PONE-D-22-20226Understanding the Effects of Physiological Stress on the P300 ResponsePLOS ONE

Dear Dr. Zhu,

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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Humaira Nisar

Academic Editor

PLOS ONE

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"This work was supported in part by the Australian Research Council (ARC) under discovery grant DP210101093 and DP220100803. Research was also sponsored in part by the Australia Defence Innovation Hub under Contract No. P18-650825, US Office of Naval Research Global under Cooperative Agreement Number ONRG - NICOP - N62909-19-1-2058, and AFOSR – DST Australian Autonomy Initiative agreement ID10134. We also thank the NSW Defence Innovation Network and NSW State Government of Australia for financial support in part of this research through grant DINPP2019 S1-03/09 and PP21-22.03.02."

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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: Partly

Reviewer #2: No

Reviewer #3: No

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

Reviewer #1: No

Reviewer #2: I Don't Know

Reviewer #3: No

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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: No

Reviewer #2: Yes

Reviewer #3: 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

Reviewer #2: Yes

Reviewer #3: Yes

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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: The experimental setup is interesting. However, this reviewer has some concerns which have to address during major revision.

1. The author should define "stress" in this context clearly. It would be better to replace "stress" by the word which can be measured quantitatively and/or having the standard questionnaire in identifying the level of mental or cognitive world load during interested periods. This famous related review article might help P. Consumer Grade EEG Measuring Sensors as Research Tools: A Review.

2. P300 or ERSP visualization and analysis need further improvement and exploration. The recent work with patients might help the author in the improvement of this point. This reviewer suggest the author to study more from A Pilot Study on Visually Stimulated Cognitive Tasks for EEG-Based Dementia Recognition.

3. Also the analysis of EEG to together with response or reaction times can learn from: Supervision of a self-driving vehicle unmasks latent sleepiness relative to manually controlled driving.

Looking forward to reading the revised version.

Reviewer #2: This is an interesting study on P300 responses under real and virtual height exposure. Basically, the experimental design is somewhat unbalanced (and hard to understand from the description) but it’s a good approach towards a quite timely question. And this is the main critique from my side. This is a nice pilot for a completely controlled study, but, in particular with some minor faults in the design and a sample size of 17 that additionally is divided into two groups, this is not sufficient for a publication. Along with this criticism, it should be noted that the main effect (the modulation of P3 amplitude in one of the two groups) just touches significance (Z=1.96, p=0.049) although the authors seem to have renounced to correct their data against alpha inflation.

As the authors claim in the final section, they were kind of surprised by the interindividual differences in behavior. Those are, besides the fact that mobile EEG is used in order to investigate more realistic behavior, an interesting point of this study, however, the authors nowhere provide evidence that there are really two distinct groups. They simply divided their sample by a split half on one RT parameter, which was for sure not bimodally distributed. The reader might have the impression that they searched for significance based on different analyses strategies and this one turned out to provide a p value below .05.

The entire MS appears to be intended in another context as presented. The introduction tlks a lot about BCIs, which are nowhere touched experimentally. This topic does not return before the final conclusion. The entire discussion section focuses on stress and cognition, which in my mind is the central issue here.

Minors

• The P300 paradigm is poorly described and not a standard for the P300 literature. Maybe this approach is usual in connection with PCIs, but the majority of visual oddball tasks uses much shorter stimulus presentations, longer interstimulus intervals which are normally additionally jittered.

• Why was the SAM presented before the trials? Would not a survey after performing the task better reflect stress?

• Removing any component that is less than 85% classification for brain component (done with ICLabel?), is a hard criterion. How many ICs remained in the data then?

• The authors appear to make separate tests for within group effecs and between groups effects on the same parameter. Was any correction against alpha error inflation applied?

• statistical parameters are missing in many places. Also not significant effects should be presented with their numbers.

• Another evidence for effect searching: In the introduction they speak about Alpha and Theta (“276 pected P300 ERSP response is the alpha and theta synchronisation during the onset (before) of the P300 peak and a gradual beta desynchronisation during the P300 peak”), in the result section they present an effect in Beta power, and in the conclusion they added gamma activity. When I look at the time frequency plots, I rather see Mu responses in Group 2, which might reflect encapsulation due to the balance task.

Reviewer #3: This study tries to address the influence of stress under a naturalistic environment on the P300 response. This research is very practical, and the paradigm design is also very interesting. However, many critical problems of statistical analysis in this manuscript make the results of this study unconvincing. Carefully checking the processes of statistical analysis and data processing are necessary before resubmitting the manuscript.

Major concerns:

1. In the pipeline of EEG preprocessing, the CAR can’t be done before the ICA. As warned by the tutorial of the EEGLab and the theory of ICA, the common averaged reference will reduce the rank of data. The sequence of preprocessing processes needs to be carefully examined.

2. What is the reference for choosing the time interval from 300ms to the reaction time for peak amplitude extraction? Please provide a further explanation of the temporal analysis window.

3. Please give a further illustration of the “median split” of dividing participants into two groups. Moreover, as the split is performed “for difference in RT between GG and PH”, this approach will increase the type I error of the following statistical analysis. If such a split is necessary, the split based on objective conditions such as baseline conditions is more acceptable.

4. In the paradigm design, four tasks, GG, PG, PP, and PH, are set. Why are there no GP and GH tasks?

5. In addition, how is the order of the actual scene and the virtual scene determined? Why is the actual scene placed before the virtual scene? Or in other words, why use PG instead of GP?

6. There should be four kinds of experiments in Randomized Walking Conditions in Figure 2, and only 1 kind of GG is drawn in this figure.

7. The authors need to supplement the questionnaire used by SAM. Was this questionnaire filled out before and after the experiment or just after the experiment for each condition?

8. The authors divided the subjects into 2 groups according to RT. Please add the RT result of each subject.

9. The authors conducted a correlation analysis between SAM and RT, but this paper wants to study the relationship between Physiological Stress and P300. Why do you not perform a correlation analysis between SAM and the amplitude or power of P300 in the two groups of subjects?

10. It can be seen from Figure 6 that Pz is not the strongest in the EEG scalp map of Group 2, so is it reasonable to use only Pz to calculate the P300 amplitude in Figure 7?

11. There are many problems in the statistical test. First, a major problem is using the paired test for multi-level comparison. As mentioned in the manuscript, “the RT, SAM, P3 peak and frequency power band data were not normally distributed”, and some non-parameter methods like the Friedman test can be used to determine whether multiple conditions are significantly different. The paired test can not determine the effect of a factor with multi-level. This problem influences the statistical test of RT, SAM, ERP amplitude, and the comparison between two groups. Second, the correction of the p-value is ignored when multi-comparisons are made. The manuscript doesn’t give an illustration of the correction method when RT, SAM, and ERSP are compared among four conditions (i.e. GG, PG, PP, and PH). Third, the objects of statistical analysis are not uniform. In some analyses, the object is the mean value of each subject, while in other analyses, the object is the value of each trial. Please choose a uniform object and ensure that the object meets the assumptions of the statistical test method.

5. The specific details of ERSP need further explanation. For example, the method of doing time-frequency decomposition, the window length of ERSP, the baseline correction method of ERSP, and the epoch range of data used by ERSP are needed to determine the effectiveness of ERSP results.

Minor issues

1. On page8 line 299, there should be repeated measures ANOVA instead of “repeated measure ANOVA”.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

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PONE-D-22-20226R1Understanding the Effects of Physiological Stress on the P300 ResponsePLOS ONE

Dear Dr. Zhu,

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 submit your revised manuscript by Apr 21 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,

Steve Zimmerman, PhD

Associate Editor, PLOS ONE

Additional Editor Comments: 

Thank you for your careful revision to your manuscript. Unfortunately two of the three original reviewers were unable to review your revised submission, therefore we sought out one additional reviewer. This reviewer (reviewer 4) still has some concerns about the the definition of physiological stress used, and the differentiation between groups (see attached file).

Could you please carefully revise the manuscript to address all comments raised?

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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

Reviewer #4: (No Response)

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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

Reviewer #4: Partly

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

Reviewer #1: Yes

Reviewer #4: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #4: Yes

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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 #4: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #4: More information is required to qualify claims made concerning the measures taken linking to the definition of stress used (including in the title). The study is a sound basis as a pilot for a more comprehensive study addressing points made in attached reviewer feedback about the differentiation of groups on which the main conclusions are based.

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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: No

Reviewer #4: No

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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.

Attachments

Attachment

Submitted filename: PONE-D-22-20226R1_review DG_GW.docx

PONE-D-22-20226R2Understanding the Effects of Stress on the P300 ResponsePLOS ONE

Dear Dr. Zhu,

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 submit your revised manuscript by Sep 10 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,

Valentina Bruno

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.

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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 #4: (No Response)

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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 #4: Partly

**********

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

Reviewer #4: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #4: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #4: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #4: The authors have resubmitted the manuscript: “Understanding the Effects of Stress on the P300 Response” following reviewer feedback that raised some concerns based on the previous draft for submission. A primary issue highlighted previously centred on the title and contention that the study was evaluating physiological aspects of stress in the experimental scenario employed. Questions were also raised in relation to the Self Assessment Mannikin measures used: specifically only measuring the arousal dimensions (and not the valence and dominance components of the SAM). Objections were raised as to whether the experimental set-up was investigating physiological or psychological aspects of stress, and that by only capturing data on the arousal dimension, this might provide an incomplete picture of the model of stress being alluded to. Further comments questioned claims regarding ability to cope with stress and P300, despite no effects being observed when groups were split.

The authors have addressed key points including specific allusions to ‘physiological’ stress, removing reference to this. They also explained that the logistics of the experimental scenario meant that incorporating all of the dimensions of the SAM would have interfered with the naturalistic context in which the participant was situated. They provide a rationale on why the arousal dimension is valid to include as a measure in itself, as this has been shown to have a close relationship to threat perception. This reviewer recognizes the unique challenge that exists when running experiments that preserve a naturalistic context in which to elicit behaviours of interest that occur in real world situations, whilst also serving to employ robust measures that are usually collected in more contrived laboratory scenarios. This addresses the concerns around definitions of stress and the reasoning for using the subjective measures (SAM).

However, with respect to the analysis and assertions made about how the P300 relates to ability to cope with stress, the authors were unable to split participants based on a marker of stress. Rather, the split between groups was done on the basis of reaction times. As mentioned previously, splitting by reaction times can involve confounds unrelated to stress. This should be acknowledged in the manuscript, or risk overstating claims made. Whilst the authors now provide the distribution of trials, it remains unclear if any speed-accuracy trade-off was apparent, or if there were any statistical differences between groups regarding error rates.

On the basis of the points addressed but with respect to the basis for splitting the data, it is recommended that minor revisions of the manuscript are made, modifying the discussion and conclusions to reflect the possibility that differences due to the split on RTs may not be specifically due to stress, and downplaying claims made about markers of ability to cope with stress. Such minor revisions would in this reviewer’s view satisfy criteria for publication.

One additional suggestion concerns the title. It may be useful to emphasise the nature of the stressor/scenario from which the effects of stress are being derived, for specificity. For instance highlighting that this work is seeking to novelly extend research into the stress response/P300 into a more naturalistic environmental context, which is a particular strength of the work. This could prove fruitful for future lines of enquiry that differentiate ‘stress’ according to varied contexts beyond height exposure.

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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 #4: 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.

PONE-D-22-20226R3Understanding the Effects of Stress on the P300 Response During Naturalistic Simulation of Heights ExposurePLOS ONE

Dear Dr. Zhu,

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 submit your revised manuscript by Jan 19 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

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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,

Valentina Bruno

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.

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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 #2: (No Response)

Reviewer #4: All comments have been addressed

Reviewer #5: 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 #2: (No Response)

Reviewer #4: Yes

Reviewer #5: Yes

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

Reviewer #2: (No Response)

Reviewer #4: Yes

Reviewer #5: Yes

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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 #2: (No Response)

Reviewer #4: Yes

Reviewer #5: Yes

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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 #2: (No Response)

Reviewer #4: Yes

Reviewer #5: Yes

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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 #2: (No Response)

Reviewer #4: The authors have resubmitted the manuscript: “Understanding the Effects of Stress on the P300 Response During Naturalistic Simulation of Heights Exposure” following additional reviewer comments recommending minor revision to the previous revised draft.

The reviewer feedback concerning the split between groups has been more or less addressed with respect to potential confounds and limitations (eg. RTs perhaps being affected by factors other than ‘stress’ per se), including a reference to the accuracy scores. Although in the limitations section there is emphasis placed on RTs with respect to utility as a metric for gauging/estimating susceptibility to stress in conjunction with brain behaviour.

Other comments have been addressed from earlier revisions concerning definition of stress, although this reviewer still has some reservations about the qualification of stress in the experiment.

“One group performed worse when stressed” - statements like this make presumption that a stress response has been elicited rather than the contention that the cognitive-affective-arousal state markers may be indicative of so-called ‘stress’ - this may sound like splitting hairs but nevertheless feels poignant to the argument being made in the paper about potential indicators of stress identified via the P300 and cognitive-arousal markers of performance...Indeed, the authors refer to ‘stress levels’ when they are talking about the different conditions. These would perhaps be more objectively described as ‘potential stress-inducing conditions’ - isn’t the point of the experiment to examine whether, and to what degree, these conditions do actually elicit changes in ‘stress levels’?

Another point of contention rests in the defining of stress, in absolute, as a negatively valenced state. Whilst the authors reference the circumplex model, with stress generally associated with high arousal/negative valence, this generalisation does not reveal the whole picture. Further subdivision of stress into negative and positive dimensions (as is increasingly occurring in the literature – eg. Bak et al., 2022) - ‘distress’ and ‘eustress’ - paints a more complex and varied picture of stress. This may ultimately contribute to resolution of inconsistency in findings in the literature with respect for instance, to alternately, cognition-impairing and cognition-enhancing effects of exposure to ‘stress’. It is a relevant point, particularly as research extends further into more naturalistic experimental scenarios territory, as the current study valiantly attempts. This is alluded to somewhat in the section: “Understanding Yerkes-Dodson Law and Participant’s Performance when Stressed” in discussion of Group 2 vs Group 1 experiencing ‘more optimal stress level’.

[Bak, S., Shin, J., Jeong, J. (2022). Subdividing Stress Groups into Eustress and Distress Groups Using Laterality Index Calculated from Brain Hemodynamic Response. Biosensors (Basel). 12(1):33.]

“In summary, the two groups we found exhibited seemingly contradictory yet rational and supported by literature, P300 behaviours” - this sentence doesn’t read clearly.

In this reviewer’s opinion the points raised from previous review have been somewhat satisfactorily addressed, but note the above comments with respect to wider debate about what constitutes ‘stress’ and how the label is employed with respect to outlining the experimental conditions and asserting that a state of stress is variably elicited.

On the basis of the revisions made this reviewer recommends that the paper is fit for submission.

Reviewer #5: This is a very interesting study, will contribute much for understanding the neural correlates of Stress. I appreciate this work on the idea and manipulations in the experiments, and also have some suggestions for clear state the findings and implications.

1. How to clearly distinguish the effect of emotional and stress during the procedure of experiment?

2. Is it possible to consider the link between stress and neural response in line with a curve model? That is, why participants performed better in some specific not all the conditions? How to reasonably to explain it?

3. What's the meanness to discriminate the real and virtual environment?

4. Did you control the daily stress and stress sensibilities in the data analysis?

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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 #2: No

Reviewer #4: No

Reviewer #5: Yes: yes

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[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.

PONE-D-22-20226R4Understanding the Effects of Stress on the P300 Response During Naturalistic Simulation of Heights ExposurePLOS ONE

Dear Dr. Zhu,

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 submit your revised manuscript by Apr 05 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Valentina Bruno

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 #5: 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 #5: Partly

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

Reviewer #5: 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 #5: Yes

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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 #5: 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 #5: Thanks for reviewing the paper again, the quality of the article has improved much. The authors answered the questions accurately and rigiously, I have one question that How to difine the virtual and real enviorment in the present study. How to exclude the effect of experiences from real environment in the virtual environment condition?

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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 #5: Yes: Fanchang Kong

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[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.

Understanding the Effects of Stress on the P300 Response During Naturalistic Simulation of Heights Exposure

PONE-D-22-20226R5

Dear Dr. Zhu,

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

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at http://www.editorialmanager.com/pone/ and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Valentina Bruno

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 #5: 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 #5: Yes

**********

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

Reviewer #5: 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 #5: 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 #5: 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 #5: The authors have answered my suggestions well, I think, I have no new questions for them. The current state of MS should be accepted to publish.

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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 #5: Yes: Fanchang Kong

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