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Glucoraphanin - an Indirect Antioxidant Found Naturally in Broccoli That Supports Cell Integrity and Protects Against Free Radical Damage by Jeremy Bartos, Ph.D

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Glucoraphanin - an Indirect Antioxidant Found Naturally in Broccoli That Supports Cell Integrity and Protects Against Free Radical Damage by Jeremy Bartos, Ph.D GENERAL INFORMATION TM Glucoraphanin - An Indirect Antioxidant Found Naturally In Broccoli That Supports Cell Integrity And Protects Against Free Radical Damage By Jeremy Bartos, Ph.D. Scientific & Regulatory Affairs Manager Glanbia Nutritionals Glanbia Nutritionals | truebroc® White Paper | June 2015 Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com TM introduction Today’s health-conscious consumers are more informed and want to WHAT MAKES TRUEBROCTM A PREMIER understand what they are eating as well as the benefits of the products ANTIOXIDANT they consume. They seek natural ingredients that have no additives or preservatives and are botanical or organic in nature. Currently, there are a large volume of products on the market that have an antioxidant health > truebrocTM boosts Phase II Enzymes, enhancing the claim. In 2015 alone almost 1000 new products with an antioxidant body’s own removal of free radicals and overall position have been released1, but how well do they really work? Recent detoxification of cellsNon-burning and non-irritating scientific research on broccoli has revealed a “next generation” antioxidant to the stomach that will potentially change the way we choose and utilize our ingested antioxidants. truebrocTM glucoraphanin, a naturally derived phytonutrient extracted from broccoli seeds, is a rechargeable antioxidant that works > Standardized to 13% glucoraphanin – highest with the body’s own cellular protection system. Currently, the most widely concentration available accepted source of ingested antioxidants are short-term “one and done” antioxidants, such as Vitamins C and E and polyphenols such as > Made from selected natural broccoli seeds grown resveratrol (grapes and wine) and EGCG (green tea) that last for only a in California and water extracted in Canada short time in the body. The effects of truebrocTM have the benefit of lasting several days in the body – long after the “one and done” antioxidants have • Complete traceability from field to product dissipated. truebrocTM glucoraphanin, the next generation of antioxidants, • Produced under cGMP activates a sustainable antioxidant system within the body that is > Patent Protected – source and process rechargeable and meets the demands of today’s health conscious consumers who are looking for “clean label” options with clearly stated benefits. > Self-Affirmed GRAS PRODUCTS WITH ANTIOXIDANT > Recommended dosage: CLAIMS ON THE RISE • Supplements – 30mg of glucoraphanin (230mg of truebrocTM) • Foods – 10-15mg of glucoraphanin (77- 115mg of truebrocTM) > Applications: • Tablets • Capsules • Powders • Functional Beverages • Functional Foods Glanbia Nutritionals | truebroc® White Paper | June 2015 These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease. 1 Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com TM THE IMPORTANCE OF ANTIOXIDANTS Everyone has heard about antioxidants and knows that they are good for them, but does everyone really know how antioxidants work in our bodies? The main purpose of antioxidants is to protect the cells in the body from damage caused by oxidative stress, namely free radicals, which are unstable compounds missing an electron (a negative charge) on their outside layer2. Free radicals are both created as a byproduct of normal cellular respiration as well as environmental byproducts of pollution, sunlight, etc. Because of their instability they “steal” electrons from any source they can find in the body to help stabilize them. The most readily available source of electrons in the body is the phospholipid bilayer that surrounds each cell. Over time, the chronic exposure of the cell walls to free radicals will break down the walls, which can have a variety of negative effects, both acute and chronic. In some cases it is a major cause of a disease, such as seen in Alzheimer’s3. In others, it simply makes an already existing condition worse, as in asthma or Rheumatoid Arthritis4,5,6,7. By definition, antioxidants are electron donors; they act as sacrificial lambs by offering free radicals another source of electrons besides the cell wall. They can give their electrons to free radicals, neutralizing them and thereby preventing them from stealing any more electrons and doing further damage. The antioxidants can thus help prevent or minimize the build-up of damage over time. Unfortunately, free radical damage is also closely linked with inflammation because the body recognizes the oxidative damage as a threat8. To answer this, the body sends out an army of soldiers in the form of immune cells. These immune “soldiers” fight against anything they perceive to be foreign and once they are done, like casualties in any battlefield, there will be a mass of dead cells. This in itself sends signals for an influx of more immune cell soldiers. What results is a cycle of oxidation - damage – immune cells - more damage. Increasing antioxidant potential in the body provides a solution by helping to prevent a lot of the oxidative damage in the first place and thus also helps minimize localized inflammation, for example in the muscles after rigorous exercise9, 10. Without antioxidants, our cells could be easily damaged by reactive oxygen species, DNA damaging electrophiles, inflammation and radiation. The body has several ways of removing free radicals before they can cause damage. The method best known to consumers is to ingest antioxidants through food or supplements, such as Vitamin C and polyphenols11,12. Slightly less known are native antioxidants and enzymes produced by the body, such as glutathione and superoxide dismutase13,14,15. And finally there is the Phase II Enzyme System; enzymes that act to increase the body’s native antioxidant and detoxifying pathways. The Phase II Enzyme System is activated by Nrf2, a transcription factor that, when induced, increases the production of the specific antioxidant enzymes16,17,18. LONG-LASTING ANTIOXIDANT EFFECTS Most dietary antioxidants, such as vitamins and polyphenols, act by donating electrons to free radicals. Once they do this, their antioxidant function is depleted: they work in a “one-and- done” fashion. The production of free radicals continues and if the supply of one-and-done antioxidants runs out then those free radicals will start stealing again, which can lead to oxidative damage. Figure 1 demonstrates the “one and done” effect of various amounts of ingested Vitamin C; unless the body gets constant replenishment (as often as 4 to 6 times a day) of these ingested antioxidants, the antioxidant protection quickly wanes in a matter of hours19. Glanbia Nutritionals | truebroc® White Paper | June 2015 These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease. 2 Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com TM Native antioxidants such as glutathione and superoxide dismutase are “rechargeable” antioxidants; once they have donated electrons they can be “recharged” by the members of Phase II Enzyme system, thus they can be reused over and over again20,21. One way to ensure this occurs is by activating the Phase II Enzyme system; without it, native antioxidants are also just “one and done”. The major regulator of the genes that code the Phase II Enzymes is nuclear factor erythroid 2-related factor 2 (Nrf2), which binds the antioxidant response element (ARE) region of the gene promoter. This in turn leads to the expression of antioxidant and Phase II Enzymes, demonstrating that Nrf2 is the key to start this Phase II Enzyme system22. Therefore, compounds that can activate Nrf2 can indirectly activate the entire Phase II Enzyme system23. Once activated, the Phase II Enzymes can remain operational for as long as 72 hours, thereby ensuring long-lasting systemic antioxidant effects24. In addition to Phase II Enzyme regulation, Nrf2-ARE binding regulates the expression of more than 100 other genes involved in cellular antioxidant and anti-inflammatory defense such as heat shock proteins and ferritin, pro- and anti-inflammatory enzymes such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and heme oxygenase-1 (HO-1)25,26. It has been shown that down- regulation of Nrf2 can have a variety of negative effects, from increased oxidative stress load and environmental toxin levels to localized inflammation and aberrant mitochondrial biogenesis27,28. Keeping this important pathway up and running is critical to maintaining healthy cellular processes and staving off chronic maladies. DISCOVERY: A NEW ANTIOXIDANT COMPOUND FOUND IN BROCCOLI In the early 1990’s, scientists at Johns Hopkins University identified a compound derived from broccoli called sulforaphane29,30. Sulforaphane and its precursor glucoraphanin (sulforaphane glucosinolate) act as part of the plant’s defense system; in fact, sulforaphane is responsible for the characteristic sulfur smell/taste of broccoli. Glucoraphanin belongs to a category of compounds called glucosinolates, which are naturally found in cruciferous vegetables. It is converted into the isothiocyanate sulforaphane by an enzyme found in broccoli called myrosinase or in the body’s gut microflora31. In the broccoli plant, myrosinase
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