PONE-D-22-13009Association of polycyclic aromatic hydrocarbons in moss with blood biomarker among nearby residents in Portland, OregonPLOS ONE

Dear Dr. Burstyn,

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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Fung-Chi Ko, PhD

Academic Editor

PLOS ONE

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Additional Editor Comments:

Reviewer’s comments:

Although inconclusive, the manuscript entitled "Association of polycyclic aromatic hydrocarbons in moss with blood biomarker among nearby residents in Portland, Oregon," is interesting and may be useful to other researchers. Yet some gaps need to be addressed before publication. One issue which needs to be addressed stems from surveys of volunteers asking about possible exposure within 48 h hours. This short period is not explained or discussed, although PAHs-adduct half-lives are estimated to be 3-4 months. The authors state that half-lives of PAHs in moss are unknown. However, one would expect, based on PAH stability, they will be at least on a month scale that is much longer than 48h hours.

It appears the authors omitted the description of sample preparation and analysis of DNA adducts and which specific adducts were analyzed. Alternatively, the reference to the lab performing the work should be provided if this was done as a service. Furthermore, some of the material reported in the results belong to the experimental section. The results section is surprisingly short, extended by the lengthy, simply formatted table. Nevertheless, the authors further narrowed it by focusing on only three representatives PAHs.

The major revision of the manuscript needed to address these issues and detailed comments below.

Detailed comments

Line 19. Description of PAHs – which is a full class of compounds as "component" is awkward; please rephrase perhaps as follows "Polycyclic aromatic hydrocarbons (PAHs) are air pollutants.".

The direct speech in the abstract using "we" pronoun is refreshing but somewhat overused. To make articles easier to read, some sentences should be rewritten. This is also the case for other sections of the paper.

Line 30. It should be "adducts" in the plural.

Line 76. In this section, the authors carefully describe IRB protocol and data collection. The questionnaire included information of possible exposure to PAHs for 48h. Yet the question is if this is sufficient time for correlations made. The PAHs are quite stable lipophilic compounds that can bioaccumulate. Thus investigation to relationships with longer times seems to be more appropriate. Perhaps authors need to add some explanation for this or can include in the discussion how their approach is appropriate.

Lines 84, 90. The symbols for degree Celsia are inconsistent; please ensure correct symbols are used everywhere. Usually, "space" is in front of the degree symbol.

Line 93. Use N2 for nitrogen gas.

Line 96. "Mass selective detector" does not need to be capitalized. Also, provide mass range and ionization – presumably electron ionization.

Line 101. Consider switching the order of 1st two sentences – The current presentation of the 1st sentence without specification of type of statistical analysis is confusing.

The experimental section does not address analysis of the PAH-adduct markers. Some material from Lines 118-126 including figure 1 belongs to the experimental section.

Lines 128-131 also belong to the experimental section, supporting the study's quality but not being actual results.

Line 137-139 describes how the study was narrowed, yet correlation for other PAHs could be performed and included in the expanded figure or supplementary materials. Such data would add strength to the paper.

Line 140. The authors mention that specific values for concentration are not informative due to the lack of data on PAHs in moss. It seems it would be useful to compare these results to papers referred in the introduction, justifying the use of this approach.

Table 1 need significant revision to achieve a professional look, but also concerning content. PAHs naming nomenclature needs to be revised (there are numerous excellent resources on correct naming). The alphabetic order of PAHs is confusing; it would be better to organize PAH based on volatility or the number of rings and also provide their totals. Correlations with totals of these groupings could be insightful and give a different look at attempted correlation.

[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: No

Reviewer #2: Partly

**********

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

Reviewer #1: No

Reviewer #2: Yes

**********

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

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

Reviewer #1: No

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: The possible relationship between atmospheric PAHs and ones in human blood. This is not clear yet, therefore this study did not conclude any relationship between bioindicator and human blood. This result is common and expected. If the authors show evidences for this unclear circumstance, it would be orijinal for the readers.

Reviewer #2: Although inconclusive, the manuscript entitled "Association of polycyclic aromatic hydrocarbons in moss with blood biomarker among nearby residents in Portland, Oregon," is interesting and may be useful to other researchers. Yet some gaps need to be addressed before publication. One issue which needs to be addressed stems from surveys of volunteers asking about possible exposure within 48 h hours. This short period is not explained or discussed, although PAHs-adduct half-lives are estimated to be 3-4 months. The authors state that half-lives of PAHs in moss are unknown. However, one would expect, based on PAH stability, they will be at least on a month scale that is much longer than 48h hours.

It appears the authors omitted the description of sample preparation and analysis of DNA adducts and which specific adducts were analyzed. Alternatively, the reference to the lab performing the work should be provided if this was done as a service. Furthermore, some of the material reported in the results belong to the experimental section. The results section is surprisingly short, extended by the lengthy, simply formatted table. Nevertheless, the authors further narrowed it by focusing on only three representatives PAHs.

The major revision of the manuscript needed to address these issues and detailed comments below.

Detailed comments

Line 19. Description of PAHs – which is a full class of compounds as "component" is awkward; please rephrase perhaps as follows "Polycyclic aromatic hydrocarbons (PAHs) are air pollutants.".

The direct speech in the abstract using "we" pronoun is refreshing but somewhat overused. To make articles easier to read, some sentences should be rewritten. This is also the case for other sections of the paper.

Line 30. It should be "adducts" in the plural.

Line 76. In this section, the authors carefully describe IRB protocol and data collection. The questionnaire included information of possible exposure to PAHs for 48h. Yet the question is if this is sufficient time for correlations made. The PAHs are quite stable lipophilic compounds that can bioaccumulate. Thus investigation to relationships with longer times seems to be more appropriate. Perhaps authors need to add some explanation for this or can include in the discussion how their approach is appropriate.

Lines 84, 90. The symbols for degree Celsia are inconsistent; please ensure correct symbols are used everywhere. Usually, "space" is in front of the degree symbol.

Line 93. Use N2 for nitrogen gas.

Line 96. "Mass selective detector" does not need to be capitalized. Also, provide mass range and ionization – presumably electron ionization.

Line 101. Consider switching the order of 1st two sentences – The current presentation of the 1st sentence without specification of type of statistical analysis is confusing.

The experimental section does not address analysis of the PAH-adduct markers. Some material from Lines 118-126 including figure 1 belongs to the experimental section.

Lines 128-131 also belong to the experimental section, supporting the study's quality but not being actual results.

Line 137-139 describes how the study was narrowed, yet correlation for other PAHs could be performed and included in the expanded figure or supplementary materials. Such data would add strength to the paper.

Line 140. The authors mention that specific values for concentration are not informative due to the lack of data on PAHs in moss. It seems it would be useful to compare these results to papers referred in the introduction, justifying the use of this approach.

Table 1 need significant revision to achieve a professional look, but also concerning content. PAHs naming nomenclature needs to be revised (there are numerous excellent resources on correct naming). The alphabetic order of PAHs is confusing; it would be better to organize PAH based on volatility or the number of rings and also provide their totals. Correlations with totals of these groupings could be insightful and give a different look at attempted correlation.

**********

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.]

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-22-13009R1Association of polycyclic aromatic hydrocarbons in moss with blood biomarker among nearby residents in Portland, OregonPLOS ONE

Dear Dr. Burstyn,

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.

Your manuscript revision has now been reviewed, and the reviewers still proposed a few amendments.  Having looked at your paper myself, I agree with the reviewers that it needs some minor work on the nomenclature of PAHs and restructuring by increasing the discussion regarding the no correlation of PAH levels in moos and PAH-DNA adducts in human blood, which seems to make more sense, and would likely attract a wider readership.

You are requested to consider these carefully and to prepare a revised version of the paper.  After your amendments, please return the manuscript to me via the system, and we will reconsider it for publication.  Please ensure you provide a revised version of the manuscript along with any Figures and Tables.

Please submit your revised manuscript by Jan 02 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,

Fung-Chi Ko, PhD

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.

Additional Editor Comments:

Reviewers' comments:

Reviewer #1:  PAHs are important air pollutants with very common sources. Many studies are being conducted to establish a relationship with the personal exposures of these pollutants. While it is difficult to establish a statistical relationship between PAHs and human disease even in active sampling and personal exposure studies, it is much more difficult to demonstrate this relationship with bioindicators. (if possible, more discussion is strongly recommended to reveal the relationship).

Reviewer #2:  The authors addressed the majority of comments, I have just a couple of comments:

I still disagree with naming nomenclature. The EPA is agency providing regulations, limits, and guidelines, not nomenclature. The letters in brackets should be in italics, and the brackets should be square. For details, see NIST reference https://nvlpubs.nist.gov/nistpubs/Legacy/SP/nistspecialpublication922.pdf

Line 101.I presume it should be “Restek” column.”

Reviewer #3:  Algae are used to determine PAH levels in outdoor air, but not enough to compare this with human DNA. As you mentioned, it is costly to examine the external environment with classical methods. For this reason, it is being investigated which plant species to use or not to use to determine the atmospheric levels of persistent organic pollutants. The results section is very short.

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 #3: (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 #2: Yes

Reviewer #3: (No Response)

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

Reviewer #2: Yes

Reviewer #3: (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 #2: Yes

Reviewer #3: (No Response)

********** 

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: Yes

Reviewer #3: (No Response)

********** 

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: The authors addressed the majority of comments, I have just a couple of comments:

I still disagree with naming nomenclature. The EPA is agency providing regulations, limits, and guidelines, not nomenclature. The letters in brackets should be in italics, and the brackets should be square. For details, see NIST reference https://nvlpubs.nist.gov/nistpubs/Legacy/SP/nistspecialpublication922.pdf

Line 101.I presume it should be “Restek” column.”

Reviewer #3: Algae are used to determine PAH levels in outdoor air, but not enough to compare this with human DNA. As you mentioned, it is costly to examine the external environment with classical methods. For this reason, it is being investigated which plant species to use or not to use to determine the atmospheric levels of persistent organic pollutants. The results section is very short. For this reason, I could not find a result that could change my initial thought. The authors described their work poorly. For this reason, I do not consider it a work worth publishing.

********** 

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 #3: 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.

Association of polycyclic aromatic hydrocarbons in moss with blood biomarker among nearby residents in Portland, Oregon

PONE-D-22-13009R2

Dear Dr. Burstyn,

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

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

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

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

Kind regards,

Fung-Chi Ko, PhD

Academic Editor

PLOS ONE