PONE-D-19-31936

Anti-inflammatory activity of the dietary supplement Houttuynia cordatafermentation product in RAW264.7 cells and Wistar rats

PLOS ONE

Dear Dr. Senawong,

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PLOS ONE

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a) Please provide details of animal welfare (e.g., shelter, food, water, environmental enrichment)

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d) Please describe the frequency of monitoring and the criteria used to assess animal health and well-being.

3) We also ask that you please provide in your Methods section, additional details regarding the source of the Houttuynia cordata fermentation product used in this study. Please provide the product number, lot number, and full ingredient list to ensure reproducibility of the analyses.

4) Please also clarify whether your animal ethics committee specifically approved this study.

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 [This study was supported by the Research and Researcher for Industry (RRi) project, Thailand Research Fund (TRF), which was cooperated with the Prolac (Thailand) Co., Ltd., Lamphun Province, Thailand.].               

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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: Yes

Reviewer #3: Yes

Reviewer #4: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: 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: No

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: 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: No

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: 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: Title: “Anti-inflammatory activity of the dietary supplement Houttuynia cordatafermentation product in RAW264.7 cells and Wistar rats”

Corresponding Author: Thanaset Senawong

In the paper entitled “Anti-inflammatory activity of the dietary supplement Houttuynia cordatafermentation product in RAW264.7 cells and Wistar rats” by Khanutsanan Woranam et al ., had showed Houttuynia cordatafermentation acts as an Anti-inflammatory RAW264.7 cells and Wistar rats . Overall, the results of this study are of some values in revealing its role in Anti-inflammatory activity.

Below are some examples of issues in which the authors might want to work on.

Many grammar, format and language mistakes are present throughout the manuscript. Manuscript need to have a review by English editor.

Material and method:

Nitrite determination was performed by spectophotmetry or any other method if so could you please explain this why spectrofluorimetric was not used as this method is considered as attractive due to its facility availability, high sensitivity and selectivity, low limits of detection and low-cost.

Primer sequences placed in method section should be added as supplementary table.

Was time course study performed when incubated with LPS (1 μg/mL) in case of RNA isolation and protein extraction.

Result section:

How many sets of experiments were performed for the cell viability assay? Could you add cell pictures before and after treatment?

In figure 2C the expression level of IL-1β when treated HCFP methanolic extract

At 12 (μg/mL) showed decreased level of expression while in quantitativeresults figure 2D showed increased level. Could you please explain which method was used for the quantitative analysis?

Discussion

The discussion part is poorly described it need to be revised and latest references should be added.

The manuscript is not acceptable in the present format. It needs to be extensively revised.

Reviewer #2: The manuscript is providing some nice conclusions about a potent dietary supplement, however, I have a concern about the conclusion. This needs to be rewritten according to the results obtained and should't be so general. Further not a single reference is quoted from the literature published in 2019. Hence, the latest references should be added.

Reviewer #3: The paper is very interesting and designed diliginty in a well professional manner. The paper entitled"Anti-inflammatory activity of the dietary supplement Houttuynia cordatafermentation product in RAW264.7 cells and Wistar rats" is considered a new approach in combining effect of Houttuynia cordatafermentation product both invivo and invitro by carrying out well designed experiments and performing cell viability test by explanation in-details and the authors reported all the kits sources and also the approval number of animal ethics in their university, the statistical analysis was performed professionally , the discussion part is brilliant in explaining the causes of inflammation and the efficacy of the used extract .

So i recommend publication of this paper which deserve publication and will be of great benefit for readers.

Reviewer #4: Thank you to give me the opportunity to review this paper

I found this is interesting.

My comments:

1. I am warried that the HCFP has a degraded the protein or has cytotoxicity on the cells.

2. It suggested to do another viability assay to confirm the result

Thanks

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: Yes: Dr. Reham Zakaria Hamza

Reviewer #4: Yes: Abdullah M Alkahtani

[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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Anti-inflammatory activity of the dietary supplement Houttuynia cordata fermentation product in RAW264.7 cells and Wistar rats

PONE-D-19-31936R1

Dear Dr. Senawong,

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.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and 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 enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and 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.

With kind regards,

Walid Elfalleh, Ph.D

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 #4: 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: Partly

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: thank you for the answering the comments. In my opinion this manuscript is fulfilling the requirements technically for the publication in PLOSone. My comments are accept this manuscript publication.

Reviewer #4: I think this work is nice and the data is publishable and make sure about the most important massages of this paper

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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: Yes: Abdullah M Alkahtnai