity of the findings: 1) The data should be robust, statistically sound, and controlled.

No comments.

2) The data on which the conclusions are based must be provided or made available in an acceptable discipline-specific repository.

No comments.

3) The conclusions should be appropriately stated, should be connected to the original question investigated, and should be limited to those supported by the results.

Excellent

4) Speculation is welcomed, but should be identified as such.

No comments.

5) Decisions are not made based on any subjective determination of impact, degree of advance, novelty, being of interest to only a niche audience, etc. Replication experiments are encouraged (provided the rationale for the replication, and how it adds value to the literature, is clearly described); however, we do not allow the ‘pointless’ repetition of well known, widely accepted results.

No comments.

6) Negative / inconclusive results are acceptable.

No comments
Additional comments: This is an excellent paper, well written, with, at least, two different and useful results:
1) The validation of a kinematic measurement system based on a drone (UAV) that carries a high speed camera. This is a good example about how low-cost new technologies can be employed to produce amazing science.
2) The kinematic gait analysis of giraffes in field. Even if there is a lot of footage of free ranging animals available, in more or less public repositories (BBC, Arkive, You tube), the quality of those videos is often questionable and problematic.
The accurate analyses of the data provided new interesting information about the locomotion of these amazing species. Giraffes are anomalous rotary gallopers, “slow gallopers”, as the authors reported. From a visual inspection, their gallop appears different to that of a cheetah or a gazelle, despite of the same footfall pattern. These sound quantitative results fill a long-standing knowledge gap.


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 RUNNING KINEMATICS OF FREE-ROAMING GIRAFFES, MEASURED USING A LOW COST UNMANNED AERIAL VEHICLE (UAV)
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This was an interesting and useful paper to read on the validity of using UAV's to measure biomechanics in the field. I think many researchers will find this to be of general interest. The information gathered on giraffe locomotion is also of interested to a broard range of biomechanists.

The paper itself was very well written, clearly describing the steps undertaken. I have only a few minor comments, the most important being the influence of focal distance on speed accuracy estimates, and why so few test distances were used.

Abstract
Line 18: Reference to the GPS + inertial sensors seems out of place here, makes more sense in the intro.
Line 30: The gold standard was a bit ambiguous in this sentence, maybe define more clearly how it was measured and report % errors.

Methods

Line 109: The mean, max and min focal distances used in the field should be reported. This could also be included as a random factor in a linear mixed effect model. (possibly along side subject).

Line 137: Video calibration. Was any form of data smoothing used to reduce digitizing error? Was this source of error measured?
Line 203: ‘’user-based determination of footfalls’’  not really sure what is meant by this. Does it mean no one limb was used to define the start of a footfall sequence?
Line 285: How much lower were speeds estimated by UAV’s? Was this reduction reflected in the percent error reported below? Would be good to get an estimate of how much lower speeds were (i.e. speeds were 90% of treadmill speed). Also good to know how repeatable this was (i.e. 90% + / - 5% lower). This would be useful for anyone using the speed estimates derived from UAV’s – i.e. they might consider increasing the reported speed by the error reported.

Line 288: 8% error at 8m focal distance. How would this translate to the 40+ m focal distance used during field trials. Surprised to see so few test distances are used here considering that these are relatively easy to collect in a lab setting. Further these tested distances do not seem to include the actual distances used in the field. Do we expect error to consistently be reduced with distance? Or will it flatten out and become constant at some distance?


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: NATION-SCALE PRIMARY PREVENTION TO REDUCE NEWLY INCIDENT ADOLESCENT DRUG USE: THE ISSUE OF LAG TIME
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is a well written paper that examines whether there is a lagged effect of important alcohol and smoking control policies, the Minimum Legal Drinking Age and Minimum tobacco purchasing age on drinking and smoking behavior. They document that the increase of drinking incidence among 21 year olds was not seen until approximately 10 years after the policy change. Declines in adolescent tobacco smoking incidence occurred at least eight years after the Synar amendment.  I recommend a major revision and hope that the authors will successfully address the methodological problems that I address below.

The abstract of the paper is difficult to follow for the general audience. In the methods section the author may want discuss that they use the Meta analysis. In their methodology section, they should discuss why the meta analysis is the best practice to measure the lagged effect of these laws.

The Authors claim that “ The effects of United States (US) nation-level policy changes on the incidence of alcohol drinking and tobacco smoking are not fully understood. “ However, there is an extensive literature on the impacts of alcohol prevention policies on alcohol related outcomes. Furthermore, the effects of tobacco policies have also been discussed.  (Wagenear and Toomey (2002), Carpenter et al. (2007))

The authors report that the increase of drinking incidence among 21 year olds was not seen until approximately 10 years after the policy change.  There does not seem to have been any 346 instant effect of NMDAA and Synar Act on the onset of drinking or smoking. However, there are large number of studies that show the immediate effect of the minimum legal drinking age on several alcohol related outcomes. When the young adults turn 21, there is a sharp increase in the probability of drinking. (See for example, Carpenter and Dobkin (2009)

The authors may also investigate the effects of these laws especially the minimum legal drinking age (MLDA) and minimum legal tobacco (MLTPA) purchasing age on young adults who belong to different demographic groups or who are subject to different MLDAs and MLTPAs such as 18 and 19.

The majority of states in the United States requires that a couple be 18 in order to marry without parental permission. Marital status may significantly affect smoking behavior of youths. It is not clear how the authors address this problem.


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: NATION-SCALE PRIMARY PREVENTION TO REDUCE NEWLY INCIDENT ADOLESCENT DRUG USE: THE ISSUE OF LAG TIME
Review round: 1
Reviewer: 2

Output:
Basic reporting: IMPORTANT ACTIONS:
i. Restructure text to provide focus to the introduction and discussion as highlighted below
ii. Reconsider Tables and Figures 1 and 2 – do Tables/Figures provide different messages? Do either enable a focussed insight into the research question?
iii. Address demographics of sample

Further comment:

‘Uptick’ not in common usage outside the US. Reconsider?

Introduction lines 114 – 130 better suited in limitations and strengths discussion, by repositioning more emphasis would be put on the research question. 324-330 is main finding but somewhat buried in the discussion, better placed up front in discussion.

Methods could mention the source of the NSDUH data – SAMHDA. Authors indicate sample size ranges in different years, though stipulating the total sample size would also be beneficial to understand the scale of the work (as provided in table and figure headings).

Some demographic information would be beneficial to the manuscript, such as gender and income group. If not available, then clarification over the sampling method NSDUH employs would provide confidence that demographic bias is minimised.

Figures have been generated with quality statistical software
-	It isn’t made clear what the vertical lines in graph indicates, though it is assumed to be the year of the policy. Annotation would clarify.
Tables are comprehensive, but do not particularly offer an interpretable outcome.
-	Im not certain what message Tables 1 and 2 offer that Figures 1 and 2 do not provide. As the figures enabling an immediate interpretation of results, the tables could be supplementary.
-	I would like to see clarification of the parenthesised numbers. I assume these to be 95%CI of the sample proportion reflecting the true population, but this isn’t defined. Similarly 95%CI for mean difference should be stipulated.
-	Table 3 shows many differences with periods 1-3 compared to period 0. However we know there are sample size issues with period 0.
      o	Can periods be given more intuitive names as defined in the methods. As Periods 0-3 are repeated for alcohol and tobacco, but the years subtly change, it would be clearer if the descriptive name was provided.
      o	It seems the real crux of the research question rests in table 3 comparing period 3 to either period 2 or 1. The importance of this comparison, in context of the hypothesis, is lost in the complex table.
      o	Summative figure would help highlight the differences according to time relative to policy, rather than the granular detail of age.
-	Whilst the authors mention a second hypothesis around a rise in incidence at age 21, no figure or table specifically addresses this (though apparent in findings). Address or remove to avoid confusion, can be discussion point.
Experimental design: IMPORTANT ACTIONS
i. Results need to focus on research question rather than granularity by age
ii. Methods have minimal statistical analysis, use more advanced approaches

Further comment:

Research question at times a bit convoluted, but interesting, important and study approach addresses gap in knowledge.
-	Question seemed to focus on adolescents, and whether there was a change in incidence 10 years post policy. However the granular results make it difficult to see wider patterns. Whilst sub/sensitivity analyses could address whether there are differences between e.g. 12, 13, 14 year olds (as focussed on here), it seems this was not the main intention of the research question, and pooling age data (even over more intuitive age groups? 12-14, 15-17, 18-21, 22+) despites its limitations, may allow a more focussed statistical analysis of the change in incidence over time. The authors already take an approach to combine data in periods of study in relation to the policy implementation.

Large datasets analysed in a simple manner
-	Analysis feels unfocussed, matrices of proportions are presented. Results seem unfinished, those presented do not wholly address the research question(s) in a way that tests the hypothesis laid out in the introduction.
-	I am unsure why the authors have not performed a form of time series regression, such as interrupted time series, which would address the impact of the policy intervention. Regression analyses would also allow adjustment for age, enabling a clearer message. A statistical approach beyond a descriptive comparison of proportions would be appropriate.
-	Rather than a focus on the granularity of the data, the authors could compare variation in annual incidence (all years, or periods) between the ages – it appears younger ages would show less variation whilst older teens would show more variation. Steps should be taken to ensure clear messages emerge from the results, requiring minimal deciphering.
-	Previous publications from the authors show robust statistical approaches to derive a clear message, including meta-analyses. Similar approach needed.

Table 3 comes the closest to addressing the research question posed in the introduction, however this could be refined and built upon. Considering the focus of 10 years, compared to studies that look earlier, the periods defined in table 3 appear a pertinent foundation to continue the analysis. Results presented prior to table 3 are informative description of the dataset, yet somewhat peripheral, taking focus away from the objective of the research.
Validity of the findings: IMPORTANT ACTIONS
i. Address NSDUH statement that 2002+ data should not be compared to earlier data for differences across time.
ii. Refocus the manuscript and results on the research question so the added value compared to previous publications is evident. Avoid needless replication of published findings.
iii. Address limitations as outlined below

Further comment:

-	The authors have recently published a number of manuscripts using the data source from 2002 onwards. The authors clearly have expertise with the data set. I am concerned that the secondary hypothesis defined in the introduction has already been answered and published by the same authors: although I do not have access, the abstract would suggest so
-	“Looking across age strata, we found rising age-specific drinking incidence rates across adolescence to a plateau at age 16–18 years and made a new discovery of a statistically robust and highly reproducible dip in incidence at age 19–20 years, followed by the major peak at age 21 years, with sharply reduced incidence thereafter.” https://doi.org/10.15288/jsad.2016.77.405
-	I am also unsure how the tables presented here offer value further to the above publication (other than addition of tobacco results).
-	The hypothesis focuses on time-lag, and I feel the authors should focus their analysis on the time-lag in order to offer new insight. I don’t feel that Table 3 adequately supports or rejects the hypothesis, but makes a good foundation for further figures and analysis.

The authors have nobly identified limitations, though emphasis could be redirected.
-	Observational study (no causation) – good, but could other techniques identify a strength of an association that offers more grounds for causation?
-	Various multi-level policies in addition to NMDAA and Synar. – Yes, relevant.
-	Methodological changes in collection pre1991 – Perhaps more emphasis needed on the sampling methods pre2001
-	Small sample sizes in earlier years – Yes, well stated but contextualise, could this over or underestimate the differences according to periods?
-	Restricted years from 1979 onwards – Is this important? more methodological than discussion, and sample size was low pre1990 anyway, would extension to 1970 or 1960 add further value?
-	Survey question change in 1994 – you have evidence in the literature that suggests you can reduce the emphasis of this limitation
-	First tobacco cigarette smoking – this is an excellent point and demonstrates a strength of the study, consistent approach. Perhaps more could be stated about the potential impact of e-cigarettes on incidence, how this is recorded, or how future study should address it?

Limitations i would like to see addressed
-	How weighting in the survey may influence sample, or whether unweighted data needs to be adjusted with reference to education, deprivation.
-	How sample bias may influence sample, NSDUH incentive scheme of $30
-	“Therefore the data from 2002 and later should not be compared with data collected in 2001 or earlier to assess changes over time.” Notes on NSDUH site provided in supplementary. The strategy appears to ignore this issue, discussion of its limitation should be much more transparent.

Discussion has addressed possible reasons for increases in tobacco incidence following policy. Speculation could be offered with regard to reasons for a lag, particularly as the manuscript title and research question focus on this. The relevance of adolescent risk-taking behaviour to the results could also be described here.
Additional comments: Thank you for the opportunity to review your manuscript. I take a consistent approach to reviewing all manuscripts that I accept, and offer a full critique. The length of the review is not a reflection of the quality of the manuscript; I always provide what I hope to be sufficient feedback for authors to address and enhance their manuscript. I feel it fair that you receive in-depth feedback to acknowledge the time, money and effort invested in your work.


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

Input:
Title: NATION-SCALE PRIMARY PREVENTION TO REDUCE NEWLY INCIDENT ADOLESCENT DRUG USE: THE ISSUE OF LAG TIME
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: NATION-SCALE PRIMARY PREVENTION TO REDUCE NEWLY INCIDENT ADOLESCENT DRUG USE: THE ISSUE OF LAG TIME
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Thank you for your responses, the manuscript is a well presented study that clearly defines its research goal; the methodology you have taken (for others to reproduce or alternate) and the limitations are very well outlined. The changes made offer focus on the take home message, which adds value to the community. This manuscript now offers excellent foundations for further studies into policy changes and lag-time, highlighting the possible issues with previous efforts and examples of what must be overcome in future projects. You have considered your response appropriately and have produced an insightful report. Good work.


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

Input:
Title: OVEREXPRESSION OF OSFTL10 INDUCES EARLY FLOWERING AND IMPROVES DROUGHT TOLERANCE IN ORYZA SATIVA L.
Review round: 2
Reviewer: 1

Output:
Basic reporting: 1) Please explain more about functions and encoding protein of genes introduced in the manuscript. It is hard to see, for example, what “GF14c” (line 250) is even when reading the corresponding position in the manuscript. Describing functions of GI, Hd1 Ehd1 and MADS51 against flowering time (induction of flowering or suppression?) as well as their encoding proteins would help readers to understand the content more clearly.

2) The OsFTL10 overexpressors exhibits early flowering whereas expression of Ehd1, a suppressor of flowering under LDs, seems to be induced in the lines. It would be good to discuss this in the discussion part.

3) In the overexpressor Hd3a expression is induced whereas RFT1 is repressed. It would be good to discuss this observation.

4) Is the increased tolerance against drought in the OsFTL10 overexpressor lines due to increased expression of Ehd1? It would be good if you explain a bit more about the possible mechanism by which OsFTL10 overexpression mediates drought-stress tolerance in the transgenic lines.
Experimental design: Overall I am satisfied with the experimental design.
Validity of the findings: As for graphs representing expression of DREB2, bZIP23 and NAC1 (Fig. 6I, J, K), positions of asterisks are inconsistent with the figure legend. Figures look to indicate significant difference in comparison between time 0 and other times in each line and not between WT and transgenic lines. Please correct positions of asterisks in the figures comparing expression levels in transgenic lines to those in WT, and also change the corresponding sentences accurately describing the results.
Additional comments: This manuscript describes a study on a mechanism that controls the timing of flowering and its linkage to the drought stress response in rice. Authors create transgenic lines in which mRNA of a flowering-time gene OsFTL10 is overexpressed, or accumulation of its endogenous mRNA is suppressed, to examine its functions to control flowering and the drought-tolerance responses. Through a series of phenotypic analyses authors demonstrate that OsFTL10 induces flowering also increasing tolerance to drought. They also show through split-YEP and yeast two-hybrid assays that OsFTL10 physically binds to rice 14-3-3 proteins, also representing through mRNA analyses using transgenic lines that OsFTL10 affects expression of genes that control flowering. Also, they represent data that the overexpression increases some drought-stress inducible genes such as DREB2.

The experiments overall look carefully performed, and represented data also include clear messages. Therefore, I feel that this manuscript is suitable for publication to Peer J. However, I found several minor issues that would need to be addressed prior to submission.


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

Input:
Title: OVEREXPRESSION OF OSFTL10 INDUCES EARLY FLOWERING AND IMPROVES DROUGHT TOLERANCE IN ORYZA SATIVA L.
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript reads well except for a few language issues (e.g lines 86-89; 374; quirky passive voice structures). A quick check by a language editor should be sufficient to correct those.

* The experiments with CaMV35S::Hd3a and analysis of the cis-elements (lines 349, 374) are not mentioned anywhere before the discussion. These data should be described in the M&Ms and Results sections.

* Fig. 1 Higher resolution photos of the GUS stainings are required if available. In the current version, I couldn't see the staining signal in pollen.

* Fig. 6 The meaning of the asterisks here does not seem to match the legend. Is it testing significances against a 0 time-point or against a previous time point? The significance levels between the WT, transgenic and knockdown plants should also be tested to support the claims.

* What are the whiskers on the bar graphs?

* For some statements, literature references are missing (lines 67, 337).
Experimental design: The manuscript describes a coherent set of experiments aimed to characterize the FTL10 gene - one of the previously uncharacterized FT-like paralogs in cultivated rice, and understand its role in regulating flowering time and drought tolerance. The research question is relevant and understandable but not well defined in the Introduction (lines 106-108). I.e. you did not investigate the environmental response of the florigens Hd3a and RFT1 in this manuscript.

M&Ms

* For the qRT-PCR experiments specify the sampling scheme - which tissue, how many replicates, what time of the day. For all the experiments, specify the number of replicates sampled.

* line 146 - Specify the traits.

* lines 188 - 189 Why did you use a different duration of stress for RNAi and ox lines?
Validity of the findings: The experiments reported here revealed the dual role of FTL10 in regulating flowering time and drought tolerance in rice.

* Most of the claims are supported by the presented data and the experiments include the necessary controls. If feasible, in addition to MADS14 and 15, I suggest measuring the expression of the remaining MADS genes downstream of FT - MADS18 and 34 (as described in the Introduction), which will support or expand the statement on lines 278-280. What was the evidence of the direct interaction of FTL10 and MADS15 as you formulated it here?

* By looking at the bar charts, it is challenging to assess whether the analyses and claimed differences are statistically sound without referring to the raw data (included in the supplement) and re-analysing them. Adding the individual measurement data points to all the bar charts or better re-plotting them as box plots (also with the individual data points) would solve this problem and greatly help future readers.

Discussion

* The hypotheses of why the oxFTL10 lines flowered later than the oxHd3a plants are interesting but rather speculative at this point. If you decide to leave them in the revised version, I suggest illustrating them as an additional figure describing the candidate model.

* Involvement of FTL10 in drought tolerance is an intriguing finding. I believe discussing what is known about the interplay between the flowering and drought tolerance pathways in other plant species (and it is not much) will strengthen the discussion. For example, Habte et al. (Plant, Cell & Environment, 2014) also found cis-elements of the drought-related genes in the promoters of the light signalling genes in barley and vice versa.
Additional comments: Minor issues.

* line 85 - spell out the gene names
* line 92 - spell out GA3
* line 95 - drought escape is not only an acceleration but also a delay of flowering.
* lines 103, 111 - Do you mean drought escape or indeed tolerance here?
* lines 106 - 108 Revise  Here, you investigated another homolog of the rice FTLs - FTL10 but not environmental responses of Hd3a and RFT1.
* line 174 - A single solution containing the mixture of these compounds?
* lines 175 - Specify times.
* line 198 - "polypeptide protein"?
* line 198-199 Revise wording to be specific what you did. Here and throughout the text, avoid formulations such as "the function of FTL10 on drought stress was analyzed" (line 225). Write specific, e.g. "we studied the effect of drought on the expression of ..." etc.
* line 202 Specify in which tissues.
* line 207 Specify in which tissues.
* lines 235 - 239 Move to M&Ms
* line 249 How does GF14c come into play? It is not mentioned anywhere in the introduction.
* line 317 “a transcription that..” → “a transcription factor that..”
* Fig. 3. “Performance of the transgenic plants in field ..” contradicts M&M line 145 “...grown under greenhouse conditions ...”.


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

Input:
Title: ENGINEERING OF CHO CELLS FOR THE PRODUCTION OF VERTEBRATE RECOMBINANT SIALYLTRANSFERASES
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. In figure 2 to 5, uniform name of samples should be used.
2. In two bars in figure 5(A), which one is Expt 1? Please add some explanations.
3. Statistical analysis (t-test) should be included in Figure 2 and 5.
Experimental design: The research question was defined as expression and characterization of a range of sialyltransferases. But I’m not convinced that expressed proteins were well characterized due to low protein yields, 1-33 micro-g, line 331 and Figure 3.
1. Expression system should be reconsidered to prepare recombinant sialyltransferases in better yield, and some experiments and results should be added to quantitatively characterize the expressed transferases, in terms of enzymatic parameters and protein stability.
2. In Figure 5, the authors performed enzyme-linked lectin assay to evaluate the enzymatic activities to glycoprotein. To ensure the activities, MALDI-TOF-MS or HPLC analysis of sugar chains from glycoproteins should be included.
Validity of the findings: I fail to understand the properties of expressed sialyltransferases through a manuscript.
1. Although recombinant proteins showed enzymatic activities, acceptors differed among experiments, LacNAc, lactose and asialofetuin (glycoproteins) for Figure 2, 4 and 5, respectively. I think it is better to describe the meaning of different acceptors.

2. Although expressed enzymes showed sialyltransferase activities (Figure 2 and 5), they seemed to take a few activities in Figure 4, and bands in WB analysis (Figure 3) were not clearly observed even in purified proteins. Because of these inconsistencies, I may fail to understand the properties of expressed sialyltransferase.
Additional comments: In this manuscript, the authors prepared a range of sialyltransferases and tried to characterize their enzymatic activity. They have a good attempt at assessing non-mammalian sialyltransferase. While this study represents a massive effort, prepared enzymes were not well characterized due to low protein yield (1-33 micro-g, line 331 and Figure 3). I think the author needs to reconsider the expression system to improve the yield, and add some experiments.
I hope that my comment is useful.