Changes in lipoproteins associated with lipid-lowering and antiplatelet strategies in patients with acute myocardial infarction

Zahra Lotfollahi; Ana P. Q. Mello; Francisco A. H. Fonseca; Luciene O. Machado; Andressa F. Mathias; Maria C. Izar; Nagila R. T. Damasceno; Cristiano L. P. Oliveira; Antônio M. F. Neto

doi:10.1371/journal.pone.0273292

Peer Review History

Original SubmissionJuly 3, 2021
5 Apr 2022 Decision Letter - Salvatore De Rosa, Editor PONE-D-21-17809Changes in lipoproteins associated with pharmacological strategies in patients with acute myocardial infarctionPLOS ONE Dear Dr. Fonseca, 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 read carefully all comments bt the external expert reviewers and address them one by one. Please submit your revised manuscript by May 20 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. [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: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: No 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: Yes Reviewer #2: No 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: Yes 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: PONE-D-21-17809: statistical review SUMMARY. This is a prospective, open label study of the effects of combined therapies on the quality of low-density lipoprotein (LDL) particles in subjects with acute myocardial infarction. Groups seem well randomized. Correctly, cross-sectional comparisons are made by unpaired t-tests or Mann-Whitney U tests, while longitudinal comparisons are made by paired t-tests or Wilcoxon tests. I however list below some specific points that should be addressed. SPECIFIC POINTS 1. Follow-up duration (6 months) is not motivated; could results change under different durations? Can the authors motivate such follow-up duration? 2. This is an open label study. As such, it is liable to ascertainment bias. This limitation should be remarked in the final discussion. 3. In this paper, all the statistical tests are two-tailed (as stated in the statistical analysis section). However, one-tail tests seem more appropriate here as the alternative hypothesis is often one-directional. For example, on page 10, the authors say that they observed a significant “increase” of theta. In this case, the alternative hypothesis is not “theta at T0 is different of theta at T6” but rather “theta at T6 is greater than theta at T0”. I would suggest to use a two-tailed test when the alternative hypothesis is that the two parameters are different and a one-tailed test when the alternative hypothesis is that a parameter is greater than another. Reviewer #2: Research article titled (Changes in lipoproteins associated with pharmacological strategies in patients with acute myocardial infarction) is an interesting one with high clinical potential. However, the presentation of the article needs revision starting from the title. 1- Title: authors better should identify what medications followed up in their study. 2-Clear exclusion and exclusion criteria should be stated 3- Stat analysis: authors mentioned tests were applied when appropriate: please give a clear statement for application of the 2 tests. 4- Most of abbreviations are not explained at the first appearance. 5- Use appropriate abbreviations (e.g. h for hours...) Reviewer #3: Congratulations to all authors involved in this important manuscript entitled "Change in lipoproteins associated with pharmacological strategies in acute myocardial infarction” Just to make it more understandable to readers, I suggest minor changes to the text. 1- I suggest standardizing the term (STEMI) to “ST-segment elevation myocardial infarction”, Instead of "ST-segment elevation acute myocardial infarction) - line 28 2- Patients who were unsuccessful after thrombolysis and were referred for rescue angioplasty were included in this cohort? 3- Regarding the comparisons between the groups, using the non-parametric test, I suggest some considerations for a better understanding of the data The Kruskal–Wallis test by ranks, Kruskal–Wallis H test is a non-parametric method for testing whether samples originate from the same distribution. It is used for comparing two or more independent samples of equal or different sample sizes. It extends the Mann–Whitney U test, which is used for comparing only two groups. The parametric equivalent of the Kruskal–Wallis test is the one-way analysis of variance (ANOVA). The test does not identify where this stochastic dominance occurs or for how many pairs of groups stochastic dominance obtains. For analyzing the specific sample pairs for stochastic dominance, Dunn's test, pairwise Mann–Whitney tests with Bonferroni correction. Thus, in this manuscript, I did not observe the new alpha value, post Bonferroni correction (original alpha divided by the number of comparisons). In this case 0.05/4, which will be the post-test alpha, to accept or reject the null hypothesis (equality). ****** 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: Yes: Sawsan Zaitone Reviewer #3: Yes:** Henrique Tria Bianco [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.0273292.r001
Revision 1
25 Apr 2022 Author Response We revised our manuscript according to the suggestions made by the journal and reviewers. Response to reviewers Reviewer #1: PONE-D-21-17809: statistical review SUMMARY. This is a prospective, open label study of the effects of combined therapies on the quality of low-density lipoprotein (LDL) particles in subjects with acute myocardial infarction. Groups seem well randomized. Correctly, cross-sectional comparisons are made by unpaired t-tests or Mann-Whitney U tests, while longitudinal comparisons are made by paired t-tests or Wilcoxon tests. I however list below some specific points that should be addressed. SPECIFIC POINTS 1. Follow-up duration (6 months) is not motivated; could results change under different durations? Can the authors motivate such follow-up duration? Authors: The period of six months was chosen to analyze the quality of LDL after the acute inflammatory phase. The purpose was to analyze the effects of antiplatelet and lipid-lowering therapies in the longer term to reduce possible effects resulting from the greater inflammatory activity and tissue repair in the early phase of STEMI. We added in the text (see page 16, lines 29, 30) The period of six months was chosen to analyze the quality of LDL after the acute inflammatory phase of myocardial infarction. 2. This is an open label study. As such, it is liable to ascertainment bias. This limitation should be remarked in the final discussion. Authors: We added the information in the study limitations (see page 18, lines 9-11) The trial was open label, but it was randomized and the LDL quality was analyzed blindly. In addition, the study compared individuals with very similar baseline characteristics. 3. In this paper, all the statistical tests are two-tailed (as stated in the statistical analysis section). However, one-tail tests seem more appropriate here as the alternative hypothesis is often one-directional. For example, on page 10, the authors say that they observed a significant “increase” of theta. In this case, the alternative hypothesis is not “theta at T0 is different of theta at T6” but rather “theta at T6 is greater than theta at T0”. I would suggest to use a two-tailed test when the alternative hypothesis is that the two parameters are different and a one-tailed test when the alternative hypothesis is that a parameter is greater than another. Authors: Thank you for your important comment. In the captions of the figures, we have indicated which tests were performed. For comparison between theta T6 and T0 we did not use unpaired two sample t-test because theta did not have normal distribution. In this case we used a non-parametric Mann-Whitney U test. (see page 9, caption Fig 3. lines 21-22). We follow your suggestion and changed the text. (see page 10, lines 26-31). Taking into account the values of θ we can conclude that: 1) Rosuvastatin vs. simvastatin/ezetimibe Greater θ at T6 compared to T0 was observed in patients treated with simvastatin/ezetimibe; 2) Clopidogrel vs. ticagrelor Greater θ at T6 compared to T0 was observed in patients treated with clopidogrel; 3) Simvastatin/ezetimibe- clopidogrel and simvastatin/ezetimibe-ticagrelor Greater θ at T6 compared to T0 was observed in patients treated with simvastatin/ezetimibe-clopidogrel and simvastatin/ezetimibe-ticagrelor. Reviewer #2: Research article titled (Changes in lipoproteins associated with pharmacological strategies in patients with acute myocardial infarction) is an interesting one with high clinical potential. However, the presentation of the article needs revision starting from the title. 1- Title: authors better should identify what medications followed up in their study. Authors: we changed the title according to your suggestion (see page 1, line 1) Changes in lipoproteins associated with lipid-lowering and antiplatelet strategies in patients with acute myocardial infarction 2-Clear exclusion and exclusion criteria should be stated Authors: we added information for the inclusion and exclusion criteria to those already described (see page 3, lines 8-13) All included patients were submitted to pharmacological thrombolysis in the first 6 hours of STEMI and referred to Hospital Sao Paulo to perform coronary angiogram and percutaneous coronary intervention (PCI) when needed, in the first 24 h of STEMI (pharmacoinvasive strategy). Key exclusion criteria were clinical instability, use of lipid-lowering or immunosuppressant therapies, autoimmune disease, known malignancy, pregnancy, or signs of active infections. 3- Stat analysis: authors mentioned tests were applied when appropriate: please give a clear statement for application of the 2 tests. Authors: we state when the test was applied. (see page 7, lines 4-6) For comparisons between groups, unpaired two sample t-test (2-tailed) or the Mann-Whitney U test, were used for variables with normal or non-Gaussian distribution, respectively. In addition, we have indicated in each figure's caption which tests have been conducted. 4- Most of abbreviations are not explained at the first appearance. Authors: we explained the abbreviations. (in the Abstract: see page 1, lines 24 and 32, in the Introduction: pages 2, lines 18-22 and 31,32, page 4, in the Materials and methods, Blood samples, lines 21,22) low-density lipoprotein (LDL) particles. ultra violet (UV)-visible spectroscopy nuclear factor kappa B (NFkB) apolipoprotein E knockout (APOE-/-) proprotein convertase subtilisin/kexin type 9 (PCSK9) phosphate-buffered saline (PBS) ethylenediaminetetraacetic acid (EDTA) 5- Use appropriate abbreviations (e.g. h for hours...) Authors: we changed hours for h Reviewer #3: Congratulations to all authors involved in this important manuscript entitled "Change in lipoproteins associated with pharmacological strategies in acute myocardial infarction” Just to make it more understandable to readers, I suggest minor changes to the text. 1- I suggest standardizing the term (STEMI) to “ST-segment elevation myocardial infarction”, Instead of "ST-segment elevation acute myocardial infarction) - line 28 Authors: we changed as suggested. (see page 2, line 31,32) lipid-lowering strategies, commonly prescribed in subjects with ST-segment elevation myocardial infarction (STEMI), on the pattern of LDL particles assessed by complementary methods. 2- Patients who were unsuccessful after thrombolysis and were referred for rescue angioplasty were included in this cohort? Authors: few patients were excluded due to clinical instability (n=4). However, some stable patients showed occluded coronary arteries, and others despite non-reperfusion criteria, their arteries were not occluded on coronary angiography. 3- Regarding the comparisons between the groups, using the non-parametric test, I suggest some considerations for a better understanding of the data The Kruskal–Wallis test by ranks, Kruskal–Wallis H test is a non-parametric method for testing whether samples originate from the same distribution. It is used for comparing two or more independent samples of equal or different sample sizes. It extends the Mann–Whitney U test, which is used for comparing only two groups. The parametric equivalent of the Kruskal–Wallis test is the one-way analysis of variance (ANOVA). The test does not identify where this stochastic dominance occurs or for how many pairs of groups stochastic dominance obtains. For analyzing the specific sample pairs for stochastic dominance, Dunn's test, pairwise Mann–Whitney tests with Bonferroni correction. Thus, in this manuscript, I did not observe the new alpha value, post Bonferroni correction (original alpha divided by the number of comparisons). In this case 0.05/4, which will be the post-test alpha, to accept or reject the null hypothesis (equality). Authors: as was suggest by other reviewer, we added a paragraph in the statistical analysis. (see page 7, lines 4-6). Authors: for the quality of LDL analysis, we compared T0 with T6 using non-parametric test, as these data did not have Gaussian distribution. Statistical analysis was reviewed and we chose Dunn’s test after a Kruskal-Wallis test statistically significant, to determine exactly which groups are different. This analysis seems appropriate. (Reference: Dinno, A., 2015. Nonparametric pairwise multiple comparisons in independent groups using Dunn's test. The Stata Journal, 15(1), pp.292-300. 6. PLOS authors have the option to publish the peer review history of their article. If published, this will include your full peer review and any attached files. Authors: Agree. Attachments Attachment Submitted filename: Response to reviewers.docx https://doi.org/10.1371/journal.pone.0273292.r002
8 Aug 2022 Decision Letter - Salvatore De Rosa, Editor Changes in lipoproteins associated with lipid-lowering and antiplatelet strategies in patients with acute myocardial infarction PONE-D-21-17809R1 Dear Dr. Fonseca, 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, Salvatore De Rosa 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: (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: Pape r titled (Changes in lipoproteins associated with lipid-lowering and antiplatelet strategies in patients with acute myocardial infarction), thanks for addressing the reviewer's comments ****** 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: Yes:** Sawsan Zaitone https://doi.org/10.1371/journal.pone.0273292.r003
Formally Accepted
18 Aug 2022 Acceptance Letter - Salvatore De Rosa, Editor PONE-D-21-17809R1 Changes in lipoproteins associated with lipid-lowering and antiplatelet strategies in patients with acute myocardial infarction Dear Dr. Fonseca: 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. Salvatore De Rosa Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0273292.r004

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