Impact of kundalini Awakening on gene Function🔗

Health: Health is defined as not merely absence of illness, but also a sense of positive feeling in physical, physiological, psychological and emotional states.

It is a core process affecting all individual cells of the body (50 trillion cells) and their relationship with one another to function as a whole (human being). The relationship of this whole, its interaction with physical and environmental factors (society), and the influence of this interdependence on the collective consciousness (spiritual health) are likewise aspects of the domain of health.

There are four different aspects of health:

Life: The ability of any structure, whether molecule, organelle, cell or multi-cellular organism to function in its environment by actively processing signals, responding to stimuli, preserving its systems, repairing damage and reproducing itself to further propagate its species, , is known as Life. Life processes also include undergoing quantum adaptive changes to deal with a dynamic and changing environment

The primordial universe consisted initially of sub atomic particles (energy), which combined together to form atoms, the smallest organized unit of life, for example, the atom of hydrogen. Hydrogen in turn can be further combined to form molecules through chemical bonds, for example, the molecule of water and so on, which is hydrogen bonded to water.

Of all the possible combinations of elements, the formation of a specially designed molecule called DNA (Deoxy ribo nucleic acid), became the fundamental mechanism of giving life to otherwise inanimate molecules.

In human beings, DNA is a long strand measuring almost 2 meters in length, but coiled inside a cell, which is only a few microns in diameter.

Courtesy, National Human Genome Research Institute

A chromosome is a long strand of DNA, found within the nucleus of a cell.

A human being has 23 pairs of chromosomes in each cell of which 22 are autosomal (not sex-related). The last pair are the sex chromosomes, which determine our gender.

The complete set of chromosomes with its genes in an organism is called its "Genome".

The Human Genome contains between 25,000 and 30,000 genes similar to that of a mouse and less than a grain of rice, which contains 50-60,000 genes. Hence it is clear that it is not the number of genes, but what a gene can do that determines the complexity of a living organism.

Genes not only transmit information from generation to generation, but also act on a minute to minute basis, reacting to external needs, synthesizing amino acids and proteins which carry out the various functions of the body, whether structural, enzymatic or defensive mechanisms.

The human and chimpanzee genomes are 96% identical. The 4% gap makes a profound difference in the physical and psychological presentation (phenotype) of these two species.

For example, language which is unique to humans, has been tracked down to a double mutation in a gene called “FOX P2”. This single factor forms the entire basis for verbal communication in the human species

Some diseases involve single gene mutation such as sickle-cell anemia, which affects only a small subset of humanity.

Other diseases involve multiple genes. Over 100 genes are implicated in susceptibility to asthma and cancer is a disease of genomic instability often involving the mutation (change) of genes from their normally programmed behavior.

Gene-related metabolic pathways such as MAP-K, which stands for Mitogen Associated Protein Kinase, are implicated in the processing of long-term potentiation (LTP), the basis for learning and memory.

In human beings, DNA is present:

It is estimated that only 10,000 of the 30,000 human genes are expressed at any given time (redundancy). Since many genes are repeated and many parts of the chromosomes do not contain genetic material, it is estimated that only 5% of the human genome are actively expressed and are functional genes.

There is a parallel to this in the cosmos, where the sum total of all known matter makes up only 5% of the universe, the remaining 95% as yet unknown.

Gene Expression: is the process by which information from a gene is used in the synthesis of a functional gene product. The process is used by all known life to generate the macromolecular machinery for life.

For example:

2. Sleep: All humans sleep. Other animals sleep. Even worms and fruit flies sleep. Experiments in flies' demonstrated differential expression of genes during waking and sleeping states. During the waking state, genes and encoding proteins involved in detoxification and defense mechanisms are highly expressed. During sleep, nerve-related genes are expressed, which lead to consolidation of memory.

In times of shortage, certain genes or their end products (specific receptors) are up regulated or increased because of an apparent shortage of access to specific materials.

Example: In Type 2 Diabetes Mellitus, there is an up regulation of insulin receptors, in the hope of capturing passing Insulin molecules for glucose metabolism in the cell. This phenomenon is called ‘Insulin Resistance’.

Certain hormones can also down regulate or decrease the receptors in order to balance the internal environment of the cell, as happens with excess availability of substances

Several papers have studied the role of meditation (relaxation response), yoga, tai chi and other eastern techniques and its effects on human physiology.

• Glaser (1990) studied psychological stress induced modulation of IL2 receptor gene expression, published in the Archives of General Psychiatry, 47, 707-712

The current study during Inner Awakening was based on a similar study done at Harvard Medical School by Dusck, et al. (2008) on genomic counter stress changes, induced by the relaxation response, published in Public Library Of Science One, 3 (7).

Control sample collection date: April 12, 2011 Test Sample collection date: May 2, 2011 Venue: Nithyananda Dhyanapeetam, Bidadi. Contract scientific services were carried out by Leads Clin, Bangalore. Principal Investigator: Dr. Krishna Shama Rao, Yenapoya Medical University.

Forty healthy adult volunteers, male and female enrolled for the 21 day Inner Awakening program, volunteered to give blood samples with informed consent for differential gene expression study before and after IA. The objective of the study was to explore Indian traditional yoga and meditation techniques through new experimental biology and bioinformatics.

In the study, we compared gene expression profiles in whole blood between Test and Control samples. Samples collected from volunteers before program were named as BIAY samples and after the program were named as AIAY samples. BIAY were considered as controls and AIAY were considered as test samples.

Sample numbers 1-10 were pooled in as BIAY1 matched against sample numbers 1-10 of after samples as AIAY1 and studied BIAY1 vs AIAY1

Blood sample collection and RNA storing:

Five milliliters of blood was collected from volunteers in EDTA vacutainers, mixed with 25ml of Erythrocyte Lysis Buffer, Qiagen, and incubated for 10 minutes on ice then mixed by vortexing briefly 2 times during incubation and kept for 10-20 minutes and centrifuged at 4000 rpm for 10 minutes at 4°C and the supernatant was completely removed and discarded. After 10-20 minutes visible pellets were re-suspended in 10ml of solution.

RNA Precipitation Procedure:

1. Add 1/10 volume 3M NaOH, pH 5.2 and 2.5 volumes ethanol.

2. Mix and incubate at -20°C for at least 1 hour.

3. Centrifuge at ≥ 12,000 x g in a micro-centrifuge for 20 minutes at 4°C.

4. Wash pellet twice with 80% ethanol.

5. Air dry pellet. Check for dryness before proceeding.

6. Re-suspend pellet in DEPC-treated water.

Absorbance of the sample was checked in 260 and 280 nm. The ratio of A260/A280 was 1.9 which is in an acceptable range.

cDNA and cRNA Synthesis,

Hybridization, Scanning, Data Extraction

were done.

a) Single cycle labeling process with

cDNA using a T4 promoter—dT24

oligonucleotide as primer and second

strand cDNA synthesis and incubation

with T4 DNA polymerase has been done

and the products were purified using

affymetrix clean up module. Biotinylated

cRNA is made using affy IVT kit.

b) cRNA is purified using Qiagen,

quantified and then fragmented by

incubation at high temperature with

Magnesium. Biotinylated cRNA is

added to a total hybridization cocktail

of 300 microliters and 200 microliters

is hybridized to gene chip after adding

control oligo-nucleotides. overnight

c) Hybridization has been done at 45o C, 17

hours overnight with constant rotation.

The hybridization mixture is then

removed and the Gene Chips are washed,

stained with phycoerythrin-labeled

straptavidin, washed, incubated with

biotinylated anti-streptavidin and then

restrained with phycoerythrin-labeled

straptavidin to amplify the signals.

d) Arrays are then scanned using

dedicated scanner, controlled by

GCOS software. Images are examined

for defects. The Affy Micro Array Suite

5 (MAS%) algorithm analyzed the

hybridization intensity data of gene

chip expression probe arrays checked

normalized and CEL files were stored

for further analysis.

e) Microarray Data Analysis was done

using the latest version software Gene

Spring 11.5. The CEL files are imported

in Affymetrix Expression workflow using

Technology for HG-U133_Plus_2.

Summarization: We opted for MAS

5 summarization. It also provides flag

information for the probe sets. Baseline to

median of all samples was applied for this data.

Analysis: A t test unpaired was used

to identify the differentially expressing entities. Fold change analysis at the cut-off of 2 was performed. Hierarchical clustering on the output was performed. Gene Ontology Analysis was done by using fold change output. NCI to find significant pathways was used for pathway analysis.