Large-scale statistical dissection of sequence-derived biochemical features distinguishing soluble and insoluble proteins

Nguyen Huy Hoang Vu; Bao Long Nguyen

doi:10.1371/journal.pone.0344883

Peer Review History

Original SubmissionFebruary 26, 2026
25 Mar 2026 Decision Letter - Anil Bhatia, Editor PONE-D-26-09831Large-Scale Statistical Dissection of Sequence-Derived Biochemical Features Distinguishing Soluble and Insoluble ProteinsPLOS One Dear Dr. Nguyen Bao, 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 May 09 2026 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 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. 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Only the individual author can complete the verification step; PLOS staff cannot verify ORCID iDs on behalf of authors. We look forward to receiving your revised manuscript. Kind regards, Anil Bhatia, 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 If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. [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 Reviewer #3: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: 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: No Reviewer #2: Yes Reviewer #3: 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: No Reviewer #2: Yes Reviewer #3: 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: I recommend that the editor request major revisions for this manuscript. The authors should address the following concerns: 1. Incorrect reference numbering: The citation format is inconsistent throughout the manuscript. For example, on page 1, line 4, the reference is cited as [1, 6], whereas it should appear sequentially (e.g., [1, 2]). The authors are advised to use reference management software such as EndNote, Zotero, or similar tools to correct and standardize citations across the entire manuscript. 2. Lack of sequence data transparency: The authors state that a total of 36 sequence-derived biochemical features were computed for each protein. However, the manuscript does not provide the number of FASTA files used or the actual protein sequences. This information should be included or properly referenced. 3. Missing supporting information: The manuscript refers to supporting information, but no supplementary materials are provided. All supporting data should be included and clearly linked. 4. Weak conclusion section: The conclusion is not clearly articulated and should be rewritten. It should explicitly explain how the statistical analysis contributes to distinguishing between soluble and insoluble proteins and highlight the significance of the findings. 5. Introduction requires improvement: The introduction lacks clarity and coherence and should be thoroughly revised to better contextualize the study and its objectives. Reviewer #2: The few points to be improved upon are as follows: 1. To ensure no bias in the AUC and MCC comparisons, it is suggested that a clarification be provided if the reference models were evaluated on the exact same merged dataset or if the performance that is reported is taken straight from the originally published data. 2. In the discussion section, negative charge appearing more influential than positive charge can be elaborated upon more. Is this a general biophysical principle of the proteome or if it is a bias in the E.coli based recombinant expression data used ? 3. It is suggested that in the conclusions section including a comment on whether the weak signal interactions are additive or if they involve higher-order sequence patterns that classical descriptors cannot capture. 4. The ROC curves in figure 2 show a significant overlap. Adding a density plot for say like the top two orthogonal features to demonstrate the extensive overlap as mentioned in the paper would be helpful in demonstrating to the reader the difficulty of the classification process. Reviewer #3: The manuscript presents a statistical analysis of sequence-derived features associated with protein solubility. While the study is carefully conducted and the dataset appears comprehensive, the overall contribution remains limited in terms of novelty and impact. Most of the analyzed features (e.g., amino acid composition, sequence length, and charge properties) are well-established determinants of solubility and have been extensively explored in previous studies. The analytical approach, particularly the merging of training, validation, and test datasets, raises concerns regarding the robustness and interpretability of the results. In addition, the proposed composite index shows relatively modest predictive performance compared to existing methods, which limits its practical applicability. Although the manuscript provides a useful summary of known trends, it does not offer sufficient new methodological developments or biological insights to justify publication in its current form. Strengthening the methodological framework and providing deeper mechanistic or predictive advances would be necessary to improve the impact of the study. ****** 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 Reviewer #3: 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.] To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. Attachments Attachment Submitted filename: Reviewer comment.pdf https://doi.org/10.1371/journal.pone.0344883.r001
Revision 1
14 Apr 2026 Author Response LARGE-SCALE STATISTICAL DISSECTION OF SEQUENCE-DERIVED BIOCHEMICAL FEATURES DISTINGUISHING SOLUBLE AND INSOLUBLE PROTEINS Dear Reviewers and Editors of the PLOS One, We would like to thank you so much, the reviewers and editors, for your careful and thorough reading of this manuscript and for your thoughtful comments and constructive suggestions, which significantly improved its quality. Our responses are following the reviewers’ comments, and new changes are highlighted in yellow in our manuscript. Yours sincerely, Authors Dear Reviewer 1, We would like to answer your comments as follows: 1. Incorrect reference numbering: The citation format is inconsistent throughout the manuscript. For example, on page 1, line 4, the reference is cited as [1, 6], whereas it should appear sequentially (e.g., [1, 2]). The authors are advised to use reference management software such as EndNote, Zotero, or similar tools to correct and standardize citations across the entire manuscript. Our answer: Thank you very much for this valuable comment, and please be aware that we have revised the manuscript accordingly and corrected the reference numbering as suggested by the reviewer. 2. Lack of sequence data transparency: The authors state that a total of 36 sequence-derived biochemical features were computed for each protein. However, the manuscript does not provide the number of FASTA files used or the actual protein sequences. This information should be included or properly referenced. Our answer: Thank you very much for this valuable comment, and please kindly note that the revised manuscript has been updated based on the reviewers’ comments. The revised Dataset section now explicitly states that our analysis used the three original FASTA files from the benchmark dataset reported by Zhang et al. (2024). We also clarify that these FASTA files were used without modification. Furthermore, in the Data and Code Availability section, we now clearly indicate that the raw sequences and labels are publicly accessible through Zenodo. Additionally, the full feature matrix used for all statistical analyses, the processed data and analysis outputs is provided in the GitHub repository (see the Dataset subsection on page 3, and the Data and Code Availability section on pages 15–16) 3. Missing supporting information: The manuscript refers to supporting information, but no supplementary materials are provided. All supporting data should be included and clearly linked. Our answer: Thank you very much for this valuable comment, and please kindly note that the revised manuscript has been updated based on the reviewers’ comments. All supporting information has now been included and clearly linked through a separate Supporting Information document submitted together with the revised manuscript. The supplementary materials include Tables S1–S5 and Figures S1–S4 (a PDF file attached). 4. Weak conclusion section: The conclusion is not clearly articulated and should be rewritten. It should explicitly explain how the statistical analysis contributes to distinguishing between soluble and insoluble proteins and highlight the significance of the findings. Our answer: Thank you very much for this important comment, and please kindly note that the revised manuscript’s conclusion has been updated based on the reviewers’ comments. We clarify how the statistical analysis contributes to distinguishing between soluble and insoluble proteins and make significance of these findings more explicit as well (see the Conclusion section on page 14). 5. Introduction requires improvement: The introduction lacks clarity and coherence and should be thoroughly revised to better contextualize the study and its objectives. Our answer: Thank you very much for this important comment. In the revised manuscript, we clarify the introduction more clarity and coherence as follows: The revised Introduction now provides a clearer explanation on pages 1–3. Overall, please kindly note that the revised manuscript states that the purpose of this study is not to develop another high-capacity predictive model, but rather to perform a statistically rigorous large-scale analysis of 36 sequence-derived biochemical features in order to quantify their effect sizes, redundancy structure, and practical discriminatory contribution in distinguishing soluble and insoluble proteins. Additionally, this study also presents a simple baseline model intended to serve as a lower-bound reference for evaluating more complex predictive approaches in future work Dear Reviewer 2, We would like to answer your comments as follows: 1. To ensure no bias in the AUC and MCC comparisons, it is suggested that a clarification be provided if the reference models were evaluated on the exact same merged dataset or if the performance that is reported is taken straight from the originally published data Our answer: Thank you very much for this valuable comment, and please be aware that we have revised the manuscript as suggested by the reviewer. Specifically, in the Results section (page 12), we now clearly confirm that the performance metrics (AUC and MCC) were taken directly from Zhang et al. (2024) (https://doi.org/10.1093/bib/bbae404). 2. In the discussion section, negative charge appearing more influential than positive charge can be elaborated upon more. Is this a general biophysical principle of the proteome or if it is a bias in the E. coli based recombinant expression data used? Our answer: Thank you very much for this valuable comment, and please kindly note that we have revised the Discussion section accordingly based on the reviewers’ comments as follows on page 9. The observed asymmetry likely reflects both general biophysical effects and E. coli expression-system bias and the dataset is based on recombinant expression in E. coli, therefore, generalization should be made with caution. 3. It is suggested that in the conclusions section including a comment on whether the weak signal interactions are additive or if they involve higher-order sequence patterns that classical descriptors cannot capture. Our answer: Thank you very much for this valuable comment, and please kindly note that the Conclusion section has been updated accordingly to clarify that based on the reviewers’ comments as follows on page 14-15. 4. The ROC curves in figure 2 shows a significant overlap. Adding a density plot for say like the top two orthogonal features to demonstrate the extensive overlap as mentioned in the paper would be helpful in demonstrating to the reader the difficulty of the classification process. Our answer: Thank you very much for this important comment, and please kindly note that the Results section has been updated based on the reviewers’ comments as follows on page 10-11. We added Figure 4 (the density distributions of the two features). We also clarified that this overlap explains the limited univariate ROC performance and highlights the difficulty of the classification process. Dear Reviewer 3, We would like to answer your comments as follows: Reviewer’s statement: The manuscript presents a statistical analysis of sequence-derived features associated with protein solubility. While the study is carefully conducted and the dataset appears comprehensive, the overall contribution remains limited in terms of novelty and impact. Most of the analyzed features (e.g., amino acid composition, sequence length, and charge properties) are well-established determinants of solubility and have been extensively explored in previous studies. Our answer: Thank you very much for this valuable comment. Please note that the manuscript has been thoroughly revised to more explicitly highlight its innovative contribution, its necessity, and its clear distinction from previous literature/studies. The novelty of the present work does not lie in introducing new descriptors. Rather, it lies in providing a large-scale, statistically controlled quantification of their practical effect sizes, redundancy structure, and descriptive discriminative limits in a large publicly available dataset, from there establishing an interpretable baseline for what can or cannot be captured by classical global sequence-derived features. This point is now stated more explicitly in the Introduction (pp. 2–3). We have also reiterated this contribution more clearly in the Conclusion (pp. 14–15), where we clarify that the study refines the interpretation of widely accepted solubility determinants by showing that their practical effect sizes are substantially smaller than statistical significance alone may suggest. Also, the proposed framework should be understood as a transparent, lower-bound, and interpretable reference for evaluating the added value of more complex models, rather than as a competitive predictive model. We confirm that every change has been made in the revised manuscript (v1), a high level of refinement in language aspect, with yellow highlighting for easy verification. We are confident that the manuscript has been substantially improved thanks to your valuable feedback. Thank you once again for your time and constructive comments. We look forward to your positive decisions. Sincerely yours, Attachments Attachment Submitted filename: Response to Reviewers.pdf https://doi.org/10.1371/journal.pone.0344883.r002
30 Apr 2026 Decision Letter - Anil Bhatia, Editor PONE-D-26-09831R1Large-Scale Statistical Dissection of Sequence-Derived Biochemical Features Distinguishing Soluble and Insoluble ProteinsPLOS One Dear Dr. Nguyen Bao, 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 Jun 14 2026 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 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. As the corresponding author, your ORCID iD is verified in the submission system and will appear in the published article. PLOS supports the use of ORCID, and we encourage all coauthors to register for an ORCID iD and use it as well. Please encourage your coauthors to verify their ORCID iD within the submission system before final acceptance, as unverified ORCID iDs will not appear in the published article. Only the individual author can complete the verification step; PLOS staff cannot verify ORCID iDs on behalf of authors. We look forward to receiving your revised manuscript. Kind regards, Anil Bhatia, Ph.D Academic Editor PLOS One Journal Requirements: 1. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. 2. 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. [Note: HTML markup is below. Please do not edit.] 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 Reviewer #3: 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: Yes Reviewer #2: Yes Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: 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 Reviewer #3: 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 Reviewer #3: 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: All queries have been addressed carefully by the author. However, the conclusion section is overly long and lacks clarity, making it difficult to understand the key takeaways of the study. I recommend that the manuscript be accepted only after the conclusion is revised to be more concise and clearly summarize the main findings Reviewer #2: (No Response) Reviewer #3: All queries are answered adequately by the authors, hence I recommend to accept the paper in revised form. ****** 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 Reviewer #3: 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.] To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. Attachments Attachment Submitted filename: Re-review comments.pdf https://doi.org/10.1371/journal.pone.0344883.r003
Revision 2
5 May 2026 Author Response LARGE-SCALE STATISTICAL DISSECTION OF SEQUENCE-DERIVED BIOCHEMICAL FEATURES DISTINGUISHING SOLUBLE AND INSOLUBLE PROTEINS Dear Reviewers and Editors of the PLOS One, We would like to thank you so much, the reviewers and editors, for your careful and thorough reading of this manuscript and for your thoughtful comments and constructive suggestions, which significantly improved its quality. Our responses are in italics following the reviewers’ comments, and new changes are highlighted in yellow in our manuscript. Yours sincerely, Authors Dear Reviewer 1, We would like to answer your comments as follows: “All queries have been addressed carefully by the author. However, the conclusion section is overly long and lacks clarity, making it difficult to understand the key takeaways of the study. I recommend that the manuscript be accepted only after the conclusion is revised to be more concise and clearly summarize the main findings.” Our answer: Thank you very much for this valuable comment, and please be aware that we have revised the manuscript’s conclusion accordingly as suggested by the reviewer. We greatly appreciate the reviewer’s recommendation. Dear Reviewer 2, Thank you very much for this valuable comment, we greatly appreciate the reviewer’s recommendation. Dear Reviewer 3, Thank you very much for this valuable comment, we greatly appreciate the reviewer’s recommendation. We confirm that every change has been made in the revised manuscript (v2), a high level of refinement in language aspect, with yellow highlighting for easy verification. We are confident that the manuscript has been substantially improved thanks to your valuable feedback. Thank you once again for your time and constructive comments. We look forward to your positive decisions. Sincerely yours, Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0344883.r004
12 May 2026 Decision Letter - Anil Bhatia, Editor Large-Scale Statistical Dissection of Sequence-Derived Biochemical Features Distinguishing Soluble and Insoluble Proteins PONE-D-26-09831R2 Dear Dr. Bao, 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. For questions related to billing, please contact billing support. 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, Anil Bhatia, Ph.D 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: All comments have been addressed 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: Yes Reviewer #2: Yes ****** 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: The authors have adequately addressed all the queries. I recommend that the editor accept the manuscript without any further revisions. Reviewer #2: (No Response) ****** 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 ******** Attachments Attachment Submitted filename: Re-review 2 comments.pdf https://doi.org/10.1371/journal.pone.0344883.r005
Formally Accepted
Acceptance Letter - Anil Bhatia, Editor PONE-D-26-09831R2 PLOS One Dear Dr. Nguyen Bao, 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 You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days 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. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. 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. Anil Bhatia Academic Editor PLOS One https://doi.org/10.1371/journal.pone.0344883.r006

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