COS Attenuates AFB1-Induced Liver Injury in Medaka through Inhibition of Histopathological Damage and Oxidative Stress
Round 1
Reviewer 1 Report
The manuscript investigates the protective effects of chitosan on aflatoxin B1 induced liver toxicity. As a comparative study, silymarin as taken as the other natural product to compare the protective effects of chitosan with silymarin. The investigation involves usage of fish as the other model for conducting the toxicity studies.
The manuscript is not well written. It uses often wrong descriptions to describe the studies; such as reduced glutathione (GSH) and malondialdehyde (MDA) are not enzymes while at many places they are described as having the activity. This is pointed out below.
Line 217: Reduced glutathione (GSH) is not an enzyme, therefore it cannot be defined as enzymatic activity as stated in the sentence/line 217 “However, the enzymatic activity of GSH was inhibited at the high dose of…”
Similarly, Line 235-236: MDA or GSH are not enzymes and the sentence should be modified “…..GSH, and MDA enzyme activities……………………….”
Line 328-332: GSH is not an enzyme as written in these words “GSH enzymatic activity was promoted 329 in low doses of AFB1, and in high doses of AFB1, unexpected inhibition occurred in contrast to low doses of AFB1, probably due to the disruption of the antioxidant defense system already[29,30], and COS also interfered with AFB1-induced changes in GSH enzymatic activity.” These words should be changed and redrafted.
Line 334: Both MDA (and GSH) will have “levels” and not the activity. Therefore, change the word activity if you mentioned for GSH or MDA.
There are many mistakes in the manuscript. These mistakes which are listed below are not EXHAUSTIVE. Authors need to read the manuscript several times and get edited from a scientific writer for proper presentation of the investigation.
Line 40: Change the word “cancerogenesis” into “carcinogenesis”.
Modify the sentences “By preventing cytoplasmic division and lowering reactive oxygen species (ROS), COS can successfully act as an anti-inflammatory and antioxidant in injured liver tissue. It can also effectively modify the harmful tissue changes taking place in the liver, hence minimizing hepatocyte apoptosis [14,15].”
Modify the sentence: “Silymarin (SIL), a natural hepatoprotective drug, was added to the COS intervention study as a beneficial drug comparison at the 66 same time to assess the COS's healing potential.” It should be written as “Silymarin (SIL), a natural hepatoprotective drug, was included in the study for comparative purposes of COS protective effects against AFB1”.
Line 95-107: The doses should be mentioned in the form of amount/body weight. The doses are mentioned as volume/L, the doses are not added to the water but rather injected. Therefore, the doses should be mentioned in the standard format; mg/Kg or µg/K.
Describe the correct method of protein estimation, Line 130-131. “In addition, protein content was assessed by the blue staining.”
Line 134: Glutathione (GSH) should be written as reduced glutathione (GSH).
Modify the sentence: “The liver tissues were then transpar[1]120 ent in xylene and dehydrated with various ethanols, embedded in paraffin, and sectioned 121 with a microtome at a thickness of 4 μm. Hematoxylin & eosin (H&E) staining was then 122 performed on the sections.”
Line 192: What is “liver stasis”
Line 215: “significantly promoted” means “significantly increase?”
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Dear Authors, I congratulate you on the very interesting research results and the work you put into obtaining them. I have some comments to improve the work. All my remarks regarding the improvement of z have been marked in the comments on the manuscript of the work which I send in the attachment.
Comments for author File: Comments.pdf
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
There are still mistakes in the manuscript, like describing the effects in low concentration which should be "at lower concentration". It is advised that authors read seriously the manuscript and still incorporate the corrections to raise the quality of manuscript.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
