PONE-D-25-038001-methylnicotinamide attenuated inflammation and regulated flora in Necrotizing enterocolitisPLOS ONE

Dear Dr. KANG,

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PLOS ONE

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Additional Editor Comments :

Dear Authors,

the manuscript has been revised and has been found to have some concerns that should be addressed as requested by reviewers in order to consider the manuscript suitable for publication.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: The manuscript is interesting and generally well written. However, it presents several points that deserve to be improved. In particular:

Introduction:

- It deserves to be pointed out that NEC are an important risk factor for colorectal cancer (see PMID: 28586045)

- "NNMT, a cytoplasmic methyltransferase that catalyzes the N-methylation of NAM to form ........inflammation through its metabolite, 1-MNA13-1" Since this enzyme plays a key role in this manuscript, it deserves to be better introduced. In fact, it plays also a key role in the onset and progression of Gastrointestinal Neoplasms (see PMID: 36139012 )

Materials and methods: authors must provide the product code of all reagents and kits used in order to allow data reproducibility

Histologic assessment of the intestinal tract: a representative image for each score must be provided as supplementary material.

Western blot analysis: i suggest to move the antibodies used in a dedicate table. Moreover, antibodies product code and dilution used must be provided

Western blot images: authors must add the molecular weights in all western blot images

Figure 2D, 4D-F: higher magnifications are needed

Figure 3C: negative control and higher magnifications are necessary

Abbreviations must be written in full length when mentioned for the first time

Reviewer #2: Authors have focused their attention on necrotizing enterocolitis(NEC) a prevalent and destructive illness in neonates. In particular, the paper investigated the role of Nicotinamide N-methyltransferase (NNMT) and its product of enzymatic activity, 1-methylnicotinamide (1-MNA) in the development of this pathology.

The experimental design has been rightly planned and correctly carried out by authors using intestinal tract from both neonatal rats and human newborns. However, the paper requires some revisions before it can be published.

1- The indroduction needs to be better structured. The introduction should be a brief description of the current knowledge about the role of NNMT and its metabolite 1-MNA in the development of patholgies. The introduction should explain the rationale of the study, beginning with the gap in existing literature. In this case, authors mentioned in the introduction the results without explaining why they decided to focus their attention on NEC pathology.

2- The experimental protocol needs improvement. The authors should provide the product codes for the antibodies used but also their dilution in order to make readers able to repeat the experiments

3- The resolution of the figures is very low and needs improvement

4- English needs improvement. There are several grammar mistakes (eg.: 16S rRNA genewas targeted instead of 16S rRNA gene was targeted)

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Reviewer #1: No

Reviewer #2: No

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1-methylnicotinamide attenuated inflammation and regulated flora in Necrotizing enterocolitis

PONE-D-25-03800R1

Dear Dr. KANG,

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.

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Kind regards,

Roberto campagna

Academic Editor

PLOS ONE

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