Other Technical Details

1. Origin of the Proposal:

Type 2 diabetes mellitus (T2DM) is a common lifestyle endocrine disorder caused by

insulin resistance with relative or absolute deficiency in insulin, which results IN chronic
hyperglycemia. In the global epidemic of diabetes, India is in second position after China with
77 million people with diabetes which is expected to rise to over 134 million by 2045. It was
reported that 31 million people became diabetic during the window of 2019-2021. Moreover,
diabetes ranks the ninth place for the reason to death and it is estimated that 592 million people
will die globally in the year 2035 (WHO 2020, Pradeepa, and Mohan, 2021). The side effects
of type 2 diabetes mellitus are increased risk of heart attacks and strokes, reduced blood flow,
neuropathy (nerve damage) in the feet, diabetic retinopathy and kidney failure.
India is a country where yoga is practiced more than 5,000 years ago and it is the
balancing formula to interconnect mind-body relationship. Simplified Kundalini Yoga (SKY)
is an integrated wellness program for achieving health, happiness and harmony in all
spectrums of life. It is the perfect blend of powerful exercises, kundalini meditation, kayakalpa
and deep introspection practices. Yogiraj Vethathiri Maharishi after years of intense research
had systemized a series of exercise that was suitable for all climates and all sections of human
in the contemporary age. Simplified physical exercise was designed by Vethathiri Maharishi to
regulate the flows of air, heat, blood and bio magnetism leading to better health and general
wellbeing. These comprises of hand exercises, leg exercises, neuro- muscular breathing
exercises, and eye exercises kapalabhati, makarasana, acupressure, massaging and relaxation.
swamiji introduced kaya kalpa yoga (Kaya means body and Kalpa means immortal of physical
body).
The studies revealed that yoga practice is useful in the management of various lifestyle
diseases, including type 2 diabetes. Incorporation of yoga practice in daily life helps to attain
glycemic control and reduces the risk of complications in people with diabetes. In the people
intervention with yoga, psycho-neuroendocrine-immune interactions plays a major in the
beneficial effects on diabetes. Additionally, the studies so far covered only the biochemical
changes in the diabetic patients and molecular level changes are not yet reported.The study
results suggest that adding mind-body practices to an existing medication regimen strongly
associated with the control of glycemic index and effective in the management of type 2
diabetes.
The oxidative stress biomarkers such as malondialdehyde (MDA), protein oxidation
(POX), superoxide dismutase (SOD), and phospholipase A2 (PLA2), nitric oxide synthetase,
anti-inflammatory marker adiponectin, 5’ adenosine monophosphate activated protein kinase,
pro inflammatory markers like TNF-α, Interleukin-6 (IL-6), C Reactive Protein (CRP), high
sensitive CRP (hs-CRP) and glucagon like peptide-1 (GLP-1) are supporting the mechanism to
control the type 2 diabetes. Further the genes associated with T2DM are
● TCF7L2 (Transcription Factor 7-Like 2): This gene is one of the most well-established
genetic risk factors for T2DM. Variants in TCF7L2 are associated with impaired insulin
secretion.
● PPARG (Peroxisome Proliferator-Activated Receptor Gamma): This gene is involved in

1
 adipocyte differentiation and insulin sensitivity. Variants in PPARG have been linked to
T2DM.
● KCNJ11 (Potassium Voltage-Gated Channel Subfamily J Member 11): This gene encodes
a subunit of the potassium channel in pancreatic beta cells. Variants in KCNJ11 can affect
insulin secretion.
● IRS1 (Insulin Receptor Substrate 1): IRS1 is part of the insulin signaling pathway.
Genetic variants in IRS1 may influence insulin resistance.
● CAPN10 (Calpain 10): Variants in CAPN10 have been associated with insulin resistance
and T2DM susceptibility (Kurian et al., 2022)

Hence, the proposed research work focuses the biomarkers and genes related to the
mechanism of action of SKY to type 2 diabetes.

2. Review of status of Research and Development in the subject

2.1 International Status:
In 2017, around 462 million people were affected by type 2 diabetes which is 6.28% of the
world’s population. The global prevalence of type 2 diabetes reflects a continued rise
across the world. In lower-income countries, the prevalence is in a rising trend. Unless
urgent measures are initiated to reduce sedentary lifestyles, unhealthy eating, rapid
urbanization, and other factors for economic development, the burden of diabetes will
continue rising (Khan et al, 2020) which may lead to a stressful arena.
Yoga practices help in diabetic people to control their blood glucose by rejuvenating
pancreatic cells, exercising the muscles to increase the glucose uptake in muscle cells,
reducing the weight and by improving the mental health (www.diabetes.co.uk).
During the yoga practice, the activity of hypothalamic-pituitary-adrenal is reduced and
metabolic function, anti-inflammatory markers and neuro endrocrine activity is improved.
Thereby, it provides the significant effect on reducing type 2 diabetes (Chattopadhyay et
al., 2020).
The research studies strongly suggests that practicing yoga significantly improves the
glycemic control and lipid profile and quality of the life style and reduces the use of
medications. But, the long term study with diverse populations are in pipeline (Innes and
Selfe, 2016).
The short term study reported that yoga is an effective complementary treatment for type 2
diabetes, and also long term studies with large sample size are to be studied further (Chen
et al., 2022)
The adverse effects such as hypoglycemia, weight gain, gastrointestinal discomfort,
urogenital infections, discomfort at the injection site, and in some cases heart failure were
reported with the commonly used anti-diabetic drugs (Goyal et al., 2023)
The insulin related genes such as CAPN10, TCF7L2, ABCC8, PIK3R1, KCNJ11, HNF4A

2
 glucose regulatory genes such as GCGR, GCK, GLUT2, SLC2A2, and fat regulatory
genes LPL, PPARG plays a major role in maintain the blood glucose level. The CAPN10
is responsible for protein degrading enzymes, ABCC8 impair the release of insulin, where
as PIK3R1for insulin signaling. GCK coordinates the glucose metabolism, GLUT2
regulates the glucose entry into β cells, SLC2A2 plays a dual role of glucose entry and
triggers the insulin secretion. KCNJ11 is the gene for the potassium channels that trigger
the release of insulin. LPL is the enzyme lipoprotein lipase, which breaks down
triglycerides (fats). PPARG regulates fat cell differentiation (Mambiya et al., 2019).
The inslico analysis revealed that ATP-citrate lyase (ACLY) involved in the molecular
mechanism of type 2 diabetes (Zhong et al., 2019). The study suggested that that Pro12Ala
of PPARG is an important modulator associated with fat and insulin control in the body.
The amino acid change Glu23Lys in the KCNJ11 gene decreased the insulin secretion
(Laakso and Fernandes Silva, 2022).
2.2 National Status:
The yoga asanas such as Surya Namaskar, Dhanurasana, Paschimottanasana, Viparita
Karani, Bhujangasana and Shavasana repaired the insulin function and thereby reduces the
use of medication (Mahajan and Mahajan, 2023). Yoga can offer clinical benefits to
chronic disease related problems caused by stressful live events and improve overall well-
being (Venkatesh et al., 2020). Yoga down regulates the hypothalamic–pituitary–adrenal
(HPA) axis which is hyperactivated as a response to stress. Yoga may stimulate the vagus
nerve improve baroreflex sensitivity, reduce inflammatory cytokines, and thereby reduce
blood pressure and resting heart rate. It improves the functions of endothelial tissues and
reduces the risk of cardiovascular diseases in type 2 diabetes (Singh and Khandelwal,
2020).
Yoga plays a crucial role in the management of diabetes as it addresses the body’s innate
regenerative ability and rebalances the functions of the nervous and endocrine systems.
Diabetes can be largely preventable and treatable through proper diet, exercise and
lifestyle modifications. Vethathiri Maharishi`s physical exercise and Kayakalpa Yoga are
effective in preventing and treating diabetes by balancing the endocrine system, massaging
and toning the abdominal organs, stimulating the nervous and circulatory systems, and
reducing Stress. It was reported that intervention of SKY practice is an effective treatment
for reducing fasting blood sugar, helps to regulate cortisol and other stress hormones,
which affects blood pressure and blood glucose levels and improves the physical, mental
health, quality of life in type 2 diabetic people (Rajasekaran and Perumal, 2018). SKY is a
non-pharmacological adjuvant treatment for type 2 diabetes and it maintains the glycemic
control and reduces the risk of cardiovascular diseases (Raja Priya, 2020). Kapalbhati – a
breathing practice created abdominal pressure and improves the activity of β-cells which
is responsible for the insulin production. Yoga practice reduces oxidative stress and anti-
inflammatory markers malondialdehyde, interleukin 6, and leptin levels, thereby increases
the insulin activity to maintain the glucose level. The insulin resistance is also decreased
by decrease in triglycerides, low density lipoprotein cholesterol, and free fatty acids, and
improves high density lipoprotein cholesterol levels. The results showed that improvement
in high density lipoprotein cholesterol levels, cell-mediated immunity, and lung function
with the invention of yoga. The yoga modulates not only the insulin sensitivity and
resistance it maintains the physical body weight and metabolic functions resulting in

3
 healthy and quality life (Raveendran et al., 2018).
The comparison between yoga exercise and physical training exercise showed that the
former one is better than later and combined both exercises have significant improve in the
T2DM. It was observed that the group performed the yoga practices for six months receive
the significant reduction in the blood glucose levels (Chimkode et al., 2015). Intervention
of yoga practice for 40 min, 5 times in a week for six months, improves the lipid profile
and showed the protective effect on T2DM (Sharme et al., 2020).
The lacunae in the study about the effects of yoga on type 2 diabetes are high-quality,
long-term investigations in populations, biased selection of the patients, identifying proper
research methodology, variation in baseline data, and correlation of results with statistical
analysis (Dewangani et al., 2020).
The risk of T2DM is high in south Asian people when compared to other global
populations. The gene responsible for these people are TCF7L2, PPARG, IG2BP2,
adiponectin, C1Q, and collagen domain containing (ADIPOQ) and alpha-ketoglutarate-
dependent dioxygenase (FTO) (Joseph et al., 2022).
2.3 Importance of the proposed project in the context of current status
Existing methodology and its limitations:
Short term studies performed on the impact of SKY practice on the treatment of type 2
diabetes mellitus. No strong clinical evidence available and the mechanism is not yet
completely revealed on the impact of SKY practice on the treatment of type 2 diabetes
mellitus.
Research Gap:
● To develop or conduct the high-quality, long-term investigations in populations are
required to confirm the potential benefits of yoga programs with type 2 diabetes
mellitus.
● To analyse the molecular level changes on the therapeutic effect of yoga on type 2
diabetes mellitus.
Hypothesis:
Practice SKY and evaluate the changes in the expression of the genes related to type 2
diabetes mellitus with the randomized control study.
Key Questions:
• Is practicing SKY have impact on the type 2 diabetes at genetic level? Is there
rechallenge?
• What are the new insights to the assessment of biomarkers for SKY practices on type
2 diabetes mellitus?
• Is clinical evidence are mandatory for yoga practices?
Outcome:
Outcome of the project will have a scope to open new avenues for creating awareness to
the people about SKY practice for the effective management of various diseases. The
scientific validation of the SKY to type 2 diabetes patient would be the promising

4
 adjuvant therapy to increase the life span and also increase their body and mental strength.
This study provides the platform to find the alternative way to reduce the amount of drugs
used and helps to improve the economy of the country.
2.4 If the project is location specific, basis for selection of location be highlighted:
The type 2 diabetes mellitus patients from Chennai, Tamil nadu will be selected for the
study. Chennai is the second most prevalence of diabetes with 22.8 % of population and
prediabetes with 38%. The study with 12,000 residents of Delhi and Chennai for 7 years
showed that the air pollution with the particles PM 2.5 increases blood sugar levels
(http://www.theguardian.com). The health camp conducted by the government revealed
that 1 in every 5 people had diabetes or hypertension or both. Recently the health team
from Rotary District 3232 conducted glucose monitoring camp in and around Chennai to
create the awareness about diabetes (http://timesofindia.indiatimes.com).

3. Work Plan:

5
 3.1 Methodology:
SKY intervention module:
The following SKY intervention modules will be included in the training program for the
intervention groups for six months; 5 days per week. Each session will be for 60 minutes. SKY
exercises will be taught to the study group by a qualified yoga teacher.
Module SKY Practice Time Schedule
I Prayer, Hand exercise, Leg exercise, Neuro- Day 1
muscular breathing exercise, Agna meditation,
Closing prayer
II Module I, Eye exercise, Kapalapathi, Kayakalpa Day 2
yoga
III Module II, Replacing Agna meditation with Day 3
Shanthi meditation
IV Module III, Makarasana Day 4
V Module IV, Massage, Acupressure, Relaxation Day 5, 6 & 7
VI Module V, Replacing Shanthi meditation with From Day 8 to Day 15
Turiya meditation
VII Module VI, Replacing Turiya with Shanthi Continues to 2 weeks
meditation on fridays
VIII Module VII, Introspection I covering Next 4 weeks – 2nd month
Panchendriya and Pancha Bhootha Navagraha
meditation
IX Module VII, Introspection 2 covering Next 4 weeks – 3rd month
Turiyadheetha meditation, Lamp gazing practice,
Process of sublimation of mind - Neutralisation
of Anger, Eradication of worries
X Module VII, Introspection 3 covering Nine centre Next 4 weeks – 4th month
meditation, Ever Bliss meditation, Mirror gazing
practice
XI Module VII, Replacing Turiya meditation with Next 4 weeks – 5th month
advanced meditation, Introspection practice
XII Module XI Next 4 weeks – 6th month

Place of study:
This study will be conducted in the Manavalakalai Yoga Trust, Kaladipet, Tiruvottiyur,
Chennai, Tamil Nadu by recruiting patients from the outpatient departments (OPD) of
Endocrinology and Medicine, Chennai. Ethical clearance will be obtained from the

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Institutional Human Ethics Committee before commencing the study. Written and informed
consent will be collected from the both yoga and non-yoga participants.
Study design: Randomized control study

Study population:
80 Type 2 diabetes mellitus patients in and around chennai, Tamil nadu.
Inclusion criteria:
Type II Diabetes Mellitus patients, Sex- both males and females, Age group-between 25-65
years
Exclusion criteria:
Type 1 Diabetes, Gestational Diabetes, Hypertension, Thyroid disorders, Carcinomas,
Smoking/Alcoholics/Tobacco chewing, other types of yoga practices.
The blood samples will be collected at the initial day of practicing yoga and at the end of the
day after six months. The biochemical and gene expression analysis will be performed.
Anthropometric parameters analysis:
Weight, height, body-mass index (BMI) will be recorded.
Biochemical analysis:
Fasting Blood Glucose, Post Prandial Blood Glucose will be analysed by enzymatic method
(GOD-POD).
HbA1c levels will be determined by affinity chromatographic method by ELISA.
Oxidative biomarker analysis:
To analyze the level of oxidative damage, the levels of peroxide- and antioxidant-related
indicators will be measured using kit (IFCC) following the manufacturer’s instruction.

Assay of superoxide dismutase (SOD) (E.C 1.15.1.1)

The assay of SOD is based on the inhibition of the formation of NADH-phenazine
methosulphatenitroblue tetrazolium formazon. The colour formed at the end of the reaction
can be extracted into butanol and measured at 560 nm.
Reagents
1. Sodium pyrophosphate buffer (0.025 M) (pH 8.3) : 1.115 g in 100 ml of distilled water
2. Phenazonium Metho Sulphate (PMS) (186 µM): 3 mg in 10 ml of distilled water (930
µM). Then 1:5 dilutions were carried out to obtain 186 µM
3. Nitro Blue Tetrazolium (chloride) (NBT) (300 µM) (pH 8.3) : 3 mg in 10 ml of
phosphate buffer

7
 4. NADH (780 µM) (pH 8.3) : 6 mg in 10 ml of phosphate buffer
Procedure
Superoxide dismutase will be assayed by taking 0.05 ml of sample followed by addition
of 0.3 ml of sodium pyrophosphate buffer, 0.025 ml of PMS (186 µM) and 0.075 ml of NBT.
The reaction will be started by addition of 0.075 ml of NADH. After incubation at 30 °C for
90 seconds, the reaction will be stopped by addition of 0.25 ml glacial acetic acid. Then the
reaction mixture will be stirred vigorously and shaken with 2.0 ml of n-butanol. The mixture
will be allowed to stand for 10 minutes and centrifuged. 1.5 ml of n-butanol alone will be
served as blank. The colour intensity of the chromogen will be read at 560 nm. One unit of
enzyme activity will be defined as the amount of SOD capable of inhibiting 50 % of nitrite
formation under assay condition.
Estimation of Glutathione peroxidase (GPX) (E.C 1.11.1.9)
A known amount of enzyme preparation will be allowed to react with hydrogen peroxide in
the presence of GSH for a specified time period. Then the remaining reduced glutathione
(GSH) will be measured.
Reagents
1. Sodium Azide (10 mM) : 16 mg in 25 ml of distilled water
2. GSH (2 mM) : 30.732 mg in 50 ml of distilled water.
3. Hydrogen peroxide (H2O2) (1 mM) : 29 µl in 1000 ml of distilled water
4. 10 % Trichloro acetic acid (TCA) : 10 g in 100 ml of distilled water
5. K.EDTA (0.4 mM) : 16 mg in 100 ml of distilled water
6. Tris HCl Buffer (0.4 mM) : 6.304 g in 100 ml of distilled water
7. DTNB (0.6 mM) : 12 mg in 50 ml PO4 buffer.
Procedure
Glutathione peroxidase (GPX) will be assayed by taking 200 µl of tris HCl buffer, 0.4
mM K.EDTA along with 100 µl of sodium azide and 200 µl of sample and mixed well.
Thereafter, 200 µl of reduced glutathione solution followed by 0.1 ml H 2O2 were added. The
overall reaction will be arrested by adding 0.5 ml of 10 % TCA. The precipitate will be
removed by centrifugation at 4000 rpm for 10 minutes. The absorbance will be read at 412 nm
using spectrophotometer. The non-enzymatic reaction rate will be correspondingly assessed
by replacing the enzyme sample by buffer. The results are expressed as nmoles of GPx / mg
of protein (Rotruck, et al. 1973).
Estimation of lipid peroxidation (TBARS - Thiobarbituric Acid Reactive Substances)
Plasma levels of malondialdehyde (MDA), a marker of lipid peroxidation, will be
measured by thiobarbituric acid reactive substances (TBARS (nM/mL) using a fluorescence
methodology. Absolute MDA levels were calculated by regression parameters using different
concentrations of the standard (1,1',3,3'- tetramethoxypropane).
Briefly, 100 μl of sample/standard will be added to appropriately labelled tubes. 100 μl of
SDS solution will be added to the tubes and swirled to mix. 4 ml of the color agent given in

8
the kit will be added forcefully downside of each tube. The tubes were boiled vigorously for
one hour. After one hour, the tubes were immediately placed in the ice bath for 10 minutes to
stop the reaction.
After 10 minutes, the tubes were centrifuged for 10 minutes at 1600 g at 4ºc. 150 μl from each
tube will be added to the clear plate and the absorbance will be measured at 530 nm.
Estimation of Nitrite
The Griess reaction is based on the two-step diazotization reaction in which acidified
NO2- produces a nitrosating agent which reacts with sulfanilic acid to produce the diazonium
ion. This ion is then coupled to N-(1-naphthyl) ethylenediamine to form the chromophoric
azoderivative which absorbs light at 540 nm.
Reagents
1.35 % Sulphosalicylic Acid (SSA) : 35 g in 100 ml of distilled water
2.Griess Reagent : 1 g of sulphanilamide is dissolved in few ml of water. Then 2 ml of
orthophosphoric acid is added and 100 mg of naphthyl ethyl diamine. The volume will
be made up to 100 ml with distilled water.
3. Saline (0.9 %):0.9 g in 100 ml distilled water.
Procedure
Nitrite will be estimated by taking 0.2 ml of sample followed by addition of 1.8 ml of
saline and 0.4 ml of SSA for protein precipitation. The precipitate will be removed by
centrifugation at 4000 rpm for 10 minutes. To 1 ml aliquot of supernatant, 2 ml griess reagent
will be added and mixed well. The mixture will be allowed to stand for 20 minutes under dark
conditions. The colour intensity of the chromogen will be read at 540 nm. Nitrite levels were
expressed as nmole / mg of protein (Pandi prabha et al., 2022)
Anti-inflammatory biomarker analysis:
TNF-α, Interleukin-6 (IL-6) will be analysed by ELISA method as per kit manufacturer’s
instruction.
Transcriptomic analysis:
The following genes will be chosen for the gene expression studies: adipocyte differentiation
and insulin sensitivity (PPARG), insulin resistance and T2DM susceptibility (IRS1, CAPN10),
Surface marker CD81
Gene expression studies:
The RNA will be isolated using standard isolation protocol and converted to cDNA using
cDNA synthesis kit and the gene expression will be studied using the GAPDH primer as
housekeeping primer/control primer.
RNA Isolation
• Total RNA will be isolated with TRIzol reagent. The following protocol describes the
procedure starting with a T175 flask, which should give around 200 µg total RNA at the
end.
• The medium will be discarded and 1ml of TRIzol will be added (per 5- 10 × 106 cells).

9
• The cell lysate will be pressed several times using pipette.
• Sample will be then transferred to 1.5ml tube and incubated and room temperature for 5
min.
• 0.2 ml of chloroform will be added and the tube will be shaken vigorously by hand for 15
min. Then, incubated at room temperature for 3 min.
• Then, the sample will be centrifuged at 14,000 rpm for 15 min. The aqueous phase will be
transferred to fresh tube.
• The RNA obtained as pellet will be precipitated by isopropyl alcohol and incubated at
room temperature for 10 min.
• Then, again centrifuged at 14,000 rpm for 10 min. The supernatant will be removed and
the pellet will be washed with 1 ml of 75 % ethanol and centrifuged at 14,000 rpm for 10
min. 9. The RNA pellet will be dried briefly and dissolved in RNase- free water and
incubated for 10 min at 55 to 60 ºC.
Quantitative Real Time Polymerase Chain Reaction (RT- PCR) Procedure
• The first cDNA strand will be synthesized with 2 μg of total RNA and 1μM of
OligodT18primer using Reverse Transcriptase enzyme.
• Quantitative real-time PCR (qRT-PCR) amplification will be performed. The primers will
be designed to amplify specific products.
• All reactions will be run in triplicate and Data Will be expressed as mean ± standard error
of the mean (SEM). Mean difference between the groups will be analysed using one way
ANOVA followed by Tukey’s multiple comparison test in Graph Pad prism 5.0. p value
less than 0.05 will be considered to be a statistically significant criterion.
• Interpretation: The results of the transcriptomic analysis are interpreted in the context of
the research question being addressed. This may involve integrating the transcriptomic
data with other omics data or functional studies to gain a more comprehensive
understanding of the biological mechanisms being studied.
3.2 Time Schedule of activities giving milestones through BAR diagram.
S. Work Duration of
Research Activity
No Plan Research Project
Prepare the brochure for advertising SKY
practice to type 2 diabetes mellitus patients.
1. Phase I 6 Months
Contacting the endocrinologist / physician
Taking survey and segregation of groups
Slot 1: SKY practice at the Manavalakalai
2. Phase IIa 6 Months
Yoga Trust, Chennai
Slot 2: SKY practice at the Manavalakalai
Phase IIb 6 Months
Yoga Trust, Chennai

10
 3. Phase III 12 Months Biochemical and gene expression analysis
Period required for analyzing the data:
4. Phase IV 6 Months Interpretation of results and preparation of
report

3.3 Suggested Plan of action for utilization of research outcome expected from the
project.
Outcome of the project will have a scope to open new avenues for creating awareness to the
people about SKY practice for the effective management of various diseases. The scientific
validation of the SKY to type 2 diabetes patient would be the promising adjuvant therapy to
increase the life span and also increase their body and mental strength. This study provides the
platform to find the alternative way to reduce the amount of drugs used and helps to improve
the economy of the country. Based on the positive impact of the SKY practice on type 2
diabetes, the government may take initiative to motivate the people to practice yoga as a life
3.4 Environmental impact assessment and risk analysis.
Generally, one of the major risk analysis of doing yoga is the use yoga products such as yoga
mat which is made up of PVC or any other non-biodegradable material. Nylon, rubber and
foam and non-biodegradable plastic materials releases toxins, which have negative
environmental impacts. When disposed into environment, it releases toxic chemicals and
volatile organic compounds which cause undesirable health effects such as headaches,
dizziness, and respiratory problems. But in the SKY practice biodegradable cloth based bed
sheets will be used. Hence, it does not cause any damage to the environment and it reduces the
carbon foot print and provides the eco friendly approach. Hence, sustainable yoga apparels and
accessories will be used to safe the environment for future generations. The proposed research
work practices ecofriendly, non-pharmacological therapeutic method for the treatment of type
2 diabetes mellitus. Hence, it does not have any impact to environment.
4. Expertise:
4.1 Expertise available with the investigators in executing the project:

The Principal Investigator Dr. S. Pandi Prabha has established the excellent research
platform in the natural medicine mediated product development to achieve the various
therapeutic effects. My previous research work on protective effect of Hydrilla verticillata
against lead induced toxicity in fish clearly proved the detoxification effect of this medicinal
plant. The fish feed was prepared by incorporating Hydrilla dry powder and the detoxification
effect was studied by gene expression analysis of marker genes. In this work, the fish feed was
prepared by supplementation of 20 % hydrilla dry powder. In the hydrilla supplemented fish
group, there was the amelioration of the liver marker enzymes, antioxidant defense systems,
and marker enzymes for lead toxicity. Administration of supplementary feed to the lead
intoxicated protective group significantly counteracted the expressions of cytochrome P450 1A

11
 (CYP1A) and cytochrome P450 3A (CYP3A) genes, metal binding stress proteins, heat shock
proteins (HSP 60 & HSP 70), and metallothionein. This study provides ample evidence that
Hydrilla verticillata may be a good source to reduce lead induced toxicity in fish. This study
provides ample evidence that Hydrilla verticillata may be a good source to reduce lead induced
toxicity in fish. The recent research work is confirmed the anti obesity effect of Hydrilla
verticillata on 3T3 L1 cell lines. Currently her research group is working on microplastic
toxicity in the in vivo models.
Dr. J. Isaivani, Co-PI has an experience in the field of animal studies, biochemical analysis
and gene expression studies. She has the excellent research platform in the biomarker analysis.
Her research studies on diabetic nephropathy: Role of asymmetric dimethyl arginine ADMA
in the progression of diabetic nephropathy.
Dr. E. Santhi Co-PI is expertise in the field of yoga. She has been teaching yoga for over a
decade and attended and presented research papers on yoga at international and national
conferences. She is having secured accreditation from UGC and the Yoga Certification Board,
Ministry of Ayush. She wish to contribute to society through yoga by involving in scientific
research.
The Department of Biotechnology, SVCE has well established laboratory facilities, animal
house facility and expertise to establish quality research in the diversified field of
biotechnology.

4.2 Summary of roles/responsibilities for all Investigators:

S Name of the Investigators Roles/Responsibilities
.
N
o.
1. Dr. S. Pandi Prabha Principal investigator / Biochemical analysis
2. Dr. J. Isaivani Co- Principal investigator / Gene expression
analysis
3. Dr. E. Santhi Co-Principal investigator / SKY practice

4.3 Key publications published by the Investigators pertaining to the theme of the
proposal during the last 5 years

Name of Vo
S. No. Author(s) Title Page Year
Journal l
1. *Pandi Hepatotoxic effect of Drug and 45 1276– 2022
Prabha S, lead and Chemical (3) 1283
Johanna hepatoprotective Toxicology
Rajkumar, effect of Hydrilla (Taylor and
Karthik C verticillata on hepatic Francis)

12
 transcriptional and
physiological
response in edible fish
Labeo rohita
2. Karthik C, Green synthesis of Environmental 29 58121 2022
Punnaivala rGO-AgNP composite Science and –
van K A, using Curcubita Pollution 58132
Pandi maxima extract for Research
Prabha S, enhanced
Caroline D photocatalytic (Springer)
G degradation of the
organophosphate
pesticide chlorpyrifos
3. Karthik C, Multifarious global International 12 313– 2022
Punnaivala flora fabricated Nano Letters 344
van K A, phytosynthesis of
Pandi silver nanoparticles: a (Springer)
Prabha S, green nanoweapon for
Caroline D antiviral approach
G including SARS-
CoV-2
4. Karthik C, Nanochitosan Polymer 79 8009– 2022
Caroline D augmented with Bulletin 8032
G, Pandi essential oils and
Prabha S extracts as an edible (Springer)
antimicrobial coating
for the shelf life
extension of fresh
produce: a review
5. *Pandi Evaluation of International 12(1 183- 2021
Prabha S, protective effect of Journal of ) 187
Karthik C aquatic weed Hydrilla Pharmaceutical
and verticillata against Sciences and
Shanmuga lead induced acute Research
Priya G liver toxicity in fish
6. Karthik C, Synthesis, Journal of 31 2532- 2021
Caroline D Characterization of Inorganic and 2541
G, Dhanam Ag-SiO2 Organometallic
Priya M, Nanocomposite and Polymers and
Pandi Its Application in Materials
Prabha S Food Packaging (Springer)
7. *Pandi In vitro evaluation of Pharmacognos 16 498- 2020
Prabha S, Hydrilla verticillata y Magazine (5) 505
Sadhana S, for anti adipogenesis
Karthik C, activity on 3T3 L1
Caroline D cell lines

13
 G
8. Karthik, C, Green synthesized Journal of 8 (1) 10357 2020
Swathi, N, rGO-AgNP hybrid Environmental 7
Pandi nanocomposite–An Chemical
Prabha S effective antibacterial Engineering
& Caroline, adsorbent for
DG photocatalytic (Elsevier)
removal of DB-14 dye
from aqueous solution
9. Pandi Phytol – A Biocatalysis 17 736- 2019
Prabha, S, biosurfactant from and 742
Karthik, C, the aquatic weed Agricultural
Hema Hydrilla verticillata Biotechnology
Chandrika,
S H, (Elsevier)
10. *Pandi Antiosteoporotic Materials 90 657- 2018
Prabha, S, effect of Hydrilla Science and 663
Johanna verticillata against Engineering C
Rajkumar, lead induced damage
Suresh S, in bone samples of (Elsevier)
Karthikeya edible fish Labeo
nS rohita – an FTIR
approach
11. *Pandi Hepatoprotective International 8 (5) 57-60 2017
Prabha, S effect of Research
& Johanna supplementary feed Journal of
Rajkumar Hydrilla verticillata Pharmacy
against lead induced
acute liver toxicity in
fish

S. Name of Pag
Author(s) Title Vol Year
No. Journal e
1. Sundararajan, Metformin Reduces the Journal of 12(3) p.29 2023
S., Progression of Lipid and 0
*Jayachandr Atherogenesis by Atherosclerosis
an, Isaivani., Regulating the
Pandey, G.K., Sestrin2-mTOR
Venkatesan, Pathway in Obese and
S., Rajagopal, Diabetic Rats
A.,
Gokulakrishn
an, K.,
Balasubraman
yam, M.,

14
 Mohan, V.
and
Manickam, N
2. Sundararajan Decreased Sestrin Journal of 44(7) 139 2021
S, levels in patients with endocrinologic 5–
*Jayachandr type 2 diabetes and al 140
an Isaivani, dyslipidemia and their investigation 5
Subramanian association with the
S C, Anjana severity of atherogenic
R M, index
Balasubraman
yam M,
Mohan V,
Venkatesan B,
&
Manickam N
3. *Jayachandr Asymmetric Scientific 10(1) 1-17 2020
an Isaivani, dimethylarginine reports
Sundararajan (ADMA) accelerates
S, Venkatesan renal cell fibrosis
S, Paadukaana under high glucose
S, condition
Balasubraman through
yam M, OX4/ROS/ERK
Mohan V, & signaling pathway
Manickam N.
4. Sundararajan Sestrin2 regulates Journal of 120(5) 820 2018
S, monocyte activation ellular 1-
*Jayachandr through AMPK- biochemistry 821
an Isaivani, mTOR nexus under 3
Balasubraman high-glucose and
yam M, dyslipidemic
Mohan V, conditions
Venkatesan B,
& Manickam
N
5. *Jayachandr Association of Clinical 50(15) 835 2017
an Isaivani, circulatory biochemistry –
Sundararajan asymmetric 842
S, dimethylarginine
Paramasivam (ADMA) with diabetic
P, Venkatesan nephropathy in Asian
B, Indians and its
Subramanian causative role in renal
S C, cell injury
Balasubraman

15
 yam M,
Mohan V, &
anickam N

S. Author(s) Title Name of Vol Pag Year

No. Journal e
6. *Santhi E, Immediate effect of International 9 180- 2023
Subha V, Simplified kundalini Journal of (Speci 191
Jayaprakash yoga nine centre Zoological al
S meditation on brain Investigations Issue
Functionality balance 2)
and chakra alignment
among college boys
7. Rajam A. Effect of simplified International 9 203- 2023
*Santhi E kundalini yoga with Journal of (Speci 216
and without tortoise Zoological al
exercise on selected Investigations Issue
physiological variables 2)
among hypothyroid
women

4.4 Bibliography
1. Chattopadhyay, K., Mishra, P., Manjunath, N. K., Harris, T., Hamer, M., Greenfield, S. M.,
& Prabhakaran, D. (2020). Development of a yoga program for type-2 diabetes prevention
(YOGA-DP) among high-risk people in India. Frontiers in public health, 8, 548674.
2. Chen, S., Deng, S., Liu, Y., & Yin, T. (2022). Effects of yoga on blood glucose and lipid
profile of type 2 diabetes patients without complications: a systematic review and meta-
analysis. Frontiers in Sports and Active Living, 4, 900815.
3. Chimkode, S. M., Kumaran, S. D., Kanhere, V. V., & Shivanna, R. (2015). Effect of yoga on
blood glucose levels in patients with type 2 diabetes mellitus. Journal of clinical and
diagnostic research: JCDR, 9(4), CC01.
4. Dewangani, H. N., Jayawardena, B., & Wijayagunarathne, H. S. P. (2020). Yoga-based
Lifestyle Intervention for Prevention and Management of Type 2 Diabetes Mellitus and
Associated Complications: A Clinical Research Review.
5. Goyal, S., Rani, J., Bhat, M. A., & Vanita, V. (2023). Genetics of diabetes. World Journal of
Diabetes, 14(6), 656-679.
6. http://timesofindia.indiatimes.com/articleshow/105189980.cms?
utm_source=contentofinterest&utm_medium=text&utm_campaign=cppst
7. https://timesofindia.indiatimes.com/city/chennai/diabetes-monitoring-camps-to-be-held-in-
chennai/articleshow/105189980.cms

16
8. https://www.diabetes.co.uk/yoga-and-diabetes.html
9. https://www.theguardian.com/global-development/2023/nov/01/air-pollution-raises-risk-of
type-2-diabetes-says-landmark-indian-study-acc
10. https://www.who.int/news-room/fact-sheets/detail/the-top-10-causes-of-death
11. Innes, K. E., & Selfe, T. K. (2016). Yoga for adults with type 2 diabetes: a systematic review
of controlled trials. Journal of diabetes research, 2016.
12. Joseph, A., Thirupathamma, M., Mathews, E., & Alagu, M. (2022). Genetics of type 2
diabetes mellitus in Indian and Global Population: A Review. Egyptian Journal of Medical
Human Genetics, 23(1), 1-16.
13. Khan, M. A. B., Hashim, M. J., King, J. K., Govender, R. D., Mustafa, H., & Al Kaabi, J.
(2020). Epidemiology of Type 2 Diabetes - Global Burden of Disease and Forecasted
Trends. Journal of epidemiology and global health, 10(1), 107–111.
14. Kurian, J., Mohanthy, S., & Nanjumdaiah, R. M. (2022). Mechanism of action of yoga on
prevention and management of type 2 diabetes mellitus: Narrative review. Journal of
Bodywork and Movement Therapies, 29, 134-139.
15. Laakso, M., & Fernandes Silva, L. (2022). Genetics of type 2 diabetes: past, present, and
future. Nutrients, 14(15), 3201.
16. Mahajan, M., and Mahajan, Y. (2023). Role of yoga in type 2 diabetes mellitus. Indian
Journal of Clinical Anatomy and Physiology 10(1):6–9
17. Mambiya, M., Shang, M., Wang, Y., Li, Q., Liu, S., Yang, L., & Liu, W. (2019). The play of
genes and non-genetic factors on type 2 diabetes. Frontiers in public health, 7, 349.
18. Pandi Prabha, S., & Johanna Rajkumar, & Karthik, C. (2020). Hepatotoxic effect of lead
and hepatoprotective effect of Hydrilla verticillata on hepatic transcriptional and
physiological response in edible fish Labeo rohita. Drug and Chemical Toxicology, 45(3),
1276-1283.
19. Pradeepa, R., & Mohan, V. (2021). Epidemiology of type 2 diabetes in India. Indian journal
of ophthalmology, 69(11), 2932.
20. Raja Priya, S. (2020). A Comparative study on the effects of Simplified Kundalini Yoga in
Type II Diabetes Mellitus (Doctoral dissertation, PSG Institute of Medical Sciences and
Research, Coimbatore).
21. Rajasekaran, V M., and Perumal, K. (2018). Role of SKY yoga in type 2 diabetes. Review Of
Research, 7 (8): 1-7
22. Raveendran, A. V., Deshpandae, A., & Joshi, S. R. (2018). Therapeutic role of yoga in type 2
diabetes. Endocrinology and Metabolism, 33(3), 307-317.

17
23. Sharma, S., Bhardwaj, S., Jangir, S., & Gupta, B. (2020). Influence of yoga on status of lipid
indices in type 2 diabetes mellitus subjects. International Journal of Diabetes in Developing
Countries, 40, 410-415.
24. Singh, V., & Khandelwal, B. (2020). Effect of yoga and exercise on glycemic control and
psychosocial parameters in type 2 diabetes mellitus: A randomized controlled study.
International Journal of Yoga, 13(2), 144.
25. Venkatesh, H., Ravish, H., Silvia, C. W. D., & Srinivas, H. (2020). Molecular Signature of
the Immune Response to Yoga Therapy in Stress-related Chronic Disease Conditions: An
Insight. International Journal of Yoga, 13(1), 9.
26. Zhong, M., Wu, Y., Ou, W., Huang, L., & Yang, L. (2019). Identification of key genes
involved in type 2 diabetic islet dysfunction: a bioinformatics study. Bioscience
reports, 39(5), BSR20182172.
5. List of Projects submitted/implemented by the Investigators

5.1 Details of Projects submitted to various funding agencies:

S
. Cost in Month of Role as Agenc
Title Status
N Lakh submission PI/Co- PI y
o
Multiomic evaluation of 29.83 April 2023 PI CSIR submitte
polyethylene, ASPIR d
polypropylene, and E
polyethyleneterephthalate
microplastics
1.
induced cardiactoxicity
by targeting NFkB-
MAPK signaling
pathway using in vitro
and in vivo models
Role of Indian herbal 29.88 December 2023 PI SERB Submitte
medicine in SURE d
cardiotoxicity: Molecular
studies with special
2. reference to
BCL2/CAS3/ROS/TGF
Beta signaling pathways
using in vitro and in vivo
models

5.2 of Projects under implementation: Nil

S.
Titl Cost in Start End Date Role Agency
N
e Lakh Date as

18
 o PI/Co-
PI
- - - - - - -

5.3 Details of Projects completed during the last 5 years:

S. Title Cost in Lakh Start End Role as Agency

No Date Date PI/Co-PI
- - - - - - -

6. List of facilities being extended by parent institution(s) for the project

implementation.
6.1 Infrastructural Facilities
Yes/No/ Not required
S. No. Infrastructural Facility Full or sharing basis
1. Workshop Facility Yes
2. Water & Electricity Yes
3. Laboratory Space/ Furniture Yes
4. Power Generator Yes
5. AC Room or AC Yes
6. Telecommunication including e-mail & fax Yes
7. Transportation Yes
8. Administrative/ Secretarial support Yes
9. Information facilities like Internet/Library Yes
10. Computational facilities Yes
11. Animal/Glass House Yes
12. Any other special facility being provided --

Equipment available with the Institute/Group/Department/Other Institutes

for the project:

Remarks including
Accessories available
Equipment Generic Name Model, Make & Year and
available with of Equipment of purchase
current usage of
equipment
PI & his group

UV - Vis M/s. Shimadzu Biochemical analysis

19
 Spectrophotomet Analytical (India) Pvt. (Accessories available-
er Ltd. in working condition)
& UV-1700, 2010
PI's
Department
Fourier transform Agilent Technologies Functional group
infrared India Pvt. Ltd. & 2022 analysis of various
spectrophotomete samples
r (FTIR)
Microbial growth
Fermentor- 2 analysis and product
M/s. Bioengineering &
numbers (Lab formation
Ralf Plus Solo, 2010
scale) (Accessories available-
in working condition)
Microbial growth
Fermentor – 1 analysis and product
M/s. New Brunswick &
number (Lab formation
Insitu Bioflo 410, 2007
scale) (Accessories available-
in working condition)
Microbial growth
Fermentor – 2 Received from SPIC analysis and product
numbers (Lab Science Foundation, formation
scale) 2010 (Accessories available-
in working condition)
GE Healthcare Pvt. Ltd Animal cell culture
Wave Bioreactor
& Base 20/50 EHT (Accessories available-
(Lab scale)
Wave Pod, 2013 in working condition)
Removal of solvent
Rotary M/s. IKA India Pvt. Ltd. from plant extract
Evaporator & RV 10 Digital, 2017 (Accessories available-
in working condition)
Qualitative and
High
M/s. Young Lin quantitative analysis of
Performance
Instrument Co. Ltd. & biomolecules
Liquid
Acme 9000, 2008 (Accessories available-
Chromatography
in working condition)
Gene expression
M/s. Eppendorf Pvt. analysis (Accessories
Real Time PCR
Ltd., 2009 available- in working
condition)
Fast Separation of
M/s. Bio-Rad
Performance biomolecules
Laboratories India Pvt.
Liquid (Accessories available-
Ltd. & UV Optics, 2014
Chromatography in working condition)
Spray Drier M/s. Labultima Pvt. Ltd. Product polishing
& LU228 Advanced, (Accessories available-

20
 2013 in working condition)
Analysis of exhaust gas
M/s. New Brunswick &
Gas Analyzer (Accessories available-
Ex-2000, 2008
in working condition)
Separation and
Tangential Flow M/s. Millipore India purification of
Filtration System Ltd. & Model -2-230 V, biomolecules
- 2 numbers 2016 (Accessories available-
in working condition)
Maintenance of
microbial culture and
Deep Freezer (- Operon & DFU-374ce,
biomolecules
80ºC) 2006
(Accessories available-
in working condition)
To maintain the
environment for cell
M/s. Sanyo & MCO
CO2 Incubator growth
20A/C, 2017
(Accessories available-
in working condition)
Gene transfer into cells
M/s. Eppendorf Pvt.
Electroporator (Accessories available-
Ltd. 2008
in working condition)
Maintain the
environment for
Ice Flaking M/s. Pooja Lab &
biochemical reactions
Machine IFO30, 2008
(Accessories available-
in working condition)
Microbial Cell
disruption (Accessories
Sonicator M/s. Allainace, 2007
available- in working
condition)
Visualize the structure
Fluorescent
Leica & Dm 1000 & of cells
Binocular
Led, 2009 (Accessories available-
Microscope
in working condition)
Growth of
Orbital Shaking REMI & IHO 2975, microorganisms
Incubator 2010 (Accessories available-
in working condition)
Water Distilled water usage
Elga & ULXXXGEM2,
Purification (Accessories available-
2011
System in working condition)
Visualize the structure
Phase Contrast of cells
Leica & DFC 295, 2009
Microscope (Accessories available-
in working condition)

21
 Cell culture and
Biohazard Safety BCAD-BS-422-B2, microbial work
Cabinet 2009 (Accessories available-
in working condition)
Other
Institute(s) in
the region
National Identification of cells
Beckman Coulter's Epics
Institute for (Accessories available-
XL and Beckton
Research in Flow Cytometer in working condition)
Dickinson's FACS Count
Tuberculosis,
System
Chennai

7. Name and address of experts/ institution interested in the subject / outcome of the
project.

Dr. Nagaraj Manickam

Scientist
Madras Diabetes Research Foundation
Chennai
Tamil Nadu

Dr. Johanna Rajkumar

Professor
Department of Biotechnology
Rajalakshmi Engineering College
Chennai
Tamil Nadu

Dr. Luke Elizabeth Hanna

Scientist E
National Institute for Research in Tuberculosis
Chennai
Tamil Nadu

22