Immunomodulatory Properties of Polyphenol-Rich Sugarcane Extract on Human Monocytes
, Lily Stojanovska 5, Matthew Roland Flavel 6,7, Barry Kitchen 6,7 and Vasso Apostolopoulos 1,3,*Round 1
Reviewer 1 Report
The aim of this study was to prove that Polyphenol-rich sugarcane extract (PRSE), a potential interesting antioxidant, has a potential immunomodulatory effect on human monocyte cells. It's interesting to try to test a new antioxydant but I'm not convainced by the methodology and the results.
Comments :
- Introduction :
l49 : in vitro has to be in italic
Could you explain why you chose U937 cells?
- Results :
In the cytometry part, you should present histograms of the relative expression of markers with appropriate isotype control.
It was not clearly explain how you define the threshold for the dot plots.
You should use the same group name between differents part results.
- Discussion :
Experiment must be reproduced to conclude about PRSE immunomodulatory effect.
- Materials and Methods :
l209 : which provider for VitD3? Why VitD3 was used and not other molecule like PMA?
l211 : which source of LPS and which provider ?
In the cytometry part, it was surprising that you used a FACS Canto with 3 Lasers and you performed single color sample.
Have you used viability dye?
There's no materials and methods corresponding to the part 2.2 (ELISA, multiplex by cytometry?) and the part 2.3 of the results.
It is surprizing to see line 218 "three technical replicates". Technical replicates are not biological replicates. You must reproduce the experiments.
Author Response
We thank the reviewer for their insight. We have made all requested changes to the manuscript - please see the attached document for additional information.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
In the manuscript by Feehan
et al. entitled
“ Immunomodulatory properties of polyphenol-rich sugar cane extract on human monocytes”.
the authors evaluate the effects of a polyphenol-rich sugarcane extract on the inflammatory phenotype of U937 cells induced upon Vit D and LPS treatment before polyphenol exposure.
The manuscript is very interesting and it may add new pieces of information related to the role of high sugar foodstuff on health.
However, the author should specify some aspects of their work.
1) I suggest producing a more accurate description of the polyphenol-rich sugarcane extract in terms of methods of production and most of all chemical composition. It is important to know exactly the composition of the bioactive used in this work. The citation of older published data is not sufficient.
2) Please specify methods related to NF-KB activation.
3) Please specify methods related to cytokine measurements.
4) I suggest to address the effect of the extract in terms of cell vitality and influence on the proliferation rate.
5) Keeping in mind that IL-6 is more pro-inflammatory than anti-inflammatory produce a more objective judgment of the results underling the possibility that the extracts may have potentially dangerous effects. I suggest evaluating the effect of the extracts on the expression and activity of some pro-inflammatory genes such as cox-2.
Author Response
We thank the reviewer for their insightful suggestions. We have made all of the requested modifications - please see the attached document for further information
Author Response File: Author Response.pdf
Reviewer 3 Report
General comments:
The authors determined the immunomodulatory properties of sugar cane extract using flow cytometry. The article is interesting, but there is not enough data and the writing should be edited.
Specific points:
Abstract: The introduction part of the abstract is too long. Please describe the results in detail. The last sentence of the abstract should be a strong summary statement of the study.
Introduction: Please divide the last paragraph into two on line 65? End the introduction (last paragraph) by stating the hypothesis for the research, and include the specific research objectives to test your hypothesis that would logically follow the rationale for the research design.
Please review about the contents of sugar cane polyphenols in line 60. It is well documented in your ref. 26.
Results: Make certain that all footnotes for the figures and tables are correct and complete. Please make certain to include in each footnote the details of what is presented, e.g. experimental groups, values, number of samples or individuals per group and statistics.
Authors used AF488, but wrote FITC in the figure 1 and 2.
In the figure 4, please remove the outlines.
L172-179, many studies have shown that the PI3K/AKT is the upstream of NF-kB. If there is no change of NF-kB, AKT signaling doesn’t seem to be altered by sugar cane extract. Could the authors suggest another pathway?
Materials and methods: It seems that authors forgot to attach half of the part including Cytokine analysis, NFkB expression, and Statistical analysis.
L216 and 222: Authors incubated the cells with the compounds for 24 and 48h, but forgot to mention about 48h.
Reference
MDPI’s reference style: The names of journals should be abbreviated.
- Author 1, A.B.; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.
Please review your manuscript for TYPOS and grammatical errors.
L14, 18, 54, 60: anti-oxidants-> antioxidant
L50: use .-> no space
L105: IL-1-> IL-1b
L115: “Figure 4” is written as italic. Please fix it.
L189: 41.p->4.1.
L213: VitD3->VitD3
Please use the same unit including h vs. hrs
Author Response
We thank the reviewer for their suggestions, all of which have been incorporated in the manuscript - please see the attached document for further information.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
The manuscript has been properly implemented and can be accepted
Reviewer 3 Report
The authors have adequately addressed my comments/concerns. Thank you.
