Reviewer’s report
Title: 
Serum and Gene Expression Profile of Cytokines Following combination
of Yoga Training and Vitamin D Supplementation in Breast Cancer Survivors: a
randomized controlled trial
Version: 
1 
Date: 
13 Jan 2022
Reviewer's report:
I congratulate the authors for nicely conducted study titled "Serum and Gene
Expression Profile of Cytokines Following Combination of Yoga Training and
Vitamin D Supplementation in Breast Cancer Survivors".
The study has tried to evaluate the usefulness of adding Yoga to Vitamin D (VD)
in alleviating physiological and psychological shortcoming in breast cancer
survivors. This 3 arm study clearly establishes the benefits of adding Yoga even
though the Yoga intervention was only for 2 days a week. In between group
comparisons, a significant difference was seen when high dose of vitamin D was
supplemented with Yoga (YHVD) in improving QoL, IL-10 and anti-
inflammatory index (IL-10/TNF
α
). Improvements, though less pronounced, was
also seen with lower dose of VD supplemented with yoga (YLVD), but the same
could not be said for only high vitamin D (HVD) group. At the same time no
significant difference was seen in between group comparison for IL-6 and TNF
α.
Of interest was the result from correlation studies; (a) negative correlation
between IL-10 with weight & BF% and positive correlation with QoL, (b) anti-
inflammatory index negatively correlated with weight and BF% and positively
correlated with QoL, and finally (c) positive correlation seen for IL-6 & TNF
α
with BF%. All of these not only help validating the known relations between
markers but also verify the study.
Among the novel findings, IL-6's upregulation with HVD and Yoga bringing it
down (thereby the inflammation) is notable. A couple of studies have indicated
about this ability of yoga to respond to stress through toll like receptors without
activating inflammatory pathways.
At the same time, there are some areas which require clarification or description
in more detail, they are as follows:
1. 
In the 'methodology section' (page 6, line 60), it is mentioned that gene
expression along with IL-6, IL-10 and TNF
α
 was measured using ELISA kits.
Gene expression cannot be measured using ELISA, this is a major error in
writing.
2. 
In the 'participant selection' section (page 7), first 3 lines describing inclusion
criteria is not clear at all. Also, reason for excluding 3 participants not given,
only reference to previous study is provided.
3. 
Also in page 7, regarding 'randomization', the text only mentions that third
party divided the participants, the method followed for doing the same is not
mentioned.
4. 
There is no mention of Yoga protocol (reference to previous study is
provided) - a brief mention of protocol will be appreciated. Also the duration of
Yoga is mentioned as 60-90 minutes a day, which is very unscientific way of
mentioning the duration of intervention.
5. 
In page 9, blood volume collected is mentioned as 5 cc and the ensuing
protocol mentions that this was used for extracting serum. Then what sample
was used for extracting RNA required for gene expression experiments?
Also, the details of reagents used for RNA extraction, cDNA synthesis, RT-PCR
instrument are all missing. Again only reference to previous study is provided,
but a research paper warrants mention of at least brief information regarding
these aspects.
6. 
Finally, a general language check is required throughout the article.
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Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
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from apparent manipulation?
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