External validation of VO2max prediction models based on recreational and elite endurance athletes

Szczepan Wiecha; Przemysław Seweryn Kasiak; Igor Cieśliński; Tim Takken; Tomasz Palka; Beat Knechtle; Pantelis Τ. Nikolaidis; Łukasz A. Małek; Marek Postuła; Artur Mamcarz; Daniel Śliż

doi:10.1371/journal.pone.0280897

Peer Review History

Original SubmissionJune 12, 2022
10 Aug 2022 Decision Letter - Alvaro Reischak-Oliveira, Editor PONE-D-22-16633External validation of VO2max prediction models based on recreational and elite endurance athletesPLOS ONE Dear Dr. Wiecha, 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. Both reviewers agree that there is a weakness in the reasons and rationale that supports the work. What are the novelties of the manuscript? I recommend a comprehensive review covering every aspect highlighted by the reviewers. Please submit your revised manuscript by Sep 16 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. 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Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 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: I Don't Know ****** 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: General Comments The manuscript provides information on external validation of VO2max prediction models based on recreational and elite endurance athletes. Overall, the study is well-conducted, methodologically robust, and provides relevant information for the area of sports and health sciences. My concern about the study is mainly related to the introduction, methodology, and discussion sections, which could be edited. Please, see in the specific comments. Specific Comments Personal suggestion: I believe the main message of the study is that direct VO2max determination (CPET plus ergospirometry) cannot be replaced or interchangeable by predictive equations for endurance athletes based on their own results 0.30 < R2 < 0.65. If the authors agree with me, please make this message clearer in the discussion and conclusion of the study. Additionally, little is discussed about the repercussions of using predictive equations with low to moderate accuracy for the contexts of sports & performance diagnostics, clinical practice, applicability of prediction equation, training prescription and follow-up. Abstract: What is the criterion used to define models with the highest accuracy? Please, include the minimum and maximum values for underestimated and overestimated that can be obtained for indirect VO2max. Introduction: The introduction does not convince the reader of the importance of direct determination of VO2max compared to indirect determination through equations. Additionally, the authors could include epidemiological data to strengthen the argument for the importance of VO2max for the health of endurance athletes. Please make it clear in the introduction what is the main focus and secondary outcomes of the study. In addition, to include the advantages and disadvantages of each of the ways of determining VO2max for the contexts of sports & performance diagnostics, clinical practice, applicability of prediction equation, training prescription and follow-up. Lines 18-19: Please, describe more variables that can influence the variability of VO2max data in endurance athletes. Lines 33-34: Please, deepen this information, including epidemiological data, if possible, on CVDs in endurance athletes. Please, include information here about the importance of directly measured VO2max can be used as a predictive cardiometabolic risk factor. In addition, there is little in the introduction about the limitations or harm that the indirect determination of VO2max may have in relation to clinical evaluation. Please, present the risks and benefits of direct and indirect determination (prediction equations) of the VO2max data. Line 55: There is little information about why endurance athletes were chosen as a sample for the study. Methodology: Lines 79-80: Please make it clear whether data collection was performed with a breath-by-breath acquisition system and a 10-seconds filter was used for data analysis. Note that this information can lead to some confusion as to how VO2max was defined as described in line 104 “Maximal oxygen uptake (VO2max) was defined as an averaged maximum oxygen uptake during the 15-s period at the end of the CPET’. Could I think that only 1 or 2 VO2 points were analysed for the average of VO2max? Please clarify. Suggestion: Please, insert the 13 equations of selected predictions models to establish pVO2max in methodology section. Statistical Analysis: Please, describe the criterion used to classify low to moderate variability in VO2max. What is the criterion used to classify accuracy (low-moderate-high)? Please, describe. Discussion: The authors could better highlight the main findings of the study. In addition, authors should focus the discussion section on possible explanation of their main results of the present study. Suggestion: The authors could organize the discussion as follows: Describe the consequences (risks and benefits) of using VO2max prediction models with low to moderate accuracy (underestimation and overestimation values) in the following contexts: (1) Sports & Performance Diagnostics; (2) Clinical Practice; (3) Endurance Training Prescription and Follow-Up. Reviewer #2: I don't understand very well this study. If you check the correlation between a previously validated formula in very specific conditions with actual values in another different setting, it is obvious that the results will be quite different. The authors should guarantee that the comparisons are made with the same protocols (step duration, starting intensity, exhaustion criteria, etc.) and populations (age, sex, sport, training level, etc.) between the current and previous validation studies. The figures are not legible. VO2max can be tested on the field in a sport setting. An example: https://pubmed.ncbi.nlm.nih.gov/24936896/ ****** 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: Yes: Dr. Giovani dos Santos Cunha 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.0280897.r001
Revision 1
30 Sep 2022 Author Response Author’s response to reviews Dear reviewers, We are grateful for the time and effort you took to write these extensive reviews. We have done our best to incorporate all of your suggestions and send back the revised manuscript. We believe the content is much improved and more clear for readers thanks to your comments. Yours sinserely, Szczepan Wiecha on behalf of all authors. In the following, please find our answers to your comments. Title: External validation of VO2max prediction models based on recreational and elite endurance athletes Version: 1 Date: 30 Aug 2022 In the following, please find our answers to your comments. Reviewers comments: Dr. Giovani dos Santos Cunha (Reviewer 1) General Comments The manuscript provides information on external validation of VO2max prediction models based on recreational and elite endurance athletes. Overall, the study is well-conducted, methodologically robust, and provides relevant information for the area of sports and health sciences. My concern about the study is mainly related to the introduction, methodology, and discussion sections, which could be edited. Please, see in the specific comments. Specific Comments Personal suggestion: I believe the main message of the study is that direct VO2max determination (CPET plus ergospirometry) cannot be replaced or interchangeable by predictive equations for endurance athletes based on their own results 0.30 < R2 < 0.65. If the authors agree with me, please make this message clearer in the discussion and conclusion of the study. Additionally, little is discussed about the repercussions of using predictive equations with low to moderate accuracy for the contexts of sports & performance diagnostics, clinical practice, applicability of prediction equation, training prescription and follow-up . Response: Thank you for the suggestion, we have added this information to the introduction section. Abstract: What is the criterion used to define models with the highest accuracy? Response: We added information: “by assessing the transferability of the currently available prediction models based on their R2, calibration-in-the-large, and calibration slope” Please, include the minimum and maximum values for underestimated and overestimated that can be obtained for indirect VO2max. Response: Due to the limited capacity of the abstract, we are not able to provide as much detail for each of the validated equations. These details are given in the results section and the supplement Introduction: The introduction does not convince the reader of the importance of direct determination of VO2max compared to indirect determination through equations. Additionally, the authors could include epidemiological data to strengthen the argument for the importance of VO2max for the health of endurance athletes. Please make it clear in the introduction what is the main focus and secondary outcomes of the study. In addition, to include the advantages and disadvantages of each of the ways of determining VO2max for the contexts of sports & performance diagnostics, clinical practice, applicability of prediction equation, training prescription and follow-up . Response: Thank you very much for your insightful analysis of the content. We have amended the content in line with your suggestions Lines 18-19: Please, describe more variables that can influence the variability of VO2max data in endurance athletes. Response: We added additional variables as suggested Lines 33-34: Please, deepen this information, including epidemiological data, if possible, on CVDs in endurance athletes. Please, include information here about the importance of directly measured VO2max can be used as a predictive cardiometabolic risk factor. In addition, there is little in the introduction about the limitations or harm that the indirect determination of VO2max may have in relation to clinical evaluation. Please, present the risks and benefits of direct and indirect determination (prediction equations) of the VO2max data. Response: We have added more detailed information including epidemiology and clinical evaluation Line 55: There is little information about why endurance athletes were chosen as a sample for the study . Response: We have added information in this regard Methodology: Lines 79-80: Please make it clear whether data collection was performed with a breath-by-breath acquisition system and a 10-seconds filter was used for data analysis. Note that this information can lead to some confusion as to how VO2max was defined as described in line 104 “Maximal oxygen uptake (VO2max) was defined as an averaged maximum oxygen uptake during the 15-s period at the end of the CPET’. Could I think that only 1 or 2 VO2 points were analyzed for the average of VO2max? Please clarify .K Response: We have corrected this section. Measurements were taken for each breath and averaging was done at 15-second intervals. The 10-second values were entered by mistake. Many thanks for finding this inaccuracy in the text Suggestion: Please, insert the 13 equations of selected predictions models to establish pVO2max in methodology section. Response: Thank you for your suggestion but we do not want to duplicate content. A detailed table describing all included equations can be found in the manuscript supplement. In the main body of the paper, such a table would take up a considerable amount of space. Bibliographic data for the included designs are also included in the supplement Statistical Analysis: Please, describe the criterion used to classify low to moderate variability in VO2max. What is the criterion used to classify accuracy (low-moderate-high)? Please, describe Response: The criterion for determining the accuracy of the models was R2 and RMSE, as described in lines 173-183. We did not use cut-off points for these parameters, as there is no explicit definition adopted here as in, for example, correlation coefficients or effect measures. Discussion: The authors could better highlight the main findings of the study. In addition, authors should focus the discussion section on possible explanation of their main results of the present study. Suggestion: The authors could organize the discussion as follows: Describe the consequences (risks and benefits) of using VO2max prediction models with low to moderate accuracy (underestimation and overestimation values) in the following contexts: (1) Sports & Performance Diagnostics; (2) Clinical Practice; (3) Endurance Training Prescription and Follow-Up. Response: We have made amendments Reviewer #2: I don't understand very well this study. If you check the correlation between a previously validated formula in very specific conditions with actual values in another different setting, it is obvious that the results will be quite different. The authors should guarantee that the comparisons are made with the same protocols (step duration, starting intensity, exhaustion criteria, etc.) and populations (age, sex, sport, training level, etc.) between the current and previous validation studies Response: We have added appropriate explanations in the text, mainly in the introduction of the manuscript and methodology regarding the need for external validation and differences in protocols and VO2max values obtained. By following the guidelines for validation studies (such as the TRIPOD statement), we obtained a cross-sectional analysis of the available equations, which enabled us to reach appropriate conclusions. The figures are not legible. Response: This is most likely a matter of converting the document in the editorial system for review purposes. The uploaded charts are in the correct high resolution and size. In the PLOS submissions system, they are presented on A4 size, each on a separate page. In the top right corner of the manuscript there is a hyperlink to download the TIFF version. A higher resolution version is then downloaded VO2max can be tested on the field in a sport setting. An example: https://pubmed.ncbi.nlm.nih.gov/24936896/ Response: Thank you very much for your valuable publication, we have completed the information Once more, we would like to thank the Reviewers for their important comments. Attachments Attachment Submitted filename: Response to Reviewers1.docx https://doi.org/10.1371/journal.pone.0280897.r002
12 Dec 2022 Decision Letter - Alvaro Reischak-Oliveira, Editor PONE-D-22-16633R1External validation of VO2max prediction models based on recreational and elite endurance athletesPLOS ONE Dear Dr. Wiecha, 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. First of all, apologies for the time elapsed. it was somewhat difficult to get all the necessary revisions to a decision. Please pay attention to reviewer 2. Please, elaborate better on the error measures reported, the potential sources of these errors, and the differences between protocols and formulae in the discussion section. I believe that this detail is sufficient for a final decision. Please submit your revised manuscript by Jan 26 2023 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, Alvaro Reischak-Oliveira, 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. [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: 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: (No Response) Reviewer #2: Please, elaborate better on the error measures reported, the potential sources of these errors, and the differences among protocols and formulae in the discussion section. ****** 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: Yes: Dr. Giovani dos Santos Cunha Reviewer #2: Yes: Daniel Boullosa ******** [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.0280897.r003
Revision 2
16 Dec 2022 Author Response Dear Editor, Dear Reviewers, Thank you very much for the thorough analysis of our manuscript, for your valuable and helpful comments and for giving us the opportunity to revise and improve our submission. We hope that our replies and explanations, as well as the amendments to the manuscript, fully address your concerns. We keep the change tracking. In the following, please find our answers to your comments. Reviewer #2: Please, elaborate better on the error measures reported, the potential sources of these errors, and the differences among protocols and formulae in the discussion section. Response: We have added a new section to the discussion Once more, we would like to thank the Reviewer for important comments and for input on our report. We believe that all remaining concerns are now fully addressed. With regards Szczepan Wiecha, PhD Attachments Attachment Submitted filename: response to Reviewer 2.docx https://doi.org/10.1371/journal.pone.0280897.r004
11 Jan 2023 Decision Letter - Alvaro Reischak-Oliveira, Editor External validation of VO2max prediction models based on recreational and elite endurance athletes PONE-D-22-16633R2 Dear Dr. Wiecha, 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 for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, 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, Alvaro Reischak-Oliveira, Ph.D. Academic Editor PLOS ONE Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #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: (No Response) Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: (No Response) 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: (No Response) 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: (No Response) 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: (No Response) Reviewer #2: No more comments. Although the writing may be improved, I think this manuscript adds relevant information in the context of the current evidence. No more comments. Although the writing may be improved, I think this manuscript adds relevant information in the context of the current evidence. ****** 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: Yes: Dr. Giovani dos Santos Cunha Reviewer #2: Yes: Daniel Boullosa ******** https://doi.org/10.1371/journal.pone.0280897.r005
Formally Accepted
16 Jan 2023 Acceptance Letter - Alvaro Reischak-Oliveira, Editor PONE-D-22-16633R2 External validation of VO_2max prediction models based on recreational and elite endurance athletes Dear Dr. Wiecha: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. 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 plosone@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. Alvaro Reischak-Oliveira Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0280897.r006

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