PONE-D-22-04274Relationship of tissue ghrelin levels with disease activity and laboratory parameters in ulcerative colitis patients; immunohistochemical studyPLOS ONE

Dear Dr. Sahin,

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 make sure, that in the abstract as well as in the main manuscript the same definitions and patient groups are addressed (see reviewer 1). One reviewer asks for the inclusion of a long list of additional literature. Please make sure that the cited literature adds to your introduction and discussion to avoid a lengthy literature list.Reviewer 2 asks why only UC and no CD patients have been included which might be an interesting question. Please add relevant therapies into your description of the patients. Please make sure that the discussion is based on your data and avoid speculations.

Please submit your revised manuscript by Jul 13 2022 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.

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PLOS ONE

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Reviewers' comments:

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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: Partly

Reviewer #2: Partly

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

Reviewer #1: N/A

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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

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Reviewer #1: No

Reviewer #2: Yes

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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: Manuscript ID: AJGASTRO-D-21-00058

Title: Relationship between the tissue ghrelin presence, disease activity and laboratory parameters in ulcerative colitis patients; immunohistochemical study

Authors: Memduh Sahin et al.

The aim of above manuscript was to determine the relationship between the presence of ghrelin in the colon and the severity of ulcerative colitis. The topic of the study is interesting; however, it should be noted out that the manuscript contains many errors and shortcomings. For this reason, the manuscript must be corrected before possible acceptance for publication.

List of shortcomings and errors:

1. First of all, the authors check correctness of description of groups of people participating in the study and the presented results. In the abstract, Materials and Methods, and Table 2, the authors stated that group A contains the patients with ulcerative colitis without the presence of ghrelin staining in biopsy samples; whereas patients with ulcerative colitis and presence of ghrelin staining in biopsy samples were gathered in group B. In addition, the authors stated that Mayo score/disease activity index (DAI) is patients with ulcerative colitis was significantly higher in group A than group B (abstract and Table 2). Final conclusions at the end of the manuscript are in agreement with mentioned above data. On the other hand, the authors present opposite conclusion in the abstract that “Colonic ghrelin staining in UC patients seems to be associated with the increased activity of this disease”. Moreover, the authors stated that “DAI were found to be higher in UC patients with showing ghrelin positivity (Group A) (Discussion, paragraph 3). Inconsistences in the conclusions should be checked and corrected.

2. All abbreviations should be presented in full form at the place where they are used for the first time in the abstract and repeated in the main body of the manuscript.

3. Introduction. The authors should write that in the gut, the protective and/or therapeutic effect of ghrelin was demonstrated in different organs (PMID: 34638910), including among others, in the oral cavity (PMID: 24304579; PMID: 29151078), pancreas (PMID: 14726611; PMID: 17084100; PMID: 25594510), stomach (PMID: 24622834) and duodenum (PMID: 19439811).

4. The authors should write that previous studies showed that ghrelin exhibit the beneficial effects in different experimental models of colitis. Among others, the protective and/or therapeutic effect of ghrelin was observed in experimental colitis evoked by acetic acid (PMID: 26769837; PMID: 27598133), dextran sodium sulfate (PMID: 26713317)

5. Major revision of English is necessary.

Reviewer #2: It is not clear how ghrelin is implicated in UC pathogenesis. Why authors choose only UCand no CD patients. Did the patients received biological agents?

It is very simple approach to have only immunohistochemistry data.

What can be the relation of grelin expression and biological therapies

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Reviewer #1: No

Reviewer #2: No

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PONE-D-22-04274R1Correlation between the tissue ghrelin presence, disease activity and laboratory parameters in ulcerative colitis patients: immunohistochemical studyPLOS ONE

Dear Dr. Sahin,

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.

One of the reviewers still raised some comments which should be addressed before sending the final version. Please have in mind that PLOS does not provide proof reading, thus please assure to send your manuscript in proper English. I also agree with the reviewer that an exemplary image showing the staining for ghrelin is desirable; however, it is your decision to present such an image or not.

Please submit your revised manuscript by Oct 02 2022 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: https://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,

Gernot Zissel, Ph.D.

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: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: N/A

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: Yes

Reviewer #2: Yes

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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

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: Manuscript ID: AJGASTRO-D-21-00058

Title: Relationship between the tissue ghrelin presence, disease activity and laboratory parameters in ulcerative colitis patients; immunohistochemical study

Authors: Memduh Sahin et al.

The second version of the manuscript exhibits some improvement, but there are still some errors and deficiencies.

List of shortcomings and errors:

1. Authors should proofread English in their manuscript. Even in the newly introduced changes, there are errors. For example, in response to the firs comment, the authors wrote “Disease activity detected in the group with positive ghrelin staining, was found to be statistically lower than the negative stained group”. Between “than” and “the negative stained group”, the authors should add “in”.

2. The second comment was “All abbreviations should be presented in full form at the place where they are used for the first time in the abstract and repeated in the main body of the manuscript”. Authors stated that the manuscript was revised as suggested by the reviewer. However, there are still abbreviations in the abstract and next parts of the manuscript that are not presented in full name. For example, ESP, CRP. Authors should check the entire manuscript. In addition, the authors should specify in which fluids they determined albumin concentration, as well as in Material and Methods, they should present the methodology used in biochemical tests and blood counts.

3. In the previous version of the manuscript, the authors presented exemplary images showing the staining of colonic mucosa for the presence of ghrelin. There are no such images in the current version of the manuscript. They should appear in the final version of the manuscript,

Reviewer #2: the authors covered the reviewer comments. The manuscript has been improved and it can be accepted for publication

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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

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[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.

Correlation between the tissue ghrelin presence, disease activity and laboratory parameters in ulcerative colitis patients: immunohistochemical study

PONE-D-22-04274R2

Dear Dr. Sahin,

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.

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Kind regards,

Gernot Zissel, Ph.D.

Academic Editor

PLOS ONE

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