PONE-D-23-36193Nutritional, molecular and functional properties of a novel enzymatically hydrolyzed porcine plasma productPLOS ONE

Dear Dr. Solà Ginés,

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PLOS ONE

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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

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: 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

Reviewer #2: 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

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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 authors use a sound methodology and carry out the experiments with precision. The manuscript is well written and easy to follow. The results are scientifically sound and of high quality, the methods are appropriate, and the assays were well performed.

The in vivo experiments in C. elegans and mouse models of intestinal inflammation provide compelling evidence for the anti-inflammatory effects of EHPP, showing improvements in survival, motility, and attenuation of pro-inflammatory responses.

Overall, this study shows that EHPP is a promising candidate for health benefits, possibly due to its characteristic low molecular weight bioactive peptides, that show potential as immunomodulatory and antioxidant agents.

There's a particular point to highlight about bioactive peptides (BAPs) in lines 82 to 89. These peptides aren't just components of food or inactive parts of proteins in their original molecules. They can also be found on or produced by microorganisms, plants, or animals. Therefore, I recommend that the authors expand on BAP with a more comprehensive explanation in the Introduction. Therefore, I suggest that the authors include a more detailed description of BAP in the Introduction.

The comparison between EHPP and SDP shows differences in their mechanisms of action, especially in the markers IL-6 and TNF-alpha, suggesting different pathways for their beneficial effects. Do the authors have an explanation for this difference?

A paper comparing the nutritional composition of SDP and EHPP was published in December 2023, https://doi.org/10.3390/molecules28237917. I suggest that the authors compare their results with those presented in this paper.

Reviewer #2: The manuscript provides valuable insights into the nutritional and molecular properties of EHPP. However, I have some specific comments on certain sections of the manuscript. I trust that the authors will carefully consider my suggestions and make appropriate improvements.

Major Problem:

One significant concern that requires attention is the lack of clarity in the figures. While I trust the authors' interpretation of the results, the figures need improvement as they are currently unreadable.

Minor Comments and Questions:

Lines 109-114: The detailed information provided in this section appears excessive and may be more appropriately placed elsewhere in the text, perhaps in the methodology or discussion. Consider relocating this content to maintain a smoother flow in the presentation.

Lines 123-124: The statement "Crude fiber (CF) was used as an indicator of fiber digestion" requires clarification. Please explain the intended meaning.

Lines 124-125: The phrase implies that tryptophan is not an amino acid. Revise the sentence for clarity.

Methodology section: It is recommended to avoid the use of the first person in describing methods. Convert sentences to the impersonal form for a more formal tone and structure.

Table 1: Why was less methionine used in the control feed? Additionally, why was less lysine used in the EHPP diet when Table 2 indicates it has a lower tyrosine content than SDP?

Line 357: Is it appropriate to categorize the fat content as "very low"?

Discussion section: At times, navigating through the discussion becomes challenging in distinguishing between studies by other authors and the current results. Emphasizing when discussing the present findings versus referencing other studies would enhance clarity for readers.

Addressing the major concern regarding figure clarity and refining minor details will contribute to the overall improvement of the manuscript.

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Reviewer #1: No

Reviewer #2: No

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Nutritional, molecular and functional properties of a novel enzymatically hydrolyzed porcine plasma product

PONE-D-23-36193R1

Dear Dr. Solà Ginés,

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

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

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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,

Mahmoud A.O. Dawood, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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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 #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

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

Reviewer #1: Yes

Reviewer #2: 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 #2: Yes

**********

6. Review Comments to the Author

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

Reviewer #1: The authors have adequately addressed my comments. Thus, I have come to the conclusion that this manuscript is now considered suitable for publication.

Reviewer #2: (No Response)

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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 #2: Yes: Katarzyna Kazimierska

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