Presented by Patricia Conway, Ph.D. VRI BioMedical & Mark Bartlett, Ph.D . Pharmanex

1. Microflora, Probiotics, and the Gastrointestinal Defense System Presented by Patricia Conway, Ph.D. VRI BioMedical & Mark Bartlett, Ph.D. Pharmanex This material has been prepared by Pharmanex to be used in conjunction with general educational events and activities only. Alterations or modifications of this presentation without the express approval of Pharmanex is prohibited.

2. Our View of Germs...

3. Our View of Germs... • All bacteria are bad and scary... • But that’s OK. Now we have antibiotics... • We also have irradiation, and domestic antibacterials... KILL THEM ALL?

4. Gastrointestinal Microflora Friendly and unfriendly bacteria form a delicate balance Colonization begins at birth continuing through life, leading to: 400 different bacterial species 100 trillion bacteria 10x number of cells than the rest of the body 70% of human immune system localized in digestive tract Two pounds of total body weight Highly concentrated in the colon (1012) accounts for half of the volume of contents in the colon Your Digestive Defense System

5. Gastrointestinal microflora consists of friendly bacteria Aid digestion and enhance the body’s own digestive immune response Friendly bacteria create an intestinal ecosystem that inhibits the growth of undesirable bacteria compete for essential nutrients produce inhibitory substances compete for adherence to intestinal mucosa Gastrointestinal Microflora

6. Delicate balance interrupted by: stress, traveling, alcohol consumption, certain foods, and a poor diet Implications of microflora imbalance: Common, minor symptoms of gastrointestinal (GI) malaise Bowel irregularity Occasional diarrhea Other digestive and non-digestive discomforts A Delicate Balance

7. Gastrointestinal Microflora: A Delicate Balance Ideally, your flora should form a nice, thick “lawn” making it difficult for “weeds” to gain a foot-hold. • Compete for essential nutrients • Produce inhibitory substances • Compete for adherence to intestinal mucosa

8. When it Starts • At birth - digestive tract of humans is sterile. • Colonised by microbes within the first few days of life • At first, predominantly bifidobacteria (breast fed infants). • With the introduction of other foods, a diverse microbial population develops in the gastrointestinal tract. • By now, of all the cells in a human body, the overwhelming majority are non-human.

9. What’s in it for us? Fortifies Digestive Defense System • Helps control occasional GI discomfort* • Promotes digestive comfort and regularity* • Supports healthy function of urinary tract* Immunomodulation • Inhibits growth and adhesion of undesirable organisms* • Cell protection* * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

10. The Search, and Discovery of a New and Unique Probiotic Presented by Patricia Conway, Ph.D. VRI BioMedical

11. Probiotics • Definition and concept of probiotics • Not all probiotics are the same • The search for the perfect probiotic • Research

12. Probiotic: Definition “A product containingviable, defined microorganisms in sufficient numbers, which alter the microflora and exert beneficial effects in the host.” Salminen S, Bouley C, et al. (1998). • Not all strains are the same • Most common species are Lactobacillus and Bifidobacterium • Genus... Species... STRAIN

13. Administered in a form that maintains its viability Have the capacity to survive passage into the intestine Survive conditions in the intestine Attach to the intestinal wall Demonstrable biological activity Stable biological activity in the product Requirements for an Effective Probiotic

14. FAO/WHO Guidelines for Probiotics Source: Joint FAO/WHO Expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Córdoba, Argentina. October 1-4, 2001.

15. Proprietary, patent-pending strain of Lactobacillus fermentum PCC Isolated from Swedish woman with extraordinary resilience A unique strain with unique properties Survives acidic intestinal environment Ability to colonize the intestine* Clinically proven to  occasional GI complaints and increase growth of friendly organisms* Lactobacillus fermentum PCC * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

16. Of human intestinal origin Survives acid, bile, digestive enzymes Ability to colonize the intestine Inhibits growth of undesirable organisms Preferentially targets immune sites Lactobacillus fermentum PCC A Unique Strain with Unique Properties

17. L. fermentum PCC Adhesion Rate L .amylovorusL. fermentum PCC • L. fermentum PCC has superior adhesion to intestinal mucosal cells • Good adhesion indicates viability and may lead to prolonged colonization • Source: Welin A, Farthing M, Conway PL. (Colonization of ileal mucosa). Publication in preparation, 2002.

18. Comparative in vitro adhesion at various sites • Source: Welin A, Farthing M, Conway PL. (Colonization of ileal mucosa). Publication in preparation, 2002.

19. Association with Peyers Patches • Scanning electron micrograph of L.fermentum PCC-like cells on • A) Peyers Patches • B) Non-lymphoid small intestinal tissue

20. Preferential Adhesion to Peyer’s Patches • Source: Plant L, Conway PL. Association of Lactobacillus spp with Peyers Patches in mice. Clinical and Diagnostic Immunology 2002;8(2):320-324.

21. Clinical Substantiation - L. fermentum PCC • Twelve clinical studies • To evaluate stability, colonization, and clinical efficacy •  symptoms of occasional diarrhea, and occasional GI complaints including constipation, flatulence, bloating, and abdominal discomfort* • Sixteen pre-clinical and in vitro studies • Strain is stable and survives transit through digestive tract • Colonization after a single dose; average duration of colonization is 14 days • Adherence to human intestinal mucosa • Preferentially targets Peyer’s Patch *Supported by independent published research. References available upon request. * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

22. Prevalence of Common GI Disturbances • Source: Conway PL, Blomberg L. Reduction in incidence and severity of diarrhoea in soldiers consuming Lactobacillus fermentum KLD. Publication in preparation, 2002.

23. Prevalence of Traveler’s Diarrhea • Source: Conway PL, Blomberg L. Reduction in incidence and severity of diarrhoea in soldiers consuming Lactobacillus fermentum KLD. Publication in preparation, 2002.

24. Development of Common, Minor GI Disturbances • Source: Conway PL, Blomberg L. Reduction in incidence and severity of diarrhoea in soldiers consuming Lactobacillus fermentum KLD. Publication in preparation, 2002.

25. Colonizes the human digestive tract Inhibits growth of undesirable bacteria* Supports immune the response* Reduces occasional GI disturbances in clinical trials* Is stable Lactobacillus fermentum PCC Lactobacillus fermentum PCC * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

26. ProBio PCC™ Maintains a Healthy Gastrointestinal Environment* Presented by Mark Bartlett, Ph.D. Senior Scientist, New Product Development * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

27. Proprietary, patent-pending strain of Lactobacillus fermentum PCC Isolated from Swedish woman with extraordinary resilience A unique strain with unique properties Survives acidic intestinal environment Ability to colonize the intestine* Clinically proven to  occasional GI complaints and increase growth of friendly organisms* ProBio PCC™ from Pharmanex * These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

28. Clinical Substantiation • Maintains healthy intestinal microflora • promotes proliferation of desirable bacteria while inhibiting growth of undesirable bacteria • Supports mucosal immunity • supports humoral immune response • acts as a protective barrier • Decrease in occasional GI complaints, including occasional diarrhea • supports growth and adhesion of desirable bacteria • Supports urinary tract health *Supported by independent published research. References available upon request.

29. For adultswho experience occasional diarrhea, constipation, or cramps resulting fromdistressed digestive flora. For adultswho experience occasional gastrointestinal disturbances, such as gas or bloating. For adultswho are currently taking other probiotics. Directions For Use Take one capsule daily. Take continuously for best results. If you are pregnant or lactating, or taking a prescription medication, consult a physician prior to use. Complementary Products - Digestive Formula, NutriFi Recommended Usage

30. Proprietary and patent-pending Conforms to FAO/WHO guidelines Supported by clinical studies Developed according to the 6S Process Unique delivery system 2 billion CFUs at the end of shelf life Does not require refrigerationstore at or below 68°F (20°C) ProBio PCC™ - Competitive Advantage Pharmanex has obtained exclusive rights to L. fermentum PCC

31. www.pharmanex.com • product information • powerpoint presentation (What’s New) • Pharmanex Information Packet • product fact sheet • frequently asked questions • Pharmanex Product Showcase • meet the experts • interactive computer product training