Effects of PrimaVie and Exercise Training on Human Skeletal Muscle (PrimaVie)

Potential subjects will see an advertisement placed in locations around the Wexner Medical
Center with RN (research nurse) listed as the main contact. After potential subjects contact
the research nurse, he/she will use inclusion/exclusion criteria to determine potential
eligibility. If the potential subject meets all inclusion/exclusion criteria, he/she will be
scheduled for a primary visit at the Morehouse Medical Plaza or Davis Heart and Lung Research
Institute. At this initial visit, the research nurse will meet the potential subject in a
private room where the study will be fully explained in lay language, and informed consent
will be obtained. Then, appropriate demographic, biometric and health/clinical information
will be recorded on the data collection sheet, and 22.5 mL of blood will be obtained via
venipuncture in the right/ left forearm. A muscle biopsy will also be collected by a trained
physician. The blood and muscle biopsy will be used for all lab related study procedures. The
subjects will be given an 8-week supply of the PrimaVie supplement with instructions, as well
as a copy of the research protocol and informed consent form. If the supplements are mailed,
they will also have instructions, as well as a copy of the research protocol and informed
consent form mailed. The subject will be instructed to call the Research Nurse/ approved
study personnel to let them know that they have received the supplements by mail and to go
over any questions or concerns the subject may have as well as to schedule follow up visits.
All follow up visits (T1, T2, T3a and T3b) will consist of a 22.5 mL blood draw, collection
of muscle biopsy/biopsies and measurement of previously specified demographic, biometric and
health/clinical variables.

For 4 weeks of exercise treadmill training (70-75% HRmax for 20 mins; 5 min warm up and 5 min
cool down - total 30 mins a day; 3 days a week), the subjects will be doing supervised (study
personnel) exercise routine (Treadmill) at study designated facilities( located on The Ohio
State University campus).

Study Hypotheses:

- Hypothesis 1: That oral supplementation of PrimaVie twice a day for 12 weeks will
influence gene expression in the skeletal muscle of sedentary pre-obese to obese humans

- Hypothesis 2: That oral supplementation of PrimaVie will synergistically act with
treadmill exercise training to favorably impact skeletal muscle gene expression.

Aim: A non-randomized, longitudinal study to determine the effect of PrimaVie and exercise
training on human skeletal muscle adaptation in 30 pre-obese to obese volunteers.

Participants: 40 volunteers will be recruited via study advertisement, and contact the
research staff based on the study inclusion and exclusion criteria. At each visit the
following will be recorded/collected:

- 22.5 mL of blood (approximately 2 tablespoons) (venipuncture)

- Muscle biopsy/biopsies (1 muscle biopsy per visit collected at T1 and T2, and 2 muscle
biopsies are collected at T3)

- Age (years)

- Sex (M/F)

- Height (cm)

- Weight (lbs)

- BMI (Kg/m2)

- Blood pressure (mm/Hg)

- Pulse

- Body fat Percentage

- During the first visit (enrollment visit, T1), the following information will be
recorded/collected as well:

- Tobacco use history

- Current alcohol use

- Current other drug use

- Current exercise regimen

- Date of first day of last menstrual period (for females only)

- Current Medications

- Allergies

- Nonmedical information, including: address (will be used to send PrimaVie supplement via
mail, when applicable); telephone number, and email address

Participants will come for three visits:

- Visit 1 (T1) - Baseline (prior to the start of supplementation)

- Visit 2 (T2) - First 8 weeks of supplementation alone

- Visit 3 (T3) - Next 4 weeks of supplementation together with exercise (T3 consists of 2
sample collections: T3a (pre-exercise) after 12 weeks of supplementation together with 4
weeks of exercise - samples taken immediately before the final exercise routine (during
Visit 3), and T3b (post exercise) after 12 weeks of supplementation together with 4
weeks of exercise immediately after the final exercise routine (during Visit 3)

Samples collected from participants:

- Peripheral blood: At all visits, 22.5 mL of peripheral venous blood will be collected.

- Muscle biopsies: biopsies will be collected from the Vastus lateralis or Gastrocnemius
muscles. A biopsy will be collected by a board certified Physician after application of
local anesthetic to the site of biopsy.

Study Supplement: PrimaVie (Natreon Inc), is a a purified and standardized shilajit extract
for nutraceutical use. Research indicates that PrimaVie® has targeted action for increasing
energy, revitalization and anti-aging. PrimaVie® also works at the cellular level, playing a
direct role in the synthesis of ATP, a compound that supplies the body with energy, and
healthy mitochondrial function. Findings on a recent murine study revealed that PrimaVie (PV)
significantly changed the expression of genes related to muscle function (unpublished data).
Based on these facts, it was thought of interest to study the effect of PrimaVie and exercise
training on human skeletal muscle adaptation. Participants will take:

- 500 mg/day PrimaVie (Natreon Inc.) supplement, 250 twice a day for the first 8 weeks of
the study( a 250 mg capsule in the morning with food and a 250 mg capsule in the evening
with food)

- 500 mg/day PrimaVie (Natreon Inc) supplement, 250 mg twice a day for the next 4 weeks (a
250 mg capsule in the morning with food and a 250 mg capsule in the evening with food)
of the study while including supervised exercise treadmill training, which includes:

- exercise supervised treadmill training (supervised treadmill training which include a 5
minute warm up; 70-75% HRmax for 20 minutes; and a 5 minute cool down - a total 30
minutes per day for 3 days per week)

Inclusion Criteria:

- Patients 21-70 years of age

- BMI 25-34.9

Exclusion Criteria:

- BMI >35 OR < 25

- Individuals who are deemed unable to understand the procedures, risks and benefits of
the study,( i.e. Informed consent) will be excluded

- Females who are pregnant as well as individuals who are therapeutically
immuno-compromised will also be excluded in order to minimize the risk to such
individuals (and fetus); to decrease statistical variability and to minimize potential
of confounders.

- Candidates for inclusion into the study will not include individuals as defined in 45
CFR 46 Subparts B, C and D, nor from any other population which may be considered
vulnerable. Pregnant women are excluded to minimize the risk to such individuals (and
fetus); to decrease statistical variability and to minimize potential of confounders.

- History of MRSA (Methicillin-resistant Staphylococcus aureus)

- Current use of any of the following medications will result in exclusion:

- Steroids (Prednisone, etc.)

- Beta-Blockers

- Immunosuppressants

- Hydrochlorothiazide

- Statins (Crestor, Lipitor, etc.)

- Aspirin

- ACE Inhibitors

- Accutane (within the last 6 months)

- Muscle relaxants and stimulants