PONE-D-19-35694

Sex-specific associations between person and environment-related childhood adverse events and levels of cortisol and DHEA in adolescence

PLOS ONE

Dear Dr. van Dammen,

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.

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Alexandra Kavushansky, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

This is an interesting and potentially important report. There are, however, several issues to address.

A lot of important information is missing. Such as: 1. How LC-MS/MS and ELIZA analyses were performed? 2. How the normality of the data distribution was tested? 3. Was the person performing the analyses blind to the experiment (e.g. participants` groups) details? 4. How was the participants` conformity to following the instructions for gathering and preserving the saliva samples verified? How were the samples delivered to the lab? (frozen?) 5. Why only half of the initial subjects` sample was tested for the saliva measures?

There are several language errors, such as: "…significant lower levels (line 234)", " boys compared than girls (line 270)", etc.

Graphs do not show neither significant results, nor trends.

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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

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

Reviewer #1: Yes

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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

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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

**********

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: This study reports differential associations between person and environment related childhood adversity and functioning of the HPA-axis in girls and boys. The main findings indicate that in girls, person-related childhood adversity was associated with higher hair DHEA levels, while environment-related childhood adversity was not associated with any of the HPA-axis measures. In boys, person-related childhood adversity was associated with higher hair cortisol levels. The introduction gives and adequate background and clearly describes the state of art in the field. The methods are sound and relevant to the topic being researched. Results are mostly clear and well written, and the discussion is properly presented. However, some minor modifications are suggested, as follows:

1.Results Section

1.1 It would be helpful for the reader if the significant differences of table 2 were clearly indicated, along with the respective p value.

1.2 Authors use terms like similar and comparable, instead of no significant differences. Please indicate the p values to clearly indicate that comparable means not significantly different.

1.3 The term “trend” appears in the results and discussion section, but the p value is not shown in the tables or text. Please insert the p value.

2 Discussion section

2.1 Please explain the sentence below in more detail in the discussion section.

“These findings collectively indicate that different types of childhood adversity may be linked to blunted daily cortisol output, but overall prolonged elevated levels of cortisol, suggesting HPA-axis dysregulation, with possible future associations with impaired mental wellbeing.”

How can a daily blunted cortisol lead to an increase in hair cortisol? Would it not be expected that lower daily cortisol levels are associated with lower hair cortisol? How can the results of previous research focusing on non-specific childhood adversity and childhood daily cortisol output be reconciled with the higher hair cortisol shown in this study?

2.2 Are the size effects found in this study for the association of DHEA with person-related childhood adversity in girls, and for the association of hair cortisol and person-related childhood adversity in boys, of clinical relevance? Please compare with the size effect of the association of DHEA and cortisol with the pubertal stage and with other studies, if possible.

2.3 Was the association of the hair cortisol with the AUCg evaluated? If not, please explain why. If it was evaluated, please state the results clearly in the text.

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Reviewer #1: Yes: Elke Bromberg

[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.]

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PONE-D-19-35694R1

Sex-specific associations between person and environment-related childhood adverse events and levels of cortisol and DHEA in adolescence

PLOS ONE

Dear Dr. van Dammen,

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.

We would appreciate receiving your revised manuscript by May 25 2020 11:59PM. When you are 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.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter.

To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

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). This letter should be uploaded as separate file and labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'.

Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

We look forward to receiving your revised manuscript.

Kind regards,

Alexandra Kavushansky, PhD

Academic Editor

PLOS ONE

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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

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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

**********

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

Reviewer #1: 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

**********

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

**********

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: Although all the comments were addressed, some important questions were only partially answered. Please see below:

2 Discussion section

2.1 Please explain the sentence below in more detail in the discussion section.

“These findings collectively indicate that different types of childhood adversity may be linked to blunted daily cortisol output, but overall prolonged elevated levels of cortisol, suggesting HPA-axis dysregulation, with possible future associations with impaired mental wellbeing.”

How can a daily blunted cortisol lead to an increase in hair cortisol? Would it not be expected that lower daily cortisol levels are associated with lower hair cortisol? How can the results of previous research focusing on non-specific childhood adversity and childhood daily cortisol output be reconciled with the higher hair cortisol shown in this study?

The authors did not answer the questions bellow:

- How can a daily blunted cortisol lead to an increase in hair cortisol?

- Would it not be expected that lower daily cortisol levels are associated with lower hair cortisol?

- How can the results of previous research focusing on non-specific childhood adversity and childhood daily cortisol output be reconciled with the higher hair cortisol shown in this study? Note that the question is about low daily cortisol and higher hair cortisol.

Even if there is no conclusive explanation to these questions, the authors should discuss the apparent contradictory results in daily cortisol and hair cortisol. Are there other studies, with children or other developmental stages, that show increased hair cortisol and blunted daily cortisol? These questions are important because hair and daily cortisol represent different time windows, i.e., chronic versus acute cortisol levels. Moreover, the decrease in cortisol seen in this study didn’t reach statistical significance (p=0.08). Is the conclusion about blunted cortisol levels in this study perhaps overestimated?

2.3 Was the association of the hair cortisol with the AUCg evaluated? If not, please explain why. If it was evaluated, please state the results clearly in the text.

Below is the answer given by the authors:

“Yes, it was evaluated, and the results are described in the descriptive analyses section in the results section (lines 217-220).”

Please note that the question is about the association of the hair cortisol with the AUCg, whereas the answer refers to the comparison of hair cortisol samples and the separate saliva cortisol samples (not to the AUCg, as asked). The analysis of the association between AUCg and hair cortisol would perhaps be helpful to the answer of comment 2.1.

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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: Elke Bromberg

[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.

Sex-specific associations between person and environment-related childhood adverse events and levels of cortisol and DHEA in adolescence

PONE-D-19-35694R2

Dear Dr. van Dammen,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

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With kind regards,

Alexandra Kavushansky, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: