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: INVASIVE CRAYFISH DOES NOT INFLUENCE SPAWNING MICROHABITAT SELECTION OF BROWN FROGS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Line 54. The term “niche overlap” is confusing since it could refer to many dimensions: thermal niche, trophic niche… it seems to me that you specifically refer to habitat preferences.
 Line 66. “The strong selection by P. clarkii on frog tadpoles”. Do you have any reference indicating that P. clarkii selects tadpoles over another resource? Or do you mean natural selection of tadpole behaviour? Or do you simply mean “the strong predation by P. clarkii”? Be more specific.
Lines 79-83. Your detailed answer raised an important complementary point: if aquatic vegetation was absent from your ponds, then shores were the only refuges in the pond. This should appear in the text to build your hypothesis.
Lines 104-107. Based on the surveys mentioned, can you provide precise estimates on crayfish occurrence? (e.g. number of invaded ponds and total number of ponds). It would be useful to provide it here to judge of the availability of uninvaded habitats.
Line 170. Unit error?
There are again careless mistakes throughout the manuscript: typos in the text and in table column names, double parentheses in citations, wrong units throughout the manuscript and table legend.
Experimental design: In the light of your modifications of the manuscript and your point-by-point response, it seems to me that the working hypothesis of the manuscript is highly questionable.
You specified in your answer that crayfish activity (and catchability) was so reduced during frog spawning that you cannot even obtain estimates of crayfish abundances. You also stated that frog breeders stay in ponds a maximum of one day. Then how could frog breeders detect crayfish, let alone change their spawning strategy in response?
These elements should rather be a starting point to establish the working hypothesis that these frogs cannot adapt their spawning strategy to this invasive species. Note that this has already been suggested in Ficetola et al. 2011 (Divers and Distrib).
There is also a confusion between adult strategies and impact/adaptive response of tadpoles. The latter is described throughout the manuscript, but it seems to be out of the scope of the manuscript – again, frog breeders do not stay in ponds after spawning.
Validity of the findings: Lines 106-108. Are you suggesting that invaded and uninvaded ponds have different hydroperiods? This has major implications for your results.
Line 134. Pond surfaces vary from 8 to 283 m². (Note that the unit indicated in legend is cm²). Did you adapt your survey time to pond surface to homogeneize detection probabilities?
Line 180-187. Variability is huge for interclutch distances and distance to pond edges! Indeed, average measures are significantly different, but conclusions must be more elaborated than “clear differences between R. l. and R. d.”.
Line 184. What is the biological implication of such a small depth difference, even significant?
Additional comments: Investigating whether native species can adopt strategies to avoid invasive species is an important and promising field research. I encourage the authors to identify plausible mechanisms for the adaptation of particular species before testing different hypotheses.


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: INVASIVE CRAYFISH DOES NOT INFLUENCE SPAWNING MICROHABITAT SELECTION OF BROWN FROGS
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors addressed my concerns adequately, I have no further comments to make.


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: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Jin et al. aimed to examine the neurovascular coupling disturbance in hemodialysis patients by using a combination of ASL and resting-state fMRI approaches. The authors found that hemodialysis patients exhibited decreased global CBF-ALFF coupling as well as decreased CBF/ALFF ratio in the amygdala relative to healthy subjects, while no significant associations between imaging measures and clinical variables were observed. In general, the study is well performed and interesting, though more from a methodological aspect. The manuscript is clearly written and its presentation is logical. Even though the study may improve the understanding of brain function in hemodialysis, specific issues should be addressed in a potential revision.
1. The study lacks a clear hypothesis. The Introduction would benefit from giving an unequivocal hypothesis, e.g., increased or decreased coupling? the most affected regions?
2. The methodological description of blood tests should be more detailed like the MRI scanning.
3. Frame-wise displacement (FD) has a unit of mm.
4. If ALFF and fALFF are calculated, the preprocessing steps of fMRI data should not include band-pass filtered.
5. Fig. 1 should be revised to highlight the inter-group difference.
6. In Fig. 3, I suggest merging the bilateral amygdale into a brain map.
7. What do the colorbars represent in Fig. 3-5? T values?
8. Language needs attention or possibly an editor. For example, “To detect significant brain regions between two groups in CBF and ALFF value, multiple comparisons were performed with controlling age and gender as covariates…” Did the authors mean multiple comparisons were used to detect group differences?


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: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This study has revealed the neurovascular coupling impairments in hemodialysis patients with a combination method between ASL and ALFF analysis,  indicating a novel imaging phenotype of the hemodialysis patients; however, the diagnostic effect is insufficient and warrant future study. Notably, there are still some uncertain items which should be explained more clearly:
1. the comparison between hemodialysis patients and healthy controls was not enough to elucidate whether the metabolism of the amygdala was fluctuated by the kidney disease or dialysis. Actually, relevant studies have demonstrated the main disrupted brain activity pattern of the ESRD with dialysis was distributed in the core regions of the default mode network, and please give more reference as for this aberrance. Moreover, It is recommended to design one more group of the ERSD without dialysis ￼￼ or analyze at different levels according to dialysis time.
2. The discussion is too tedious to propose the main findings. Better organised explanation of the discussion would be contributed to make a suitable prediction for the underlying neuropathological mechanism of hemodialysis. ￼￼
3.￼The six months of hemodialysis duration was regarded as for inclusion criteria, which need clarifying with some standard or reference.


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: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: See attached PDF
Experimental design: See attached PDF
Validity of the findings: See attached PDF
Additional comments: See attached PDF


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

Input:
Title: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: All my concerns have been addressed. I am satisfied with the revision and this manuscript can be accepted.


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: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The revised manuscript has been improved in experimental design and discussion. I have no comment about it and look forward to further study.


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

Input:
Title: DISTURBED NEUROVASCULAR COUPLING IN HEMODIALYSIS PATIENTS
Review round: 2
Reviewer: 3

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: IDENTIFICATION OF HUB GENES AND SMALL-MOLECULE COMPOUNDS IN MEDULLOBLASTOMA BY INTEGRATED BIOINFORMATIC ANALYSES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The structure of article is well prepared, but the font is not consistent. Before starting experimental results, the rationale should be well addressed
In Introduction, Authors should summarize recent study consequence of gene expression profile in medulloblastoma. What is the unique point of the present study ??
All data in this manuscript is bioinformatics analysis. There is no any experimental confirmation. To strengthen the study quality, Authors may make more efforts on comparing their results with other published databases. Can Authors provide evidence to conclude which signaling is major to promote medullablastoma ??
Experimental design: It is appreciated that all data used for bioinformatic analysis is available, and methods were clearly described. The rationale for each experiment should be described in the beginning of results.
Validity of the findings: Actually, numerous studies are attempting to establish gene expression databases for precision medicine. In addition, most of gene expression data are available online. Hence, the novelty of this study is intensively limited.
The current study will get improved if the confirmation for gene expression in some human specimens is performed.
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: IDENTIFICATION OF HUB GENES AND SMALL-MOLECULE COMPOUNDS IN MEDULLOBLASTOMA BY INTEGRATED BIOINFORMATIC ANALYSES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well written, fluent and well structured in all its part (introduction, methods, results, discussion and conclusion)
Experimental design: The aim, methods and limitations are clearly defined.
Results are relevant and deeply commented in the discussion.
Validity of the findings: Meaningful results. I would stress the gap that still exist between these results and treatments although a better knowledge of genetic basis could reduce the gap.
Additional comments: Good paper. Nothing too add.


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: IDENTIFICATION OF HUB GENES AND SMALL-MOLECULE COMPOUNDS IN MEDULLOBLASTOMA BY INTEGRATED BIOINFORMATIC ANALYSES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The focus of this paper is the bioinformatic analysis of RNA microarray expression in medulloblastoma (MB), the most common brain cancer in children. The goal was the identification of differentially expressed genes.  After expression analysis, the authors also performed in silico identification of potential new molecules for personalized target therapy of MB.
The authors started their study from microarray expression data deposited in a public NCBI database, Gene Expression Omnibus database (GEO). From this database, they have selected four different datasets derived from four published studies. Indeed, in GEO there are many deposited datasets relative to MB microarray. It is not particularly evident how and why the authors made their choice. The authors should explain better.
From this starting point, the authors performed an excellent bioinformatic analysis. The paper is very well written with clear, unambiguous, technically correct terms. The article includes a sufficient introduction and background to demonstrate how the work fits into the broader field of knowledge. The study is very well performed following a pathway that is becoming very common for expression data analysis in cancer. A paper that uses a very similar approach to study RNA expression data in MB was recently published (Fateme Shaabanpour Aghamaleki, Behrouz Mollashahi , Nika Aghamohammadi , Nematollah Rostami , Zeinab Mazloumi , Hamidreza Mirzaei, Afshin Moradi, Mojgan Sheikhpour , Abolfazl Movafagh. Bioinformatics Analysis of Key Genes and Pathways for Medulloblastoma as a Therapeutic Target in Asian Pacific Journal of Cancer Prevention, vol 20, page 221, 2019). The authors do not mention this paper and, although their manuscript is undoubtedly superior in the analysis and original in the approach to use CMap software to identify small molecule compounds as effective drugs for MB, they should discuss the differences and the similarities with the published literature. Relevant prior research should be appropriately referenced.
The structure of the article is in an acceptable format; the figures are relevant to the content of the article, of sufficient resolution, and appropriately described and labeled. All appropriate raw data have been made available under the journal policy.
Experimental design: The study is within the Aims and Scope of the journal. The paper defines clearly the research question, which is relevant and meaningful. The investigation was conducted rigorously and with a high technical standard. Methods are described with sufficient details.
Validity of the findings: The data are robust, bioinformatically sound, and controlled. The impact of the findings is undetermined. The degree of advance is not immediately apparent. The molecules identified by CMap analysis are a potential novelty that the authors should discuss and stress better. Also, the conclusions are well stated and related to the original question investigated.
Additional comments: The authors have done an excellent and smart bioinformatic analysis of microarray expression data of MB. They used their analysis to identify in silico new compounds to test in MB therapy with a new intelligent approach. They should better discuss their interesting data and compare them to published literature to deserve publication


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

Input:
Title: IDENTIFICATION OF HUB GENES AND SMALL-MOLECULE COMPOUNDS IN MEDULLOBLASTOMA BY INTEGRATED BIOINFORMATIC ANALYSES
Review round: 2
Reviewer: 1

Output:
Basic reporting: All questions and requests were answered well. I have no further questions.
Experimental design: Clear
Validity of the findings: It is benefited after confirmation by qPCR.
Additional comments: No further requests


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: IDENTIFICATION OF HUB GENES AND SMALL-MOLECULE COMPOUNDS IN MEDULLOBLASTOMA BY INTEGRATED BIOINFORMATIC ANALYSES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors can now show the robustness of their in silico data analysis by in vivo RT-qPCR measurements that experimentally confirm the downregulation of 5 hub genes. I believe the manuscript is now in better shape and can be considered for publication.
Experimental design: The authors can now show the robustness of their in silico data analysis by in vivo RT-qPCR measurements that experimentally confirm the downregulation of 5 hub genes. I believe the manuscript is now in better shape and can be considered for publication.
Validity of the findings: The authors can now show the robustness of their in silico data analysis by in vivo RT-qPCR measurements that experimentally confirm the downregulation of 5 hub genes. I believe the manuscript is now in better shape and can be considered for publication.
Additional comments: The authors can now show the robustness of their in silico data analysis by in vivo RT-qPCR measurements that experimentally confirm the downregulation of 5 hub genes. I believe the manuscript is now in better shape and can be considered for publication.


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

Input:
Title: RARE GROUND DATA CONFIRM SIGNIFICANT WARMING AND DRYING IN WESTERN EQUATORIAL AFRICA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Good
Experimental design: Good
Validity of the findings: Good
Additional comments: This manuscript presents an impressive 34-year ground data of temperature and rainfall and short-term data of several climatic variables from Lopé National Park, Gabon, western equatorial Africa.  The seasonal and inter-annual variation, long-term trends in local weather patterns were evaluated.  The authors also investigated the effects of several natural climatic oscillations on the local weather patterns.  Lopé weather has a low-light, cool, and dry season.  There is a warming and drying trend in Lopé.  Sea surface temperatures (SSTs) of the major oceans, like ENSO and NAO, had significant effects on the inter-annual variation in Lopé

I have two primary suggestions.

First, the authors used generalized linear mixed models (GLMM) and linear mixed models (LMM) to evaluate the ocean influences on the local weather in Gabon.  However, the association between the local weather and SSTs of major oceans may be transient.  The relationships between these two systems may change through time, occurring at different time and periodicity.  Moreover, the time for the local system to response to the regional influences (the time lags between two systems) may also change through time.  Thus, it would be better if wavelet coherence analysis can be applied in this study.  Wavelet coherence analysis can deal with the problems mentioned above.

Second, there are very strong seasonal cycles within the climatic time series analyzed, and these seasonal cycles (either annual or biannual) may mask the long-term trends within the time series.  Please consider to remove the seasonal components within the time series before testing the relationships between local and regional systems.

Some minor suggestions follow:

It would be easier for readers to interpret the results if the authors report the delta AIC (the differences of AIC) between the best model and others.  The delta AIC for the best model will be zero, which will help readers identify which model is the best.

L402-403: I would suggest to use the words like ‘the signal of biannual cycle decreased’ instead of ‘lose power’.

Figure 3F: Part of the curve for the annual cycle is masked by the figure legend.  Please move the figure legend to the top left of the figure.


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: RARE GROUND DATA CONFIRM SIGNIFICANT WARMING AND DRYING IN WESTERN EQUATORIAL AFRICA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is clear, well written and easy to read. The introduction is clear and the inclusion of review of published literature on drivers of weather variability is unique and adds much value to the manuscript. However, there is the need to provide more information on the other climate variables (L33-34; absolute humidity, wind speed, solar radiation and aerosol optical depth), which was introduced in the abstract. Moreover, on L156-158 there appears to be a missing link - what may happen to these forests if warming climate push the region towards drought-adapted deciduous ecosystem? I think this portion of the introduction is incomplete. The authors have to provide the reader with potential impact/influence on the region if it dominated by drought-adapted ecosystems. Nevertheless, the figures and tables are clear and relevant (but see general comments) and the raw data have been supplied.
Experimental design: The authors have defined the research questions and set them appropriately within the context of the study. They have conducted vigorous analysis, despite the challenges with the data. However, they can improve the analysis. First, I think it will be easier to understand the GLM models used if they are represented as equations (e.g. L326-339). Further, testing whether in the different interaction terms improved the model, the authors may want to consider using a likelihood ratio test rather than depending on only the AICs to select their models (e.g. L326-339). See example in Wang et al. (2011). Second, on L175-210 the authors elaborated on the challenges they faced in collecting the long-term climate data – a problem common across Africa. Wouldn't the standardisation approach they undertook (L211-213) also introduce other biases, which may affect their findings? I also suggest the authors explain to the reader why they used a 10-day running mean of the calender year (L271-272 and others) when describing seasonality and the other trends? There is also the need for more details on how the spectral analysis was done (L278-279)
Validity of the findings: The authors have provided interpretation for the results but this should be improved. For instance, the authors analyzed seasonal trends of humidity, wind speed, solar radiation and aerosol optical depth (L34, L240-246, L365-378, Fig. 2B, Fig. 2E, Fig. 2F and Fig. 2G and 2H; Supplementary 3.1-6.1) but there was no discussion on these trends. I suggest the authors either remove the details on these variables from the entire text or provide interpretation on the seasonal patterns they observed in these climate variables. This is important because Central African forests may be susceptible/vulnerable to climate variables such as changes in solar radiation (Philippon et al. 2019). Importantly, since the data had many gaps and the fact that they only focused on minimum temperature I suggest the authors do not overstate their results in the discussion (e.g. (L508-511 'As.....in the region (as in Collins 2011’ and also L460-462). Also the link between the warming and drying observed by the authors and how they impact the survival of forests (as stated in the title) in the region was weak.
Additional comments: The authors have worked hard to get this long-term weather records in Lopé, which is notoriously difficult to obtain in most tropical African regions. This manuscript is relevant and will provide insights into the long-term climate trends and their drivers in tropical Africa. But there are other few minor issues that the authors have to consider to improve

L158: Provide reference
L454: Is the change (0.15°C) increasing or decreasing?
L479-485: Remove/delete sentence. It is a repetition of the results
L501: Remove one 'in'
L525 and 526: consistency in writing et al.
Please explain what the asterisk (*) mean (Table 4, Table 6, Table 7)

Reference
Wang et al. (2011). Modelling growth responses of individual trees to early-age thinning Eucalytus globulus, E. nitens and E. grandis plantations in northern Victoria. Australian Forestry 74: 62-72.

Philippon et al. (2019). The light-deficient climates of western Central African evergreen forests. Environmental Research Letters 14: 034007.


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: RARE GROUND DATA CONFIRM SIGNIFICANT WARMING AND DRYING IN WESTERN EQUATORIAL AFRICA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Good
Experimental design: Good
Validity of the findings: Good
Additional comments: This manuscript presents an impressive 34-year ground data of temperature and rainfall and short-term data of several climatic variables from Lopé National Park, Gabon, western equatorial Africa. Their results indicate that there is a warming and drying trends in Lopé, and the inter-annual variations in minimum temperature and rainfall were associated with ENSO and NAO. The manuscript is well-prepared and has greatly improved from previous version. However, there are some points to follow up.

1. The authors applied wavelet coherency analysis to evaluate the associations between sea surface temperatures (SSTs) and the climatic variables in Lopé. However, they did not remove the seasonal component from the time series before the wavelet analysis. There is a strong signal of seasonality (both annual and bi-annual) in SSTs and the local climatic variables, as we can see in the Figures 3 and S3. Since this seasonality comes from other external driving forces, such the movement of ITCZ zone, and may mask other signals (inter-annual variation and long-term trends). The authors may try using additive monthly factors (subtract the long-term mean of each month from the time series) or Seasonal Decomposition of Time Series by Loess (function ‘stl’ in R) to remove the seasonal component before performing the analysis of trend detecting and wavelet coherence.

2. Figure S3 is one of the main results, while Figure 4 did not provide much information. Please consider add Figure S3 to the main text and move Figure 4 to the appendix.

3. In Fig. 3E and 3F, there are no labels indicated which one is the wavelet power of rainfall time series and which one is temperature. There is no explanation in the figure caption, either. Please add the figure labels to clarify.


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: RARE GROUND DATA CONFIRM SIGNIFICANT WARMING AND DRYING IN WESTERN EQUATORIAL AFRICA
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: 1.	The approach of using models to fill up the gaps of missing data may influence results of trend analyses. Therefore, please provide information about how bad the missing data problem (e.g. how many weeks of data are missing in that 34 years) is and elucidate possible impacts of this approach on your results.
Validity of the findings: No comment
Additional comments: General comments

1.	It is better to remove “at a critical level for forest survival” from the title. To keep these words in the title, you need to show that tree mortality caused by drought stress is increasing in the last 3 decades. Please still keep sentences for forest survival in the discussion.

2.	Annual rainfall is not a good index to quantify drought stress for plants. Plants are under drought stress when water input (precipitation) is lower than water removal (evapotranspiration). Moreover, annual rainfall contributes partially to the total water input of an area with low cloud (see the paragraph below for precipitation contributed by intercepted cloud). In other words, low annual rainfall does not lead to high tree mortality resulted from drought stress if there are additional sources of precipitation and evapotranspiration is low. Compared with rainfall, the difference between actual evapotranspiration and potential evapotranspiration and the difference between the precipitation and potential evapotranspiration are much better ways to quantify drought stress for plants (e.g. Rind et al. 1990).

3.	It is better to replace “western equatorial Africa” with a small-scale location name. Balas et al. (2007) (in L604-606 of the present study) reported that rainfall varies with sites. If you want to keep “western equatorial Africa” in the title, you need to provide evidence that the trends derived from the weather data of Lopé National Park happen ubiquitously throughout western equatorial Africa.

4.	Please remove the discussions for low plant photosynthesis rates resulted from light deficit and add discussions for low evapotranspiration associated with low direct solar radiation. The main finding of the present study is the warming and drying trends in your study site. Plant survival is closely related to evapotranspiration in dry seasons (i.e. water input is low). Therefore, the later discussions should be added to infer how cloudiness interacts with drying and in turn influences forest survival. Removing the former discussions will allow readers to focus on the later discussions, otherwise they are kind of distracting.

5.	Please provide information about precipitation resulted from cloud intercepted by tree foliage. In addition to the reduction of evapotranspiration, low cloud can provide an importance source of precipitation for forests. For example, in a subtropical rainforest where annual rainfall measured with conventional rain gauges is as low as 1125 mm, additional precipitation resulted from cloud intercepted by tree foliage can provide up to 40% of annual rainfall (Hutley et al. 1996).

6.	There is hardly low-lying cloud when it is windy. The sentence in L138-139 mentions the presence of low-lying cloud in the long dry season and the sentence in L142-143 mentions that the long dry season is the windiest season. Please explain how these contradicted situations can happen in the same place and the same period of time.

7.	Please use tables to show some of your contents. For example, compared with texts in L230-260, a table with a few columns (e.g. time period, instrument manufacturer) is easier for readers to figure out how and what weather data are collected in that 34 years.

8.	Please use figures and tables of the present study to support your conclusions in the discussion. For example, the sentences in L518-519, L526-528 and L563-564.

9.	Please make sure that all cited references are listed in the references section. For example, Wu et al. (2016) (cited in L553) is not listed.

Specific comments

1.	Please mark the location of the weather station in Figure 1.
2.	Please insert space between values and their units. For example, “2000mm” in L93 should be “2000 mm” and “800m” in L229 should be “800 m”.
3.	L532, L534, L536: “°c” should be “°C”.
4.	L535-536: Add “+” before “0.26” and “0.22”.
5.	L535-536: Is the warming rate (+0.26°C) of America correct? The warming rate (+0.25°C) of your study site is actually lower than +0.26°C.
6.	Figure number is missing. For example, “Fig. 2C and D” in L441 should be “Fig. 2C and 2D”.
7.	L632: “Iit” should be “It”.

Reference
1.	Rind, D., Goldberg, R., Hansen, J., Rosenzweig, C., & Ruedy, R. (1990). Potential evapotranspiration and the likelihood of future drought. Journal of Geophysical Research: Atmospheres, 95(D7), 9983-10004.
2.	Hutley, L. B., Doley, D., Yates, D. J., & Boonsaner, A. (1997). Water balance of an Australian subtropical rainforest at altitude: the ecological and physiological significance of intercepted cloud and fog. Australian Journal of Botany, 45(2), 311-329.


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: RARE GROUND DATA CONFIRM SIGNIFICANT WARMING AND DRYING IN WESTERN EQUATORIAL AFRICA
Review round: 3
Reviewer: 1

Output:
Basic reporting: 1. Please always place a space between the numerical value and unit symbol. For example, in L564, “2.3mm” should be ”2.3 mm”.
proper:	2 m, 15 mm
improper: 2m, 15mm

For more information, please refer to the websites below.
https://www.monash.edu/about/editorialstyle/editing/numbers
https://physics.nist.gov/cuu/Units/checklist.html

2.  L571:  It is a subtropical rather than tropical forest.

3.  L492:  “+0.19Cc”?

4.  Reference formats are not uniform. I listed three inconsistencies as examples.   Please systematically double check the formats of all references and make sure they are uniform.
i.	Some references show “pp.” (L706, L717, L751, L760, L792……) but most references don’t.
ii.	Page number is missing in L863.
iii.	Please remove underscores and asterisks from L947-948.
Experimental design: NA
Validity of the findings: NA
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: AGILITY-BASED EXERCISE TRAINING COMPARED TO TRADITIONAL STRENGTH AND BALANCE TRAINING IN OLDER ADULTS: A PILOT RANDOMIZED TRIAL
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is well written, uses relevant and current references and the results reflect the hypotheses.
I suggest two minor language changes:
1: Line 86 - masked assessors is now the preferred term - rather than blinded.
2: Please consider an alternative word to "Corroboratory" on line 292.
Experimental design: The research represents a clinically relevant research question which aligns with the Aims and Scope of the journal.
The investigations is well though out and well conducted.
The methods section is well described.
1. Please consider also outlining, in the "General Design" section, the distribution of the training sessions to allow for replication. For example, 24 training sessions (3 sessions per week for eight weeks). Each session was 50 minutes in duration. (LINE 81)
2. It is noted that the Participants were excluded for "cardiologic, neurological and orthopaedic conditions" (LINE 91). Can you please expand on whether this means specific conditions or all conditions. If it means all conditions under these categories it will rule out  many older adults and therefore reduce scalability of findings. This should therefore be noted as a limitation in the results section of the paper.
Under the Interventions section please consider describing whether the difficulty was also increased by increasing resistance (not just repetitions) and if not, why not as this is usual practice for strength training programs. (LINE 125)
3. Figure 1: Please include details on the 26 excluded people. How many declined and how many didn't meet the inclusion criteria.
Validity of the findings: The underlying data has been provided and are robust, statistically sound and controlled.
I have the following questions:
1. There were large differences in the baseline measures for 6mwt and SLEO for each group. Can you please comment on whether you think this may have affected your findings.
2. Please describe your cut off score for p value significance for between group differences in change scores. They appear to be 0.1 or 0.2 in some cases and we need to agree on whether that can be determined as significant.
3. The Agility Group had a much bigger change in Perturbation measures, although the CI are wide, can you please comment on whether you think this may be clinically relevant even if not statistically so?
4. Background Conclusion: please provide references that state plantar flexion strength and trunk trunk extension RTD are falls risk factors. Otherwise it may be better to say that :agility- based training seems at least as efficacious as traditional strength and balance training in affecting selected physical performance indicators among community dwelling healthy seniors"
5. I believe that the fact that the agility based training is much more time efficient, while potentially delivering comparable outcomes may be the key finding of this study. The authors have mentioned this but may like to expand on the point. This finding is important as current best practice now recommends up to 3 hours per week to achieve a falls prevention effect (Sherrington et al, 2017). This is very hard to achieve in the real world so this pilot study contributes an important finding to push for the implementation of a larger study to test whether the use of the intervention achieves a falls prevention outcome. Such a finding has the potential to increase uptake and adherence, which would be a very important clinically.
Additional comments: Thank you for conducting this pilot study, and submitting this well written manuscript. You are to be congratulated on designing an intervention that has the potential to reduce the time commitment required by older adults to maintain or improve their physical performance and potentially reduce falls. I have made a few comments for your consideration and would be happy to review again if required.


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: AGILITY-BASED EXERCISE TRAINING COMPARED TO TRADITIONAL STRENGTH AND BALANCE TRAINING IN OLDER ADULTS: A PILOT RANDOMIZED TRIAL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Basic reporting is clear and professional throughout.
Experimental design: Well designed study but further description is required to replicate.
Validity of the findings: Findings need to be better aligned with expected change data - what is clinically important?
Additional comments: I commend the authors on an interesting an important study assessing the effect of agility training on balance and strength in older adults. I understand that the study was a randomised pilot in nature and only certain inferences can be made with a small sample size, but I believe it could be strengthened further, particularly by including more details about the interventions. Please see my specific comments attached.


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: AGILITY-BASED EXERCISE TRAINING COMPARED TO TRADITIONAL STRENGTH AND BALANCE TRAINING IN OLDER ADULTS: A PILOT RANDOMIZED TRIAL
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Thank you for your responses to my previous feedback, I believe this paper is now ready to be accepted for publication and provides a valuable contribution to falls prevention research.


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

Input:
Title: AGILITY-BASED EXERCISE TRAINING COMPARED TO TRADITIONAL STRENGTH AND BALANCE TRAINING IN OLDER ADULTS: A PILOT RANDOMIZED TRIAL
Review round: 2
Reviewer: 2

Output:
Basic reporting: Basic reporting is clear and professional throughout.
Experimental design: Well designed study with appropriate detail in revised version.
Validity of the findings: Appropriate and clinical effects discussed.
Additional comments: Well done on a well conducted novel training program. Thank you for addressing my concerns/questions - you have amended appropriately and made the paper easier to follow/replicate.


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: CATALYZING RAPID DISCOVERY OF GOLD-PRECIPITATING BACTERIAL LINEAGES WITH UNIVERSITY STUDENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors present an approach to collect a high volume of samples by engaging individuals across an entire campus.  The manuscript is well written and concise.  There a few instances where the clarity of the manuscript could be enhanced in places.  Below are suggested changes:

Line 53: The abstract states there was an analysis of “evolutionary trajectories” however in the study there was only a comparison of ‘gold gene’ sequences obtained in this study to current published sequences.  Please remove “evolutionary trajectories”.  Suggested replacement is DNA sequences.

Line 62: Add “experimental” before questions.

Line 66-67: The reference listed to support this statement (Schlaberg, Simmon & Fisher, 2012) is a paper that looks at identifying novel pathogens in clinical settings.  This doesn’t support studying “unique abilities of bacteria to everyday human life”.  Please remove this sentence.

Line 71: change the word trivial (it implies the stated procedure is unimportant)

Lines 75-77: fix indent of sentences

Line 90:  Take out “Arguably, one of the most remarkable bacterial taxa is”.  While the gold-precipitating ability of this bacteria is a great way to interest the non-science community, some in the scientific community might not find this significant enough to warrant these bacteria as the most remarkable of all bacteria.

Line 93: After obvious potential uses please state a few of the those uses.

Line 98: change taxa to species

Line 111: change ‘classes’ to course to emphasize the ability of the study to be applied to different courses

Line 118: Define CCG and NGR.

Line 132: Italicize Safety

Line 135-136: Reword sentence to include the student was wearing the disposable gloves.

Line 193: please clarify Appendix 4 is a list of all samples.

Line 224-226: Remove this sentence.  A quick Pubmed search using “Delftia” results in 373 results.  I would not conclude that these are rare bacteria, just not a ‘household name’ bacteria.

Line 243-244: What is meant by neutral?  Please clarify this statement.
Experimental design: The goals of this study were to engage the campus community, conceptualize science for students, examine the abundance of Delftia spp. in samples collected and identify novel species of Delftia using DNA sequences of the Delftia-specific gold gene.  In my opinion, the authors were highly successful in all their aims except for the last.

Unfortunately, there is a flaw in the latter portion of the experimental design that invalidates some of the conclusions drawn from the results.  Since the DNA was extracted from environmental samples it cannot be assumed that only one species of Delftia is represented and therefore sequencing of the gold gene in these samples could be considered ambiguous.  The sequences obtained could possibly be a mixture of sequences from various species/strain of Delftia obtained in a single environmental sample.  This may explain the areas of your electropherograms that show overlapping peaks.  (For example, a quick contig construction for the Forward and Reverse sequencing results for Sample 1-3 shows roughly 8% disagreement of the sequences.)

To elucidate the confounding nature of environmental samples the bacteria should first be isolated or the PCR products should be ligated into a cloning vector and transformed into competent bacterial cells, cloned and plasmids extracted and sent for sequencing.

In addition, after these clean sequences are obtained, it would be beneficial to run a phylogenetic analysis of the sequences obtain in addition to reporting the BLAST results.  This would be helpful for future readers to understand the relationship among the reference strains used in the comparison and the environmental isolates.  For example, I would like to know how similar the gold gene is in Delftia tsuruhatensis strain CM13 as compared to the gold gene of Delftia acidovorans strain ANG1, SPH-1 and RAY209.

Specific comments:

Line 54 and Line 61: This study did not discover “new lineages” of Delftia spp.  It may be more appropriate to say the aim was to assess the prevalence of Delftia spp. based on targeted metagenomic sequencing.

Line 142:  Quick note, in future studies you may also want to treat the metal scoops and scissors with DNaway between samples to ensure no free DNA carry over contamination from sample to sample.

Lines 161-168: Please include standard curve information.

Line 165: reference qPCR conditions in the supplemental material

SUPPLEMENTAL 2: The primer and probe concentrations are not listed.
Validity of the findings: The authors present an engaging citizen-science study that shows the power of involving the community in sample collection.  The distribution of Delftia spp. and relative abundance is demonstrated using two individual Delftia-specific qPCR reactions.  However, some of the specific findings and conclusions made by the authors regarding the discovery of new species cannot be supported by the data due to the complex nature of the DNA sample used as template in the gold gene sequencing reactions.

Specific comments:

Line 239-241: The Schlaberg et al. 2012 study presents species differences based on differences in 16S rRNA genes, this can not be extrapolated to individual genes, more specifically the gene gold.

Lines 251-256:  Agreed that there was engagement of students and the campus community that helped to better understand the distribution of Delftia, however the conclusion of new strains of Delftia cannot be stated based on the experimental procedure used in this study.

Line 255-262, 289-290:  New species cannot be inferred with certainty based on the issue mentioned above.
Additional comments: Overall, the premise of this paper is very interesting and the manuscript is well written.  It is impressive to learn the first author of this paper is an undergraduate researcher and, as a fellow teaching-track faculty, I commend the faculty member on the resourcefulness of recruiting sample collectors outside the classroom.  This is a great way to engage the general public and allowed for significant amounts of information to be collected regarding Delftia spp. in a short amount of time, however some of the conclusions made in the manuscript cannot be confidently supported based on the experimental protocol used.  However, the authors outline ways to better identify Delftia spp. in their samples in lines 277-287.  I suggest removing language from the current manuscript stating the discovery of new strains/species of Delftia and instead focus on the success of the campus-wide initiative and emphasize the presence and abundance of the Delftia in the samples that were collected.  (For example, how many of the total samples analyzed for positive for Delftia?  Also, is there any research regarding the gold gene copy number in a Delftia cell?  This would give an indication of the quantity of Delftia in your samples.)


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: CATALYZING RAPID DISCOVERY OF GOLD-PRECIPITATING BACTERIAL LINEAGES WITH UNIVERSITY STUDENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript introduction provides a compelling argument for the value of citizen science in helping to identify new species of bacteria by expanding the number and types of sampling sites that can be evaluated. It also provides strong case for focusing on Delftia spp as the target genus because of its unique properties that may be of general interest – the ability to precipitate gold. Overall, the manuscript is well written and some restructuring of the discussion section would be helpful to the reader as some of the information is repetitive and seems to go back and forth between topics. For example, line 262 refers to ten of seventeen samples having less than 95% or lower match and line 291 refers to 14 strains with less than 97% match. Some revision of the discussion section would be helpful for the reader to better understand the impact these results have had on our knowledge about the abundance and distribution of Delftia spp.
Experimental design: The experimental design provides clear instructions on the methods used and are technically appropriate for the project. The unique aspect of the project is the citizen science element and the recruitment of participants and the distribution of sampling kits is critical. The process is reasonably well described and additional information on the public element would be helpful. For example, on line 116 the authors mention a public event to inform the public and recruit participants and it would be helpful to have some more information on how the event was structure and how well attended. Line 120 mentions that participants were primarily recruited from introductory environmental science courses and it would be helpful to know how many participants came from the courses relative to the general public. The manuscript also emphasizes that participants came from a wide-range of disciplines as stated on line 254 and again on line 311 so examples would be helpful. Are the introductory environmental sciences courses non-majors courses that attract students from diverse majors or had these students from diverse majors come from the public event?  On line 60, it is mentioned that the results are share through a publicly accessible website and workshops and it would be helpful to have more information about the workshops – how many, how were they structures, how well attended, etc.
Validity of the findings: The results of the project regarding the discovery of potential new species are clear and well documented. The findings are solid and demonstrate the value of citizen science for discovery. The technical aspects are appropriate and readily adaptable to a variety of situations. This illustrates the potential impact of this approach on making new and valuable discoveries. The proposed expansion on these new findings by isolating new Delftia spp. should prove to be very valuable experience for expanding this project into additional courses. The many questions raised regarding the ecology of the organism should lead to opportunities for students to participate in the project and to also have the opportunity to have a critical role in designing the experiments to test the hypothesis that could be generated. I love the future plans to engage additional courses in the project, including a first-year English class. . If available, I would like to see more information about the composition of the participant population to get a better sense of how effectively the project is reaching a broader audience, particularly outside of the sciences.
Additional comments: The manuscript describes a terrific approach to integrate citizen science with course based laboratory projects. It illustrates how an effective program can lead to novel scientific discoveries by engaging the community at large in the sampling process.


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: CATALYZING RAPID DISCOVERY OF GOLD-PRECIPITATING BACTERIAL LINEAGES WITH UNIVERSITY STUDENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: This article describes an ambitious effort to engage undergraduate students in citizen science and inquiry-based laboratory modules focused on identification of Delftia in samples from a university campus. This initiative is a valuable contribution to curriculum development in undergraduate science education, and, as discussed in the article, may be adapted to engage non-STEM students. However, in its current state, this article does not present enough data to constitute a substantial contribution to the scholarly literature, and it is not currently suitable for publication in PeerJ.

In the article’s Abstract and Introduction, the authors outline two main goals: (1) “sampling, identification, and analysis of the evolutionary trajectories of a unique yet common microbe, Delftia spp.” with an emphasis on discovery of Delftia lineages able to precipitate gold and (2) enhancing undergraduate learning, specifically focused on “helping contextualize and solidify their learning of the molecular biology techniques”. In its current state, the article does not present enough data to fulfill either of these two goals.

For goal (1) focused on Delftia biology, the authors present data on Delftia presence in 135 samples. Abundance of Delftia in these samples is estimated based on qPCR of two Delftia-specific gene regions. Some analysis of genetic variation of gold precipitation genes is presented, but this is not yet substantial enough to warrant publication. To fulfill goal (1), the authors need more analysis of the gold precipitation gene cluster, expanding to include the entire region. Data are currently presented in tables and alignments, but these data are best presented via phylogenetic analyses and broader comparisons including publicly available database sequences. (For example, are there publicly available metagenomic datasets that could be mined for Delftia sequences?) More data is needed to determine the genetic diversity of Delftia on campus, which is a stated goal in the Introduction. In addition, it is unclear whether the presence of the single 1,045 bp region in the gold precipitation gene cluster is sufficient to conclude whether these Delftia can precipitate gold, which is another stated goal of the article. Overall, although the authors have presented data on sampling and identification of Delftia, there is not enough data on analysis of evolutionary trajectories and genetic variation (which could be achieved by phylogenetic analysis of an expanded gene dataset) or ability to precipitate gold (which could be achieved by analysis of the full gold precipitation gene cluster).

For goal (2) focused on enhancing learning, the authors do not present data on student outcomes. To align with standards in science education research, many inquiry-based curriculum initiatives use pre- and post-assessment instruments to measure student outcomes and determine the impact of the initiative on the targeted student population. I understand that it is not possible to go back in time to conduct these assessments, but this is an important point to consider if the authors are planning to implement this initiative in future courses with the intention of publishing. Even without these assessments, the article in its current state does not present enough data on goal (2) to warrant publication. For example, the article states that students from diverse majors were involved in the project, but there is no data on which majors are represented. One of the aims of the initiative is to “test whether students are able to aid the speed of discovery of novel lineages”, but the outcomes of this aren’t explicitly discussed.

Regarding background/context and literature references, the Introduction lacks a detailed description of the gold precipitation gene cluster in Delftia. Because this is a targeted gene region that is used as an indicator of the presence of Delftia, the authors should present more information on the genes involved in this function and the known variation among published sequences. Considering the model adopted for this initiative, the authors may also want to reference SEA-PHAGES and Tiny Earth (previously the Small World Initiative), which are inquiry-based laboratory and course curricula focused on microbiology.
Experimental design: From the Materials and Methods, it was unclear to me whether all metagenomic DNA was extracted from samples by a single undergraduate researcher (the first author) or by students as part of a course. In addition, it may be worth considering how sample storage and/or processing protocols might affect recovery of Delftia genomic DNA and estimates of abundance.

What were the quality criteria for trimming Sanger sequencing data? Currently, this is described as “stringent quality settings to match existing sequences in the NCBI database”. From the Materials and Methods, it seems that forward and reverse reads were generated. How were these assembled to generate the final consensus sequence? Was manual editing required at any positions?

As mentioned above in Basic Reporting, the analysis of sequence data is currently presented as alignments and tables of BLAST results. These data would be better presented as phylogenetic analyses, which may reveal interesting relationships for further investigation.
Validity of the findings: A major point here involves the authors’ use of sequence similarity cutoffs developed for 16S rRNA gene sequences. As described in the reference cited in this section, the cutoffs of 97% and 95% are relevant for 16S rRNA gene sequences, not necessarily other genes. The 16S rRNA gene has been widely evaluated across many diverse bacterial lineages, and, even so, the cutoffs are often debated among taxonomists. It isn’t appropriate to apply these criteria to other bacterial genes, which experience different selective pressures compared to 16S rRNA genes. With the current dataset, it isn’t possible to estimate the percentage of samples that may contain new Delftia species. It is possible to discuss variation and divergence in gold precipitation genes, which is a more appropriate interpretation of the data.
Additional comments: Overall, in its current state, this article does not present enough data and analysis to achieve the goals outlined by the authors in the Abstract and Introduction. The current manuscript is a solid foundation for a publication, but it is not currently suitable for publication. I have attempted to provide some guidance to the authors regarding further analyses. I understand that budgets and time may be limited, therefore it may not be possible to extend this project beyond the current dataset. If this is the case, the authors may want to consider publishing as a note, with some additional analyses using the existing dataset.


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 PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: It is an interesting study focusing on the prognostic value of serum CA724 in CEA-normal colorectal cancer patients. It should be revised by a native English speaker and a professional editor. There are a lot of language and spelling errors in this study (see the attached pdf file). The figures and tables were poorly prepared and should be revised.
Experimental design: Experimental design is reasonable. More details of the inclusion and exclusion of patients should be added.
Validity of the findings: The findings were basically valid.
Additional comments: 1. I suggest to use "elevated /normal CA724", not the "positive/negative CA724".
2. capitalize the first letter of a Word.
3. The conclusion is too affirmative. This is only a small retrospective study, the evidence level is low.
4. CA724 is not only a biomarker for gastric cancer, but also for gastrointestinal tract, lung, breast, and ovaries.
5. The defect of the study should be added to the DISCUSSION part.
6. The first Table shoulde be Table 1, not Table 2.
7. What's the clinical significance of Figure 3?


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 PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: It is an interesting study. In clinical practise, we meet many cases which CEA is low but CA724 is high.It is valuable to explore the  preoperative CA724 with the prognosis for CRC.
Experimental design: It is ok , but I think caculate OS and DFS of other tumor mark than CA724 is not necessary.
Validity of the findings: Though CA724 is detected for patients in clinical practise for many years , such study is interesting and valuabl.
Additional comments: In abstract  section, "the OS and DFS in patients with positive CA724 was poorer than those
with negative."  "positive" and "negative" should altered to "high" and "normal",thus they may be consistent with the graphical description.The  headline of Table 1 was written Table 2 by mistake.


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 PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article seems good, but I recommend to send the paper to professional editing.
In the introduction section will be better to reduce the first paragraph because it contains too much general information. Also will be better to discuss more widely references 4 and 5. Line 52-I recommend paraphrasing it because the concentration of these markers can increase during other different diseases and pregnancy. Please also add information about CA 19-9 in combination with CEA.
Line 100 is empty, please check it.
It is impossible to understand Table 2. Please either split this table into 2 separate tables (OS and PFS) and separately show information about circulating markers.
In figure 2 please check commas in the title.
Experimental design: A research question is ok, but novelty is limited.
In methods section please provide information about tumor whether the patient was first diagnosed with a tumor or is it a relapse. Why the amount of metastatic patients is so small? Explain it, please.
Also in the abstract, you discuss the only R but use SPSS as well, It will be better to avoid this misunderstanding.
In the discussion section please consider hypermethylation of the Septin 9 gene as a diagnostic marker.
And is it possible to divide patients based on the left or right tumor localization?
Validity of the findings: All data seem reliable. The conclusion is robust but the discussion is not so detailed.
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 PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 1
Reviewer: 4

Output:
Basic reporting: This manuscript is well written. Literature references are updated but novel information considering more recent publications on the European Group on Tumour Markers (EGTM) guidelines will enrich the introduction.
Experimental design: In this manuscript, authors evaluated the prognostic value of pre-operative serum levels of the carbohydrate antigen CA72.4 in colorectal cancer (CRC) patients with normal CEA levels. Therefore, a series of 295 colorectal cancer patients was retrospectively analysed and the serum preoperative levels of CA72.4, CA19.9, CA125 and CA242 were correlated with patients clinicopathological information, namely overall survival and disease-free survival. The authors have concluded that preoperative serum levels of CA72.4 may serve, in CRC, as a potential prognostic factor in patients with normal CEA levels.

Methods were described in a concise way but major methodologies are annoted. Ethical agreements were considered.
Validity of the findings: This study was interesting and focused on an important question. In fact, the serum presence of carbohydrate antigens have for a long time been investigated, aiming to identify novel biomarkers of disease progression and, eventually, of therapeutic response. Several attempts were performed in distinct cancer types. In CRC, in particular, the European Group on Tumour Markers (EGTM) guidelines clearly suggested the use of CEA as a prognosis marker, especially in stage II patients, as postoperative surveillance on stage II and III patients, but also to monitor therapy response in advanced disease. Nevertheless, patients with normal CEA levels (generally over 50%) have no other sufficiently strong prognostic serum marker. The use of CA19.9 and CA242 as a prognostic marker have not been consensually recommended.

Data is based on a retrospective study using a cohort of 295 patients. Statistical analysis may require some considerations and revision. Based on the analysis performed, conclusions are well stated but validation on another population would be certain an improvement, although not demanding.
Additional comments: This manuscript is well- written and analyses, in a retrospective way, a considerable cohort of 295 patients with complete clinicopathological information. Inclusion criteria and ethical considerations were  well approached.  The preoperative expression levels of the selected serum markers in patients who underwent surgery were assessed through a chemiluminescence immunoassay. The document is well ome considerations are taken in relation to the statistics.
Some points require however some considerations:
-	The authors statistically analysed the expression of the distinct markers in relation to clinicopathological features using the Chi-square and not the Friedman’s test followed by intergroup comparisons with Wilcoxon test. The authors should better explain the reasoning that led to the  choose of the selected statistical tests.
-	It would be also interesting to indicate from the 295 patients that underwent surgical interventions to indicate those that had indeed relapsed.
-	In this series reduced metastasis are accounted (approx. 3%) is there any possible explanation?
-	It would be very important to stratify the cases not only in regard of TNM but also on clinical stage (I, II, III, IV) and to verify if any association with the expression of CA72.4 is emerging
-	Knowing that colon tumors are different entities regarding their location, it would be also very important to stratify them in regard tumor location as left and right colon. Other emergent information in CRC is the immunoscore. Although this was not on the scope of the manuscript, it would be very interesting it authors could provide additional information, and discuss it in integration with the present findings, regarding the inflammatory infiltrate present, since many these carbohydrates antigens are also antigens for immune cells.

Minor comments:
1)table 1 is miss designated table 2. Please correct
2) the word “the” is repeated in the last sentence of the section survival follow-up


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 PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: It has been improved after revision.
Experimental design: It is good design.
Validity of the findings: It is feasible.
Additional comments: It is an interesting study.


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

Input:
Title: THE PROGNOSTIC VALUE OF PREOPERATIVE SERUM CA724 FOR CEA-NORMAL COLORECTAL CANCER PATIENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: I still recommend professional editing and checking for errors.
Line 64 - references.
Line 79 - capital letters.
Line 95 and so on.
Also, I advise checking manuscript again.
Experimental design: There are no major changes since the previous submission.
Validity of the findings: Conclusions look reasonable.
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: SERUM MICRORNAS AS POTENTIAL NEW BIOMARKERS FOR CISPLATIN RESISTANCE IN GASTRIC CANCER PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: the article should include sufficient introduction as well as discussion , without repeating  the  same meaning
Experimental design: methods described with sufficient details

statistical analyses are very well.so, i think this manuscript must be reviewed by professor of statistic.
Validity of the findings: the results too long, contain details and conclusion, repeating the value present in the tables.
the results must be summarize
Additional comments: * IN abstract: conclusion must be clarify.
* the last paragraph in the introduction is not the aim but it is the methods.
* lines 59-66: there are repetition around the same meaning
* MGC803 cells were cultured with DDP in a continuous stepwise fashion for 15 months (please confirm, where is the references for that )  .
* this study include only one cell line MGC803  not two as mention.
*  line 85-86: mention had received DDP-based chemotherapy as86 first-line treatment; (3) had received at least 2 cycles of chemotherapy;
then mention in line 91:Prechemotherapy serum samples of patients who received neoadjuvant chemotherapy or palliative treatment were collected
                                         there is contrast.
* no prime sequences mention.
* housekeeping gene don,t mention under statistical analysis, must be mention under methods.
* discussion:  there is repetition the results, must be   more clarify.


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: SERUM MICRORNAS AS POTENTIAL NEW BIOMARKERS FOR CISPLATIN RESISTANCE IN GASTRIC CANCER PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: 1-	Abbreviations should be given in the first metion.
2-	In Table 1, log, FC and p values should be given as 4 digits after the comma.
3-	The target genes shown in Figure 3 should be given as supplementary material. For example, in Figure 3A, 25 genes, which are common to four different databases and the potential target of miR-9, should be given as supplementary tables.
4-	The “Above all, miR-519a-5p and miR-522-5p changed consistently with sRNA-seq, but the others showed the opposite results. Besides, miR-519a-5p and miR-522-5p showed the same result, for they had the same primer.” part in the Result section is not fully open. This section should be written more clearly and in detail. Also why are miR-370-3p, miR-519a-5p and miR-522-5p miRNAs not shown in GC patient serum samples? The results should be shown even if they are insignificant.
5-	Stars in tables are given next to P values, not need to be given in table footnote.
6-	Does Table 2 refer to GC patients or GC cell lines?
7-	“miR-9-3p (AUC = 0.824, SD = 0.047, 95%CI: 0.731-0.916, p<0.0001, SE 70.6%, SP 67.5%, Figure 5A), miR-9-5p (AUC = 0.856, SD = 0.042, 95%CI: 0.773-0.939, p<0.0001, SE 76.5%, SP 72.4%, Figure 5B), miR-146a-5p (AUC = 0.799, SD = 0.052, 95%CI: 0.697-0.900, p<0.0001, SE 64.7%, SP 62.5%, Figure 5C) and miR-433-3p (AUC = 0.838, SD = 0.045, 95%CI: 0.750-0.925, p<0.0001, SE 70.6%, SP 67.5%, Figure5D)” in the result section is already given in the figure and table. There is no need to be given in the text. Data not specified in figures and tables should only be given in the text.
8-	In Figure 9A, C and E, the p values should be indicated in the graph.
9-	Figure 10A, B, C, and D belongs to which miRNA should be indicated.


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: SERUM MICRORNAS AS POTENTIAL NEW BIOMARKERS FOR CISPLATIN RESISTANCE IN GASTRIC CANCER PATIENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I think this is a research which has significant meaning, I'm impressed by the significant difference of  the AUC value between combined group and miR-9-5p group, hopefully it may be used as a marker of chemoresistance of GC. But there were some problems needs to be correct before it can be accepted:
1.  In the abstract, I think you should list the specific AUC, P value and also the 95% CI in the "results" part.(line 33-38)
2. You have mentioned that "MicroRNAs (miRNAs) are a recently discovered..." in line 51, I don't think so, miRNAs have been discovered in the last century and have been extensively studied, so "recently" is not suitable.
3. In figure 9 and 10, I can find three types of survival plots, could you unify them?


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: SERUM MICRORNAS AS POTENTIAL NEW BIOMARKERS FOR CISPLATIN RESISTANCE IN GASTRIC CANCER PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
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: NOMOGRAMS FOR PREDICTING OVERALL SURVIVAL AND CANCER-SPECIFIC SURVIVAL IN YOUNG PATIENTS WITH PANCREATIC CANCER IN THE US BASED ON THE SEER DATABASE
Review round: 1
Reviewer: 1

Output:
Basic reporting: I suggest to change the tense of the first sentence of the abstract from "The incidence of young patients with pancreatic cancer (PC) was on the rise, and there was a lack of models that could effectively predict their prognosis." to "The incidence of young patients with pancreatic cancer (PC) is on the rise, and there is a lack of models that could effectively predict their prognosis."

Line 54-55, suggest changing "inadequate" to "insufficient".

Citation format needs reviewing throughout the manuscript e.g. lines 54-55, 59, 62.

A more detailed explanation of what a nomogram is would benefit readers.

Please change "COX risk regression model"  to "Cox regression model" or "Cox proportional hazards model", e.g. line 97-98.

A citation on how the C-index is defined or how it is calculated would also be beneficial for an average reader.
Experimental design: The selection criteria and exclusions are clearly stated.

There seems to be a typo of the diagnosis time in line 93, please clarify.
Validity of the findings: Can the authors comment whether they consider patients who died from a different cause other than PC as censored? In other words, is there a reason authors didn't use a more standard competing risk strategy for their CSS analysis?

Regarding multivariate analysis, further explanation on how the authors decided which variables to include would be appreciated. Specifically, did the authors use any kind of step-wise/p-value approach?
Additional comments: The authors conducted a thorough multi-center retrospective study to identify prognostic factors for OS and CSS in young patients with PC. Moreover, the authors utilize nomograms to ease interpretation in the clinical setting and beyond. I applaud the authors' effort to provide as much details as possible. However, I believe the manuscript can be further enriched by addressing the comments/suggestions I'm raising.


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: NOMOGRAMS FOR PREDICTING OVERALL SURVIVAL AND CANCER-SPECIFIC SURVIVAL IN YOUNG PATIENTS WITH PANCREATIC CANCER IN THE US BASED ON THE SEER DATABASE
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. In Figures 3 and 4, the x-axis labels, y-axis labels, x-tick labels and y-tick labels are not clear. It is strongly suggested that the authors adjust the labels appropriately.

2. In Line 149 of the Results Section, it is not intelligible in the description that “it also can be seen from the nomogram that AJCC stage had the greatest effect on prognosis.” Which OS does the “prognosis” denote, 1-year, 3-year, or 5-year OS? Please explain more details in how to interpret the nomograms and how to evaluate the effect of the variables on the specific prognosis outcome from the nomograms directly.

3. The authors should provide the raw data and the scripts (or instructions of point and click in SPSS) used to perform all analyses, including how to generate the nomograms and calibration curves.
Experimental design: No comment
Validity of the findings: 1. In Line 95 of the Statistical Analysis Section, the authors mentioned that the whole cohort was randomly divided into the training and validation groups. Do the authors try another random split of the whole cohort and evaluate the results? How about the results of 10-fold cross-validation? Are the current results consistent to the results of cross-validation?
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: NOMOGRAMS FOR PREDICTING OVERALL SURVIVAL AND CANCER-SPECIFIC SURVIVAL IN YOUNG PATIENTS WITH PANCREATIC CANCER IN THE US BASED ON THE SEER DATABASE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Change "univariate" for "univariable" throughout the manuscript
Change "mutivariate" for "multivariable" throughout the manuscript
Page 8, line 148: change "COX" to "Cox"
Page 9, line 179: change "competing models" to "competing risks models"
Page 11, line 215: change "precious" to "previous"
Experimental design: no comment
Validity of the findings: Using only significant results in univariable models into the multivariable models is an inadequate strategy for variable selection and is not recommended. The main reason is that this selection strategy fails to include potentially meaningful predictors or even well-known confounders if deemed non-significant. Thus, a more robust or clinically-guided model is often preferred. Have the authors analyzed a more comprehensive multivariable model with most (if not all) available factors in the SEER database to evaluate the prognostic value of this "comprehensive" model? I understand the rationale of constructing a more parsimonious model, however, the approach you used for variable selection may have left out potentially meaningful predictors such as gender in the CSS analysis. I believe that given the sample size in the database, fitting a slightly more complex model won't represent a major issue.
Additional comments: I appreciate the work the authors have put to improve their work during the first round of revision. Significant changes and refinements were made as requested. Considering the explanations on how the multivariable models were constructed major 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: NOMOGRAMS FOR PREDICTING OVERALL SURVIVAL AND CANCER-SPECIFIC SURVIVAL IN YOUNG PATIENTS WITH PANCREATIC CANCER IN THE US BASED ON THE SEER DATABASE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The description of the nomogram method is clear and straightforward.

The displaying of figures and tables is proper and clear to readers.
Experimental design: The selection criteria and exclusions are clearly stated.
Validity of the findings: Can the author describe the results of 10-fold cross-validation and the calibration between the actual probabilities and the plot of the nomogram in Line 116-118?
Additional comments: The authors suggest nomograms which can help interpret the results more easily in clinical setting. I appraise the authors' effort to explore the independent factors associated to PC using different analysis methods and to provide as much details as possible.


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: CLINICAL APPLICABILITY OF THE FELINE GRIMACE SCALE: REAL-TIME VERSUS IMAGE SCORING AND THE INFLUENCE OF SEDATION AND SURGERY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Generally well executed. See specific section within general comments for grammatical errors noted. Mention of previous cat pain assessment methods, particularly those focusing on facial expressions would be useful in order to supply sufficent background/context.
Experimental design: Some limitations regarding reproducibility of methods and potential design flaws due to coding bias – see comments in following section
Validity of the findings: During RT scoring, it seems that the coder may have had knowledge of the general posture and behaviour of the cat, their level of sedation and the relative time point of their assessment (i.e. pre or post op and time since surgery), as well as their SF-UBCPS  scores. Important controls to add here would be to assess inter-coder reliability of the still images that were coded, using a properly blinded coder. The second would be to get the blinded coder to code the FGS from the videos, using the same methods as FGS-RT scoring.
Additional comments: This is a worthwhile contribution to the field of clinical pain assessment based on facial expressions in cats. However, it makes it difficult to fully evaluate its merit when the efficacy of the main tool of measurement being tested (FGS) has yet to be published, and only very limited detail about the FGS and how it was performed is provided. As such the methods are potentially not easily reproduceable in their current state. I would recommend either delaying submission until the paper referencing the FGS is published, or include sufficient detail in the methods to support the FGS scoring method.

Additionally, there is a potentially substantial confound in the way that FGS coding was performed which could call into the question the validity of the main assumptions being made within the paper. The fact that there are potentially substantial biases present during RT coding, that the same person then coded the still images, and that the authors found a difference between RT and IMG scores limits the ability of this paper to properly demonstrate the clinical applicability of the FGS in real time. These issues should be addressed - please see previous comments for suggestions.

Specific section comments:

Abstract:
“Cats were video-recorded simultaneously at baseline, 15 minutes after sedation, and at 2, 12, and 24 hours after extubation for subsequent image assessment (FGS-IMG) performed six months later” – the last part of this sentence could do with a little rewording to explain the relevance of the 6 month image assessment

Introduction:

Ln 66: It would be really useful here to provide a brief example of the confounding effects found

Ln 69: This statement is not true entirely true – see Finka et al 2019

Ln 70: ‘there’ not ‘they’

Your first hypothesis suggests a possible design flaw due to the non-blind nature of image scoring

Your second hypothesis is reasonable, although it is not fully supported by the literature discussed in the introduction. If you are going to test a specific hypothesis in this way, I  would recommend providing sufficient evidence in advance to justify this

Methods:
The method of FGS scoring and video recording of cats needs further explanation to enable it to be easily replicated. It is not sufficient to state that methods followed those detailed elsewhere and to reference a paper that is currently in submission.
Please state whether the person preforming the RT FGS was blinded to the cat’s SF-UBCPS scores and relative scoring time point at which scoring took place as these may otherwise act as potential confounds.
Please also include information detailing the way scoring took place (i.e. was the scorer told only to look at the cat’s face, or were they able to observe other features of the cat and also perform other types of observation or clinical assessment on the cat?)
The linear mixed models could do with further explanation – e.g. how may separate models were performed, and the response and fixed effects/explanatory variables in each case. I’m struggling to understand how the outputs in table 2 relate to the LMMs as currently described. I would also suggest you briefly mention how your data met the criteria/assumptions for each of the statistical tests you performed in support of their selection.
Ln 211 – is there a suitable reference to support the conclusion of 'narrow LOAs' as stated?
Ln 222: Would it not make sense to analyse and report the differences in RT and IMG FGS at different time points compared to baseline in the same way here?
Ln 242 – surely (as supported by your results) surgery did influence action units obtained via FGS in line with induction of post-op pain, although I’m not sure this is the point you are trying to make here?

Discussion:
Ln 227: It would be worth to speculate the possible reasons for the over estimation of real-time scoring in one species and the underestimation in another, rather than just state that this is the case.
Paragraph 266: Is it also not possible that the difference in scores or ‘over estimation’ may be due to the RT scoring method (i.e. the scorer is not blind to the cat’s current condition, they have examined the cat, performed other composite pain assessment test and can observe the whole of the cat and their demeanour prior to FGS scoring?)
Ln 279: It’s great that you touch upon the lack of consensus regarding LOAs for pain scores and its clinical relevance to analgesic intervention
Ln 293: taken not taking
Ln 305: If the FGS scorer was not blinded to the sedation score of the cat, can you properly conclude that sedation scores did not affect FGS?


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: CLINICAL APPLICABILITY OF THE FELINE GRIMACE SCALE: REAL-TIME VERSUS IMAGE SCORING AND THE INFLUENCE OF SEDATION AND SURGERY
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: The article meets all of the adjacent criteria.
Validity of the findings: No comment
Additional comments: The authors describe a convincing test of the applicability of the FGS in a clinical setting, comparing both real-time and image-based assessment of an impressive number of cats. The results and conclusion appear to support  a previous 'reported' study ( Evangelista et al); but unfortunately this is not yet available. In numerous places this is described as 'submitted', but according to the conclusions section the article is 'in press'. Which is it ? Otherwise, in most respects the the article is very good one, but there are occasions when greater clarity might improve it. In particular, the reviewer was unclear about the timing of recording of each action unit of the FGS. Line 137. It is not clear what is meant by 'one score per minute of observation'. Does this mean all 5 FAU's were scored during minutes 1, 2 and 3? If so, please say this another way. Likewise, line 163 says one image per minute was selected does this mean the clip was paused at 1, 2 and 3 minutes, and the image clipped? I find it hard to believe that selecting these times would have avoided times when cats were not looking at the camera.  The is no doubt a simple explanation of this given greater clarity in the description. Again, on line 168, 'the average score of the 3 minutes' is unclear.
Again, I do not see the range of selected images (line 204) could include 0 or have a maximum of 20? Why did you not choose the minimum acceptable number for each cat and select from these randomly.? Perhaps this '0' value recognizes that 160 videos were unusable. This is a worryingly high number, and that fact should be  emphasized in the discussion as a general problem in off-line data analysis using any type of grimace scale.
Introduction line 76 and Discussion line 263. It is a little confusing when you say surgery did not influence FGS scores, when presumably the whole point of the investigation was determine that it did; ie caused pain that can be detected with the FGS. Similarly, In the introduction  why would you hypothesize that FGS scores would not be elevated by 24 hours compared with baseline? This suggests you thought it unlikely that the effects of OVH would not extend beyond that time?  Why? -  This arises again on line 320 where you say OVH did not affect FGS scores, when clearly it did.  Please clarify these statements or re-state the original hypothesis on this aspect if the reviewer has misunderstood. Line 274, in contrast to what?
Overall, this article is ethically sound and well presented and the outcomes seems clear and worthy of publication after resolution of these relatively small issues. However, Please state whether all cats were successfully re-homed or what proportion had to be euthanased, if any.

Other minor points: Introduction, line line 61, this is usually called the Mouse Grimace Scale. Line 129, what is meant by weight was redistributed to the other action units? I am assuming this means if only 4 units were scored, then the combined total was divided by 4 and not 5. Please clarify.


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: CLINICAL APPLICABILITY OF THE FELINE GRIMACE SCALE: REAL-TIME VERSUS IMAGE SCORING AND THE INFLUENCE OF SEDATION AND SURGERY
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Thank you for taking the time to address the various comments - the clarity of the manuscript and its methodology is improved.

A few outstanding grammatical issues:

Ln 29 “Cats were video-recorded simultaneously at baseline, 15 minutes after sedation, and at 2, 6, 12, and 24 hours after extubation for subsequent image assessment (FGS-IMG) performed six months later by the same observer” – suggest adding in ", which was" before the word 'performed' for ease of reading

Ln 254 Ovariohysterectomy did not influence any action unit scores obtained using either FGS-RT or FGS-IMG. -  this sentence still needs attending to – wasn’t the whole point of your study to show exactly this?! I think the point you want to make is that after pain was abated, the surgical intervention in itself didn’t appear to affect FGS…this issue of wording also still needs addressing in the discussion