Increased serum caspase-1 in adult-onset Still’s disease

Haruki Matsumoto; Shuhei Yoshida; Tomohiro Koga; Yuya Fujita; Yuya Sumichika; Kenji Saito; Jumpei Temmoku; Tomoyuki Asano; Shuzo Sato; Masashi Mizokami; Masaya Sugiyama; Kiyoshi Migita

doi:10.1371/journal.pone.0307908

Peer Review History

Original SubmissionMarch 7, 2024
5 Apr 2024 Decision Letter - Yoshito Nishimura, Editor PONE-D-24-06426 Increased serum caspase-1 in adult-onset Still’s disease PLOS ONE Dear Dr. Migita, 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. In particular, multiple concerns were raised by reviewer 2. The authors are encouraged to review the comments, and appropriately address the concerns for acceptance. Please submit your revised manuscript by May 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, Yoshito Nishimura, MD, PhD, MPH 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 provide additional details regarding participant consent. In the ethics statement in the Methods and online submission information, please ensure that you have specified (1) what type of consent you obtained (for instance, written or verbal, and if verbal, how it was documented and witnessed). If your study included minors, state whether you obtained consent from parents or guardians. If the need for consent was waived by the ethics committee, please include this information. 3. Thank you for stating the following financial disclosure: "The study was supported by the Japan Grant-in-Aid for Scientific Research (20K08777)." Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 4. In the online submission form you indicate that your data is not available for proprietary reasons and have provided a contact point for accessing this data. Please note that your current contact point is a co-author on this manuscript. According to our Data Policy, the contact point must not be an author on the manuscript and must be an institutional contact, ideally not an individual. Please revise your data statement to a non-author institutional point of contact, such as a data access or ethics committee, and send this to us via return email. Please also include contact information for the third party organization, and please include the full citation of where the data can be found. 5. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. 6. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. [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: Yes Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: No ****** 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: No ****** 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: Thank you for the opportunity to review this article. Some English grammar was not typically used in scientific articles. You could proofread it again before submission. You wrote in the conclusion of the study, "Our findings revealed that serum caspase-1 was elevated in AOSD patients and that elevated caspase-1 was correlated with systemic scores and inflammatory cytokines including IL-18 in AOSD patients." This should be in the conclusion section or omitted. In scientific articles, you might not want to explain the results in the figure caption. Instead, you could include the results in the main text. In Figure 1 caption, what does it mean by "the four groups (p<0.001)"? Also, you could explain which statistical analysis you have used in the methods. In Figure 3, you wrote "Japanese," it should be changed to "English." Would you include baseline characteristics of HCs and RA patients in tables, or have you omitted them? Differences in baseline characteristics are also significant limitations. You could explain briefly why the each limitations you suggest are important. Reviewer #2: This study was to assess the level of caspase-1 in AOSD patients for the diagnosis and monitoring disease activity. While there are several major issues that should be take care of. 1.Are there any data about the behaviors of caspase-1 in the event of a flare (other than MAS) in patients with AOSD? 2.Is the kinetics of caspase-1 increase/decrease different from that of ferritin or CRP or ESR? 3.I have not found any information about the ongoing treatment of AOSD patient (and controls) included in the study. Besides，clinical manifestations related to Pouchot score should be provided in table 1. 4.How do the levels of caspase-1 compare between patients with MAS and non-MAS, as well as between those with active and inactive disease, considering the potential role of Caspase-1 to access disease activity. 5.The article suggests that caspase-1 can be used for diagnosing AOSD. What are the values for the area under the ROC curve (AUC), specificity, and sensitivity of caspase-1 in distinguishing between AOSD and RA? 6.Among the 71 serum inflammatory factors tested, what specific information was obtained from AOSD patients, and how many cases were included in the analysis? 7.Given the presence of numerous albumin and immunoglobulins in serum, it is necessary to remove these proteins before further testing by precipitating the proteins. Additionally, Western blot analysis for serum caspase-1 lacks internal controls. ****** 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.0307908.r001
Revision 1
12 Jun 2024 Author Response 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 →Thank you for your comment. We confirmed that our manuscript meets PLOS ONE`s style requirements. 2. Please provide additional details regarding participant consent. In the ethics statement in the Methods and online submission information, please ensure that you have specified (1) what type of consent you obtained (for instance, written or verbal, and if verbal, how it was documented and witnessed). If your study included minors, state whether you obtained consent from parents or guardians. If the need for consent was waived by the ethics committee, please include this information. →Thank you for your comment. We described that “The institutional Review Board waived the need for written informed consent from participants due to the non-interventional design of the retrospective study” in part of patients and study design. 3. Thank you for stating the following financial disclosure: "The study was supported by the Japan Grant-in-Aid for Scientific Research (20K08777)." Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. →Thank you for your comment. We added the following text to the cover letter: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." 4. In the online submission form you indicate that your data is not available for proprietary reasons and have provided a contact point for accessing this data. Please note that your current contact point is a co-author on this manuscript. According to our Data Policy, the contact point must not be an author on the manuscript and must be an institutional contact, ideally not an individual. Please revise your data statement to a non-author institutional point of contact, such as a data access or ethics committee, and send this to us via return email. Please also include contact information for the third party organization, and please include the full citation of where the data can be found. →Thank you for your comment. We revised the data statement to a non-author institutional point of contact (Contact information for the representative of our medical office managing the data set). 5. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. →Thank you for your comment. We revised the Patients and study design part and added the following sentence: “This study was conducted in accordance with the principles of the Declaration of Helsinki and approved by the institutional review boards of Fukushima Medical University (No. 2021-290; the date of approval: 16 - April 2023) and the National Center for Global Health and Medicine (NCGM-G-003472; the date of approval: 19 - April 2023). ” 6. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. →Thank you for your comment. We added the following sentence: “We submitted gel image as the supporting information.” Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: Thank you for the opportunity to review this article. Some English grammar was not typically used in scientific articles. You could proofread it again before submission. →Thank you for your comment. We proofread the final version of the manuscript again in English. You wrote in the conclusion of the study, "Our findings revealed that serum caspase-1 was elevated in AOSD patients and that elevated caspase-1 was correlated with systemic scores and inflammatory cytokines including IL-18 in AOSD patients." This should be in the conclusion section or omitted. →Thank you for your comment. In the introduction, we omitted the following sentence: “Our findings revealed that serum caspase-1 was elevated in AOSD patients and that elevated caspase-1 was correlated with systemic scores and inflammatory cytokines, including IL-18 in AOSD patients.” In scientific articles, you might not want to explain the results in the figure caption. Instead, you could include the results in the main text. In Figure 1 caption, what does it mean by "the four groups (p<0.001)"? Also, you could explain which statistical analysis you have used in the methods. →Thank you for your comment. “the four groups (p<0.001)" was a mistake. The correct description is shown in Figure legend as follows; The AOSD group had significantly higher caspase-1 levels for both HC and RA groups (p<0.001). In Figure 3, you wrote "Japanese," it should be changed to "English." →Thank you for your comment. We revised the Figure 3 you pointed out. Would you include baseline characteristics of HCs and RA patients in tables, or have you omitted them? Differences in baseline characteristics are also significant limitations. You could explain briefly why the each limitations you suggest are important. →Thank you for your comment. Demographic data of HC and patients of RA were attached in Supplemental Table1 and following sentence was added to the result section; “24 of the HCs were female, and the average age was 41 years [IQR]; 28–52 years. They had no medical history or ongoing medications, and no abnormalities were noted in recent physical examinations. The demographic data of patients with RA and HCs were attached in Supplemental Table 1.” Characteristic baseline differences between AOSD patients, RA patients, and HC are described in the limitation section; “Additionally, characteristic baseline differences between AOSD patients, RA patients, and HC differ in terms of age and gender, so a background-aligned study is necessary in further.” Reviewer #2: This study was to assess the level of caspase-1 in AOSD patients for the diagnosis and monitoring disease activity. While there are several major issues that should be take care of. 1.Are there any data about the behaviors of caspase-1 in the event of a flare (other than MAS) in patients with AOSD? →Thank you for your comment. Data at the time of a flare other than MAS were not collected and considered in this study. 2.Is the kinetics of caspase-1 increase/decrease different from that of ferritin or CRP or ESR? →Although ESR data are lacking, no correlation was found between CRP and Caspase-1 (r=0.143, p=0.356). The correlation with Ferritin is shown in Fig. 3. The following sentence was added to the result. “However, no correlation was found between CRP and caspase-1 (r=0.143, p=0.356) (data was not shown in figure)” 3.I have not found any information about the ongoing treatment of AOSD patient (and controls) included in the study. Besides，clinical manifestations related to Pouchot score should be provided in table 1. →Thank you for your comment. We added the information of details of treatment and clinical manifestations related to Pouchot`s score. 4.How do the levels of caspase-1 compare between patients with MAS and non-MAS, as well as between those with active and inactive disease, considering the potential role of Caspase-1 to access disease activity. Comparison of caspase-1 between MAS and non-MAS showed no significant difference at p=0.189. This was added in the result section as follow; “Additionally, the comparison of caspase-1 between MAS and non-MAS showed no significant difference (p=0.189).” Since many of the active and inactive data we collected were for non-MAS (6/8 patients had non-MAS, and 2/8 patients had MAS), it was not possible to compare active and inactive data for MAS and non-MAS, respectively. 5.The article suggests that caspase-1 can be used for diagnosing AOSD. What are the values for the area under the ROC curve (AUC), specificity, and sensitivity of caspase-1 in distinguishing between AOSD and RA? →Thank you for your comment. We showed the values for the area under the ROC curve in Fig 2 and added the following sentence in the result section; “We analyzed the cut-off value for the caspase-1 in distinguishing between AOSD and RA by obtaining the ROC curve, which revealed 26.9 ng/mL (Sensitivity 100.0% and Specificity 93.9%). Area under the ROC curve was 0.987. (Figure 2).” 6.Among the 71 serum inflammatory factors tested, what specific information was obtained from AOSD patients, and how many cases were included in the analysis? →Cytokines were analyzed in all 51 AOSD patients. The comprehensive analysis of cytokines was performed only in AOSD, and we did not search which cytokines were specifically elevated compared to RA and HC. However, IL-12/IL-23, IL-5, 7, and 13 were detected with sensitivity or higher in only about half of all AOSD cases, suggesting that these cytokines may have little relation to the inflammatory pathogenesis of AOSD among 71 cytokines. This point was described in the limitation section as follows; “Second, multiple serum cytokines were measured in AOSD patients only, and not in control patients (RA) or HCs.” 7.Given the presence of numerous albumin and immunoglobulins in serum, it is necessary to remove these proteins before further testing by precipitating the proteins. Additionally, Western blot analysis for serum caspase-1 lacks internal controls. →Thank you for your comment. As you pointed out, fragmentation of the serum IgG into IgG light chains (25 kDa, respectively) was detected by the secondary antibody used for western blot. To avoid these interfering IgG-derived bands, sera were pre-absorbed using protein G beads and subjected anti-cleaved caspase-1 western blot. As shown in Figure 6B, single cleaved caspase-1 bands (20kDa) were exclusively detected in the sera from untreated AOSD patients. Additionally, as internal controls, we confirmed that serum transferrin was detected to the same extent in untreated and treated AOSD patients (Figure 6C). These results were added in the results section. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0307908.r002
15 Jul 2024 Decision Letter - Yoshito Nishimura, Editor Increased serum caspase-1 in adult-onset Still’s disease PONE-D-24-06426R1 Dear Dr. Migita, 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, Yoshito Nishimura, MD, PhD, MPH 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 #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 #1: Yes Reviewer #3: Yes Reviewer #4: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes 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 #1: Yes 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 #1: Yes 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 #1: Thank you very much for the opportunities reviewing your manuscript all issues I pointed out seems to be addressed. Reviewer #3: All my queries were addressed. I have no further comments on the manuscript. In this form, manuscript can be published. Reviewer #4: In this study, the authors evaluated the potential of serum caspase-1 levels as a useful inflammatory biomarker in patients with adult-onset Still's disease (AOSD). They measured serum caspase-1 levels in 51 consecutive patients diagnosed with AOSD based on Yamaguchi criteria using enzyme-linked immunosorbent assay and compared them with controls, rheumatoid arthritis (RA) patients, and healthy controls (HC). The results showed that serum caspase-1 levels were significantly elevated in AOSD patients than in RA and HC patients. Serum caspase-1 showed a significant positive correlation with disease activity score, serum ferritin level, and the levels of inflammatory cytokines such as IL-18. Immunoblot analysis detected a truncated form of caspase-1 (p20) in sera from untreated AOSD patients but not from inactive AOSD patients receiving immunosuppressive treatment. They also performed a cluster analysis of serum cytokines in AOSD patients. From the above, the authors conclude that caspase-1 is a useful biomarker for the diagnosis and monitoring of AOSD. The authors appropriately addressed the reviewer's comments and revised the paper. ****** 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 #3: No Reviewer #4: No ******** https://doi.org/10.1371/journal.pone.0307908.r003
Formally Accepted
19 Jul 2024 Acceptance Letter - Yoshito Nishimura, Editor PONE-D-24-06426R1 PLOS ONE Dear Dr. Migita, 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. Yoshito Nishimura Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0307908.r004

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 .