Document prepared by:

Associate Professor Alison Heather BMedSci, BMath, DipArts, PhD

1996-1999 Post doctoral Scientist, Royal Prince Alfred Hospital 1999-2004 Post doctoral Fellow, Discipline of Medicine, University of Sydney 2004- Gene Regulation Group Leader, Heart Research Institute, Sydney 2009- Associate Professor, University of Technology, Sydney for the School of Medical and Molecular Biosciences, Faculty of Science. Member of the Australian Atherosclerosis Society (AAS) Member of the Cardiac Society of Australia and New Zealand (CSANZ) Member of the Anti-Doping Research Program (ADRP)

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Formulation Hiper’s main active ingredients are pine bark extract, aloe vera, papaya extract and honey.

Pine Bark Extract is the primary active ingredient

There are many studies published in the scientific literature showing the effects of Pine Bark Extract (PBE) on the inflammatory system, oxidative stress system and its capacity to generate the vasodilator molecule, Nitric Oxide (NO).

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eNOS

eNOS is able to take arginine and convert it into nitric oxide (NO) using the following chemical equation.

eNOS

BH4

L-arginine NO + L-citrulline + NAD (P)+

If a healthy diet is consumed, there will be ample arginine in the diet. What is in limited amounts in the body is the enzyme, eNOS. Therefore, only so much nitric oxide can be generated at any one time because the enzyme is in rate limiting amounts. When Hiper sports supplement is consumed, the Pine Bark Extract acts on the endothelial cells in blood vessels to switch on the synthesis of eNOS (Nishioka et al, 2007). As there is more eNOS then it is possible to produce more nitric oxide. Moreover, PBE activates eNOS- it makes the enzyme work faster so nitric oxide is generated more rapidly. This is very important for the active athlete.

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Pine Bark Extract is a Nitric Oxide generator

Nitric oxide is a powerful vasodilator (dilates the blood vessels in the body) but it also has a number of other very beneficial effects on skeletal muscles. Nitric oxide is produced from arginine, an amino acid obtained from the diet from dairy products, beef, pork, poultry, gelatin, seafood, wheat germ, oatmeal, granola, peanuts and other nuts, seeds, chick peas and soybeans. Arginine is converted to nitric oxide by an enzyme called nitric oxide synthase. In our blood vessels, the enzyme is called endothelial nitric oxide synthase, abbreviated to eNOS.

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Nitric Oxide (NO) has the ability to dilate blood vessels. Dilated blood vessels allow more efficient nutrient delivery to active muscles and other tissues. More efficient blood supply will also allow higher levels of oxygen to be transported at a quicker rate to our tissues, including our active muscles. Dilated blood vessels also include our venous system which will allow the faster elimination of toxic waste products from the body. Nitric oxide is important during exercise because it is able to boost the function of mitochondria in cells, including those in skeletal muscle cells. Mitochondria are the energy generating units in cells. The substrate that goes into mitochondria are simple sugars such as glucose and these are converted into the energy unit, ATP. Nitric oxide boosts the speed at which glucose is converted to ATP and it facilitates the entry of glucose into the cell. The faster glucose can be taken up by cells, the faster it can be converted into ATP.

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Nitric Oxide is a natural stimulant of muscle growth and repair. This will benefit the active athlete increasing strength and endurance.

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Hiper Sports Supplement contains Pine Bark Extract which will boost Nitric Oxide generation and this will

help the athlete in many ways.

Hiper Sports Supplement will be effective pre-exercise, during exercise and post-exercise.

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Pine Bark Extract as an anti-oxidant and anti-inflammatory

Inflammation is a natural consequence of endurance exercise. The production of pro-inflammatory cytokines such as IL-1β, TNF-α, IFN-γ, is increased during intense and prolonged exercise (MacKinnon, 1999). The inflammatory cytokines will cause damage within skeletal muscle (Malm, 2001). It has been demonstrated that if blood is sampled 30 min after an Ironman event, TNF-α levels rise 3.5-fold, IFN-γ levels rise by 16.5-fold and IL-1β levels increase by 7-fold and all of these cytokines stay high for the next 48 hours, damaging muscle fibres. Pro-inflammatory cytokines increase the stress hormones, catecholamines and corticosteroids. This is best demonstrated by the dramatic rise in cortisol levels by endurance exercise.

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A cytokine involved in co-ordinating glucose homeostasis (eg. glycogen re-synthesis) and anti-inflammatory responses, IL-6, increases during endurance exercise. It has been measured 24 hours after an Ironman when levels are high and these remain high 5 days after the Ironman event (Neubauer et al, 2008). The increase in inflammatory cytokines is, in part, due to the trauma caused to skeletal muscles during and after endurance exercise. The leading causes of trauma are (a) eccentric muscle contractions, as in the long distance triathlon, (b) the impact of the extremities against the ground and (c) the number of repetitions of the same movement (Bansil et al 1985). The damage to the muscles may lead to a phenomenon of delayed onset muscle soreness (DOMS) that develops 24-48 hours post-endurance exercise that is accompanied by prolonged muscle strength loss, a reduced range of motion, and high levels of creatine kinase activity in the blood (Milles and Clarkson, 1994).

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Muscle swelling and DOMS is all consistent with muscle inflammation that is mediated by infiltrating neutrophils and macrophages that produce inflammatory cytokines but also prostaglandin E2 (PGE2), and this is a key mediator of pain 24-48 hours after an exercise session (Volek and Rawson, 2004). Inflammatory activity within skeletal muscles can also be driven by local endothelial cells and smooth muscle cells. The cytokine inflammatory response persists for at least 5 days post-recovery as a low-grade systemic inflammatory response. For those that start training again, even at a level 1/5th of the training prior to an Ironman race, the low-grade systemic inflammatory response can persist for longer. The initial increase in pro-inflammatory cytokines is followed by elevated levels of anti-inflammatory cytokines. These anti-inflammatory cytokines such as IL-10 will act to dampen the pro-inflammatory response and prevent the overshooting of inflammation.

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Well-trained athletes are able to balance the acute exercise-induced inflammation (Fehrenbach and Schneider, 2006) by increasing their levels of IL-10. Increased IL-10 levels are associated with better performance. A recent study reported that in Ironman triathletes there is a strong compensatory anti-inflammatory cytokine response (Suzuki et al 2006). A study reported that subjects that drop out of Ironman races due to self-reported fatigue symptoms have much higher pre-race CRP and IL-6 levels than those athletes that completed the race. The levels in the athletes that dropped out of the race were 316 to 1442% higher than the average pre-race concentration for those athletes that completed the race. Overtraining/prolonged training may lead to a prolonged inflammatory condition and lead to underperformance syndrome (Robson-Ansley et al 2007; Robson 2003; Smith 2000).

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Aloe vera, papaya extract and honey as an anti-oxidant and anti-inflammatory

Aloe vera, papaya extract and honey are all reported natural anti-inflammatory agents. Like Pine Bark Extract they have been reported in scientific studies to reduce pro-inflammatory cytokine levels in the blood of subjects that have elevated cytokines for a variety of reasons. Papaya extract and honey contain polyphenols that also have antioxidant effects. All of these Hiper components will complement and synergise with the effects of Pine Bark Extract.

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Hiper Sports Supplement has anti-inflammatory effects

A laboratory study has shown that Hiper Sports Supplement can reduce plasma concentrations of the inflammatory marker, acute phase reactant, C Reactive Protein (CRP). Athletes were asked to complete a long run (15-20km+) or long ride (90-160km+) on two separate occasions, 3-weeks apart. Blood sampling was performed prior to exercise and within 15 minutes post-exercise. On one of the occasions, Hiper supplement was ingested, prior to the training run or ride. On the other occasion a placebo supplement was ingested. The athletes were blinded to the supplement that they ingested each time. ELISA tests showed that CRP levels were markedly decreased in the blood of the athletes post-training after Hiper, whereas there was no effect of the placebo supplement (slight increase in CRP levels after exercise). A decrease in CRP reflects a decrease in systemic inflammation caused by endurance exercise training. In turn, this may reduce muscle damage due to inflammation.

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It was next tested if Hiper Sports Supplement has direct anti-inflammatory actions in human cells. To test this, human coronary artery endothelial cells (the cells that line the arteries that supply blood to the tissues of the body) were exposed to Hiper Sports Supplement at concentrations equivalent to a 50-, 25-, 12.5- and 6.25- ml dose of Hiper Sports Supplement. It was shown that Hiper Sports Supplement was able to suppress inflammation caused by the pro-inflammatory cytokine, TNFα, in human coronary artery endothelial cells as assessed by measuring an inflammatory marker in the cells called VCAM-1.

HIPER blocks VCAM-1 levels from rising in response to the pro-

inflammatory cytokine

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In the experiment on the previous page, Hiper Sports Supplement was added directly to the cells from the bottle. However, in the body Hiper Sports Supplement is digested and components are absorbed into the blood stream where they pass through the liver and undergo type II metabolism. Therefore, the original molecules in the Hiper Sports Supplement may be modified and it is these modified components that will cause the cells in the body to respond.

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To test if the Hiper Sports Supplement modified components within the blood stream are effective as anti-inflammatory molecules, subjects were asked to consume 100 ml of Hiper Sports Supplement. Blood sampling was completed immediately before Hiper consumption and 3 hours after Hiper consumption. From the blood collected, the plasma was isolated (this is the part of the blood that contains all of the nutrients, gases, etc and therefore where the Hiper molecules will be). The plasma was then used to treat human coronary artery endothelial cells. Cells were treated for 24 hours with the plasma and then the cells were exposed to an inflammatory stimulus, pro-inflammatory cytokine, TNF-α. We then measured the extent of inflammation by monitoring VCAM-1 levels.

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Summary

There is compelling evidence that Hiper Sports Supplement has anti-inflammatory effects that will result in less tissue damage and help with both performance and recovery.

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References

Bansil CK, Wilson GD (1985) Role of prostaglandins E and F2a in exercise-induced delayed muscle soreness. MedSci Sports Exerc 17:276-280. Fehrenbach E, Schneider ME (2006) Trauma-induced systemic inflammatory response versus exercise-induced immunomodulatory effects. Sports Med 36(5):373-384. Mackinnon LT (1999) Advances in exercise immunology. Human Kinetics, Champaign, IL. Malm KT (2001) Exercise-induced muscle damage and inflammation: fact or fiction? Acta Physiol Scand 171:233-239. Milles MP, Clarkson PM (1994) Exercise-induced muscle pain, soreness, and cramps. J Sports Med Phys Fitness 34:203-206. Neubauer O, Konig D, Wagner K-H (2008) Recovery after an Ironman triathlon: sustained inflammatory responses and muscular stress. Eur J Appl Physiol 104:417-426. Nishioka K, Hidaka T, Nakamura S, Umemura T, Jitsuiki D, Soga J, Goto C, Chayama K, Yoshizumi M, Higashi Y (2007) French maritime pine bark extract. Augments endothelium-dependent vasodilation in humans. Hypertension Research 30:775-780. Robson-Ansley PJ, Blannin A, Gleeson M (2007) Elevated plasma interleukin-6 levels in trained male triathletes following an acute period of intense interval training. Eur J Appl Physiol 99(4):353-360. Robson P (2003) Elucidating the unexplained underperformance syndrome in endurance athletes: the interleukin-6 hypothesis. Sports Med 33(10):771-781. Smith LL (2000) Cytokine hypothesis of overtraining: a physiological adaptation to excessive stress? Med Sci Sports Exerc 32(2):317-331. Suzuki K, Peake J, Nosaka K, Okutsu M, Abbiss CR, Surriano R, Bishop D, Quad MJ, Lee H, Martin DT, Laursen PB (2006) Changes in markers of muscle damage, inflammation and HSP70 after an Ironman triathlon race. Eur J Appl Physiol 98:525-534. Volek JS, Rawson ES (2004) Scientific basis and practical aspects of creatine supplement for athletes. Nutrition 20:609-614.

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