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Article
Peer-Review Record

MARCKS Inhibition Alters Bovine Neutrophil Responses to Salmonella Typhimurium

Biomedicines 2024, 12(2), 442; https://doi.org/10.3390/biomedicines12020442
by Haleigh E. Conley 1,2, Chalise F. Brown 1, Trina L. Westerman 3, Johanna R. Elfenbein 3 and M. Katie Sheats 1,2,*
Reviewer 1: Anonymous
Reviewer 2:
Biomedicines 2024, 12(2), 442; https://doi.org/10.3390/biomedicines12020442
Submission received: 12 January 2024 / Revised: 8 February 2024 / Accepted: 12 February 2024 / Published: 16 February 2024
(This article belongs to the Special Issue Neutrophilic Inflammation: Molecular Mechanisms and Drug Discovery)

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The study by Conley et al. is very complex, well-written and conducted, providing solid proof of concept data on Myristoylated Alanine Rich C-kinase Substrate as a potential therapeutic  target for Salmonella enterocolitis. Overall, the introduction sections are well-written and nearly cover the context and importance of the study. The "Materials and Methods" section is comprehensive, ensuring the study's reproducibility and offering a clear understanding of the techniques used. The presented results add credibility to the study. The discussion section effectively interprets the results and provides a deeper understanding of their significance.

Therefore, I found the manuscript data important in its field, and I support its further processing after appropriate minor modifications as outlined below:

L2: MARCKS – please avoid the direct use of acronyms in the title. Revise!

L17, L20: “We hypothesized” “our data”, etc. – please avoid the personal mode verb formulations, may sound unprofessional. Revise this concern throughout the manuscript!

L29: “…oxygen species (ROS).” – In the introduction section, please ensure the using of appropriate citations after each statement. Furthermore, please mention and enhance the documentation about the importance of Salmonella enterica for human medicine.

L64-66: according to the MDPI journal requirement, when you present reagents or devices please uniformly mention their production company name, city, and country throughout the materials and methods section.

L89-90: “All cows were considered healthy.” – This is a very general statement, considering the study aim. Can you provide some details on how was this issue evaluated? The same concern for line 110 “10 healthy”

 

L540: please provide a separate Conclusion section and mention the study limitations.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The data are intriguing but provide no mechanistic insights.

The authors show that a myristoylated-MARKS peptide attenuates Salmonella Typhimurium (STm)-mediated adhesion and migration while increasing phagocytosis and production of reactive oxygen species (ROS). 

 

General comment: It is a well-written manuscript. The data are intriguing but provide no mechanistic insights. The fact that MAMS peptide reduces cell adhesion could introduce a bias. Can the authors repeat the phagocytosis assay under synchronized conditions (centrifugation at 170 g for 5 min)?

 

Specific criticisms:

 

Why not use IL-8 as a control to determine whether MAMS peptide reduces neutrophil migration? Of course, IL-8 will induce neutrophil migration. However, mixing IL-8 and Salmonella Typhimurium (STm) adds very little insight into the mechanism through which MAMS inhibits adhesion.

Figure 4 should include the phagocytosis data at 15 min. The observation that MANS can promote STm phagocytosis is intriguing. Is the effect of MANS restricted to serum-opsonized STm or enhanced phagocytosis observed with other opsonized particles (IgG-opsonized E. Coli, serum-opsonized zymosan)?

Figure 5. The authors must include a quantitative assessment of the STm-positive phagosome to support their conclusion.

Is STm-induced ROS production signification inhibited by DPI?

 

The supplemental figure 1 requires a figure legend.

Author Response

Please see attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

I have no further comments.

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