Biomedical Diagnostic Biomedical Diagnostic Research LaboratoryResearch Laboratory

Arthur F. Banné, Ph.D. Director

Chesterland, Ohio

Presented by LifeSpan™ International, Incorporated

BDRLBDRL

CLIA Licensed Reference Laboratory The only laboratory in the U.S.A. validated

and authorized to perform the Serum Thiol Test commercially by Dr. Pero and the University of Lund, Sweden.

The Serum Thiol test is Internationally patent protected.

United States Patent 5,925,571 Pero July 20, 1999 Method of testing immune

competency

Abstract A method of testing the immune competency of an individual by

determining, from a sample of the blood of the individual, a value for total plasma/serum thiols including both protein thiols and non-protein thiols, and comparing the value so determined with a reference value of total plasma/serum thiols to ascertain whether the determined value is higher or lower than the reference value, a determined value lower than the reference value being indicative of impaired immune function.

Inventors: Pero; Ronald W. (Lund, SE) Assignee: Oxi-Gene, Inc. (Boston, MA)

Appl. No.: 847664Filed: April 22, 1997Current U.S. Class:436/120; 435/974; 436/64; 436/86; 436/119; 436/164 Intern'l Class: G01N 033/48; G01N 033/49; G01N 033/50Field of Search: 436/119,120,86,64,164 435/974

Wellness Testing in the

Twenty-First Century

DNA Repair in DNA Repair in Health & Risk Health & Risk AssessmentAssessment

Dr. Ronald PeroDr. Ronald Pero

A world class scientist specializing in genetic ecotoxicology, Dr. Pero has been involved in mainstream cancer research in the field of DNA repair for over 30 years.

His development of DNA Repair testing led to him being selected to direct the biological blood blank for the Malmo Diet and Cancer Study as outlined in Journal of Internal Medicine (Vol.233 No.1 Jan 1993)

Serum Thiol is defined asSerum Thiol is defined as

A test to monitor human health status Such a test was once thought impossible to

provide to the general public It is now possible due to the genius and efforts of

Dr. Ronald Pero, geneticist

What is a Thiol?What is a Thiol?

Thiols are a part of amino acids (AA) which are polymerized into large molecular proteins that in turn signal cells to carry out their functions

Thiol structures are extremely sensitive to oxidatve stress because they can react with free radicals as: protein(AA)-SH SH(AA)-protein oxidizes to protein(AA)-S-S-(AA)-protein + H20

Thus oxidized thiols modulate protein structure which in turn profoundly down regulates cell function that relates to disease

What is a Thiol? What is a Thiol? (Cont’d.)(Cont’d.)

Thiols are the sulfur analog of alcohols. Thiols are a good estimate of endogenous

oxidative stress in vivo as they reflect reduction / oxidation throughout the body due to peripheral circulation of blood to all tissues.

A component of the blood, which is sensitive to changes in the individual’s health

When do you use the Test?When do you use the Test?

There is no clinical disease indication such as: infection or disease

Evaluates underlying biochemical events leading to disease susceptibility

Therefore it can be used any time a health assessment is desired

What is Unique about the Test?What is Unique about the Test?

It will assess the individual’s present health status

In risk assessment, it will provide a measure related to mortality or longevity

It enables the health provider with a means of evaluating the success of current treatment.

Evidence of Thiol Value Evidence of Thiol Value ImplicationsImplications

Serum Thiols as a Surrogate Estimate of DNA Repair Correlates to Mammalian Life Span, RW Pero, C Hoppe, Y Sheng, Journal of Anti-Aging Medicine, Vol.3 No.3 Fall 2000 pp.241-249 (source: www.liebertpub.com)

Low Serum Thiol levels predict shorter times-to-death among HIV-infected injecting drug users , Michael Marmor et al,AIDS 1997, Vol. 11 No.11(source: PubMed)

Side BenefitSide Benefit

Serum Thiol has been validated as a Prognostic Indicator for HIV infection

Predicts clinical outcome more strongly than Beta-2-microglobulin

It is also less expensive and easier to use

Enhancement of DNA RepairEnhancement of DNA Repair

C-MED-100® / AC-11®

Nature’s Magical Ingredient

Joins Man’s Modern Science

Linear Regression Chiropractic StudyLinear Regression Chiropractic Study

y = 16.076x + 24.304

-20

0

20

40

60

80

100

120

140

160

180

0.5 1.5 2.5 3.5

Group

Th

iol

-4

-3

-2

-1

0

1

2

3

4

0.8 1.3 1.8 2.3 2.8

Group

Sta

nd

ard

ized

res

idu

als

How can one product have How can one product have multiple effects?multiple effects?

Because it doesn’t treat specific conditions, but acts by reducing Oxidative Stress (OS), the major cause of human disease

By functioning as a molecular soap, it prepares the cell membrane to absorb nutrients

DNA RepairThe Ultimate Antiaging Tool

by: Ron Pero and Marcia Zimmerman

Reverse Aging – DNA RepairReverse Aging – DNA Repair

The Process of Aging

DNA: The Origin of Life

DNA Damage – Root of All Evil

Nutrition Solutions to Aging

Cat’s Claw Miracle Herb from Peru

Rounding Out The Antiaging Program

Getting Results – What to Expect from Natural DNA Repair

Testing Results – Serum Thiol Test

C-Med-100 Source

Human Genome ProjectHuman Genome Project

Molecular Medicine

Antiaging Medicine

What is Aging?What is Aging?

“. . . the accumulation of random damage to the building blocks of life - especially to DNA, certain proteins, carbohydrates and lipids (fats) – that begins early in life and eventually exceeds the body’s self repair capabilities”

Leonard Hayflick, PhD., UCSF

S. Jay Olshanky, PhD., UICBruce Carnes, PhD., UIC

Scientific American, June 2002; 286(6):92-95

Why Do We Age?Why Do We Age?

Oxidative Stress

Damage to DNA (nucleus & mitochondria)

Poor DNA Repair

Aging and Aging Conditions

Results of Poor Results of Poor DNA RepairDNA Repair

DNA (gene) chemistry

Double stranded helical polymer

DNA damage caused by toxins, diet,

environment, radiation

Repair enzymes / Protein Processes

PARP, endonuclease, exonuclease, polymerase,

ligase

GOOD repair

BAD repair

GOOD

No disease

BAD

BAD REPAIR CASUES ALTERED GENE EXPRESSION, LEADS TO

CHRONIC DISEASES

Cancer, autoimmune, aging cardiovascular, inflammatory

BAD REPAIR CASUES ALTERED GENE EXPRESSION, LEADS TO

CHRONIC DISEASES

Cancer, autoimmune, aging cardiovascular, inflammatory

DNA Damage – Health DNA Damage – Health ConsequencesConsequences

DNA DAMAGEDNA DAMAGE

Mutagenic Diseases

e.g. cancer

Mutagenic Diseases

e.g. cancer

Toxic Disease

e.g. Inflammation

Toxic Disease

e.g. Inflammation

Two possible health consequences

The major cause of human disease has been identified as DNA damage by reactive oxygen species. This has been reviewed as a contributing factor for the origin and progression of a number of conditions including aging, autoimmunity, HIV infection and cancer.

Cellular Control of DNA RepairCellular Control of DNA Repair

DNA repair of damageDNA repair of damage

No diseaseNo disease

Mutaganic and toxic disease

Mutaganic and toxic disease

Incomplete Complete

Gene Deficiency

Biochemical Inhibition

Review of the cellular control of DNA damage

7 STEPS IN AGING7 STEPS IN AGING

YOUR ENVIRONMENT

(Poor diet, stress, accidents generate free

radicals)

OXIDATIVE STRESS

GOOD GENE REPAIR

equals Antiaging

GENE DAMAGE

equals Aging

ALTERED GENE

EXPRESSION

Hobbled Proteins

(Glycation)

Wrong soup mix

(Mitochonria function)

Misfiring genetic switches

(Methylation and acetylation)

Immune glitches

(Inflammation)

Faulty cell communication

(Hormone imbalance)

Aging Effects of Oxidative StressAging Effects of Oxidative Stress

Oxidative stress

Reduces apoptosis

Increases cell survival

Increases TNFalpha

Inflammation

anti-infection

NF-kB

Tumor toxicity

Decreases cell survivalal

Increases apoptosis

Decreases TNFalpha

Anti-inflammation

Enhances immune cells Increases

differentiation/function

Anti-oxidation

Increases DNA repair

Decreases DNA damage

Enhances immune cellsPro-oxidant

Increases DNA damage

Inhibits immune cells

ActivationInhibition

C-MED-100

Control OxidationControl Oxidation

Diet

Exercise

Supplements

Mitochondrial DNAMitochondrial DNA

A single ATP molecule must be recycled within a mitochondrion approximately

1,000 times per day for the body to maintain its energy supply

Quenching Reactive Oxygen Quenching Reactive Oxygen Species in the MitochondrionSpecies in the Mitochondrion

About 10% of the air we breathe is converted into free radicals in the mitochondria of our cells to produce the energy of life (ATP).

Hydrogen peroxide, hydroxyl radical, superoxide, lipid peroxide and singlet oxygen are metabolic mistakes of ATP production.

Mitochondria -Primary source of free radical production in cells.

Savengered by Vitamin E, glutathione, co-enzyme Q10, lipoic acid, carnitine and taurine.

NF-kB / DNA repair-- control NF-kB / DNA repair-- control DNA damage and your healthDNA damage and your health

Second primary source of free radicals is inflammation

NF-kB inhibition reduces this source Regardless the source, DNA repair

removes DNA damage These processes are controlled by

dietary factors

Enhance DNA RepairEnhance DNA Repair

STEP 1.. DNA damage detected. Enzymes uncoil and unzip DNA. (NFk-B inhibition works here also by stopping free radicals that damage DNA)

STEP 2.. Uncoiled structure gives access to repair enzymes.

STEP 3.. Damaged or misplaced nucleotides are cut out by enzymes. (Active genes are repaired first, then less active genes)

STEP 4. New segments are moved into place and the seams are sealed by other enzymes.

STEP 1.. DNA damage detected. Enzymes uncoil and unzip DNA. (NFk-B inhibition works here also by stopping free radicals that damage DNA)

STEP 2.. Uncoiled structure gives access to repair enzymes.

STEP 3.. Damaged or misplaced nucleotides are cut out by enzymes. (Active genes are repaired first, then less active genes)

STEP 4. New segments are moved into place and the seams are sealed by other enzymes.

#4

#1

##2#2

#3

Antioxidants and DNA DamageAntioxidants and DNA Damage Without the entire complement of antioxidants toxic intermediate Without the entire complement of antioxidants toxic intermediate free radicals build upfree radicals build up

Free radical, radiation, drugs, chemicals,

endotoxic metabolites

Ascorbate (Vit. C)

Bioflavonoids

Tocopherols, Tocotrienols

Carotenoids

Glutathione, lipoic acid

Cysteine, selenium, niacin, copper, zinc,

manganese

C-MED-100C-MED-100®® / (AC-11 / (AC-11®®)) The name says it allThe name says it all “C” stands refers to the Campa Indians of the

Amazon basin who have used the bark of the herb called uncaria tomentosa, the source of C-MED-100™ (AC-11®)

“MED” implies not only a clinical effect, but also that this has been developed with the same methods used to develop standard medicines

“100” refers to the 100% bioavailability of product due to the active ingredient CAE (Carboxyl Alkyl Ester) which strongly affects membrane integrity

C-Med-100 / (AC-11C-Med-100 / (AC-11®®))

Water soluble extract of Uncaria tomentosa; 8% CAE’s*, less than 0.05% oxindole alkaloids

*CAE’s examples are quinic acid lactone analogs

C-Med-100 / C-Med-100 / ((AC-11AC-11®®)) primary bioactive primary bioactive ingredient has been identifiedingredient has been identified Found in Shayman natural extractions of UncariaFound in Shayman natural extractions of UncariaQuinic acid/Quinic lactone 5 times more bioactive when Quinic acid/Quinic lactone 5 times more bioactive when purified from C-Med-100 / purified from C-Med-100 / ((AC-11AC-11®®))

C-Med-100 / AC-11® – C-Med-100 / AC-11® – FDA Registered ClaimsFDA Registered Claims

C-Med-100 / (AC-11®) helps enhance natural immunity

C-Med-100 / (AC-11®) helps enhance natural DNA repair process

Nutrients to Enhance C-Med-100 / Nutrients to Enhance C-Med-100 / AC-11® AC-11®

Niacin (niacinamide): a substrate for DNA repair enzymes

Zinc: attaches to DNA repair enzymes, activating them

Carotenoids: scavenge oxy-radicals and enhance immune response

Edible mushroom extracts

C-Med-100 / AC-11® research validates it’s C-Med-100 / AC-11® research validates it’s use to treat DNA damage via NF-kB use to treat DNA damage via NF-kB

inhibition and DNA repair enhancementinhibition and DNA repair enhancement

Induction of apoptosis and tumor inhibition; Anticancer Res., 18:1998

Enhanced DNA repair, immune function, reduced toxicity; J. Ethnopharm., 69:2000

Enhanced antibody response to vaccination; Phytomed., 8:2001

Human DNA repair enhancement; Phytomed., 8:2001

DNA repair enhanced by combination with niacin, carotenoids and zinc; Can. Detect Prev., 22:1998

C-Med-100 / AC-11® Research, C-Med-100 / AC-11® Research, cont’dcont’d..

Serum thiols to estimate DNA repair; J. Antiaging Med., 3:2000

WBC elevation with no pathology or loss of immune function – proof of concept in mouse; Phytomed. In press, 2002

DNA repair as a primary molecular target for antiaging therapies; Antiaging Med., in press, 2002

Deep Cell EffectsDeep Cell Effects

C-MED-100™/ (AC-11®) affects control genes which are the subject of intense drug research for antitumor and anti-inflammatory properties and therefore represent sub-cellular activity.

ResearchResearch C-MED-100™ / (AC-11®) has been tested with

findings published in peer reviewed journals such as Journal of Ethnopharmacology, Anticancer Research and Phytomedicine

It has been shown to suppress tumor growth by inducing apoptosis, the process which forces cells to differentiate or die, without toxic effects to the body

Research Cont’dResearch Cont’d It has enhanced DNA Repair and modulated

immune system function It has successfully improved drug-induced

leukopenia and rendered the pneumococcal vaccine effective

Product ProtectionProduct Protection

Patent protection (US patents 6,039,949 and 6,238,675)

Trademark AC-11® / (C-MED-100™) (10-6-98)