PONE-D-19-33766

Perinatal factors affecting platelet parameters in late preterm and term neonates.

PLOS ONE

Dear Dr. Hayato Go,

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 address the comments identified by the two reviewers at your earliest convenience

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Anna Palatnik, M.D.

Academic Editor

PLOS ONE

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Additional Editor Comments (if provided):

Reviewer 1:

Authors have chosen interesting subject for research.

However, there are many unanswered questions in the mansucript-

1. The study is retrospective in nature; what were the indications for doing all the studied platelet parameters in these late preterm and term neonates at the first place? Is that a routine practice in their unit to screen all babies for platelet issues? Is it justified ethically?

2.All late preterms and term neonates admitted during the study period (491 total) were included in the study and there is no exclusion.How is the data managed in the unit?

3. The data shown in table 1 describes platelet parameters between late PT and term neonates. The given data is highly inadequate. There is no mention whether it is mean or median and no standard deviation or IQR is shown.Further except for GA, BW and PCT , other values are exactly the same which further raise queries on data collection and analysis.

4.Again in table 2 and 3, there is no mention of measures of variation.In multivariate analysis , odds ratio should have been shown instead of just p value.

5.Authors mention use of cord blood or venous blood at admission for study.What site of cord blood was used whether maternal side or neonatal side?Further the mean sampling time was under 2 hours , the studied parameters might reflect maternal characteristics. It would have been clinically relevant if postnatal changes also studied simultaneously.

6.Authors rightly acknowledged their limitations about sepsis and its effect on platelet parameters, which is a major factor which affect platelets

7.Typographical error in the introduction first line -"....in various disease diseases"; remove disease.

Reviewer 2:

General Comments:

This manuscript attempts to provide a cross sectional observation of the platelet’s morphology parameters associated with late preterm and term births. While compelling, the manuscript is lacking in both writing and grammatical issues as well as is lacking in some important analysis. Overall if these issues are addressed this could be an important contribution to the base of knowledge in neonatal intensive care.

The most obvious omission is the lack of connection between the existing base of knowledge on this topic om platelet function and how this would attribute to a clinical outcome. What is the hypothesis? How would a change in platelet number or size influence clinical care?

Better definition of clinical variables needs to be presented. The AGA and SGA descriptors need to be updated. It is important to note in the demographic table 1, what is the proportion of SGA’s in the term and late preterm population.

Specific comments follow below:

Page: 12

1. The opening sentence of the Introduction is very vague and not compelling. Please be more specific and deliberate in your writing.

2. What is your hypothesis? It’s not clear what you are adding to the existing body of knowledge.

3. What factors? What comparisons? What are the variables you are interested in studying? At this point, this sounds like an observational study, so please state that explicitly.

4. Why was analysis on preterm babies not performed? That data is discussed in the introduction, but you do not provide a rationale for why it is not included in this study. Why did you limit to term and late preterm?

Page 13:

1. When were these samples collected? Beginning of Day of Life 1? At the end? Any time in the first 24 hours? This will confound your results significantly.

2. One of the factors you have failed to examine is how many babies required resuscitation at birth with epinephrine? Epinephrine has been reported to cause platelet aggregation as well as recruitment into the circulation. See Kjeldsen SE, Weder AB, Egan B, Neubig R, Zweifler AJ, Julius S. Effect of circulating epinephrine on platelet function and hematocrit. Hypertension. 1995;25(5):1096–1105.

3. Furthermore, reactivity to epinephrine via ADP and thromboxane differs between neonates and adults. Rajasekhar D, Barnard MR, Bednarek FJ, Michelson AD. Platelet hyporeactivity in very low birth weight neonates. Thromb Haemost. 1997;77(5):1002–1007.

4. Therefore, it is important to note what the exposure to resuscitation and epinephrine is in these subjects.

Page 14:

1. It would have been nice if platelet morphology was examined via microscopy on peripheral smears

Page 15:

1. Here it would be very important to separate the term and late preterm subjects by AGA and SGA status. So, have four columns instead of two.

Page 17:

1. What is your definition of SGA?

Page 18:

1. You have omitted definitions for several of these variables. Is CAM clinical or culture proven? What is the definition of clinical chorio at your institution?

Page 24:

1. Timing of sampling needs to be clearly stated in the methods.

2. You also need to differentiate which values were collected from cord and which were from venous samples. Were these values compared to one another? If not, it would be vital to know if there is a difference in parameters from these two methods of sampling.

Page 25:

1. Determining exposure to epinephrine and resuscitation may help understand this better. See comment above.

[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: No

**********

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: No

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:

This manuscript attempts to provide a cross sectional observation of the platelet’s morphology parameters associated with late preterm and term births. While compelling, the manuscript is lacking in both writing and grammatical issues as well as is lacking in some important analysis. Overall if these issues are addressed this could be an important contribution to the base of knowledge in neonatal intensive care.

The most obvious omission is the lack of connection between the existing base of knowledge on this topic om platelet function and how this would attribute to a clinical outcome. What is the hypothesis? How would a change in platelet number or size influence clinical care?

Better definition of clinical variables needs to be presented. The AGA and SGA descriptors need to be updated. It is important to note in the demographic table 1, what is the proportion of SGA’s in the term and late preterm population.

Specific comments follow below:

Page: 12

1. The opening sentence of the Introduction is very vague and not compelling. Please be more specific and deliberate in your writing.

2. What is your hypothesis? It’s not clear what you are adding to the existing body of knowledge.

3. What factors? What comparisons? What are the variables you are interested in studying? At this point, this sounds like an observational study, so please state that explicitly.

4. Why was analysis on preterm babies not performed? That data is discussed in the introduction, but you do not provide a rationale for why it is not included in this study. Why did you limit to term and late preterm?

Page 13:

1. When were these samples collected? Beginning of Day of Life 1? At the end? Any time in the first 24 hours? This will confound your results significantly.

2. One of the factors you have failed to examine is how many babies required resuscitation at birth with epinephrine? Epinephrine has been reported to cause platelet aggregation as well as recruitment into the circulation. See Kjeldsen SE, Weder AB, Egan B, Neubig R, Zweifler AJ, Julius S. Effect of circulating epinephrine on platelet function and hematocrit. Hypertension. 1995;25(5):1096–1105.

3. Furthermore, reactivity to epinephrine via ADP and thromboxane differs between neonates and adults. Rajasekhar D, Barnard MR, Bednarek FJ, Michelson AD. Platelet hyporeactivity in very low birth weight neonates. Thromb Haemost. 1997;77(5):1002–1007.

4. Therefore, it is important to note what the exposure to resuscitation and epinephrine is in these subjects.

Page 14:

1. It would have been nice if platelet morphology was examined via microscopy on peripheral smears

Page 15:

1. Here it would be very important to separate the term and late preterm subjects by AGA and SGA status. So, have four columns instead of two.

Page 17:

1. What is your definition of SGA?

Page 18:

1. You have omitted definitions for several of these variables. Is CAM clinical or culture proven? What is the definition of clinical chorio at your institution?

Page 24:

1. Timing of sampling needs to be clearly stated in the methods.

2. You also need to differentiate which values were collected from cord and which were from venous samples. Were these values compared to one another? If not, it would be vital to know if there is a difference in parameters from these two methods of sampling.

Page 25:

1. Determining exposure to epinephrine and resuscitation may help understand this better. See comment above.

Reviewer #2: Authors have chosen interesting subject for research.

However, there are many unanswered questions in the mansucript-

1. The study is retrospective in nature; what were the indications for doing all the studied platelet parameters in these late preterm and term neonates at the first place? Is that a routine practice in their unit to screen all babies for platelet issues? Is it justified ethically?

2.All late preterms and term neonates admitted during the study period (491 total) were included in the study and there is no exclusion.How is the data managed in the unit?

3. The data shown in table 1 describes platelet parameters between late PT and term neonates. The given data is highly inadequate. There is no mention whether it is mean or median and no standard deviation or IQR is shown.Further except for GA, BW and PCT , other values are exactly the same which further raise queries on data collection and analysis.

4.Again in table 2 and 3, there is no mention of measures of variation.In multivariate analysis , odds ratio should have been shown instead of just p value.

5.Authors mention use of cord blood or venous blood at admission for study.What site of cord blood was used whether maternal side or neonatal side?Further the mean sampling time was under 2 hours , the studied parameters might reflect maternal characteristics. It would have been clinically relevant if postnatal changes also studied simultaneously.

6.Authors rightly acknowledged their limitations about sepsis and its effect on platelet parameters, which is a major factor which affect platelets

7.Typographical error in the introduction first line -"....in various disease diseases"; remove disease.

**********

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 to be viewed.]

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 us at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: Summary of Comments on PONE-D-19-33766_reviewer_Bhatti comments.pdf

Attachment

Submitted filename: Summary of Comments on PONE-D-19-33766_reviewer_Bhatti comments.docx

PONE-D-19-33766R1

Perinatal factors affecting platelet parameters in late preterm and term neonates.

PLOS ONE

Dear Dr. Go,

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 justify why the sample size was reduced to 400 from the original 491 in the first submission.

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. We recommend that authors seek independent editorial help before submitting a revision. These services can be found on the web using search terms like “scientific editing service” or “manuscript editing service.”

Please submit your revised manuscript by Dec 05 2020 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Kelli K Ryckman

Academic Editor

PLOS ONE

[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: (No Response)

**********

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: Partly

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: (No Response)

**********

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: No

**********

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 for addressing the comments/reviews you received. Most issues have been adequately addressed. My only remaining comment is that the results section is still difficult to follow with the abundance of abbreviations being used. Table 2 is also very confusing. Perhaps by reversing the X and Y axis, it will be easier to read and follow. This also applies to Table 3 and 4.

Reviewer #2: 1.Although , the authors explain that sampling was done in order to exercise caution , it is still unnecessary to do samplings routinely in newborn and the practice is to avoid unless indicated in order to avoid sampling induced blood loss and consequent anemia.

2. The flow chart has probably typographical error with regards to defining premature birth as > /= 35 weeks .Needs clarification on below which gestation they have defined as premature birth.

3.Regarding the statistical analysis , the authors need to compile the tables more compressed and crisp so that it is reading friendly. Emphasis should be given to what does it imply . The number of tables should be restricted to 2 or 3 maximum.Rest statistical tables can be added as supplementary material

4.Odds ratio may be preferred with 95% CI for interpretation of regression analysis

5.If a complete blood count has also been done along with platelets , then including TLC and ANC as a proxy for sepsis and correlating it with platelet parameters could be of value.

**********

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

[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.

Perinatal factors affecting platelet parameters in late preterm and term neonates.

PONE-D-19-33766R2

Dear Dr. Go,

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.

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Kind regards,

Kelli K Ryckman

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: