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Revised manuscript is acceptable in the present form.
# PeerJ Staff Note - this decision was reviewed and approved by Keith Crandall, a PeerJ Section Editor covering this Section #
The following two issues should be fixed for final consideration:
English editing and a minor issue of the sentence presentation in the discussion section (see detailed comments).
English text needs revision by a native or by a specialized company in this kind of service.
The manuscript was significantly improved after the first review. However, minor changes should be made before it can be accepted.
English text needs revision by a native or by a specialized company in this kind of service;
The sentence in the discussion “In contrast several reports have been inconsistent, wherein no significant association was observed with respect to smoking and alcohol consumption in breast cancer patients (Byrne, Rockett & Holmes, 2002; Allen et al., 2009; Gathani et al., 2017).” should be placed along with the next paragraph, where this matter is discussed.
The text has been revised for continuity and additional references have been included as requested by reviewers.
The details of experimental design have now been explained.
The authors present a case control study which opens up direction for further investigation.
The authors have satisfactorily addressed the concerns raised in the review report and have made the appropriate changes
Authors need to provide literatures on CYBA (930 A/G and 242 C/T) polymorphism in introduction section. Need language editing for better flow. Oxidative stress should be precisely measured. High quality figures with full legends should be provided. Statistical analysis should be precisely written. Authors should carefully survey literatures on link of diets and smoking with breast cancer and cite in the manuscript. Please carefully read reviewers comments and modify manuscript accordingly.
poorly written manuscript
good experiment design and original research
data meet the criterion
Association of CYBA (-930 A/G and 242 C/T) gene polymorphisms with Oxidative stress in Breast Cancer: a case-control study
Mohini A Tupurani, Chiranjeevi Padala, Kaushik Puranam, Rajesh Galimudi, Keerthi Kupsal, Nivas Shyamala, Srilatha Gantala, Ramanjaneyulu Kummari, Sanjeeva Chinta, Surekha R Hanumanth
In this paper authors introduced with relationship between common functional polymorphisms of the p22phox gene (-930A > G and +242C > T) and breast cancer as a result of oxidative stress in Indian population. As authors reported here, this polymorphism is highly expressed in breast cancer patients. Furthermore, authors demonstrated that this polymorphism would increase oxidative stress by measuring MDA level in blood samples. They also tried to correlate other factors such as smoking and alcoholism with polymorphism and breast cancer. Author’s case-control study in this paper proves their hypothesis but errors in paper reduced the enthusiasm.
1. Authors could not introduce CYBA (930 A/G and 242 C/T) polymorphism in introduction section. They must add existing research information on this polymorphism.
2. Authors used telegraphic English and sometime sentence looks incomplete, for example Line 50. Results and discussion sections were not scholarly written. They look like a case report not a journal article.
3. Authors used MDA quantification to determine oxidative stress. MDA is not sole product of lipid peroxidation, and on the other hand, thiobarbituric acid (TBA) in thiobarbituric acid reactive substance (TBARS) assay dose not determine only MDA. MDA are a measure of oxidative damage on lipids, not of "oxidative stress" per se. Oxidative stress is actually not truly measurable with only one marker: it is a state when oxidant attacks exceed antioxidant defenses, so you need different markers to get the whole picture if you insist on OS.
4. Major problem I am facing in this manuscript is figures and their legends. Legends are incomplete and does not have information about figures. PDF file I downloaded shows all figures were stretched and quality of some figure was too poor to read.
Correcting these major errors will make the author’s paper look more professional, and certainly should have to be corrected before being accepted for publication.
Materials and methods description.
There is some missing information, such as:
1-) The period of time in which the study was conducted;
2-) The inclusion and exclusion criteria for entry or not in the study;
3-) The statement informing that this study follows the ethical principles according to Helsinki declaration;
4-) Authors should give more details about the blood collection, such as the patient's position during collection if the patient needs to be in fasting condition, time or period of the day when the blood was collected and what was the vein in which the blood was collected.
5-) Authors also should better describe the MDA assessment in the blood samples of the patients, such as: if there was some previous preparation of the samples before evaluation, including the reagents and the concentrations used in each stage of the analysis. If it was used some specific kit, please, specify the trademark and the local of manufacturing.
6-) Please, include the student´s t-test in the description of statistical analysis, that is missing.
The study of Tupurani et al. aims to investigate the association between the CYBA gene polymorphisms with oxidative stress in breast cancer initiation and development. The study was well designed, the statistical analysis was properly conducted and the data support the conclusions. However, some minor changes need to be made.
The authors can shorter the background section in the abstract and insert the aim of the study, as stated at the end of the introduction.
The authors should briefly describe for what the Insilco analysis was employed in the abstract instead of “Insilco analysis was performed using appropriate tools.”
The sentence that defines oxidative stress is lacking reference(s) – Lines 42-45.
Authors are encouraged to point out and briefly discuss the potential limitations of the study. As an example, the last sentence in the conclusions (lines 226-229) authors cited some limitations but they should be in the discussion.
In the current manuscript, the authors conducted a study on 300 breast cancer patients with a comparison to 300 healthy control volunteers to evaluate the association of SNPs: -930 A/G and 242 C/T in the CYBA gene to increased markers of oxidative stress.
In the results section, the authors begin with making a significant claim of the association of non-vegetarian diet, smoking and alcohol consumption with increased risk of breast cancer. However, there have been several such studies with a larger sample size that have reported no risk associated with non-vegetarian diets and smoking with breast cancer. The association of smoking with breast cancer risk is an issue of debate. It appears that the current study is underpowered and has a small sample size to make such claims. Further, this result is not within the scope of the proposed hypothesis.
Below are listed some of the competing research that refute the claim drawing associations of non-vegetarian diet, smoking and alcohol consumption with increased risk of breast cancer.
1. Lifelong vegetarianism and breast cancer risk: a large multicentre case control study in India
Gathani et al, BMC Womens Health. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5241933/
2.Active smoking and breast cancer risk: original cohort data and meta-analysis. Gaudet MM, Gapstur SM, Sun J, Diver WR, Hannan LM, Thun MJ. J Natl Cancer Inst. 105(8):515-25, 2013 and Lawlor DA, Ebrahim S, Davey Smith G.
3.Smoking before the birth of a first child is not associated with increased risk of breast cancer: findings from the British Women's Heart and Health Cohort Study and a meta-analysis. Br J Cancer. 91(3):512-8, 2004
4. Moderate alcohol intake and cancer incidence in women. J Natl Cancer Inst. 2009 Mar 4;101(5):296-305. doi: 10.1093/jnci/djn514. Epub 2009 Feb 24.
It is recommend that the authors prepare the manuscript to address the counter arguments present in the literature.
With respect to the C242T polymorphism, a previous study has reported that this variation leads to reduced NAD(P)H oxidase activity in human blood vessels and therefore a decreased production of ROS and vascular oxidative stress. (Xu Q, Yuan F, Wu J. "Polymorphisms of C242T and A640G in CYBA Gene and the risk of coronary artery disease: A meta-analysis." PLoS One. 2014; 9(1): e84251). This opposes the hypothesis of association with increased ROS production as reported in the current manuscript.
The insilico analysis reveals interesting results with mRNA structure prediction, transcription factor binding and 3D protein structure prediction which will need further validation by in vitro experimentation.
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