PONE-D-21-07877

A study of sodium carboxymethyl starch with high substitution degree in defecation

PLOS ONE

Dear Dr. Cao,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Kind regards,

Zongxin Ling

Academic Editor

PLOS ONE

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2.Thank you for stating the following in the Financial Disclosure * (delete as necessary) section:

"This research was financially supported by Lipont Pharmaceutical Co. (China). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: No

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: I have not seen before such application of starch. seems interesting. Revision is suggested. Please see my important comments below

1-ToC graphical abstract could be added to improve the manuscript.

2-Figures (e.g., Fig. 3, 4, 5, 6, 8B): Please do not use Time new Roman font. Please use more readable fonts such as Helvetica and Arial. Please modify the figures throughout the manuscript. Also, some fonts may need to be enlarged to be more readable.

3-The color of fig 7 and 8 A are not visible well. The graph would be better to be colorful + having pattern. Therefore, it will be suitable for both B/W and colorful printing.

4- Fig 1 should be deleted. instead, A schematic illustration is needed on the reaction/preparation of the system to help the readers to understand the process.

5-References: There are some old references. Please remove/replace the outdated references. Some of them are listed below but please replace the other references.

Meijer-Severs GJ, Cats A, Verschueren RCJ, Santen EV, Kleibeuker JH. Anaerobes and their fermentation products in faeces of patients with familial adenomatous -

polyposis before and after subtotal colectomy and ileorectal anastomosis. European journal of clinical investigation. 1993 Jun; 23(6):356-60. https://doi.org/10.1111/j.1365-2362.1993.tb02036.x. PMID: 8393794. eng.

Richard K LL, Brown IL, Hu Y, Morita T, Esterman A, Young GP. Effect of dietary resistant starch and protein on colonic fermentation and intestinal tumourigenesis

in rats. Carcinogenesis. 2007 Feb; 28(2):240-245. https://doi.org/ 10.1093/carcin/bgl245 PMID: 17166881. Epub 2006/11/13.eng.

Englyst HN, Cummings JH. Digestion of the polysaccharides of some cereal foods in the human small intestine. Am J Clinic Nutr. 1985 Nov; 42(5):778-787.

https://doi.org/10.1093/ajcn/42.5.778 PMID: 2998174. Epub 1985/11/01.eng.

Meijer-Severs GJ, Cats A, Verschueren RC, Santen EV, Kleibeuker JH. Anaerobes and their fermentation products in faeces of patients with familial adenomatous polyposis before and after subtotal colectomy and ileorectal anastomosis. Europe J Clinic Invest. 1993 Jun; 23(6):356-60. https://doi.org /10.1111/j.1365-

2362.1993.tb02036.x PMID: 8393794.eng

6-There are many errors in the references.

7-Introductio needs to be revised carfully. It was not written well, the aim of it is not apparent. What is the problem and potential disease of defecation? It should be added. Also no need of use abbreviation of Resistant starch (RS). updated ref should be added into introduction, e.g., Maspes et al, Gels 7 (1), 6, 2021;; Makvandi et al, Environmental Chemistry Letters, 1-29, 2021;;

8-Conc,usion should be elaborated.

Reviewer #2: This paper describes the effect and potential mechanism of CMS Na in prevention and treatment. The main problem is to clarify the relationship between the causes and results of different animal models. At the same time, it needs further improvement in writing and grammar.

1. Page 1, the title of this manuscript should more accurately reflect the core content of the research.

2. Page 2, “Food additive, Sodium carboxymethyl starch” should be “Food additive, sodium carboxymethyl starch”. “gas chromatogra- phy” should be “gas chromatogra- phy”. “the proportion of Alloprevotella” should be “the relative abundance of Alloprevotella”, so were those in the other parts of this manuscript. Which kind of the biodiversity should be pointed out.

3. Page 2, from the current data, the author cannot get the conclusion “by regulating Alloprevotella and Lactobacillus in the colon, increasing short-chain fatty acids and promoting the synthesis of Tph1 and 5-HT”. The mechanism may be only related to the above observations.

4. Page 4, Figure 1 should be integrated into other figures. Figure 2 is unscientific and should be deleted.

5. In the method and results section, why did the author build the same model with rats and mice? It is more important to clarify why different experiments of the same study, such as water content, fecal pH, short chain fast acid, 16S rDNA sequence analysis TPH1 and 5-HT in serum, were performed in rats or mice, rather than on the same animal. This makes the whole article more confusing, and it is difficult to analyze the correlation between the results.

6. In the whole result section, the author should make the result title more accurately reflect the important content of each section, rather than a general description.

7. Page 10, The following description is not clear. “The first 2,000,000 reads of the document were counted. Reads with a length > 75 BPwere truncated to 50 BP for statistical duplication.” It is suggested to modify the description and put it into the method. In addition, it is better to describe how many samples have been tested and how many clean reads each sample has.

8. Page 11, if there is no significant change at the phylum level, it is unnecessary to describe. It is suggested to add the description of the changes of the Family level. The increase or decrease should be described as those of the relative abundance of a particular bacterial taxonomy. Family, genes should be italicized.

9. Page 15, this conclusion is not accurate. The effect of CMS Na alleviates con stitution is only related to that described later such as regulating the proportion of the anaerobic bacteria Alloprevotella and Lactobacillus in the colon, increasing short-chain fatty acids, and promoting the synthesis of Tph1 and 5-HT., and its causality has not been verified.

10. In the figures，such figure 5 and 10, don't mark A when there is only one panel.

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Reviewer #1: No

Reviewer #2: No

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PONE-D-21-07877R1

The effect of carboxymethyl starch sodium with high substitution degree on defecation

PLOS ONE

Dear Dr. Cao,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Aug 14 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Zongxin Ling

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: (No Response)

Reviewer #2: No

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The quality of the figures seems low. Some fonts are too big or in some case small.

The authors claimed to address the references, but it seems that the references were not implemented.

Reviewer #2: The authors have adequately addressed our comments raised in a previous round of review. However, before be published, this manuscript should be carefully checked for grammatical errors. Furthermore, the clarity of all figures needs to be improved. Usually, authors should provide pictures (300DPI) in tiff format.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

The effect of sodium carboxymethyl starch with high degree of substitution on defecation

PONE-D-21-07877R2

Dear Dr. Cao,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Zongxin Ling

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: (No Response)

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #2: The author has made detailed modifications to the paper. Before publishing, you need to carefully calibrate full text. For example, the “aim” inside the summary is written as “arm”.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Pooyan Makvandi

Reviewer #2: No