PONE-D-24-46321Polymorphisms in rs2069845 are associated with IL-6 and soluble IL-6 receptor levels during total joint replacementPLOS ONE

Dear Dr. Ross,

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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: Yes

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

Reviewer #1: Yes

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: No

Reviewer #2: No

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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

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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: The manuscript entitled “Polymorphisms in rs2069845 are associated with IL-6 and soluble IL-6 receptor levels during total joint replacement” is a well-written, interesting topic, some modifications are needed.

1- The language/grammar/syntax needs to be edited in several areas especially the introduction and some of the methods as it was hard to understand in some areas.

2- Please italicize the gene throughout the manuscript.

3- What was the biological rationale for the selection of these SNPs?

4- Please insert a paragraph of sample size in the methodology section, please include the study power and a short description of how it was calculated.

5- How much is the MAF? Please include it in the method section

6- The Hardy-Weinberg equilibrium should be performed considering the control and subjects population

7- In discussion, please provide focused description on the effects of the SNPs

8- Please, move the paragraph of ethical approval with the IRB number from the acknowledgment section to the methodology section.

Reviewer #2: Manuscript Number PONE-D-24-46321

Authors of the manuscript entitled “Polymorphisms in rs2069845 are associated with IL-6 and soluble IL-6 receptor levels during total joint replacement” have duly evaluated the three SNPs present in the promoter region of IL-6 gene and checked the dynamics with circulating IL-6 and sIL-6r level in pre- & post-operative conditions. Overall, the manuscript seems technically good. However, I have the following comments on the manuscript

Introduction:

1) Line no. 62 “several cytokines …… late-stage biomarkers” authors should mention the other cytokines.

2) why rs2069845 has been used by the author (reported for leprosy)?

Materials and method:

1) What sample size was predicted when the authors performed power analysis? Please mention the test details, sample size, and version of the Quanto power calculation tool.

2) Three SNPs are mentioned in the line no. 85 of the introduction, but it has been changed to four SNPs in line no. 103.

3) Why age and gender-matched control has not been taken.

4) Line no. 116 - initials of Lead investigators are not necessary.

5) Rewrite IL6-R (line no. 132) as IL-6r

6) The author should include either an abbreviation table in the beginning or expand the acronyms used in the manuscript wherever it is mentioned for the First time for example, TKR (Line no. 157); and THR (Line no. 157).

General comments:

1) Are there SNPs reported from other cytokine genes related to bone resorption after TJR?

2) What are the other biomarkers/proteins that influence IL6/IL6 receptor binding?

3) Besides implant material, what other possible factors that cause intrinsic sensitivity of patients also need to be mentioned?

4) Legends for figures 1 and 2 should have the p-value.

5) Provide footnote for tables 2,3, 4, and 5

6) Authors need to give details of statistical tests in Tables for clarity

7) Limitations of the study should be incorporated after the discussion.

8) The authors need to improve the resolution of all the figures

9) Discussion and conclusion require a systematic writing

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Reviewer #1: No

Reviewer #2: No

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PONE-D-24-46321R1Polymorphisms in rs2069845 are associated with IL-6 and soluble IL-6 receptor levels during total joint replacementPLOS ONE

Dear Dr. Ross,

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 Apr 13 2025 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,

Nan Jiang

Academic Editor

PLOS ONE

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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: All comments have been addressed

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: Partly

**********

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

Reviewer #1: Yes

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

**********

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: Yes

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:  Thanks alot for your efforts, in doing all required modifications

All comments have been addressed, and the manuscript is valid or publication in the current form

Reviewer #2:  All necessary corrections have been addressed by the authors. Nonetheless, I have a few additional suggestions that will further improve the manuscript.

1. In line 153, there is a typo that needs to be corrected. Please change "bdSNP" to "dbSNP.".

2. The minor allele frequency in line 154 should be rechecked using the 1000 Genomes database or dbSNP for the European population rather than the global population, followed by SNP analysis. For instance, the ref. allele “C” of rs4537545 has a frequency of 0.592611 in the European population (rs4537545 RefSNP Report - dbSNP - NCBI), whereas in figure-3, very few individuals are homozygous CC, with low mean pre-operative IL-6 level. Therefore, normally, SNP-based studies or analyses should not be done with a low sample size.

3. What tests are used for Hardy-Weinberg equilibrium analysis of SNPs in R-Studio? Please mention the test name in the methodology section.

4. The SNPs discussed in the article are located within introns (rs4537545 RefSNP Report - dbSNP - NCBI ; rs2069845 RefSNP Report - dbSNP - NCBI) and missense variant (rs2228145 RefSNP Report - dbSNP - NCBI), therefore, to strengthen these findings, the authors should conduct in-silico analyses to demonstrate how these SNPs regulate IL6 levels.

5. Authors can utilize various online tools available on the UCSC browser for this purpose. This approach would also differentiate their manuscript from previous studies focusing on these variants (e.g., reference 25).

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Reviewer #1: Yes:  Sally Mohammed El-Hefnawy

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.

Polymorphisms in rs2069845 are associated with IL-6 and soluble IL-6 receptor levels during total joint replacement

PONE-D-24-46321R2

Dear Dr. Ross,

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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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,

Nan Jiang

Academic Editor

PLOS ONE

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 #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 #2: Yes

**********

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

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 #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 #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 #2: Authors have made an effort to include all the suggested comments. The Manucsript is now technically sound and meets the standards of Plos One journal, so 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 #2: No

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