DNA damage can have genotoxic and cytotoxic effects on the cell (Tuteja, Singh, Misra, Bhalla, &Tuteja, 2001). DNA damage has been shown to either promotes anti-cancer, anti-aging response or enhance development of premature aging pathologies including obesity, hypertension, diabetes, show disintegration of DNA repair mechanisms, resulting in accumulation of damaged DNA. With continuous accumulation of DNA damage, the body resorts to maintenance mode, where the growth will cease which is seen in premature aging pathologies and senescence (Li, 2008).

The other triggering factors are mutagens, radiation and even inevitable replication process of DNA. DNA damage from all endogenous sources give rise to some 20,000 lesions per day per cell. Calculation considering all possible processes causing DNA damage reports that there can be as much as 3 x 10¹⁷ DNA damaging events per hour in a human body. Strictly, DNA molecule can be said to be damaged when there is any abnormality from regular double stranded structure, (Drabløs et al., 2015). DNA damages include covalent changes in DNA structures and noncovalent anomalous structures, including base pair mismatches, loops, and bubbles arising from a string of mismatches (Li, 2008). Change in structure can happen because of one of the following, (1) single base change, (2) structural distortion or (3) DNA backbone damage. Single base changes occur due to deamination, alkylation or oxidation of the bases resulting in the formation of transversion of bases from GC to AT. Most common structural alteration is caused by the cross-linked Thymine dimmers while DNA backbone damage, which is the most severe type of DNA damage, is seen when there are double strand breaks.

Stability of the genome is supported by an intricate machinery of repair, damage tolerance and checkpoint pathways that counteracts DNA damage. Response of the cell to DNA damage can be, bypassing it, reversing it or replacing the damaged part. Bypassing mechanism is taken care mostly by different kinds of DNA polymerase and the reversal is done by methyltransferases. In damage removal process of managing DNA damage there are four mechanisms, base excision repair, mismatch repair, nucleotide excision repair and double-strand break repair involving a wide variety of proteins is a well-established link between disintegration of DNA repair mechanisms.

to oxidative stress which effect a series of metabolic event within the cells that leads to activation of nuclease enzymes which damage the backbone of DNA (Halliwell & Aruoma, 1991). Oxidative stressresults when reactive oxygen species are not adequately removed. This can happen if antioxidants are depleted or if the formation of reactive oxygen species is increased beyond the ability of the defenses to cope with them which can result in severe metabolic dysfunctions, including peroxidation of membrane lipids, depletion of nicotinamide nucleotides, rises in intracellular free Calcium ions, cytoskeleton disorder and DNA damage (Halliwell & Aruoma, 1991).

Commonly produced free radicals are superoxide (O), nitric oxide (NO⁺) and OH radicals. These are not highly reactive but under certain conditions it generates toxic and cause irreversible/reversible damage in nucleic acid, amino acid, lipid, protein, lipoprotein, carbohydrates and connective tissues. This types of free radical are formed due to inflammation, aging, induced by exercise, cancer and many other traumatic disease (Michalsen, A et al., 2012).

Figure 1: DNA damage response reactions in mammalian cells (Sancar& Lindsey-boltz, 2004).

Twice of kriya sessions will be given in a week.

Table 1: Yoga based lifestyle routine under IAYT intervention

Assessments

The assessment was done before and after intervention. The only assessment done in this study was to assess the DNA damage assessed through comet assay also called single cell gel electrophoresis (SCGE), developed by N.P. Singh, combines the simplicity of biochemical techniques for detecting DNA single strand breaks (frank strand breaks and incomplete excision repair sites), alkali-labile sites and crosslinking with the single cell approach typical of cytogenetic assays. Following is the protocol in brief as described by N.P. Singh (Dhawan et al., 2003).

Protocol in brief

1. Slides are prepared by coating 1% Normal Melting Agarose (NMA) solution.
2. To the coated slide, add 75 μL of Low Melting Point Agarose (LMPA) (0.5%; 37ºC) mixed with ~10,000 lymphocytes in ~5-10 μL of whole blood and a third layer of LMPA is poured and allowed to solidify by placing at 4^o
3. After about 2 hrs in alkaline buffer (pH >13) place the slides in the electrophoresis tank and the electrophoresis done 24V and 300mA for 30 mins.
4. After electrophoresis gently lift the slide and stain with 1% ethidium bromide for 5 mins.

between pre & post were done using these averages. The tail length (used as a measure of DNA damage) was seen both increasing and decreasing, signifying that the extent of DNA damage both increased and decreased respectively. In 19 out of 30 participants the DNA damage decreased while it increased in the remaining 11 participants (Figure 2). Also, the data showed the normal distribution pattern for all the parameters measured, one for tail length is shown below (Figure 3).

Table 2: Means of pre & post intervention 

The chart shows the pre-post DNA tail length changes. Out of the total sample (n=30) 63% (samples 1-19) have shown decrease in tail length while 37% (samples 20-30) have shown an increase in tail length. Table also shows pre & post tail lengths in arbitrary units.

Figure 3: Normal distribution curve

The extent of DNA damage decreasing was more significant than the increase in damage. Sixty three percent of the participants with diabetes showed 38% decrease in the extent of DNA damage, while 37% participants showed a 19% increase in DNA damage (Table 3 & 4 and Figure 4 & 5).

Table 3: Positive change of tail length

Table is showing a reduction in the tail length after the intervention. The result is from 19 subjects. There was significant reduction in the scores from 113.95±42.56 to 57.96±36.40 with 38 % of change.

Table 4:  Negative change of tail length 

Table showing an increase in the tail length after the intervention. The result is from 11 subjects. There was significant increase in the scores from72.60±43.21 to 106.50±52.83 with -19 % of change.

Figure 6: Comparison of percentage of DNA in the head and tail.

This bar chart shows the logic of increase and decrease in tail length. Decrease in tail length after intervention means decrease in tail DNA% and increase in headDNA%. Likewise, increase in tail length after intervention means increase in tailDNA% and decrease in headDNA%.