Chromosomal Translocations in Chandigarh Police

Definition of the problem: Evaluate the molecular impact of yoga on DNA damage response and repair on security guards of Chandigarh Police

Objectives:

1. Analysis of molecular markers related to DNA damage response and repair before and after the practice of yoga on Chandigarh Police
2. To examine the molecular mechanism by which yoga may modulate DNA damage response and repair.

Recruitment of individuals: Department of Police, Chandigarh has agreed to help the investigators on this aspect. Policemen working for Chandigarh Police will be chosen based on the inclusion and exclusion criteria.

Subjects: Those security guards (age: >20 and \<60) who are willing to consent to participate in this study will be screened for moderate and higher stress on Cohen's PSS, according to inclusion criteria and randomly divided between Intervention (Yoga Group) and Control Group.

Sample Size: The sample size of 206 was calculated for 99% confidence level (reliability coefficient = 2.58), at the alpha level of 0.05, a power of 0.95, and a relative precision of 0.2, derived from a pilot study done on breast cancer patients and yoga practitioners (Ram et al, 2013). Assuming a 10% drop out rate, a total of 229 subjects will be recruited.

Design of the study In a randomized control trial (RCT), participants are allocated randomly between intervention and control groups to compare the treatments, which enables statistical control over the influences imparted by the aforementioned treatments. In the present study, the policemen who satisfy the study's eligibility criteria and willing to consent to participate in this study will be enrolled, grouped randomly in intervention and control groups, and assessed for the dependent variables of the study before and after the Integrated Yoga intervention period.

Variables Socio-demographic details - age, diet, alcohol consumption, etc. Evaluation of stress level on Cohen's Perceived Stress Scale (pre and post intervention) (Cohen, Kamarck \& Mermelstein, 1983) Evaluation of stress level using vital parameters: Blood pressure and resting heart rate of participants will be recorded pre and post intervention.

Evaluation of stress level using molecular parameters: (pre and post intervention) 10 ml of blood will be withdrawn from the subjects by trained phlebotomists, and coded. WBCs and plasma will be separated using standard protocols and stored at -80℃. Plasma will be utilized to assess the levels of cortisol.

Assessment of chromosomal translocations: (pre and post intervention) Genomic DNA will be extracted from WBCs isolated, and tested for chromosomal translocations using PCR assay. Isolated genomic DNA will be subjected to nested PCR using primers specifically designed to check for t(14;18) and t(11;14) translocations.

Assessment of DDR and DNA repair:

Immunofluorescence: Isolated PBMCs will be subjected to DDR analysis by immunofluorescence. For this, these cells will be fixed and stained with various DDR markers like γH2AX, 53BP1, pATM.

Gene expression analysis: Total RNA will be isolated from these cells; cDNA will be synthesized and the expression of following DDR and DNA repair genes will be performed using real-time PCR: RAD51, MRE11, RAD50, NBS1, KU70, LIGASE IV, PARP1 Comet assay: Cells will be used to perform both the neutral as well as alkaline comet assay to assess for the number of single- and double-strand breaks.

Telomeric length: Relative telomere length (TL) using quantitative polymerase chain reaction (qPCR) will be performed on genomic DNA isolated from control and group blood samples and will be compared.