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The previous reviewers did not respond, but I have evaluated the revisions myself. All issues pointed by the reviewers were adequately addressed and the manuscript was revised accordingly. Therefore, the amended manuscript is acceptable now.
Please address concerns of both reviewers and amend manuscript accordingly.
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Since salusins are secreted by inflammatory cells, what is the purpose of measuring their concentrations in functional intestinal diseases? Why are blood concentrations of this protein lower in healthy individuals compared to those with irritable bowel syndrome (IBS)? Furthermore, why do salusins not differentiate between the various forms of IBS?
From a prognostic standpoint, comparing IBS results with those from inflammatory bowel diseases (such as microscopic colitis, ulcerative colitis, and Crohn's disease) and examining correlations with other inflammatory biomarkers would be useful. Additionally, changes in the concentration of this protein have been reported in many diseases, evaluating correctly diagnosed IBS complex.
Numerous tests are required to rule out other diseases in individuals over 50. This involves expanding exclusion criteria, performing histological evaluations of the intestinal mucosa, and examining the intestinal microbiome.
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- Abstract
1. Methodology: The term enzyme-linked immune-sorbent assay (ELISA) should be used when describing ELISA for the first time in the manuscript.
2. Conclusion: salusins instead of salusines.
- Literature Review
1. The introduction provides a reasonable overview of IBS and its associated pathophysiology. However, it is very concise and does not explain the rationale of the study precisely. Also elaborate on why studying D- and C- type IBS separately is important.
2. The introduction about salusins could be better integrated, emphasizing why their role in IBS is of particular interest, given their established associations with inflammation and oxidative stress in other diseases.
3. Authors should also explain in the introduction the gaps or voids in the current knowledge that are being filled with your study conducted.
4. Line 17: Authors have written “Middle-aged women are more likely to have this disease”. Why is the case? Also explain in the manuscript.
5. Line 18: “The etiology of IBS is multifactorial”. Try to justify this line with some factors responsible.
- Methodology
6. There are several grammatical errors and phrases undermine from its readability. For example, "Participants diagnosis of IBS using Rome IV criteria were participated in the study" (line 42) is poorly constructed. A thorough language edit is necessary.
7. The abbreviated words should be explained for the first time appearing in the manuscript. e.g., line 46: BUN.
- Results
7. Results for different parameters need to be explained under different sub-headings. The current state is very brief, which makes it difficult to apprehend the results.
8. Line 75, 78, 80: αlfa and βeta should be written as α and β respectively.
9. Table 1; Age: decimal point instead of comma.
- Discussion
10. The initials of first name from in-text citations must be removed and year must be mentioned.
11. Add a discussion on the potential mechanisms linking salusins with IBS, supported by more recent literature.
12. Clarify the clinical relevance of current findings. How salusin levels might aid in diagnosing or managing IBS in practice?
13. Add a more comprehensive comparison with your results rather than stating literature about IBS and salusin.
14. If an increase in salusin-β is linkd to inflammation, then justify the decrease in its levels in your results.
15. Compare and discuss the differences between levels of salusin-α and salusin-β.
1. The inclusion and exclusion criteria are adequate, though more details on the clinical severity of IBS cases (e.g., duration of symptoms, previous treatments) would add depth to the analysis.
2. The gender distribution is mentioned, but its relevance to the study outcomes is not analyzed or discussed.
3. Blood collection and processing procedures are described in sufficient detail. However, the specific ELISA kit and its validation for salusin measurement in human samples should be elaborated on to ensure reproducibility.
4. The study employs a reasonable design, including both IBS subtypes and a control group. However, the sample size (n=85) is relatively small, especially given the heterogeneity of IBS.
- Statistical analysis
5. The manuscript does not report effect sizes or confidence intervals, which are crucial for interpreting the clinical relevance of findings.
6. The significant differences should be interpreted with asterisk in tables and figures.
1. The finding that salusin-α levels are lower in IBS-D compared to controls is intriguing. However, the study does not adequately address the role of confounding factors such as dietary habits, psychological stress, or coexisting conditions that may influence salusin levels.
2. The lack of significant differences between IBS-C and IBS-D groups diminishes the impact of the results. The authors should explore potential explanations for this observation.
3. The potential for circadian variation in salusin levels, given the fixed morning blood collection time, should be addressed.
1. The years of in-text citations are not mentioned in the whole manuscript.
2. The manuscript requires substantial language editing and restructuring for clarity and precision.
3. Support the facts with the already present literature. The citations are very few in the whole manuscript.
4. The manuscript explores a novel area of research, and its findings are a valuable contribution to understanding IBS pathophysiology. However, its impact is undermined by methodological gaps and a lack of contextual discussion.
5. The authors can perform subgroup analyses to explore possible interactions (e.g., gender, age) affecting salusin levels.
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