Application of attenuated total reflection–Fourier transform infrared spectroscopy in semi-quantification of blood lipids and characterization of the metabolic syndrome

Tz-Ping Gau; Jen-Hung Wen; I-Wei Lu; Pei-Yu Huang; Yao-Chang Lee; Wei-Po Lee; Hsiang-Chun Lee

doi:10.1371/journal.pone.0316522

Peer Review History

Original SubmissionApril 14, 2024
8 May 2024 Decision Letter - Muhammad Iqhrammullah, Editor PONE-D-24-11405Application of Attenuated Total Reflection–Fourier Transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of the metabolic syndromePLOS ONE Dear Dr. Lee, 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 improve the novelty statement and sampling handling explanation. ============================== Please submit your revised manuscript by Jun 22 2024 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, Muhammad Iqhrammullah, Ph.D Academic Editor PLOS ONE Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Please note that PLOS ONE has specific guidelines on code sharing for submissions in which author-generated code underpins the findings in the manuscript. In these cases, all author-generated code must be made available without restrictions upon publication of the work. Please review our guidelines at https://journals.plos.org/plosone/s/materials-and-software-sharing#loc-sharing-code and ensure that your code is shared in a way that follows best practice and facilitates reproducibility and reuse. 3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. 4. In this instance it seems there may be acceptable restrictions in place that prevent the public sharing of your minimal data. However, in line with our goal of ensuring long-term data availability to all interested researchers, PLOS’ Data Policy states that authors cannot be the sole named individuals responsible for ensuring data access (http://journals.plos.org/plosone/s/data-availability#loc-acceptable-data-sharing-methods ). Data requests to a non-author institutional point of contact, such as a data access or ethics committee, helps guarantee long term stability and availability of data. Providing interested researchers with a durable point of contact ensures data will be accessible even if an author changes email addresses, institutions, or becomes unavailable to answer requests. Before we proceed with your manuscript, please also provide non-author contact information (phone/email/hyperlink) for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If no institutional body is available to respond to requests for your minimal data, please consider if there any institutional representatives who did not collaborate in the study, and are not listed as authors on the manuscript, who would be able to hold the data and respond to external requests for data access? If so, please provide their contact information (i.e., email address). Please also provide details on how you will ensure persistent or long-term data storage and availability. Additional Editor Comments: Please improve the novelty statement and sampling handling explanation. [Note: HTML markup is below. Please do not edit.] 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: Partly Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: No Reviewer #2: Yes ****** 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 ****** 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: Yes ****** 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: 1. After carefully reviewing the introduction section, I did not find a clear and explicit statement highlighting the novelty or newness of the study. The authors do not appear to directly state what makes their work novel compared to previous research in the field. 2. The authors emphasize the advantages of their ATR-FTIR method, such as rapid assay times and the need for small blood samples (0.5 μL). These benefits make the method potentially better for frequent monitoring and clinical applications compared to established techniques. Unfortunately, the authors lack a comprehensive comparison of the accuracy, sensitivity, and specificity of their approach against established methods. A deeper analysis of the performance metrics and potential limitations of their technique, in relation to existing gold standards, would better support their case for the usefulness and innovation of their method. 3. The conclusion should explicitly reiterate the potential impact of early detection and management of MetS on improving patient outcomes and quality of life, as highlighted in the introduction. 4. What do you mean by the brackets in table 2 as standard deviation (SD)? 5. However, there are a few minor points that could be improved in the reporting of the SHAP analysis: The authors could provide more details on how exactly the SHAP values were calculated (e.g., using the TreeExplainer for tree-based models like GBT) and what they represent (e.g., the contribution of each wavelength to the model's output relative to the average prediction). They could also discuss any potential limitations or caveats of using SHAP for model interpretation in this specific context. 6. From a purely statistical perspective, an R2 value of 0.419 indicates that only 41.9% of the variance in LDL-C levels can be explained by the ATR-FTIR spectral data using the HGBT model. This suggests that the model's predictions for LDL-C are not as accurate as those for other lipid parameters like TG (R2 = 0.854) or HDL-C (R2 = 0.758). In many scientific contexts, an R2 value below 0.5 would be considered poor model performance. I'm uncertain about the authors' argument, which claims that despite the low R² value, the mean absolute error (MAE) for LDL-C prediction was between 9 and 12 mg/dL. Reviewer #2: Interesting work that discusses the application of ATR-FTIR in biological samples. The study is needed and the use of AI algorithms added value. Just some comments are needed to clarify: -How were the samples stored? -How did the authors ensure stability of samples? -Was any heating of the sample applied prior to measurement? To prevent water interference? -Comment on transferability of the models to other matrices in the discussion ****** 6. 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. https://doi.org/10.1371/journal.pone.0316522.r001
Revision 1
28 Jul 2024 Author Response 28-July-2024 Editor-in-Chief PLOS ONE Dear Editors: We would like to thank all the reviewers and editors for their comments on the manuscript (PONE-D-24-11405) entitled “Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndrome”. Following the reviewers’ comments, we revised the manuscript and provided responses to describe the manuscript amendments. Point-to-point responses have been included in this submission. We apologize that the grant information we provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. One of our authors forgot to add a Financial Disclosure Statement before the primary submission. We received funding to support this study and updated the Funding Information on the Submission page as follows: National Health Research Institutes (NHRI-EX107-10724SC, NHRI-EX108-10724SC, NHRI-EX109-10724SC, and NHRI-EX110-10724SC), Kaohsiung Medical University Hospital (KMUH111-1R07 and KMUH110-0R11), Taiwan Ministry of Science and Technology/National Sciences, and Technology Council Grants (MOST 109-2314-B-037-111-MY3) NSTC 112-2314-B037-069. Please change the online submission form on our behalf. Thank you. Regarding raw data sharing, all authors have decided on a data-sharing plan, and our entire data will be made freely accessible if our manuscript is accepted for publication. We will adhere to your open-data policy. To meet your requirement that all data presented in the study be made publicly available at or before the time of acceptance, the file named “encoded_data” is uploaded to the Submission System as a Supporting Information. We believe that, after incorporating the valuable reviewers’ and editors’ opinions, the manuscript is worthy of consideration for publication in PLOS ONE. Thank you for your consideration. Sincerely yours, Hsiang-Chun Lee, MD, PhD Professor, College of Medicine, Kaohsiung Medical University Attending Physician, Division of Cardiology, Department of Internal Medicine, Kaohsiung Medical University Hospital 100 Tzyou 1st Rd, Kaohsiung 807, Taiwan Tel.: +886-7-3121101#7741 Fax: +886-7-3234845 E-mail: hclee@kmu.edu.tw Attachments Attachment Submitted filename: Specific Responses to Reviewers.docx https://doi.org/10.1371/journal.pone.0316522.r002
5 Nov 2024 Decision Letter - Rajesh Kumar Singh, Editor PONE-D-24-11405R1Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndromePLOS ONE Dear Dr. Lee, 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 Dec 20 2024 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, Rajesh Kumar Singh, 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. 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 #3: All comments have been addressed Reviewer #4: 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 #3: Yes Reviewer #4: (No Response) ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #3: Yes Reviewer #4: 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 #3: Yes Reviewer #4: 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 #3: Yes Reviewer #4: 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 #3: The study based on Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndrome is highly significant. However I would suggest to authors please expand this study by using molecular methods. Reviewer #4: • Provide additional context on ATR-FTIR: Add a brief explanation about how ATR-FTIR spectroscopy works, specifically for readers less familiar with the technique. This could improve the accessibility of the study’s technical approach. • Add a visual comparison: Consider adding a figure that visually compares the ATR-FTIR spectrum with traditional spectra of individual lipid classes or a figure showing predicted vs. actual lipid levels for better visual interpretation. ****** 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 #3: No Reviewer #4: 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. https://doi.org/10.1371/journal.pone.0316522.r003
Revision 2
10 Dec 2024 Author Response PONE-D-24-11405R1 Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndrome PLOS ONE Comments to the Author Reviewer #3: The study based on Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndrome is highly significant. However I would suggest to authors please expand this study by using molecular methods. Response: Thank you for your comments. We agree that molecular methods are also very important for applying non-destructive ATR-FTIR spectrometry in serum analysis, particularly for lipids. The following studies demonstrate the use of ATR-FTIR spectrometry to analyze blood samples. Bel’skaya et al. published their work on using model solutions in applying FTIR spectroscopy for the quantitative analysis of blood serum.[32] Portaccio et al. published their study results on ATR-FTIR spectra of commercial lipid samples and demonstrated specific fingerprint regions in ATR-FTIR spectra for phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol) from 3500 to 2800 cm−1 and 1800 to 600 cm−1), sphingolipids (ceramide, sphingosine 1-phosphate, ceramide 1-phosphate, and sphingomyelin) from 3500 to 2800 cm−1 and 1800 to 600 cm−1), and cholesterol (3150–2800 cm−1) [33]. Their study proved the usefulness of FTIR spectra for analyzing complex lipids, and this application can ultimately contribute to lipid research and its implications for human health. (Line 319-328) Based on the known advantage of FTIR spectra in quantitative analysis and sensitivity to differentiate various lipids, our study approach was directed toward linkage to clinical application. Consistent with the aforementioned literature, our results revealed the reliability of ATR-FTIR for semi-quantitative lipid analysis of human serum samples. Unlike those publications, our study emphasizes the predictive capability of machine learning models. The core concept is to uncover the correlations between FTIR spectra, clinically derived lipid values, and diagnostic classifications. Since this approach centers on data-driven correlations rather than direct molecular causation, it inherently lacks molecular specificity, which remains a limitation of ML-based predictions. In future work, we may explore causation-focused models to investigate molecular associations further and validate these predictions. However, given the study’s current scope, our ML approach offers valuable insights into broad lipid level trends without requiring the extensive molecular characterization that lies beyond this study’s objectives. (Line 380-384) Two new citations have been added. 32. Bel’skaya LV, Sarf EA, Solomatin DV. Application of FTIR Spectroscopy for Quantitative Analysis of Blood Serum: A Preliminary Study. Diagnostics. 2021;11(12):2391. PubMed PMID: doi:10.3390/diagnostics11122391. 33. Portaccio M, Faramarzi B, Lepore M. Probing Biochemical Differences in Lipid Components of Human Cells by Means of ATR-FTIR Spectroscopy. Biophysica. 2023;3(3):524-38. PubMed PMID: doi:10.3390/biophysica3030035. Reviewer #4: • Provide additional context on ATR-FTIR: Add a brief explanation about how ATR-FTIR spectroscopy works, specifically for readers less familiar with the technique. This could improve the accessibility of the study’s technical approach. Response: Thank you for your comments. We have added a paragraph to the Introduction and Methods section to briefly explain how ATR-FTIR spectroscopy works. The context is as follows. ATR-FTIR spectroscopy is a technique for analyzing the chemical composition of materials by detecting their characteristic absorption of mid-infrared light. In ATR-FTIR, molecules absorb mid-infrared light at specific wavenumbers (or frequencies), depending on their chemical bonds and structure. Each type of bond (e.g., C=O and C–H) has a unique absorption pattern, creating a molecular fingerprint. (Line 77-81) This study applied a small sample volume (0.5 µL to a Ge ATR crystal, as shown in Figure 1(A). A modulated mid-infrared beam was directed into the Ge crystal, generating an evanescent wave at the crystal surface, allowing the light to interact with the sample. The sample absorbs light at characteristic wavenumbers, and the remaining light is detected using a liquid nitrogen-cooled mercury cadmium telluride (MCT) infrared detector. The resulting spectrum shows peaks corresponding to specific molecular bonds, providing a unique chemical profile of the sample. (Line 122-127) Figure 1(A). The schematic depicts the attenuated total reflection for acquiring the FTIR spectra of a blood sample. The mid-infrared light is modulated using the Michaelson interferometer of an FTIR spectrometer. (Line 142-144) • Add a visual comparison: Consider adding a figure that visually compares the ATR-FTIR spectrum with traditional spectra of individual lipid classes or a figure showing predicted vs. actual lipid levels for better visual interpretation. Response: Thank you for your comments. To illustrate the ability of spectral values to quantify traditionally categorized lipid classes (e.g., cholesterol and triglycerides), we categorized lipid values into five equally divided groups from maximum to minimum to examine potential spectral differences among varying lipid levels (see Figures below). The X-axis represents the input ATR-FTIR wavenumbers, the Y-axis represents the input ATR-FTIR spectral values, and the color group corresponds to the measurement group obtained from conventional biochemical assays. The average ATR-FTIR spectrum for each group is shown as a line, with the shaded regions representing plus one standard deviation. This visualization highlights the trends in spectral differences across lipid groups. However, although these trends suggest potential relationships, they do not allow for direct quantitative conclusions or inferences. Overlapping or minimal differences in FTIR spectral absorption across lipid ranges further suggest that nonlinear associations may exist that are not directly observable in raw spectral data. By applying machine learning models, we can efficiently capture and interpret these complex patterns, offering precise and meaningful lipid-level predictions beyond what is visually discernible. All figures are listed in Appendix 2. Grouped Spectrum with Specified Lipid Value Ranges (Line 211-223) In response to your suggestion, we have also included Figure 2 to illustrate the comparison of predicted versus actual lipid levels. In this figure, the x-axis represents the lipid values predicted by the model based on the input ATR-FTIR spectral data, whereas the y-axis shows the actual measured lipid values. Additionally, we included a visualization of the distribution of error values to further elucidate the prediction accuracy and deviations of the model. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0316522.r004
13 Dec 2024 Decision Letter - Rajesh Kumar Singh, Editor Application of attenuated total reflection–Fourier transform Infrared spectroscopy in semi-quantification of blood lipids and characterization of metabolic syndrome PONE-D-24-11405R2 Dear Dr. Lee, 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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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, Rajesh Kumar Singh, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0316522.r005
Formally Accepted
Acceptance Letter - Rajesh Kumar Singh, Editor PONE-D-24-11405R2 PLOS ONE Dear Dr. Lee, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. 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. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Rajesh Kumar Singh Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0316522.r006

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .