PONE-D-19-24305

Vernonia polysphaera Baker: anti-inflammatory activity in vivo and inhibitory effect in LPS-stimulated RAW 264.7 cells

PLOS ONE

Dear Dr. Almeida-Souza,

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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Partha Mukhopadhyay, Ph.D.

Academic Editor

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: In the present study, the authors evaluated the anti-inflammatory effects of hydroalcoholic extract of a plant Vernonia polysphaera Baker in mouse peripheral and central inflammatory lesion models. The authors performed comprehensive analysis in both in vivo and in vitro effects of said plant extract. The experiments are carried out with balanced designs and results are properly interpreted. There are a few minor details that requires addressing:

Methods: Please indicate type of culture medium used to reconstitute the extract for in vitro and in vivo use (Line 101). The extract was prepared in 3 different ways: in DMSO, in culture medium and/or in PBS. For culture medium and PBS prepared extract, which was used in what experiments? I suggest including the info in respective paragraphs of experiments. Same goes when mentioning control or standard reagents for measurements, e.g. nitrite standards prepared in culture medium if that is the case.

Table 1: include two-tailed p-values for each extract-treated group to control group.

Reviewer #2: In this manuscript, the authors described the action by which organic extracts from Vernonia polysphaera Baker reduces inflammation both in vivo and in vitro. The study was well presented and performed with care. I have the following minor points, mostly relating to the model presented in figure 6.

1) Does Vernonia polysphaera extracts reduce both NFkB activation and pro-inflammatory cytokine production by independent actions as shown in Figure 6? Or is the reduction of pro-inflammatory cytokine a result of reduced NFkB activity?

2) Does Vernonia polysphaera extracts inhibit NFkB signal by interfering with the processing of NFkB precursor or its generation? Ideally, a simple set of experiments that should be included is western blotting for both the precursor and processed NFkB subunits in Vernonia polysphaera treated vs. non-treated groups.

I am a traditionalist concerning quantifying signaling molecules using PCR method and would like to see measurable protein changes. I leave it up to the editor to decide whether this condition needs to be met.

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Reviewer #1: No

Reviewer #2: No

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Vernonia polysphaera Baker: anti-inflammatory activity in vivo and inhibitory effect in LPS-stimulated RAW 264.7 cells

PONE-D-19-24305R1

Dear Dr. Almeida-Souza,

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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With kind regards,

Partha Mukhopadhyay, Ph.D.

Section 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

**********

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: After reading through the revised manuscript, the previous comments are all addressed.

I hope the authors would describe their revisions in the manuscript in response letter as well, as changes in text especially in large paragraphs could result in shifting line numbers and difficulties in tracing back those changes.

Reviewer #2: The authors have addressed all concerns in the revised manuscript and I have no further questions.

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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: No