1. Beyond Pathological Calcification: Strategies for Coping with the Effects of Glycation, Oxidation and Inflammation on Cellular Aging

2. David B. Wood, ND BSc. Univ of WA (Microbiology) 1977 ND. Bastyr Univ. 1983 Cofounder, Vice President, CMO: BioGenesis Nutraceuticals, Inc.(2000 - ) Cofounder, President: Trinity Family Health Clinic, PS (1984 - ) Functional Medicine Forum - 2006

3. Factors Affecting Aging Chronic Inflammation Excess or pathological Calcification Glycation Methylation Deficit Mitochondrial Energy Depletion Hormone Imbalance Fatty Acid Imbalance

4. DNA Mutation Immune Dysfunction Non-Digestive Enzyme Imbalance (intracellular) Digestive Enzyme Deficit Excitotoxicity Circulatory Deficit Oxidative Stress

5. In this lecture I will discuss how specific nutraceutical interventions can help offset the aging effects of glycation (AGEs) and its concomitant effects on oxidation and inflammation on our cellular structure, glands and organs. Benefits positively affecting health, vitality and longevity can be achieved with a healthy lifestyle (diet, nutrition, exercise, stress reduction) and specific nutraceutical intervention.

7. Advanced Glycation Endproducts (AGE) When proteins are exposed to elevated levels of glucose the following series of non-enzymatic chemical reactions occur Glycation of proteins and formation of both Amadori adducts and AGE compounds can have biological consequences Glycation Rearrangement Cross-linking hours days weeks/months GlucoseGlycated Amadori +proteins AdductsAGES Protein-NH 2 (Schiff Base) Cell Activation Tissue Structural Changes

8. Glycation Occurs in everyone, but at a faster rate in diabetics Has devastating effects on the health of our tissues Most evident in: Senile dementia Stiffening of the arterial system Degenerative diseases of the eye

9. Some outward examples of AGEs Some outward examples of AGEs Not us!

10. Negative Aspects of AGEs Wrinkles Collagen and Elastin lose their suppleness CataractsArthritisErectile Dysfunction Trigger Inflammatory Reactions Chemokines, Cytokines and Adhesion Molecoles toxic to neurons Key role in development of Cognitive decline and Alzheimers Dz Oxidize Tau proteins Neurofibrillary tangles assoc w/Alzheimers Skeletal muscle Carnosine content s 63% from age 10 to age 70 Good News! Good News! When added to living cell cultures, carnosine extends their life span. When added to decrepit aged cells, it rejuvenates them.

11. AGEx (Advanced Glycation Endproduct inhibitor) Each 4 capsules contain: L-Carnosine 1000mg Galega officinalis (50% guanidine with negligible content of galegine)500mg L-Arginine300mg DMAE (dimethylaminoethanol)300mg Ascorbic acid100mg PABA (para aminobenzoic acid)100mg Vit E (d-alpha tocopheryl succinate) 200IU Thiamine HCl 50mg Alpha R Lipoic acid 50mg Pyridoxal 5 phosphate 35 mg Take 2 capsules two times per day without food or as directed. Recommendation:Take AGEx with Glucostat multivitamin/mineral and a diet made with low glycemic functional foods and dietary foods. 120ct

12. L-Carnosine (B-alanyl-L-histidine) Naturally occurring di-peptide Found in muscle, brain, innervated tissues, lens and other tissues Powerful antioxidant Singlet oxygen, hydrogen peroxide, peroxyl and hydroxyl radicals Inhibits free radical induced damage from iron, copper and zinc Powerful anti-glycation agent Activates myofibillar – ATPase enhancing muscle contractions

13. Increases cellular energy by enhancing mitochondrial oxidative energy production (ATP) Average dietary intake: 50 – 250 mg from one serving of beef, pork or chicken (3-4 ounces) Therapeutic intake (supplemental): 1000+ mg QD Protects SOD from oxidation Prevents accumulation of age-related free radicals

14. L-Carnosine (B-alanyl-L-histidine) May protect against oxidative stress associated w/Alzheimers Dz Protects neuronal and endothelial cells from damage Has anti-glycating properties Improves memory in Alzheimers Improves cognition in Alzheimers Protects against malondialdehyde toxicity Provides protection to cells and molecules from free radical damage

15. Delays aging in human cells Protects against toxic aldehydes. Thus offers protection from diabetes complications, inflammatory ailments, and ETOH related liver disease Positive affect on healthy protein metabolism Positive affect on cellular homeostasis Prevents development of senility features Aids in wound healing (Its degradation product, B- Alanine, enhances collagen production)

16. Enhances the immune system Reduces lactic acid accumulation Promotes muscle recovery, enhancing athletic performance Anti-hypertensive effects Reduces lipid peroxide production and inhibits LDL – C oxidation Source - Life Extension – Carnosine overview www.lef.org

17. Impaired reverse cholesterol transport Impaired esterbation of cholesterol Toxicity to endothelium Facilitation of oxidation

18. Calcification often accompanies glycation!

19. L-Carnosine (B-alanyl-L-histidine) Stvolinskii SL, Fedorova TN, Yuneva MO, Boldyrev AA. Protective effect of carnosine on Cu, Zn-superoxide dismutase during impaired oxidative metabolism in the brain in vivo. Institute of Neurology, Russian Academy of Medical Sciences, Moscow. Bull Exp Biol Med. 2003 Feb;135(2):130-2. Dukic-Stefanovic S, Schinzel R, Riederer P, Munch G. AGES in brain aging: AGE-inhibitors as neuroprotective and anti-dementia drugs? Pysiological Chemistry 1, Biocenter, Univ of Wurzberg, Germany. Biogerontology 2001;2(1):19-34. Forster MJ, Dubey A, Dawson KM, Stutts WA, Lal H, Sohal RS. Age- related losses of cognitive function and motor skills in mice are associated with oxidative protein damage in the brain. Dept. of Pharmacology, Univ. of N. Texas Health Science Center, Fort Worth, TX. Proc Natl Acad Scid. 1996 May 14;93(10):4765-9.

20. Gulyaeva NV, Dupin AM, Levshina IP. Carnosine prevents activation of free-radical lipid peroxidation during stress. Bull Exp Biol Med. 1989; 107(2):148-152. Horning MS, Blakemore LJ, Trombley PQ. Endogenous mechanisms of neuroprotection: role of zinc, copper, and carnosine. Brain Res 2000 Jan 3;852(1):56-61. Our results demonstrate that carnosine can rescue neurons from zinc- and copper-medicated neurotoxicity and suggest that one function of carnosine may be as an endogenous neuroprotective agent Boldyrev A, Song R, Lawrence D, Carpenter DO. Carnosine protects against excitotoxic cell death independently of effects on reactive oxygen species. Neuroscience. 1999;94(2):571-7.

21. Pluripotent protective effects of carnosine, a naturally occurring dipeptide Selected quotes from this study: Evidence will be presented to suggest that carnosine, in addition to antioxidant and oxygen free-radical scavenging activities, also reacts with deleterious aldehydes to protect susceptible macromolecules. Our studies show that, in vitro, carnosine inhibits nonenzymic glycosylation and cross-linking of proteins induced by reactive aldehydes (aldose and ketose sugars, certain triose glycolytic intermediates and malondialdehyde (MDA), a lipid peroxidation product).

22. Hipkiss AR, Preston JE, Himsworth DT, Worthington VC, Keown M, Michaelis J, Lawrence J, Mateen A, Allende L, Eagles PA, Abbott NJ. Pluripotent protective effects of carnosine, a naturally occurring dipeptide. Ann NY Acad Sci. 1998 Nov 20;854:37-53. We propose that carnosine (which is remarkably nontoxic) or related structures should be explored for possible intervention in pathologies that involve deleterious aldehydes, for example secondary diabetic complications, inflammatory phenomena, alcoholic liver disease, and possibly Alzheimers disease.

23. Sztanke K, Pasternak K. The Maillard reaction and its consequences for a living body. Ann Univ Mariae Curie Sklodowska [Med] 2003;58(2):159-162. Wautier JL, Schmidt AM. Protein glycation: a firm link to endothelial cell dysfunction. Circ Res. 2004 Aug 6;95(3):233-8. Loeser RF, Jr. Aging cartilage and osteoarthritis – whats the link? Sci Aging Knowledge Environ. 2004 Jul 21;2004(29):e31. Seidler NW, Yeargans GS, Morgan TG. Carnosine disaggregates glycated alpha-crystallin: an in vitro study. Arch Biochem Biophys. 2004 Jul 1;427(1):110-15. Dukic-Stefanovic S, Schinzel R, Riederer P, Munch G. AGES in brain ageing: AGE- inhibitors as neuroprotective and antidementia drugs? Biogerontology. 2001;2(1):19-34. Betty Crocker Slow-CookOven

24. Beyond Carnosine Compounds that Inhibit Sugar attachment to Protein (Glycation) Pyridoxal 5 phosphate ASA (Aspirin) Compounds that Inhibit or Block formation of Crosslinks (Goats Rue – Galega officinalis) Guanidine (Goats Rue – Galega officinalis) Aminoguanidine Compounds that Trap reactive Carbonyl Intermediates (C=O compounds lead to AGEs) (Galega)Carnosine, L-Arginine Guanidine (Galega) /Aminoguanidine, Carnosine, L-Arginine

25. Chelating Agents that inhibit conversion of Schiff Bases to Amadori Products EDTA (Liposomal oral EDTA, rectal suppository or IV) Penicillamine Antioxidant Agents that inhibit conversion of Schiff Bases to Amadori Products Vitamin C Vitamin E Lipoic Acid

26. Botanical Antioxidants that may inhibit conversion of Schiff Bases to Amadori products Green Tea, Hawthorn, Grape Seed, Milk Thistle, Ginger Root, Ginkgo Aboca Aboca provides these botanicals as organic cold processed, freeze-dried whole phytocomplex concentrates! Compounds that Inhibit formation of Amadori Products (Galega) Guanidine (Galega) /Aminoguanidine

27. GlucoseGlycated Amadori +proteins AdductsAGES Protein-NH 2 (Schiff Base) Pyridoxal 5 Phosphate Aspirin Galega Guanidine Aminoguanidine EDTA Penicillamine Vit C, E, ARLA Galega Guanidine Aminoguanidine Galega Guanidine Aminoguanidine Carnosine L-Arginine Reactive Intermediates C=O

28. Pyridoxal 5 phosphate Bioactive form of Vitamin B6 Significantly reduces non-enzymatic glycation of proteins Inhibits AGE formation Exceeded only by aminoguanidine

29. Thiamine Vitamin B1 or thiamine is the parent compound for the development of a promising new compound ALT-711. UNDUE This compound shows promise in being able to UNDUE existing crosslinked proteins. Thiamine is an effective crosslink breaker Thiamine is an effective crosslink breaker (Pearson and Shaw) Breaking existing crosslinks has been shown to improve arterial elasticity.

30. Galega officinalis (Goats Rue) Historical use for Diabetic treatment for centuries Contains Guanidine which results in its glucose/insulin regulating properties Insulin sensitizing, glucose lowering Safe Galega should have negligible content of galegine. Galegine may cause nasal discharge and blood pressure lowering. Metformin (Glucophage) antidiabetic biguanide derived from Galega officinalis.

31. Alpha R-Lipoic Acid Strong antioxidant protection and enhanced antioxidant recycling Enhanced biological energy production Fat and water soluble Natural form Claims that lipoic acid slows aging of the brain and has anti-aging benefits seem to be related to is potent antioxidant properties Reduces production of Amadori adducts –Packer L, Tritschler HJ, Wessel K. Neuroprotection by the metabolic antioxidant alpha-lipoic acid. Free Rad Biol Med. 1997; 22:359-378.

32. Antioxidants and Neuroprotection Vitamin E Alpha R Lipoic Acid Coenzyme Co Q10 Antioxidants reduce formation of Amadori adducts

33. Vitamin E and Cognitive Decline in Older Persons There was a 36% reduction in the rate of decline among persons in the highest quintile of total vitamin E intake (-4.3 x 10(-2) standardized units per year) compared with those in the lowest quintile (-6.7 x 10(-2) standardized units per year) (P =.05), in a model adjusted for age, race, sex, educational level, current smoking, alcohol consumption, total calorie (energy) intake, and total intakes of vitamin C, carotene, and vitamin A. We also observed a reduced decline with higher vitamin E intake from foods (P =.03 for trend). There was little evidence of association with vitamin C or carotene intake. CONCLUSION: Vitamin E intake, from foods or supplements, is associated with less cognitive decline with age. Arch Neurol 2002 Jul;59(7):1125-32

34. DMAE: Dimethyl amino ethinol Alleviates behavioral problems and hyperactivity associated with Attention Deficit Disorder (ADD) Made naturally in the brain Increases attention span Decreases aggression Improves learning ability Occassionally shows increase in IQ (70% of ADD patients)

35. DMAE Inhibits and reverses cross-linking of proteins Inhibits and reverses cross-linking of proteins Facilitates removal of lipofuscin from neurons and skin (age spots) Facilitates removal of lipofuscin from neurons and skin (age spots) Increases alertness Alleviates anxiety Reduces apathy and increases motivation Improves interhemispheric flow of information in the corpus callosum thereby improving creativity and verbal fluency Improves behavior and mental function in Downs syndrome children Inhibits and reverses cross-linking of proteins Inhibits and reverses cross-linking of proteins Facilitates removal of lipofuscin from neurons and skin (age spots) Facilitates removal of lipofuscin from neurons and skin (age spots) Increases alertness Alleviates anxiety Reduces apathy and increases motivation Improves interhemispheric flow of information in the corpus callosum thereby improving creativity and verbal fluency Improves behavior and mental function in Downs syndrome children

36. DMAE Improves memory and learning Elevates mood Reduces sleep need by ~ 1 hour after 6 weeks of use Dreams become more lucid (vivid) Sleep is sounder with clearer head on waking and more refreshed Enhances Acetylcholine levels within the brain Increases RNA in the brain (rat research) Increases Choline levels within the brain due to DMAEs superior ability to cross the Blood-Brain Barrier Improves memory and learning Elevates mood Reduces sleep need by ~ 1 hour after 6 weeks of use Dreams become more lucid (vivid) Sleep is sounder with clearer head on waking and more refreshed Enhances Acetylcholine levels within the brain Increases RNA in the brain (rat research) Increases Choline levels within the brain due to DMAEs superior ability to cross the Blood-Brain Barrier

37. DMAE References: Coleman N, et al. DMAE in the treatment of hyperactive children. Psychosomatics 17:68-72, 1976. Oettinger L. The use of DMAE in the treatment of disorders of behavior in children. J Pediatrics. 53:671-675, 1958. Cedar G, et al. Effects of 2-Dimethylaminoethanol (DMAE) on the metabolism of choline in plasma. J Neurochemistry. 30:1293- 1296, 1978. Zs-Nagy, I, et al. On the role of cross-linking of cellular proteins in aging. Mech Agi Dev. 14:245- 251, 1980. Zs-Nagy, I, et al. On the role of cross-linking of cellular proteins in aging. Mech Agi Dev. 14:245- 251, 1980. Hochschile R. Effect of dimethylaminoethanol on the life span of senile male A/J mice. Exp Gerontol, 1973, 8: 4, 185-191. Hochschile R. Effect of dimethylaminoethanol on the life span of senile male A/J mice. Exp Gerontol, 1973, 8: 4, 185-191.

38. Also present in:

39. PABA (para amino benzoic acid) Antioxidant Quenches singlet oxygen Provides protection against: –Ozone –Smoking –Other air pollutants Anti-Crosslinking Agent Appears to slow or reverse crosslinking in protein connective tissue structures (ie. Collagen)

40. PABA Anti-Aging Promotes greater flexibility Useful in Tx of vitiligo (a depigmenting disease) Helps prevent graying of hair or restore normal hair color in 10-25% of patients Reduces fibrotic processes: –Peyronnies disease –Dupuytrens contracture –Scleroderma

41. Some PABA references Zarafonetis C. Darkening of gray hair during para-amino- benzoic acid therapy. J Invest Derm, 399-401. Allen JM. Rapid Reaction of Singlet Oxygen with P- Aminobenzoic Acid (PABA) in Aqueous Solution. Biochem Biophys Res Commun, July 1995. Bjorksten J. Crosslinkage and the aging process, in: Theoretical Aspects of Aging, by Morris Rockstein (ed), Academic Press, NY, 1974. Zarafonetis C. Antifibrotic Therapy with POTABA. Am J of Med Sci, 1964, 248:550-561 Dean W. DMAE and PABA, An Alternative to Gerovital (GH3), the Romanian Youth Drug. Vit Res Prod. (vrp.com)

42. Additional References Carpenter D. Correction of biological aging. Rejuvenation, 1980,7:31-49 Bjorksten J. Possibilities and limitations of chelation as a means for life extension. Rejuvenation, 1980, 8:67- 72. Zinsser J, Butt EM, Leonard I. Metal content correlation in aging aorta. J Am Geriatrics Soc, 1957, 5:20-26. Chappell LT, Stahl JP, Evans R. EDTA Chelation treatment for vascular disease: A Meta-Analysis using unpublished data. J Adv Med, 1994, 7: 3, 131-142.

43. BioGenesis products to consider for Anti-Aging: AGE x Liposomal EDTA CogniFactors Aboca botanicals: Aboca botanicals: Green Tea, Hawthorn, Grape Seed, Milk Thistle, Ginger Root, Ginkgo Vitamin E, Alpha R Lipoic Acid, Coenzyme CoQ10, Seleno ExCell, Oxy ATP, Cardio Complete, High ORAC Berry Blend

44. Contacts for Dr. David Wood dwoodnd@bio-genesis.com dwoodnd@trinityclinic.com May your practices and your patients be blessed by your care