A New Concept in Nutrition

There are proven benefits from compounds called proanthocyanidins (OPCs) that are highly concentrated in grape seeds.

"Free Radicals have been implicated in over a hundred disease conditions in humans"

...including arthritis, hemorrhagic shock, atherosclerosis, advancing age, ischemia and reperfusion injury of many organs, Alzheimer and Parkinson's disease, gastrointestinal dysfunctions, tumor promotion and carcinogenesis, and AIDS.Antioxidants are potent scavengers of free radicals and serve as inhibitors of neoplastic processes." (Toxicology 2000 Aug 7;148(2-3):187-97)

"Evidence is accumulating that most of the degenerative diseases that afflict humanity have their origin in deleterious free radical reactions."
(Florence TM, Centre for Environmental and Health Science Pty Ltd, Sydney, NSW. "The role of free radicals in disease", ust N Z J Ophthalmol 1995 Feb;23(1):3-7)

Antioxidants

are important supplements required in our diet to neutralize, scavenge, and eliminate from the body, free radical molecules before they can do serious damage to our tissues and organs. Free radicals are unstable molecules which attack DNA and mitochondria, the basic building blocks of all tissues, thereby impairing the functional health of membranes and organs. This damage impedes the replication of healthy cellular material throughout the body and leads to premature aging. Free radicals are implicated in the causation of over 60 degenerative diseases.

~ Omega Biotech


Studies

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The Power of Grape Seed Extract
By Michael Murray, N.D. http://my.webmd.com/content/dmk/dmk_article_5462836

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Journal of the Science of Food and Agriculture
Volume 80, Issue 7, 2000. Pages: 1094-1117

Proanthocyanidins and tannin-like compounds - nature, occurrence, dietary intake and effects on nutrition and health
(Special Issue: Nutritional Enhancement of Plant-based Food in European Trade (NEODIET). Issue Edited by DG Lindsay, MN Clifford.)
"...As metal ion chelators, they influence the bioavailability of several minerals... As reducing agents, they may participate in the prevention of cancers, both of the digestive tract and inner organs. They may also protect LDLs against oxidation and inhibit platelet aggregation and therefore prevent cardiovascular diseases."\


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Altern Med Rev 2000 Apr;5(2):144-51
Oligomeric proanthocyanidin complexes: history, structure, and phytopharmaceutical applications.
Fine AM

International Clinical Research Center, Scottsdale, AZ 85260, USA.
"...OPCs are primarily known for their antioxidant activity. However, these compounds have also been reported to demonstrate antibacterial, antiviral, anticarcinogenic, anti-inflammatory, anti-allergic, and vasodilatory actions. In addition, they have been found to inhibit lipid peroxidation, platelet aggregation, capillary permeability and fragility, and to affect enzyme systems including phospholipase A2, cyclooxygenase, and lipoxygenase. Based on these reported findings, OPCs may be a useful component in the treatment of a number of conditions."

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J Gerontol
 1980 Jan;35(1):45-56
Aspects of free radical reactions in biological systems: aging.
Leibovitz BE, Siegel BV

"...Free radicals are produced during mitochondrial respiration, during the autooxidation of a variety of biological molecules and chemicals, during irradiation damage, and are found as environmental pollutants. Free radicals induce lipid peroxidation which results in membrane damage, increased disulfide/sulfhydryl ratios, and accumulation of aging pigments. Superoxide dismutase, glutathione peroxidase, vitamin E, vitamin C, and selenium are of importance with respect to free radical and lipid peroxide quenching. During aging, the levels of vitamin C appear to decline in the human, guinea pig, and the mouse. Synthetic antioxidants, added to the diets of mice, have been noted to extend the lifespan and mean half-survivale times.


ar5 Toxicology 2000 Aug 7;148(2-3):187-97
Free radicals and grape seed proanthocyanidin extract: importance in human health and disease prevention.
Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, Joshi SS, Pruess HG

Department of Pharmaceutical and Administrative Sciences, Creighton University School of Pharmacy & Allied Health Professions, 2500 California Plaza, Omaha, NE 68178, USA.

Free radicals have been implicated in over a hundred disease conditions in humans, including arthritis, hemorrhagic shock, atherosclerosis, advancing age, ischemia and reperfusion injury of many organs, Alzheimer and Parkinson's disease, gastrointestinal dysfunctions, tumor promotion and carcinogenesis, and AIDS. Antioxidants are potent scavengers of free radicals and serve as inhibitors of neoplastic processes.
These experiments demonstrated that GSPE is highly bioavailable and provides significantly greater protection against free radicals and free radical-induced lipid peroxidation and DNA damage than vitamins C, E and beta-carotene. GSPE was also shown to demonstrate cytotoxicity towards human breast, lung and gastric adenocarcinoma cells, while enhancing the growth and viability of normal human gastric mucosal cells... GSPE provided significantly better protection as compared to vitamins C and E, singly and in combination. GSPE also demonstrated excellent protection against acetaminophen overdose-induced liver and kidney damage ... GSPE demonstrated excellent protection against myocardial ischemia-reperfusion injury and myocardial infarction in rats... Topical application of GSPE enhances sun protection factor in human volunteers, as well as supplementation of GSPE ameliorates chronic pancreatitis in humans.

ar6 J Basic Clin Physiol Pharmacol 1995;6(3-4):205-28 
The role of free radicals in toxicity and disease.
Stohs SJ

Free radicals...are responsible for the tissue damaging effects as lipid peroxidation, and DNA and protein damage. Oxidative stress associated with production of reactive oxygen species is believed to be involved not only in ... the pathophysiology of aging, and various age-related diseases, including cataracts, atherosclerosis, neoplastic diseases, diabetes, diabetic retinopathy, chronic inflammatory diseases of the gastrointestinal tract, aging of skin, diseases associated with cartilage, Alzheimer's disease, and other neurologic disorders.


ar7
Free radicals, antioxidants and preventive geriatrics.
ar7Ward J

Aged Care Services, Eastern Sydney Area Health Service, Pagewood, New South Wales.
Free radical damage seems likely to be significant in the pathophysiology of atherosclerosis, ischaemia-reperfusion injury, Parkinson's disease, cataract, some cancers and rheumatoid arthritis. Evidence to suggest a protective effect from antioxidant vitamins exists for ischaemic heart disease, cataract and some cancers.

ar8
Int J Clin Lab Res
 1999;29(2):49-55 
An update on the role of free radicals and antioxidant defense in human disease.
Vendemiale G, Grattagliano I, Altomare E

Department of Internal and Occupational Medicine, University of Bari - Medical School, Piazza G. Cesare, II, I-70124 Bari, Italy.
evidence indicates that free radicals play
ar8c important roles in many physiological and pathological conditions. The wider application of free radical measurement has increased awareness of functional implications of radical-induced impairment of the oxidative/antioxidative balance. In the following review, the role of oxygen free radicals in some human and experimental pathological conditions is described, with particular emphasis on the mechanisms by which they produce oxidative damage to lipids, proteins, and nucleic bases. The role of free radicals and the activation of the antioxidant systems in arteriosclerosis and ageing, diabetes, ischemia/reperfusion injury, ethanol intoxication, and liver steatosis is discussed.

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Am Pharm 1994 Sep;NS34(9):26-35 
Oxygen free radicals and antioxidants: a review.
Barber DA, Harris SR

Department of Surgery, Mayo Clinic & Foundation, Rochester, Minn.
two of every three people in this country die from either cardiovascular disease or cancer. Based on both experimental and epidemiological evidence, investigators believe that free radicals play a critical role in the development of both diseases. Low levels of antioxidants, which increases free radical activity, are clearly associated with an increased risk of these diseases. This link has led to the conclusion that use of antioxidant vitamin supplements to scavenge free radicals could potentially decrease the risks of cancer and cardiovascular disease
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Ceska Slov Farm 1996 Nov;45(6):296-301 
Natural substances with antioxidant activity
Spilkova J, Dusek J
Katedra farmakognozie Farmaceuticke fakulty Univerzity Karlovy, Hradec Kralove.
substances of phenolic nature (flavonoids, hydroxycinnamic acids, tannins), in which antioxidant effects and ability to quench free radicals have been demonstrated. A number of substances seem to be perspective in the treatment of cardiovascular diseases, cancer and liver diseases.

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Journal of the Science of Food and Agriculture
Volume 80, Issue 7, 2000.
 Pages: 1094-1117
Proanthocyanidins and tannin-like compounds - nature, occurrence, dietary intake and effects on nutrition and health
Proanthocyanidins ... form stable complexes with metal ions and with proteins and are, like other polyphenols, good reducing agents. Many of their biological effects of nutritional interest derive from these properties. As metal ion chelators, they influence the bioavailability of several minerals. The nutritional significance of the non-specific complexation of proteins is less clear. As reducing agents, they may participate in the prevention of cancers, both of the digestive tract and inner organs. They may also protect LDLs against oxidation and inhibit platelet aggregation and therefore prevent cardiovascular diseases.

ar12J Agric Food Chem 1999 May;47(5):1892-7 
Increase of antioxidative potential of rat plasma by oral administration of proanthocyanidin-rich extract from grape
seeds.
Koga T, Moro K, Nakamori K, Yamakoshi J, Hosoyama H, Kataoka S, Ariga T

Noda Institute for Scientific Research, Chiba, Japan.
"...Proanthocyanidins... increases the resistance of blood plasma against oxidative stress and may contribute to physiological functions through their in vivo antioxidative ability."

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Phytotherapy Research
Volume 12, Issue 8, 1998.
 Pages: 568-571
Hydrogen peroxide-induced modulation of intracellular oxidized states in cultured macrophage J774A.1 and neuroactive PC-12 cells, and protection by a novel grape seed proanthocyanidin extract
D. Bagchi 1, C. Kuszynski 2, J. Balmoori 1, M. Bagchi 1, S. J. Stohs 1 *
1Departments of Pharmaceutical and Administrative Sciences, and Pharmacology, Creighton University Health Sciences Center, Omaha, NE 68178, USA
2University of Nebraska Medical Center, Omaha, NE 68198, USA
grape seed proanthocyanidin extract (GSPE) with respect to their scavenging abilities against biochemically generated free radicals in both in vitro and in vivo models. The results demonstrated that GSPE is a significantly more potent oxygen free radical scavenger compared with vitamins C and E and -carotene. GSPE has been reported to exhibit a wide range of biological and pharmacological activities including free radical scavenging, antibacterial, antiviral, antiinflammatory, antiallergic and vasodilator actions.
GSPE can significantly protect against hydrogen peroxide-induced oxidative stress in cultured J774A.1 macrophage and neuronal PC-12 cells.

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Nutr Clin Pract 1995 Feb;10(1):19-25 
Role of antioxidants in health maintenance.
Sardesai VM

Free radicals are produced in the body as by products of normal metabolism and as a result of exposure to radiation and some environmental pollutants. Because they are highly reactive, they can damage cellular components and are implicated in a variety of diseases. Free radicals are normally neutralized by efficient systems in the body that include the antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) and the nutrient-derived antioxidant small molecules (vitamin E, vitamin C, carotenes, flavonoids, glutathione, uric acid, and taurine). In healthy individuals, a delicate balance exists between free radicals and antioxidants. In some pathologic conditions such as diabetes, and in critically ill patients, oxidative stress causes the level of antioxidants to fall below normal. Antioxidant supplements for such conditions are expected to be of benefit.

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Free radical tissue damage: protective role of antioxidant nutrients.
Machlin LJ, Bendich A

Clinical Nutrition, Hoffmann-La Roche Inc., Nutley, New Jersey 07110.
Highly reactive molecules called free radicals can cause tissue damage by reacting with polyunsaturated fatty acids in cellular membranes, nucleotides in DNA, and critical sulfhydryl bonds in proteins. Free radicals can originate endogenously from normal metabolic reactions or exogenously as components of tobacco smoke and air pollutants and indirectly through the metabolism of certain solvents, drugs, and pesticides as well as through exposure to radiation. There is some evidence that free radical damage contributes to the etiology of many chronic health problems such as emphysema, cardiovascular and inflammatory diseases, cataracts, and cancer.
Based on the growing interest in free radical biology and the lack of effective therapies for many of the chronic diseases, the usefulness of essential, safe nutrients in protecting against the adverse effects of oxidative injury warrants further study.


ar16 Mater Med Pol 1993 Jan-Mar;25(1):37-43 
Free radicals and antioxidant systems.
Erenel G, Erbas D, Aricioglu A

Department of Physiology, Medical Faculty, Gazi University.
Free radicals are undesirable peroxides that have long been implicated in several deleterious effects in our body including aging process.


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Nutritional support, free radicals, and antioxidants
Faintuch J, Aguilar PB, Dias MC, Nadalin W, Pinotti HW
Antioxidants and free radical scavengers are molecules endowed with the ability of neutralizing reactive oxygen species that may accumulate in the organism during various pathologic processes. In circumstances of peroxidation of lipids and damage to enzymatic chains, cell membranes and other structures may be followed by functional losses and even cell death.

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Role of free radicals, oxidative stress and antioxidant systems in liver diseases
Jakus V, Lopuchova M

Dpt of Medical Chemistry, Biochemistry and Clinical Biochemistry, Medical Faculty, Comenius University, Bratislava, Slovakia. jakus@fmed.uniba.sk
Recent experimental findings suggest that free radicals and oxidative stress play an important role in the pathogenesis of alcoholic and toxic liver diseases and viral hepatitis. The presented review summarizes knowledge on the pathomechanism of free radical reactions in liver diseases and the results of experimental observations of antioxidant systems and adjuvant antioxidant pharmacotherapy. Some of the hepatoprotective drugs have antioxidant activity and can produce such beneficial effects as membrane stabilisation, neutralization of free radicals and immunomodulation. Some liver diseases can be successfully prevented or treated by supplementation with antioxidant active substances of even plant origin.

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Free radical scavenging by brain homogenate: implication to free radical damage and antioxidant defense in brain.
Mori A, Liu J, Wang X, Kawai M

Department of Neuroscience, Okayama University Medical School, Japan.
To study the mechanisms of free radical-induced brain damage and the antioxidant defense in the brain
The ability of brain homogenate to scavenge free radicals implies that brain damage can be induced by free radicals since they are known to react virtually with any type of molecule such as nucleic acids, membrane lipids, and proteins in the brain
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Nutr Rev
 1994 Aug;52(8 Pt 1):253-65 
Free radicals and antioxidants: a personal view.
Halliwell B

Neurodegenerative Disease Research Center, University of London King's College, UK.
The reactivity of different free radicals varies, but some can cause severe damage to biological molecules, especially to DNA, lipids, and proteins. Antioxidant defense systems scavenge and minimize the formation of oxygen-derived species,
but they are not 100% effective. Hence, diet-derived antioxidants may be particularly important in diminishing cumulative oxidative damage and helping us to stay healthier for longer. Repair systems exist to deal with molecules that have been oxidatively damaged. Damage to DNA by hydroxyl radicals appears to occur in all aerobic cells, and might be a significant contributor to the age-dependent development of cancer. Lipid peroxidation probably contributes significantly to the development of atherosclerosis.

ar21 Indian J Med Sci 1997 Sep;51(9):319-36 
The free radicals--the hidden culprits--an update.
Ansari KN

M.L.N. Medical College, Allahabad.
Free radicals are capable of killing bacteria, damage biomolecules, provoke immune response, activate oncogens, cause atherogenesis and enhance ageing process. However, in healthy conditions nature has endowed human body with enormous antioxidant potential. Subtle balance exists between free radical generation and antioxidant defence system to cope with oxidative stress by various enzymes and vitamins at cellular level which prevent the occurrence of disease. However, factors tilting the balance in favour of excess free radicals generation lead to widespread oxidative tissue damage and diseases. Therefore, trouble starts when there is an excess of free radicals and the defence mechanism lags behind. Overwhelming production of free radicals in response to exposure to toxic chemicals and ageing may necessitate judicious antioxidant supplement to help alleviate free radical mediated damage.

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Orv Hetil 1993 Mar 28;134(13):693-6 
The role of free radical scavengers in gastrointestinal
diseases
Feher J, Pronai L

Semmelweis Orvostudomanyi Egyetem, Budapest, II., Belklinika.
Be it ever so complex the natural antioxidant scavenger system of the organism, the protection provided by it seems to be inappropriate in certain diseases of the gastrointestinal mucus membrane, such as chronic inflammation, immune- and malignant diseases of the bowel. Despite the wide range of therapeutic attempts, the treatment of these diseases has not yet been solved, and the anti-inflammatory-, immunomodulatory-, cytostatic drugs currently used, have several side effects. Therefore, the so called natural- and synthetic antioxidants have been more widely employed for additional and adjuvant treatment. It also has become clear that the direct free radical scavenging effect and/or the membrane protection play an important role in the action mechanism of several old-established drugs. The recent report lists those natural and synthetic antioxidants which have been successfully employed in the clinical treatment of bowel diseases.


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ust N Z J Ophthalmol
 1995 Feb;23(1):3-7 
The role of free radicals in disease.
Florence TM

Centre for Environmental and Health Science Pty Ltd, Sydney, NSW.

Evidence is accumulating that most of the degenerative diseases that afflict humanity have their origin in deleterious free radical reactions. These diseases include atherosclerosis, cancer, inflammatory joint disease, asthma, diabetes, senile dementia and degenerative eye disease. The process of biological ageing might also have a free radical basis. Most free radical damage to cells involves oxygen free radicals or, more generally, activated oxygen species (AOS) which include non-radical species such as singlet oxygen and hydrogen peroxide as well as free radicals. The AOS can damage genetic material, cause lipid peroxidation in cell membranes, and inactivate membrane-bound enzymes. Humans are well endowed with antioxidant defences against AOS; these antioxidants, or free radical scavengers, include ascorbic acid (vitamin C), alpha-tocopherol (vitamin E), beta-carotene, coenzyme Q10, enzymes such as catalase and superoxide dismutase, and trace elements including selenium and zinc. The eye is an organ with intense AOS activity, and it requires high levels of antioxidants to protect its unsaturated fatty acids. The human species is not genetically adapted to survive past middle age, and it appears that antioxidant supplementation of our diet is needed to ensure a more healthy elderly population.



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Am J Clin Nutr 1991 Apr;53(4 Suppl):1050S-1055S 
Protective role of vitamin E in biological systems.
Packer L

Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Vitamin E is well accepted as nature's most effective lipid-soluble, chain-breaking antioxidant, protecting cell membranes from peroxidative damage. Free-radical-mediated pathology has been implicated in the development over time of degenerative diseases and conditions. This paper reviews the current research on the protective role and requirements for vitamin E and the other antioxidants in preventing or minimizing free-radical damage associated with specific diseases and lifestyle patterns and processes, including cancer, aging, circulatory conditions, arthritis, cataract, pollution, and strenuous exercise. While awaiting results of further human studies, research evidence suggests that an adequate intake of vitamin E and the other antioxidants can provide protection from the increasingly high free-radical concentrations caused by air pollutants and current lifestyle patterns.

 
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Proc Nutr Soc 1994 Jul;53(2):251-62 
Vitamin E: molecular and biological function.
Burton GW

Steacie Institute for Molecular Sciences, National Research Council of Canada, Ottawa, Ontario.
There is a growing body of evidence implicating free radicals in a wide variety of medical diseases and conditions, especially the diseases of ageing, such as cancer and cardiovascular disease, which appear to be ultimate expressions of long-term, cumulative and sustained cellular damage. Vitamin E is an excellent lipid-soluble, chain-breaking antioxidant in the presence of other co-operative antioxidants such as vitamin C or ubiquinol, but it can act as a pro-oxidant in their absence. Epidemiological findings and animal studies support the belief that vitamin E is protective against cardiovascular disease and possibly cancer. The wide range of symptoms associated with vitamin E deficiency is consistent with a loss of antioxidant protection in those long-lived cells in which there is sufficient opportunity for accumulation of free radical damage. The cellular damage is proposed to arise from the generation of free radicals during normal aerobic metabolism. Some susceptible tissues may have enhanced levels of radicals that are produced, for example, by the action of cytochrome P-450 enzymes in steroidogenic tissues, or by the generation of NO in neural tissues.


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Recenti Prog Med 1991 Jan;82(1):39-44 
Free radicals in human pathology
Casaril M, Corso F, Corrocher R

Istituto di Patologia speciale medica, Universita, Verona.
The role of free radicals has been suggested in many different diseases; the molecular mechanisms of radical-induced damage have been widely investigated: the main effects on cellular components are lipid peroxidation, protein denaturation and DNA damage causing alteration in membrane functions, impaired enzyme activity and genetic alterations, including cancer. Since oxidative metabolism produces some radicals, aerobic organisms acquired a complex defensive system against radical attack, based on localization of oxidative reactions, enzymes that scavenge free radicals or their products and antioxidant vitamins. Diseases may arise from increased exposure to radicals or from impaired efficiency of protective systems. The role of oxygen radicals in cancerogenesis, alcoholic liver disease and the aging process is also briefly discussed.


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Int J Sport Nutr 1994 Sep;4(3):205-20 
Free radicals, exercise, and antioxidant supplementation.
Kanter MM

Gatorade Sport Science Institute, Quaker Oats Company, Barrington, IL 60010.
Free radicals have been implicated in the development of diverse diseases such as cancer, diabetes, and cataracts, and recent epidemiological data suggest an inverse relationship between antioxidant intake and cardiovascular disease risk. Data also suggest that antioxidants may delay aging. Research has indicated that free radical production and subsequent lipid peroxidation are normal sequelae to the rise in oxygen consumption with exercise. Consequently, antioxidant supplementation may detoxify the peroxides produced during exercise and diminish muscle damage and soreness. Vitamin E, beta carotene, and vitamin C have shown promise as protective antioxidants. Other ingestible products with antioxidant properties include selenium and coenzyme Q10. The role (if any) that free radicals play in the development of exercise-induced tissue damage, or the protective role that antioxidants may play, remains to be elucidated. Current methods used to assess exercise-induced lipid peroxidation are not extremely specific or sensitive; research that utilizes more sophisticated methodologies should help to answer many questions regarding dietary antioxidants.


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Br J Cancer Suppl 1987 Jun;8:153-7 
Lipid antioxidants: how they may act in biological systems.
Niki E

Department of Reaction Chemistry, Faculty of Engineering, University of Tokyo, Japan.
Chain breaking antioxidants scavenge the chain carrying oxygen radicals and suppress the peroxidation of liposomal and biological membranes in aqueous dispersions. Vitamin E scavenges peroxyl radicals rapidly and its lateral diffusion is suggested to be fast, but its antioxidant efficiency in the liposomal and bio-membranes appears to be considerably smaller than in homogeneous solution. Water soluble chain breaking antioxidants, such as uric acid, cysteine, glutathione, and vitamin C, scavenge radicals in the aqueous region and suppress the peroxidation. However, they cannot scavenge the peroxyl radicals within the lipid region of the membranes. Nevertheless, vitamin C can interact with vitamin E radical, probably at membrane-water interface, and regenerate vitamin E.


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Oxidative injury in diseases of the central nervous system: focus on alzheimer's disease.
Pratico D, Delanty N

Department of Pharmacology and Center for Experimental Therapeutics, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
[Medline record in process]

Alzheimer's disease is one of the most challenging brain disorders and has profound medical and social consequences. It affects approximately 15 million persons worldwide, and many more family members and care givers are touched by the disease. The initiating molecular event(s) is not known, and its pathophysiology is highly complex. However, free radical injury appears to be a fundamental process contributing to the neuronal death seen in the disorder, and this hypothesis is supported by many (although not all) studies using surrogate markers of oxidative damage. In vitro and animal studies suggest that various compounds with antioxidant ability can attenuate the oxidative stress induced by beta-amyloid. Recently, clinical trials have demonstrated potential benefits from treatment with the antioxidants, vitamin E, selegiline, extract of Gingko biloba, and idebenone. Further studies are warranted to confirm these findings and explore the optimum timing and antioxidant combination of such treatments in this therapeutically frustrating disease.

PMID: 11063960, UI: 20519007


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Compr Ther 1995;21(1):41-5
Levels of plasma lipid peroxide products and antioxidant status in rheumatoid arthritis.
Kajanachumpol S, Vanichapuntu M, Verasertniyom O, Totemchokchyakarn K, Vatanasuk M

Research Center, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
Oxygen free radicals have been implicated as mediators of tissue damage in patients with rheumatoid arthritis.
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Res Commun Mol Pathol Pharmacol 1997 Feb;95(2):179-89 
Oxygen free radical scavenging abilities of vitamins C and E, and a grape seed proanthocyanidin extract in vitro.
Bagchi D, Garg A, Krohn RL, Bagchi M, Tran MX, Stohs SJ

School of Pharmacy, Creighton University, Omaha, NE 68178, USA.
Proanthocyanidins, a group of polyphenolic bioflavonoids, have been reported to

exhibit a wide range of biological, pharmacological and chemoprotective properties against oxygen free radicals. We have assessed the concentration-dependent oxygen free radical scavenging abilities of a grape seed proanthocyanidin extract (GSPE), vitamin C and vitamin E succinate (VES) as well as superoxide dismutase, catalase and mannitol against biochemically generated superoxide anion and hydroxyl radical using a chemiluminescence assay and cytochrome c reduction. A concentration-dependent inhibition was demonstrated by GSPE. At a 100 mg/l concentration, GSPE exhibited 78-81% inhibition of superoxide anion and hydroxyl radical. Under similar conditions, vitamin C inhibited these two oxygen free radicals by approximately 12-19%, while VES inhibited the two radicals by 36-44%. The combination of superoxide dismutase and catalase inhibited superoxide anion by approximately 83%, while mannitol resulted in an 87% inhibition of hydroxyl radical. The results demonstrate that GSPE is a more potent scavenger of oxygen free radicals as compared to vitamin C and VES.

PMID: 9090754, UI: 97246155


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C R Acad Sci III 1998 Jan;321(1):31-8 
High protection by grape seed proanthocyanidins (GSPC) of polyunsaturated fatty acids against UV-C induced peroxidation.
Bouhamidi R, Prevost V, Nouvelot A

Laboratoire de neurosciences, UMR 6551 du CNRS, universite de Caen, France.
The antioxidative effects of grape seed proanthocyanidins (GSPC) were studied in three in-vitro models in which polyunsaturated fatty acids (PUFAs) in aqueous solution and mice liver or brain microsomes were used as oxidative substrates, and UV-C irradiation as the pro-oxidant system. Analysis of UV-C induced lipid peroxidation was carried out by two methods: gas liquid chromatography of residual PUFAs and release of thiobarbituric acid-reactive substances (TBARs) measured by TBA reaction. Results indicate that PUFAs are more radiosensitive when incorporated in single component micelles than in mixed component micelles or microsomes. In every case, PUFA peroxidation was inhibited by low concentrations of GSPC (2 mg/L) while epigallocatecin (EGC) and epigallocatechin gallate (EGCG) monomers, at an equivalent level of epicatechin, exhibited no efficacy in our experimental conditions. This latter effect might be explained by a synergistic action of flavan-3-ol monomers, dimers and oligomers contained in the grape seed extract.

PMID: 9759355, UI: 98431519


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Gen Pharmacol 1998 May;30(5):771-6 
Protective effects of grape seed proanthocyanidins and selected antioxidants against TPA-induced hepatic and brain lipid peroxidation and DNA fragmentation, and peritoneal macrophage activation in mice.
Bagchi D, Garg A, Krohn RL, Bagchi M, Bagchi DJ, Balmoori J, Stohs SJ

Creighton University School of Pharmacy, Omaha, Nebraska, USA.
1. The comparative protective abilities of a grape seed proanthocyanidin extract (GSPE) (25-100 mg/kg), vitamin C (100 mg/kg), vitamin E succinate (VES) (100 mg/kg) and beta-carotene (50 mg/kg) on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced lipid peroxidation and DNA fragmentation in the hepatic and brain tissues, as well as production of reactive oxygen species by peritoneal macrophages, were assessed. 2. Treatment of mice with GSPE (100 mg/kg), vitamin C, VES and beta-carotene decreased TPA-induced production of reactive oxygen species, as evidenced by decreases in the chemiluminescence response in peritoneal macrophages by approximately 70%, 18%, 47% and 16%, respectively, and cytochrome c reduction by approximately 65%, 15%, 37% and 19%, respectively, compared with controls. 3. GSPE, vitamin C, VES and beta-carotene decreased TPA-induced DNA fragmentation by approximately 47%, 10%, 30% and 11%, respectively, in the hepatic tissues, and 50%, 14%, 31% and 11%, respectively, in the brain tissues, at the doses that were used. Similar results were observed with respect to lipid peroxidation in hepatic mitochondria and microsomes and in brain homogenates. 4. GSPE exhibited a dose-dependent inhibition of TPA-induced lipid peroxidation and DNA fragmentation in liver and brain, as well as a dose-dependent inhibition of TPA-induced reactive oxygen species production in peritoneal macrophages. 5. GSPE and other antioxidants provided significant protection against TPA-induced oxidative damage, with GSPE providing better protection than did other antioxidants at the doses that were employed.

PMID: 9559333, UI: 98220087


ar34
Free Radic Res 1998 Oct;29(4):351-8
Antioxidant properties of catechins and proanthocyanidins: effect of polymerisation, galloylation and glycosylation.
Plumb GW, De Pascual-Teresa S, Santos-Buelga C, Cheynier V,
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Williamson G

Biochemistry Department, Institute of Food Research, Norwich Research Park, Colney, UK. geoff.plumb@bbsrc.ac.uk
A range of catechins and oligomeric procyanidins was purified by high performance liquid chromatography (HPLC) from grape seed, apple skin, lentil and almond flesh. Catechins, galloylated epicatechin, glycosylated catechin, procyanidin dimers, galloylated dimers, trimer, and tetramer species were all identified, purified and quantified by HPLC, LC-MS and NMR. The antioxidant properties of these compounds were assessed using two methods: (a) inhibition of ascorbate/iron-induced peroxidation of phosphatidylcholine liposomes; (b) scavenging of the radical cation of 2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate) (ABTS) relative to the water-soluble vitamin E analogue Trolox C (expressed as Trolox C equivalent antioxidant capacity, TEAC). Antioxidant activity in the lipid phase decreased with polymerisation in contrast with antioxidant action in the aqueous phase which increased from monomer to trimer and then decreased from trimer to tetramer. Galloylation of catechin and dimeric procyanidins decreased lipid phase and increased aqueous phase antioxidant activity. Glycosylation of catechin demonstrated decreased activity in both phases.

PMID: 9860050, UI: 99075393


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Am J Clin Nutr 2000 Feb;71(2 Part 2):621S-629S 
Oxidative stress and Alzheimer disease(1).
Christen Y

Fondation Ipsen, Paris.
Research in the field of molecular biology has helped to provide a better understanding of both the cascade of biochemical events that occurs with Alzheimer disease (AD) and the heterogeneous nature of the disease. One hypothesis that accounts for both the heterogeneous nature of AD and the fact that aging is the most obvious risk factor is that free radicals are involved. The probability of this involvement is supported by the fact that neurons are extremely sensitive to attacks by destructive free radicals. Furthermore, lesions are present in the brains of AD patients that are typically associated with attacks by free radicals (eg, damage to DNA, protein oxidation, lipid peroxidation, and advanced glycosylation end products), and metals (eg, iron, copper, zinc, and aluminum) are present that have catalytic activity that produce free radicals. beta-Amyloid is aggregated and produces more free radicals in the presence of free radicals; beta-amyloid toxicity is eliminated by free radical scavengers. Apolipoprotein E is subject to attacks by free radicals, and apolipoprotein E peroxidation has been correlated with AD. In contrast, apolipoprotein E can act as a free radical scavenger and this behavior is isoform dependent. AD has been linked to mitochondrial anomalies affecting cytochrome-c oxidase, and these anomalies may contribute to the abnormal production of free radicals. Finally, many free radical scavengers (eg, vitamin E, selegeline, and Ginkgo biloba extract EGb 761) have produced promising results in relation to AD, as has desferrioxamine-an iron-chelating agent-and antiinflammatory drugs and estrogens, which also have an antioxidant effect.

ar36
Miner Electrolyte Metab 1991;17(2):124-32 
Oxygen free radicals in acute renal failure.
Greene EL, Paller MS

University of Minnesota, Minneapolis.
Renal ischemia injures the renal tubular cell by disrupting the vital cellular metabolic machinery. Further cell damage is caused by restoration of blood flow when oxygen free radicals are produced. Cellular sources of oxygen free radicals include the electron transport chain, the microsomal electron transport chain, oxidant enzymes (xanthine oxidase, cyclo-oxygenase), phagocytes, and cellular auto-oxidation of Fe2+ and epinephrine. Oxygen radicals cause lipid peroxidation of cell and organelle membranes, disrupting the structural integrity and capacity for cell transport and energy production. Studies in models of acute renal failure have yielded convincing evidence that oxygen free radical production occurs during ischemia/reperfusion. More than a dozen reports have demonstrated the ability of exogenous antioxidants to ameliorate renal injury in vivo. Direct demonstration of increased oxygen free radical production during reoxygenation following hypoxia has been shown in cultured renal epithelial cells. Oxygen free radicals also play a role in toxic acute renal failure. The therapeutic usefulness of free radical scavengers remains to be tested.


ar37
Ren Fail 1999 Nov;21(6):627-34 
Protective effect of a bioflavonoid proanthocyanidin-BP1 in glycerol-induced acute renal failure in the rat: renal stereological study.
Avramovic V, Vlahovic P, Mihailovic D, Stefanovic V

Institute of Histology and Embryology, Faculty of Medicine, Nis, Yugoslavia.
Renal ischemia as well as oxygen metabolites play an important role in renal injury during myoglobinuric acute renal failure (ARF). On the other hand, flavonoids, a diverse group of constituents naturally occurring in plants, have a strong antioxidative activity, and have been implicated in vascular relaxation. In this study the protective effect of a new bioflavonoid proanthocyanidin-BP1 (BP1), extracted from seeds of grapes, was evaluated in glycerol-induced ARF in rats. Stereological methods were used to quantify changes in renal morphology associated with ARF. Volume density of tubular lumen and intratubular cast formations, nuclear parameters (area, diameter, volume) of epithelial cells in the cortical proximal tubules, and glomerular parameters (surface area, diameter, volume, perimeter) were estimated on kidney sections of rats treated either with 50% glycerol (8 mL/kg i.m.) alone. BP1 (20 mg/kg i.p.) in addition to glycerol, or BP1 alone. It was noted that the volume density of tubular lumen and cast formations were significantly lower (p < 0.001) in kidneys of the rats treated with BP1 in addition to glycerol, compared with those treated with glycerol alone. There were no significant differences in glomerular and nuclear parameters between glycerol treated, and BP1 in addition to glycerol treated rats. Renal function was significantly improved in rats treated with BP1 in addition to glycerol. The results suggest that BP1 is a protective agent in glycerol model of ARF. This effect is probably due to the antioxidative activity of BP1 and reduced toxicity of myoglobin in renal tissue. Moreover, it is possible that the ability of BP1 to protect the kidney is dependent upon renal vascular relaxation. The potential beneficial effects of bioflavonoid-BP1 demonstrated in experimental ARF could be considered in therapy of myoglobinuric ARF.

PMID: 10586425, UI: 20052872

ar38
J Mol Cell Cardiol 1999 Jun;31(6):1289-97 
Cardioprotective effects of grape seed proanthocyanidin against ischemic reperfusion injury.
Sato M, Maulik G, Ray PS, Bagchi D, Das DK

University of Connecticut School of Medicine, Farmington, Connecticut 06030-1110, USA.
There is increasing evidence to indicate cardioprotective effects of red wine consumption. Such cardioprotective properties of wine have been attributed to certain polyphenolic constituents of grapes. The purpose of this investigation was to examine whether proanthocyanidins derived from grape seeds possess cardioprotective properties. Rats were randomly divided into two groups: grape-seed proanthocyanidin was administered orally to one group of rats (100 mg/kg/day) for 3 weeks while the other group served as control. After 3 weeks, rats were killed, hearts excised, mounted on the perfusion apparatus and perfused with Krebs-Henseleit bicarbonate (KHB) buffer. After stabilization hearts were perfused in the working mode for baseline measurements of contractile functions. Hearts were then subjected to 30 min of global ischemia followed by 2 h of reperfusion. Coronary perfusates were collected to monitor malonaldehyde formation, a presumptive marker for oxidative stress development. At the end of each experiment, the heart was processed for infarct size determination. Peroxyl radical scavenging activity of proanthocyanidin was determined by examining its ability to remove peroxyl radical generated by 2,2'-azobis (2-amidinopropane) dihydrochloride while hydroxyl radical scavenging activity was tested with its ability to reduce 7-OH.-coumarin-3-carboxylic acid. The results of our study demonstrated that proanthocyanidin-fed animals were resistant to myocardial ischemia reperfusion injury as evidenced by improved recovery of post-ischemic contractile functions. The proanthocyanidin-fed group revealed reduced extent of myocardial infarction compared to the control group. Fluorimetric study demonstrated the antioxidant property of proanthocyanidin as judged by its ability to directly scavenge peroxyl radicals. Taken together, the results of this study showed that grape seed-proanthocyanidins possess a cardioprotective effect against ischemia reperfusion injury. Such cardioprotective property, at least in part, may be attributed to its ability to directly scavenge peroxyl and hydroxyl radicals and to reduce oxidative stress developed during ischemia and reperfusion. Copyright 1999 Academic Press.

PMID: 10371703, UI: 99301913


ar39
Conn Med 1995 Oct;59(10):579-88 
(gsx7) Free radicals, oxidative stress, oxidized low density lipoprotein (LDL), and the heart: antioxidants and other ar39c
strategies to limit cardiovascular damage.
Sinatra ST, DeMarco J

Manchester Memorial Hospital, USA.
The heart is the most susceptible of all the organs to premature aging and free radical oxidative stress. Clinical research has clearly documented the role of free radical damage and the progression of numerous degenerative diseases, particularly cardiovascular disease. This may be the result of acute ischemia-reperfusion injury, endothelial damage of hyperhomocysteinemia, as well as chronic oxidative damage secondary to lipid peroxidation. Fortunately, although highly responsive, and therefore vulnerable to the effects of oxidative stress, the heart is also receptive to the benefits of targeted phytonutrients, antioxidants, and nutritionals. The effects of antioxidant nutrients have been extensively evaluated in epidemiological, population, and clinical studies. Phytonutrients such as the natural flavonoids and carotenoids found in fresh fruits and vegetables or vitamins C, E, and beta-carotene have powerful antioxidant effects. In addition, minerals like selenium and nutrients such as coenzyme Q10 will minimize free radical risk and optimize a favorable outcome from the ubiquitous presence of oxidative stress on the cardiovascular system. The B complex, particularly folic acid, B12, and B6 are also essential in the prevention of hyperhomocysteinemia, another major risk factor for the circulatory system. Measures to minimize accumulation of heavy metals in the body, especially iron and copper, which are capable of initiating adverse free radical reactions, will also help to assuage oxidative stress. Thus, the combination of a healthy diet supplemented with antioxidants and phytonutrients may be useful in the prevention and promotion of optimum cardiovascular health.


ar40
Ann Thorac Surg 1989 Jun;47(6):939-45 
Free radicals and myocardial protection: a surgical viewpoint.
Menasche P, Piwnica A

Department of Cardiovascular Surgery, Hopital Lariboisiere, Paris, France.
Oxygen-derived free radicals are now considered important contributors to tissue injury associated with ischemia and reperfusion. Transition metals, primarily iron, greatly enhance the generation of these active species, which can destroy a large variety of biomolecules, in particular the lipid components of cell membranes. This review tries to demonstrate why cardiopulmonary bypass and aortic cross-clamping are situations that predispose to oxygen free radical production, and how "anti-free radical" agents such as enzymatic scavengers, antioxidants, and iron chelators may prove to be useful therapeutic adjuncts in the clinical setting of open heart surgery.

ar41
Free Radic Biol Med 1988;4(1):15-24
Evidence of direct toxic effects of free radicals on the myocardium.
Burton KP

Department of Physiology, University of Texas Health Science Center, Dallas 75235.
The hypothesis that oxygen-derived free radicals do indeed play a role in myocardial ischemic and reperfusion injury has received a lot of support. Experimental results have shown that free radical scavengers can protect against certain aspects of myocardial ischemic injury and that on reperfusion the heart approaches a level that is more normal than those hearts not receiving additional scavenging agents.

ar42
Basic Res Cardiol 1982 Sep-Oct;77(5):465-85 
The role of lipid peroxidation in pathogenesis of ischemic damage and the antioxidant protection of the heart.
Meerson FZ, Kagan VE, Kozlov YuP, Belkina LM, Arkhipenko YuV

A working hypothesis on pathogenesis of ischemic heart damage has been proposed. According to this hypothesis, a crucial role in conversion of reversible damage into irreversible damage is played by cardiomyocyte membrane destruction caused by the so-called "lipid triad". The latter comprises activation of lipid peroxidation, activation of phospholipases, and the detergentlike action of excessive amounts of free fatty acids and lysophospholipids. Marked activation of lipid peroxidation in experimental myocardial infarction, as well as reoxygenation following transitory ischemia, have been demonstrated. The proposed hypothesis and experimental data underly successful application of synthetic free radical scavengers (antioxidants) for heart protection against experimental myocardial infarction, transitory ischemia, and emotional, painful stress.

PMID: 7181828, UI: 83100260


ar43
J Mol Cell Cardiol 1999 Jun;31(6):1289-97 
Cardioprotective effects of grape seed proanthocyanidin against ischemic reperfusion injury.
Sato M, Maulik G, Ray PS, Bagchi D, Das DK

University of Connecticut School of Medicine, Farmington, Connecticut 06030-1110, USA.
There is increasing evidence to indicate cardioprotective effects of red wine consumption. Such cardioprotective properties of wine have been attributed to certain polyphenolic constituents of grapes. The purpose of this investigation was to examine whether proanthocyanidins derived from grape seeds possess cardioprotective properties. Rats were randomly divided into two groups: grape-seed proanthocyanidin was administered orally to one group of rats (100 mg/kg/day) for 3 weeks while the other group served as control. After 3 weeks, rats were killed, hearts excised, mounted on the perfusion apparatus and perfused with Krebs-Henseleit bicarbonate (KHB) buffer. After stabilization hearts were perfused in the working mode for baseline measurements of contractile functions. Hearts were then subjected to 30 min of global ischemia followed by 2 h of reperfusion. Coronary perfusates were collected to monitor malonaldehyde formation, a presumptive marker for oxidative stress development. At the end of each experiment, the heart was processed for infarct

size determination. Peroxyl radical scavenging activity of proanthocyanidin was determined by examining its ability to remove peroxyl radical generated by 2,2'-azobis (2-amidinopropane) dihydrochloride while hydroxyl radical scavenging activity was tested with its ability to reduce 7-OH.-coumarin-3-carboxylic acid. The results of our study demonstrated that proanthocyanidin-fed animals were resistant to myocardial ischemia reperfusion injury as evidenced by improved recovery of post-ischemic contractile functions. The proanthocyanidin-fed group revealed reduced extent of myocardial infarction compared to the control group. Fluorimetric study demonstrated the antioxidant property of proanthocyanidin as judged by its ability to directly scavenge peroxyl radicals. Taken together, the results of this study showed that grape seed-proanthocyanidins possess a cardioprotective effect against ischemia reperfusion injury. Such cardioprotective property, at least in part, may be attributed to its ability to directly scavenge peroxyl and hydroxyl radicals and to reduce oxidative stress developed during ischemia and reperfusion. Copyright 1999 Academic Press.

PMID: 10371703, UI: 99301913


ar44
Nutr Metab Cardiovasc Dis 2000 Feb;10(1):38-44 
Dietary antioxidants for cardiovascular prevention.
Giugliano D

Dipartimento di Gerontologia, Geriatria e Malattie del Metabolismo, Seconda Universita di Napoli, Italy.
The generation of reactive oxygen species (ROS) is associated with life in aerobic conditions. ROS are thought to be implicated in the pathogenesis of various human diseases since they are capable of damaging biological macromolecules such as DNA, carbohydrates and proteins. The organism maintains defense against ROS, including enzymes and low molecular-weight antioxidants. An important source of antioxidants is diet which contains numerous compounds exhibiting antioxidant activity. A shortage of antioxidants in the diet might promote coronary heart disease through accumulation of oxidized LDL in macrophages. However, antioxidants may also influence endothelial functions, smooth muscle cell proliferation, thrombosis and plaque rupture.


ar45
Drugs Exp Clin Res 1999;25(2-3):115-20 
Cardioprotection of red wine: role of polyphenolic antioxidants.
Das DK, Sato M, Ray PS, Maulik G, Engelman RM, Bertelli AA, Bertelli A

University of Connecticut School of Medicine, Farmington, CT 06030-1110, USA.
Epidemiological studies suggest that the consumption of wine, particularly of red wine, reduces the incidence of mortality and morbidity from coronary heart disease. This has given rise to what is now popularly termed the "French paradox". The cardioprotective effect has been attributed to antioxidants present in the polyphenol fraction of red wine. Grapes contain a variety of antioxidants, including resveratrol, catechin, epicatechin and proanthocyanidins. Of these, resveratrol is present mainly in grape skin while proanthocyanidin is present in the seeds. In this report, we provide evidence that red wine extract as well as resveratrol and proanthocyanidins are equally effective in reducing myocardial ischemic reperfusion injury, which suggests that these red wine polyphenolic antioxidants play a crucial role in cardioprotection.

PMID: 10370873, UI: 99299011


ar46
J Cardiovasc Pharmacol 1995 Jul;26(1):90-5
"Endothelium-dependent vasorelaxation caused by various plant extracts."
Fitzpatrick DF, Hirschfield SL, Ricci T, Jantzen P, Coffey RG
In a previous study (Am J Physiol 1993;265: H774-8), we found that certain red wines and other grape products caused endothelium-dependent vasorelaxation. In the present study, aqueous extracts of a variety of vegetables, fruits, teas, nuts, herbs, and spices were tested for their endothelium-dependent relaxing ability in vitro. Rings of rat aorta, with or without an intact endothelium, were mounted in tissue baths, contracted with phenylephrine, and then exposed to diluted plant extracts. Many, but not all, extracts exhibited endothelium-dependent relaxations that were reversed by NG-monomethyl-L-arginine, a nitric oxide synthase inhibitor, which suggested involvement of nitric oxide, the endothelium-derived relaxing factor in the response. Furthermore, extracts that caused relaxation also increased tissue levels of cyclic GMP, the mediator of nitric oxide-induced vascular smooth-muscle relaxation. These results may lend further support to mounting evidence that plant foods contain compounds that, if absorbed intact and in sufficient quantities, could conceivably be beneficial in prevention of cardiovascular disease.


ar47
Orv Hetil 1997 Sep 7;138(36 Suppl 2):2283-7
The role of oxidative stress and the preventive effect of free radical scavengers in arteriosclerosis

Feher J, Blazovics A, Somogyi A, Lengyel G
Semmelweis Orvostudomanyi Egyetem II. Belgyogyaszati Klinika, Budapest.
The role of oxidative stress in the development of arteriosclerosis is well established. This pathogenetic explanation unificates in itself the lipid and thrombotic theories. The authors summarize the most substantial literary data in this relation, they discuss in details those therapic methods, in which the natural and synthetic antioxidants are involved as preventive drugs in the development and consequences of arteriosclerosis. Thus the effects of the dihydroquinoline type antioxidants as well as those of Vitamins A, C and E are discussed partly in experimental, partly in clinical studies. The authors conclude on the basis of own and literary data that the application of antioxidants could decrease the blood vessel alterations produced by arteriosclerosis, as well as the pathological tissue alterations developed in the consequences of ischaemia.

ar48
Atherosclerosis 1999 Jan;142(1):139-49 
Proanthocyanidin-rich extract from grape seeds attenuates the development of aortic atherosclerosis in cholesterol-fed rabbits.
Yamakoshi J, Kataoka S, Koga T, Ariga T

Research and Development Division, Kikkoman Corporation, Noda City, Chiba Pref, Japan.
The aim of this study was to evaluate the antiatherosclerotic effect of proanthocyanidin-rich extracts from grape seeds in cholesterol-fed rabbits. Proanthocyanidin-rich extracts (0.1% and 1% in diets [w/w]) did not appreciably affect the changes in serum lipid profile of cholesterol-fed rabbits. The level of cholesteryl ester hydroperoxides (ChE-OOH) induced by 2,2'-azobis(2-amidinopropane-dihydrochloride (AAPH) were lower in the plasma of rabbits fed proanthocyanidin-rich extract plus cholesterol than in the plasma of rabbits fed cholesterol alone, but not in the low-density lipoprotein (LDL). Aortic malondialdehyde (MDA) content decreased in rabbits fed proanthocyanidin-rich extract. Feeding proanthocyanidin-rich extracts (0.1 and 1% in the diet) to rabbits significantly reduced severe atherosclerosis in the aorta. Immunohistochemical analysis revealed a decrease in the number of oxidized LDL-positive macrophage-derived foam cells in atherosclerotic lesions in the aorta of rabbits fed proanthocyanidin-rich extract. When proanthocyanidin-rich extract was administered orally to rats, proanthocyanidin was detected in the plasma by Porters method but not in the lipoproteins (LDL plus VLDL). In an in vitro experiment using human plasma, proanthocyanidin-rich extract added to the plasma inhibited the oxidation of cholesteryl linoleate in LDL, but not in the LDL isolated after the plasma and the extract were incubated in advance. These results suggested that proanthocyanidins, the major polyphenols in red wine, might trap reactive oxygen species in aqueous series such as plasma and interstitial fluid of the arterial wall, thereby inhibiting oxidation of LDL and showing an antiatherosclerotic activity.

Comments:

*Comment in: Atherosclerosis 1999 Aug;145(2):421-2PMID: 9920515, UI: 99117243


ar49
Minerva Cardioangiol 1999 Jan-Feb;47(1-2):39-46 
Clinical and capillaroscopic evaluation of chronic uncomplicated venous insufficiency with procyanidins extracted from vitis vinifera
Costantini A, De Bernardi T, Gotti A

Istituto di Chirurgia Vascolare e Angiologia, Universita degli Studi, Milano.

Minerva Cardioangiol 1999 Jan-Feb;47(1-2):39-46

[Clinical and capillaroscopic evaluation of chronic uncomplicated venous insufficiency with procyanidins extracted from vitis vinifera].Costantini A, De Bernardi T, Gotti A Istituto di Chirurgia Vascolare e Angiologia, Universita degli Studi, Milano. BACKGROUND: The pharmacological treatment of non-complicated chronic venous insufficiency is a current and well-debated topic. The introduction of new products with action on the venous system, improved knowledge on the physiopathology of venous insufficiency and the possibility provided by new analytical instruments, have given new impulse to the consolidation of the clinical value of phlebotonics in this indication. METHODS: In light of this, 24 patients with non-complicated chronic venous insufficiency were treated with oral administration of Oligomeric Proanthocyanidins (Pycnogenols-OPC) 100 mg/day. To evaluate the therapeutic efficacy of the treatment, an instrumental evaluation by optical probe capillaroscope was employed in addition to the traditional subjective clinical parameters: swelling, itching, heaviness and pain.

The videocapillaroscope examination was performed at the lower third of the leg and the first toe. Edema in the capillaroscopic field, the number of observable capillaries and the capillary dilatation were the parameter chosen to evaluate the efficacy of treatment. All patients completed the study with no reports of adverse events during the period of observation. RESULTS: The results obtained show a positive clinical response (improved or absent symptoms) in over 80% of patients, with significant improvement of symptoms already evident after the first 10 days of treatment. The mechanism of action of the OPCs explains the rapid reduction of the swelling of the lower limbs and correlated with this are the other evaluable symptoms: heaviness and itching. Particularly striking results were observed for itching and pain which completely disappeared during the course of therapy in 80% and 53% of the patients respectively. Noteworthy is the good correlation between the clinical and instrumental data, with improvement in a total of 70% of patients. CONCLUSIONS: The results obtained in the course of this clinical experience, with evident improvement already during the first weeks of treatment, the absence of adverse events added to the benefit of a once-a-day administration, justify the use of OPC in the treatment of non-complicated chronic venous insufficiency. PMID: 10356940, UI: 99285428


ar50
Free Radic Biol Med 1996;21(4):505-11 
Effect of vitamin E on human aortic endothelial cell responses to oxidative injury.
Martin A, Wu D, Baur W, Meydani SN, Blumberg JB, Meydani M

Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA 02111, USA.
Reactive oxygen species produced by the cells present in the arterial wall may cause oxidative damage to cellular components altering endothelial cell (EC) function. Changes in the EC function appear to play a key role in the pathogenesis of atherosclerosis. Human aortic endothelial cells (HAEC) were employed to investigate the protective role of vitamin E upon exposure of endothelial cells to oxidative stress in vitro. HAEC assimilate d-alpha-tocopherol from the media in a dose-dependent manner. Exposure of HAEC to 16.5 mM of the free radical generator 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) for 16 h decreased cell viability (assessed by trypan blue exclusion) from 90 to 28%. HAEC preincubated with vitamin E at 15, 30, and 60 microM prior to the AAPH exposure resulted in a dose-dependent increase in resistance to oxidative stress and increased cell viability by 37, 66, and 85%, respectively. An increase in prostacyclin (PGI2) production by HAEC in response to AAPH exposure was correlated positively with cell damage and negatively with vitamin E concentration. Interleukin (IL)-1 production also increased in parallel with cell damage induced by AAPH. Vitamin E treatment significantly reduced IL-1 production after AAPH exposure. This modulatory role of vitamin E on HAEC function following exposure to an oxidative stress may reflect its antioxidant protection against lipid peroxidation.

PMID: 8886801, UI: 97041530


ar51
Ann Med 1994 Dec;26(6):435-41
Antioxidants and carcinogenesis.
Rautalahti M, Huttunen J
Cancer Prevention Unit, National Public Health Institute, Helsinki, Finland.

It is established beyond doubt that free radicals in tissues and cells can damage DNA, proteins, carbohydrates and lipids. These potentially deleterious reactions are at least partly controlled by antioxidants capable of scavenging free radicals. It is widely believed that a proper balance between free radicals and antioxidants is essential for the health of an organism. A vast body of observational epidemiological studies has suggested that high intake of dietary or supplemental antioxidants protects against ischaemic heart disease, various types of cancer and several other diseases.


ar52
Compr Ther 1995;21(1):41-5 
Antioxidants protection against cancer and other human diseases.
Uddin S, Ahmad S

Department of Medicine, Stritch School of Medicine, Loyola University of Chicago, Maywood, Illinois, USA.
Antioxidants are of primary value biologically by restricting the damage that reactive free radicals can do to the cell and the cellular components. Antioxidants may afford varying degrees of protection against the cell damage caused by these reactive oxygen free radicals. The antioxidant(s) is/are much more likely to be effective against chemical carcinogens that are metabolically activated to the

reactive free-radical intermediates that are self-propagating than in situations where the metabolic activation results in nonradical reactive intermediates, such as epoxides or carbonium ions. Free-radical disturbances may be of primary and major significance in some important human diseases and intoxication, administration and/or intake may turn out to be of substantial value in the treatment of such cases.

ar53
Free-radical processes in multistage carcinogenesis.
Borek C

Department of Radiation Oncology, Columbia University New York, N.Y. 10032.
Rodent and human cells in culture, transformed in vitro by radiation or chemicals into malignant cells, afford us the opportunity to probe into early and late events in the neoplastic process at a cellular and molecular level. Transformation can be regarded as an abnormal expression of cellular genes. The initiating agents disrupt the integrity of the genetic apparatus altering DNA in ways that result in the activation of cellular transforming genes (oncogenes) during some stage of the neoplastic process. Events associated with initiation and promotion may overlap to some degree, but in order for them to occur, cellular permissive conditions prevail. Permissive and potentiating factors include free radicals, and thyroid hormone, and inadequate antioxidants. Protective factors which suppress the carcinogenic process include enzymatic and dietary antioxidants. These are constitutive under normal circumstances and can be induced under conditions of oxidative stress produced by a wide range of carcinogens.


ar54
Mol Cell Biochem 1999 Jun;196(1-2):99-108 
The cytotoxic effects of a novel IH636 grape seed proanthocyanidin extract on cultured human cancer cells.
Ye X, Krohn RL, Liu W, Joshi SS, Kuszynski CA, McGinn TR, Bagchi M, Preuss HG, Stohs SJ, Bagchi D
Creighton University School of Pharmacy and Allied Health Professions, Omaha, NE 68178, USA.
Grape seed proanthocyanidins are natural antioxidants which possess a broad spectrum of chemoprotective properties against free radicals and oxidative stress. In this study, we have assessed the cytotoxicity of a novel IH636 grape seed proanthocyanidin extract (GSPE) against MCF-7 human breast cancer cells, A-427 human lung cancer cells, CRL-1739 human gastric adenocarcinoma cells and K562 chronic myelogenous leukemic cells at 25 and 50 mg/lit concentrations for 0-72 h using cytomorphology and MTT cytotoxicity assay. In addition, we compared the effects on normal human gastric mucosal cells and normal J774A.1 murine macrophage cells with the effects on the cancer cell lines. Concentration- and time-dependent cytotoxic effects of GSPE were observed on the MCF-7 breast cancer, A-427 lung cancer and gastric adenocarcinoma cells. Following incubation of the MCF-7 cells with 25 mg/lit of the GSPE approximately 6.5, 30 and 43% inhibitions in cell growth were observed at 24, 48 and 72 h of incubation, respectively, while incubation of the MCF-7 cells with 50 mg/lit of the GSPE resulted in 11, 35 and 47% inhibition in cell growth at these same points, respectively. Similar results were observed in the A-427 and gastric adenocarcinoma cells. GSPE exhibited no cytotoxicity toward the neoplastic K562 myelogenous leukemic cells. However, GSPE enhanced the growth and viability of the normal human gastric mucosal cells and J774A.1 murine macrophage cells. These data demonstrate that GSPE exhibited cytotoxicity towards some cancer cells, while enhancing the growth and viability of the normal cells which were examined.

PMID: 10448908, UI: 99376138


ar55
Anticarcinogenic Effect of a Polyphenolic Fraction Isolated From Grape Seeds in Human Prostate Carcinoma DU145 Cells: Modulation of Mitogenic Signaling and Cell-Cycle Regulators and Induction of G1 Arrest and Apoptosis

Chapla Agarwal 1 *, Yogesh Sharma 1, Rajesh Agarwal 1 2
1Center for Cancer Causation and Prevention, AMC Cancer Research Center, Denver, Colorado
2University of Colorado Cancer Center, University of Colorado Health Sciences Center, Denver, Colorado
*Correspondence to Chapla Agarwal, Center for Cancer Causation and Prevention, AMC Cancer Research Center, 1600 Pierce Street, Denver, CO 80214.

(gsx4)

Abstract

There is an increasing interest in identifying potent cancer preventive and therapeutic agents against prostate cancer (PCA). In a recent study, we showed that a polyphenolic fraction isolated from grape seeds (hereafter referred to as GSP) that is substantially rich in antioxidant procyanidins exerts exceptionally high preventive effects against tumorigenesis in a murine skin model. In the present study, we investigated the anticarcinogenic effect of GSP against PCA by employing DU145 human prostate carcinoma cells. GSP treatment (10-100 g/mL doses for 2-6 d) of cells resulted in a highly significant (P < 0.01-0.001) inhibition of cell growth in both dose- and time-dependent manner. Compared with the vehicle, 2 d of GSP treatment resulted in 27, 39, and 76% growth inhibition at 50, 75, and 100 g/mL doses, respectively, whereas 28-97% and ar55c

12-98% inhibition was evident at 10-100 g/mL doses of GSP after 4 and 6 d of treatment, respectively. These doses of GSP also resulted in dose- and time-dependent cell death (6-50%, P <0.1-0.001) that was later characterized as apoptotic death. In molecular mechanistic studies, treatment of DU145 cells with GSP at 25-75 g/mL doses for 24, 48, and 72 h resulted in 77-88%, 65-93%, and 38-98% reduction, respectively (P < 0.001), in phospho-extracellular signal-regulated protein kinase (ERK) 1 and 78%, 19-76%, and 63-71% reduction (P < 0.1-0.001) in phospho-ERK2 levels, respectively. In other studies, similar doses of GSP showed up to 1.9-fold increases in Cip1/p21 and a significant (P < 0.001) decrease in cyclin-dependent kinase (CDK) 4 (up to 90% decrease), CDK2 (up to 50% decrease), and cyclin E (up to 60%decrease). GSP treatment of DU145 cells also resulted in a significant (P < 0.001) G1 arrest in cell-cycle progression in a dose-dependent manner. The growth-inhibitory and cell-death effects of GSP were also observed in another human PCA line, LNCaP. Together, these results suggest that GSP may exert strong anticarcinogenic effect against PCA and that this effect possibly involves modulation of mitogenic signaling and cell-cycle regulators and induction of G1 arrest, cell-growth inhibition, and apoptotic death. Mol. Carcinog. 28:129-138, 2000. © 2000 Wiley-Liss, Inc.

 


ar56
Potent inhibitory action of red wine polyphenols on human breast cancer cells
Athina Damianaki, Efstathia Bakogeorgou, Marilenna Kampa, Anastassia Hatzoglou, Claudia Gemetzi, Elias Castanas
Laboratory of Experimental Endocrinology, University of Crete, School of Medicine and University Hospital, Heraklion GR-71110, Greece
George Notas, Elias Kouroumalis
Laboratory of Gastroenterology, University of Crete, School of Medicine and University Hospital, Heraklion GR-71110, Greece
Simone Panagiotou, Pierre-Marie Martin
Laboratory of Experimental Cancerology, University of Marseille, F-13916, France

Abstract: Breast cancer (one of the most common malignancy in Western societies), as well as esophagus, stomach, lung, bladder, and prostate cancer, depend on environmental factors and diet for growth and evolution. Dietary micronutriments have been proposed as effective inhibitory agents for cancer initiation, progression, and incidence. Among them, polyphenols, present in different foods and beverages, have retained attention in recent years. Red wine is a rich source of polyphenols, and their antioxidant and tumor arresting effects have been demonstrated in different in vitro and in vivo systems. In the present study, we have measured the antiproliferative effect of red wine concentrate, its total polyphenolic pool, and purified catechin, epicatechin, quercetin, and resveratrol, which account for more than 70% of the total polyphenols in red wine, on the proliferation of hormone sensitive (MCF7, T47D) and resistant (MDA-MB-231) breast cancer cell lines. Our results indicate that polyphenols, at the picomolar or the nanomolar range, decrease cell proliferation in a dose- and a time-dependant manner. In hormone sensitive cell lines, a specific interaction of each polyphenol with steroid receptors was observed, with IC50s lower than previously described. Interaction of polyphenols with steroid receptors cannot fully explain their inhibitory effect on cell proliferation. In addition, discrete antioxidant action on each cell line was detected under the same concentrations, both by modifying the toxic effect of H2O2, and the production of reactive oxygen species (ROS), after phorbol ester stimulation. Our results suggest that low concentrations of polyphenols, and consecutively, consumption of wine, or other polyphenol-rich foods and beverages, could have a beneficial antiproliferative effect on breast cancer cell growth. J. Cell. Biochem. 78:429-441, 2000. © 2000 Wiley-Liss, Inc.


ar57
Anticancer Res 1995 Jul-Aug;15(4):1183-9 
Ability of m-chloroperoxybenzoic acid to induce the ornithine decarboxylase marker of skin tumor promotion and inhibition of this response by gallotannins, oligomeric proanthocyanidins, and their monomeric units in mouse epidermis in vivo.
Chen G, Perchellet EM, Gao XM, Newell SW, Hemingway RW, Bottari V, Perchellet JP

Anti-Cancer Drug Laboratory, Kansas State University, Manhattan 66506-4901, USA.
m-Chloroperoxybenzoic acid (CPBA) was tested for its ability to induce the ornithine decarboxylase (ODC) marker of skin tumor promotion. In contrast to benzoyl peroxide, dicumyl peroxide, and 2-butanol peroxide, 5 mg of CPBA applied twice at a 72-h interval induce ODC activity at least as much as 3 micrograms of 12-O-tetradecanoylphorbol-13-acetate (TPA). ODC induction peaks 36 h after a single CPBA treatment but is maximal 5 h after two applications of CPBA at a 48-h interval. The ODC-inducing activity of CPBA is dose dependent and sustained after chronic treatment. In contrast to TPA, two CPBA treatments at 12-24 h intervals produce no refractory state against ODC induction. The mechanism of ODC induction by CPBA is iron dependent. Various hydrolyzable tannins, condensed tannins (CTs) and their monomeric units remarkably inhibit the ODC response to multiple CPBA treatments. At 12 mg, gallic acid, Aleppo gall tannic acid (TA), catechin, and loblolly pine bark CT inhibit the most CPBA-induced ODC activity. Aleppo gall TA is even effective when applied several hours before CPBA. The tumor-promoting activity of CPBA and its inhibition by plant tannins remain to be evaluated.

PMID: 7653998, UI: 95382557


ar58
Cancer Lett 1999 Jan 29;135(2):151-7
Inhibition of TPA-induced tumor promotion in CD-1 mouse epidermis by a polyphenolic fraction from grape seeds.
Bomser JA, Singletary KW, Wallig MA, Smith MA

Department of Food Science and Human Nutrition, University of Illinois, Urbana 61801, USA.The anti-tumor promoting activity of a polyphenolic fraction from grape seeds (GSP) was examined in CD-1 mouse skin epidermis. Specifically, the ability of this fraction to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced tumor promotion and two markers of promotion in mouse skin, ornithine decarboxylase (ODC) and myeloperoxidase (MPO) activities, was evaluated. Pretreatment of mouse skin with 5, 10, 20 and 30 mg of GSP resulted in a dose-dependent reduction in TPA-induced epidermal ODC activity of 27, 37, 48 and 70%, respectively, compared to controls. In addition, pretreatment of mouse skin with 1, 5, 10 and 20 mg of GSP resulted in a significant 43, 39, 54 and 73% inhibition of MPO activity, respectively, compared to controls. In 7,12-dimethylbenz[a]anthracene (DMBA)-initiated CD-1 mice, biweekly treatment of mouse skin with 5, 10, and 20 mg of GSP 20 min prior to TPA application resulted in a 30, 40, and 60% inhibition of final skin tumor incidence, respectively, compared to controls. In addition, the final number of tumors per mouse in the 5, 10 and 20 mg GSP-treated animals was decreased 63, 51, and 94%, respectively, compared to controls. These studies indicate that GSP possesses anti-tumor promoting activity when applied to CD-1 mouse skin prior to treatment with TPA. The mechanism of this tumor inhibition is due, in part, to a GSP-associated inhibition of TPA-induced epidermal ODC and MPO activities. Thus, GSP warrants further evaluation as a skin cancer chemopreventative agent.

PMID: 10096423, UI: 99194087

ar59
Molecular Carcinogenesis
Volume 28, Issue 3, 2000. Pages: 129-138

Anticarcinogenic Effect of a Polyphenolic Fraction Isolated From Grape Seeds in Human Prostate Carcinoma DU145 Cells: Modulation of Mitogenic Signaling and Cell-Cycle Regulators and Induction of G1 Arrest and Apoptosis
Chapla Agarwal 1 *, Yogesh Sharma 1, Rajesh Agarwal 1 2
1Center for Cancer Causation and Prevention, AMC Cancer Research Center, Denver, Colorado
2University of Colorado Cancer Center, University of Colorado Health Sciences Center, Denver, Colorado
There is an increasing interest in identifying potent cancer preventive and therapeutic agents against prostate cancer (PCA). In a recent study, we showed that a polyphenolic fraction isolated from grape seeds (hereafter referred to as GSP) that is substantially rich in antioxidant procyanidins exerts exceptionally high preventive effects against tumorigenesis in a murine skin model. In the present study, we investigated the anticarcinogenic effect of GSP against PCA by employing DU145 human prostate carcinoma cells. GSP treatment (10-100 g/mL doses for 2-6 d) of cells resulted in a highly significant (P < 0.01-0.001) inhibition of cell growth in both dose- and time-dependent manner. Compared with the vehicle, 2 d of GSP treatment resulted in 27, 39, and 76% growth inhibition at 50, 75, and 100 g/mL doses, respectively, whereas 28-97% and 12-98% inhibition was evident at 10-100 g/mL doses of GSP after 4 and 6 d of treatment, respectively. These doses of GSP also resulted in dose- and time-dependent cell death (6-50%, P <0.1-0.001) that was later characterized as apoptotic death. In molecular mechanistic studies, treatment of DU145 cells with GSP at 25-75 g/mL doses for 24, 48, and 72 h resulted in 77-88%, 65-93%, and 38-98% reduction, respectively (P < 0.001), in phospho-extracellular signal-regulated protein kinase (ERK) 1 and 78%, 19-76%, and 63-71% reduction (P < 0.1-0.001) in phospho-ERK2 levels, respectively. In other studies, similar doses of GSP showed up to 1.9-fold increases in Cip1/p21 and a significant (P < 0.001) decrease in cyclin-dependent kinase (CDK) 4 (up to 90% decrease), CDK2 (up to 50% decrease), and cyclin E (up to 60% decrease). GSP treatment of DU145 cells also resulted in a significant (P < 0.001) G1 arrest in cell-cycle progression in a dose-dependent manner. The growth-inhibitory and cell-death effects of GSP were also observed in another human PCA line, LNCaP. Together, these results suggest that GSP may exert strong anticarcinogenic effect against PCA and that this effect possibly involves modulation of mitogenic signaling and cell-cycle regulators and induction of G1 arrest, cell-growth inhibition, and apoptotic death. Mol. Carcinog. 28:129-138, 2000. © 2000 Wiley-Liss, Inc.


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Phytotherapy Research
Volume 12, Issue 8, 1998.
 Pages: 568-571
Hydrogen peroxide-induced modulation of intracellular oxidized states in cultured macrophage J774A.1 and neuroactive PC-12 cells, and protection by a novel grape seed proanthocyanidin extract
D. Bagchi 1, C. Kuszynski 2, J. Balmoori 1, M. Bagchi 1, S. J. Stohs 1 *
1Departments of Pharmaceutical and Administrative Sciences, and Pharmacology, Creighton University Health Sciences Center, Omaha, NE 68178, USA
2University of Nebraska Medical Center, Omaha, NE 68198, USA
We have previously compared selected antioxidants including vitamins C and E, -carotene and a novel IH636 grape seed proanthocyanidin extract (GSPE) with respect to their scavenging abilities against biochemically generated free radicals in both in vitro and in vivo models. The results demonstrated that GSPE is a significantly more potent oxygen free radical scavenger compared with vitamins C and E and -carotene. GSPE has been reported to exhibit a wide range of biological and pharmacological activities including free radical scavenging, antibacterial, antiviral, antiinflammatory, antiallergic and vasodilator actions. In this study hydrogen peroxide-induced oxidative damage was assessed in macrophage J774A.1 and neuroactive adrenal pheochromocytoma PC-12 cells, and the concentration-dependent ability of GSPE to protect these cells. Laser scanning confocal microscopy was used to determine intracellular oxidized states. Approximately 5.8- and 4.5-fold increases in fluorescence intensity were observed following incubation of macrophage J774A.1 and PC-12 cells with 0.50 mM H2O2 for 24 h, respectively. Pretreatment of the J774A.1 cells with 50 mg/L and 100 mg/L GSPE decreased H2O2-induced fluorescence intensity by 36% and 70%, respectively, while under these same conditions approximately 50% and 70% decreases in fluorescence intensities were observed in PC-12 cells. The results indicate that hydrogen peroxide significantly increases oxidative stress in these cells as demonstrated by the increase in intracellular oxidized states. Furthermore, GSPE can significantly protect against hydrogen peroxide-induced oxidative stress in cultured J774A.1 macrophage and neuronal PC-12 cells. Copyright © 1998 John Wiley & Sons, Ltd.


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Environmental and Molecular Mutagenesis
Volume 35, Issue 2, 2000.
 Pages: 86-98
Differential modulation of the genotoxicity of food carcinogens by naturally occurring monomeric and dimeric polyphenolics
Fenton Catterall 1, Jean-Marc Souquet 2, Veronique Cheynier 2, Sonia de Pascual-Teresa 3, Celestino Santos-Buelga 3, Michael N. Clifford 1, Costas Ioannides 1 *
1School of Biological Sciences, University of Surrey, Guilford, Surrey, United Kingdom
2Unité de Recherche Biopolymères et Arômes, INRA, Montpellier Cedex, France
3Nutricion y Bromatologia, Facultad de Farmacia, Campus Miguel de Unamuno, Salamanca, Spain
Naturally occurring dimeric polyphenolics and their gallate esters were isolated from grape seeds, almond fruits, and apple skin, and their ability to modulate the mutagenicity of food carcinogens was studied in the Ames test, and compared to that of the monomeric green tea flavonols, (+)-catechin and (-)-epicatechin. Neither the monomeric nor the dimeric polyphenols and their galloylated derivatives influenced the mutagenic activity elicited by the indirectly acting food carcinogens benzo[a]pyrene and 2-amino-3-methylimidazo-[4,5-f]quinoline (IQ), in the presence of a hepatic activation system derived from Aroclor 1254-treated rats; the only exception was the B7 dimer, which, at concentrations above 1 M, suppressed the mutagenicity of IQ. None of the polyphenolics modulated the mutagenic activity elicited by the directly acting carcinogen N-methyl-N-nitro-nitrosoguanidine (MNNG). In contrast, all the dimeric polyphenols and the galloylated metabolites, at concentrations over 1 M, potentiated the mutagenic activity induced by the indirectly acting carcinogen N-nitrosopyrrolidine, in the presence of an activation system derived from isoniazid-treated rats. In conclusion, dimeric polyphenols and galloylated derivatives of plant origin are unlikely to influence the initiation stage of the carcinogenicity of chemicals through mechanisms that involve inhibition of their cytochrome P450-mediated bioactivation or scavenging of the reactive, genotoxic intermediates. Environ. Mol. Mutagen. 35:86-98, 2000 © 2000 Wiley-Liss, Inc.


ar62
Journal of the Science of Food and Agriculture
Volume 80, Issue 1, 2000. Pages: 91-101

Modulation of the mutagenicity of food carcinogens by oligomeric and polymeric procyanidins isolated from grape seeds: synergistic genotoxicity with N-nitrosopyrrolidine
Fenton Catterall 1, Jean-Marc Souquet 2, Veronique Cheynier 2, Michael N Clifford 1, Costas Ioannides 1 *
1School of Biological Sciences, University of Surrey, Guildford, Surrey GU2 5XH, UK
2Unité de Recherche Biopolymères et Arômes, INRA, Place Viala, F-34060 Montpellier Cedex, France
Oligomeric and polymeric procyanidins were isolated from grape seeds, and their antimutagenic potential against food carcinogens was evaluated in the Ames test. Both procyanidins suppressed the mutagenicity of IQ and benzo[a]pyrene but did not modulate the mutagenic activity of MNNG. At the concentrations where antimutagenic activity was expressed, the oligomeric and polymeric procyanidins inhibited the hepatic O-dealkylation of methoxy- and ethoxyresorufin. It is concluded that the antimutagenic activity exhibited by oligomeric and polymeric procyanidins is the consequence of inhibition of CYP1A-mediated bioactivation. In contrast with these findings, oligomeric and polymeric procyanidins potentiated the mutagenicity of N-nitrosopyrrolidine; the monomeric tea flavanols (+)-catechin and (-)-epicatechin also elicited the same effect. Both the flavanols and procyanidins, at the concentrations studied, failed

to elicit a mutagenic response in the Ames test, either in the presence or absence of an activation system. Incorporation of catalase and superoxide dismutase to the activation system failed to prevent the synergistic effect between (+)-catechin and the nitrosamine. The mutagenic activity of N-nitrosopyrrolidine was much higher when the bacteria were grown in nutrient broth supplemented with (+)-catechin compared with bacteria grown in nutrient broth alone. It may be cautiously inferred that the synergistic genotoxicity between polyphenolics and N-nitrosopyrrolidine involves interaction of (+)-catechin with bacterial DNA, facilitating the covalent binding of the ultimate carcinogens of the nitrosamine to the DNA.

© 2000 Society of Chemical Industry



ar64
J Invest Dermatol 1999 Mar;112(3):310-6 
Procyanidin oligomers selectively and intensively promote proliferation of mouse hair epithelial cells in vitro and activate hair follicle growth in vivo.
Takahashi T, Kamiya T, Hasegawa A, Yokoo Y

Tsukuba Research Laboratories, Kyowa Hakko Kogyo, Ibaraki, Japan.
We have previously reported that proanthocyanidins extracted from grape seeds possess growth-promoting activity toward murine hair epithelial cells in vitro and stimulate anagen induction in hair cycle progression in vivo. This report constitutes a comparison of the growth-promoting activity of procyanidin oligomers and the target cells of procyanidins in the skin. Results show that procyanidin dimer and trimer exhibit higher growth-promoting activity than the monomer. The maximum growth-promoting activity for hair epithelial cells with procyanidin B-2, an epicatechin dimer, reached about 300% (30 microM) relative to controls (= 100%) in a 5 d culture. Optimum concentration of procyanidin C-1, an epicatechin trimer, was lower than that of procyanidin B-2; the maximum growth-promoting activity of procyanidin C-1 was about 220% (3 microM). No other flavonoid compounds examined exhibit higher proliferative activities than the procyanidins. In skin constituent cells, only epithelial cells such as hair keratinocytes or epidermal keratinocytes respond to procyanidin oligomers. Topical application of 1% procyanidin oligomers on shaven C3H mice in the telogen phase led to significant hair regeneration [procyanidin B-2, 69.6% +/- 21.8% (mean +/- SD); procyanidin B-3, 80.9% +/- 13.0%; procyanidin C-1, 78.3% +/- 7.6%] on the basis of the shaven area; application of vehicle only led to regeneration of 41.7% (SD = 16.3%). In this paper, we demonstrate the hair-growing activity of procyanidin oligomers both in vitro and in vivo, and their potential for use as agents to induce hair growth.

PMID: 10084307, UI: 99181798



ar65
Acta Derm Venereol 1998 Nov;78(6):428-32 
Proanthocyanidins from grape seeds promote proliferation of mouse hair follicle cells in vitro and convert hair cycle in vivo.
Takahashi T, Kamiya T, Yokoo Y

Tsukuba Research Laboratories, Kyowa Hakko Kogyo Co., Ibaraki, Japan.
For the purpose of discovering natural products which possess hair growing activity, we examined about 1000 kinds of plant extracts concerning growth-promoting activity with respect to hair follicle cells. After an extensive search, we discovered that proanthocyanidins extracted from grape seeds promote proliferation of hair follicle cells isolated from mice by about 230% relative to controls (100%); and that proanthocyanidins possess remarkable hair-cycle-converting activity from the telogen phase to the anagen phase in C3H mice in vivo test systems. The profile of the active fraction of the proanthocyanidins was elucidated by thiolytic degradation and tannase hydrolysis. We found that the constitutive monomers were epicatechin and catechin; and that the degree of polymerization was 3.5. We demonstrated the possibility of using the proanthocyanidins extracted from grape seeds as agents inducing hair growth.

PMID: 9833041, UI: 99050235


ar66
J Invest Dermatol 1997 Apr;108(4):495-500 
Hair cycle stage of the mouse vibrissa follicle determines subsequent fiber growth and follicle behavior in vitro.
Robinson M, Reynolds AJ, Jahoda CA

Department of Biological Sciences, University of Durham, United Kingdom.
The establishment of culture models representative of all aspects of in vivo hair follicle behavior is an important goal for theoretic and analytic studies. Rodent vibrissa follicles have regular, predictable, and relatively short growth cycles. In this investigation, we took advantage of these properties; we classified mouse vibrissa follicles according to different phases in the hair cycle and then compared fiber growth and morphologic changes in culture. Follicles isolated in the early phase of the growth cycle produced fine growing fibers with an average growth that exceeded 3 mm over 15 d. Even when hair growth had slowed or halted subsequently, histology showed that these follicles retained an anagen-like morphology. By contrast, follicles isolated toward the end of the growing cycle produced thicker fibers for much shorter periods, after which growth ceased and the fibers lifted up from the base of the follicle. Internally, these specimens resembled their telogen counterparts in situ. Follicles isolated in mid-growth demonstrated intermediate fiber growth characteristics. In organ culture, mouse vibrissa follicles therefore closely reflect their in vivo origin in growth characteristics and cycle timing. These data provide new opportunities for studying hair growth cycle mechanisms in vitro, but present a caveat for quantitative studies because there may be a greater growth cycle-related variation than has previously been assumed.

PMID: 9077480, UI: 97232205


ar67
Exp Dermatol 1999 Aug;8(4):317-9 
In vitro maintenance of isolated hair follicles: current status and future development.
Philpott M

Department of Anatomy, St Bartholomew's and the Royal London School of Medicine and Dentistry.

ar68
Planta Med 1980;Suppl:84-90 
Bioassay of crude drugs for hair growth promoting activity in mice by a new simple method.
Tanaka S, Saito M, Tabata M

PMID: 7454882, UI: 81101527

ar69
Rev Med Liege 1997 Oct;52(10):671-4 
Hair follicles and hair growth cycles: recent considerations
Pierard-Franchimont C, Pierard GE

Service de Dermatopathologie, Universite de Liege.

ar70
Arch Klin Exp Dermatol 1966;227(1):390-409 
Histology and anatomy of the hair follicle in the course of the hair cycle

Bandmann HJ, Bosse K


ar71
Pharmacol Appl Skin Physiol 2000 May-Aug;13(3-4):133-42 
Several selective protein kinase C inhibitors including procyanidins promote hair growth.
Takahashi T, Kamimura A, Shirai A, Yokoo Y

Tsukuba Research Laboratories, Kyowa Hakko Kogyo Co., Tsukuba, Ibaraki, Japan. tomoya.takahashi@kyowa.co.jp
We have previously reported that procyanidin oligomers selectively promote growth of murine hair epithelial cells in vitro and stimulate anagen induction in vivo. We report here the possible relationship between the protein kinase C-inhibiting activity of procyanidins and their hair-growing activity. Of the procyanidins, procyanidin B-2 and procyanidin C-1, which selectively inhibit protein kinase C, intensively promote hair epithelial cell proliferation in vitro and stimulate anagen induction in vivo. On the other hand, procyanidins, which inhibit

both protein kinase C and A, showed relatively low activity in in vitro and in vivo evaluations. We also found that calphostin C, which is a selective inhibitor of protein kinase C, possesses hair epithelial cell growth-promoting activity in vitro and anagen phase-inducing hair-growing activity in vivo. Other selective protein kinase C inhibitors, such as hexadecylphosphocholine, palmitoyl-DL-carnitine chloride, and polymyxin B sulfate, also show marked anagen phase-inducing hair-growing activity in vivo. Nonselective protein kinase inhibitors, such as staurosporine and K252a, inhibit the growth of hair epithelial cells. 1,2-Dioctanoyl-sn-glycerol, a protein kinase C activator, dose-dependently decreases the growth of hair epithelial cells. Forskolin, an adenylate cyclase activator, promotes hair epithelial cell growth and boosts the growth-promoting effect of procyanidin B-2. It is speculated that the hair-growing activity of procyanidins is related to their protein kinase C-inhibiting activity. Copyright 2000 S. Karger AG, Basel

PMID: 10859531, UI: 20319197


ar72
Hepatogastroenterology 1994 Aug;41(4):343-8 
Role of free radicals in liver diseases and hepatic fibrosis.
Britton RS, Bacon BR

Department of Internal Medicine, St. Louis University Health Sciences Center, Missouri.
An increased production of free radicals in the liver has been implicated in a variety of liver diseases. Free radicals can damage cellular macromolecules and, therefore, may participate in hepatocellular injury when produced in excess. Strong evidence exists for hepatic free radical production in animal models of iron and copper overload, ethanol consumption, and ischemia-reperfusion. Although less is known about the situation in humans with liver diseases, the available evidence is consistent with the findings in animal experiments. Treatments that reduce free radical production and/or levels have protective effects in hepatic ischemia-reperfusion. Free radical-initiated lipid peroxidation may play a role in hepatic fibrogenesis, perhaps through an effect of aldehydic peroxidation products on Kupffer cells and lipocytes. This hypothesis is supported by the observation that dietary supplementation with vitamin E has a protective effect on carbon tetrachloride-induced hepatic fibrosis. While cellular damage in human liver diseases is probably multifactorial, free radicals may play important roles in initiating and/or perpetuating this damage.
ar73
Arch Biochem Biophys 1999 Sep 1;369(1):42-58 
A novel proanthocyanidin IH636 grape seed extract increases in vivo Bcl-XL expression and prevents acetaminophen-induced programmed and unprogrammed cell death in mouse liver.
Ray SD, Kumar MA, Bagchi D

Department of Pharmacology, Toxicology & Medicinal Chemistry, Arnold & Marie Schwartz College of Pharmacy and Health Sciences, Brooklyn, New York, 11201, USA.
Several molecular events in the apoptotic or necrotic death of hepatocytes induced by acetaminophen (AAP) now appear to be well defined. Recent studies also indicate that select expression of bcl-Xl is possibly modified during AAP-induced liver injury. The purpose of this study was several-fold: (i) to examine the hepatoprotective ability of short-term (3-day) and long-term (7-day) exposures of a grape seed proanthocyanidin extract (GSPE) on AAP-induced liver injury and animal lethality; (ii) to monitor effects of GSPE on one of the prime targets of AAP, i.e., hepatocellular genomic DNA and associated apoptotic and necrotic death; and (iii) to unravel changes in the pattern of expression of an antiapoptotic gene, bcl-Xl in the liver. In order to investigate these events, male ICR mice (30-40 g) were administered nontoxic doses of GSPE (3 or 7 days, 100 mg/kg, po), followed by hepatotoxic doses of AAP (400 and 500 mg/kg, ip), and sacrificed 24 h later. Serum was analyzed for alanine aminotransferase activity (ALT) and the liver for histopathological diagnosis of apoptosis/necrosis. The ability of AAP to promote apoptotic DNA fragmentation and its counteraction by GSPE in the liver was also evaluated quantitatively (by a sedimentation assay) and qualitatively (by agarose gel electrophoresis). Portions of livers were also subjected to Western blot analysis (27,000g fraction of liver homogenates) to examine the pattern of expression of cell death inhibitory gene bcl-Xl. Results indicate that 7-day GSPE preexposure induced dramatic protection and markedly decreased liver injury and animal lethality culminated by AAP, when compared to a short-term 3-day exposure. Abrogation of toxicity was also mirrored in DNA fragmentation. Histopathological evaluation of liver sections showed remarkable counteraction of AAP-toxicity by this novel GSPE and substantial inhibition of both apoptotic and necrotic liver cell death. Agarose gel electrophoresis revealed that 7-day GSPE preexposure prior to AAP administration completely blocked Ca(2+)/Mg(2+)-Ca(2+)/Mg(2+)-dependent-endonuclease-mediated ladder-like fragmentation of genomic DNA and significantly altered the bcl-Xl expression. The most dramatic changes observed in this study were: (i) substantial increase in the expression of bcl-Xl in the liver by 7-day GSPE exposure alone; (ii) significant modification bcl-Xl expression by AAP alone; and (iii) dramatic inhibition of AAP-induced modification of bcl-Xl (phosphorylation?) expression by GSPE. In summary, these observations demonstrate that GSPE preexposure may significantly attenuate AAP-induced hepatic DNA damage, apoptotic and necrotic cell death of liver cells, and, most remarkably, antagonize the influence of AAP-induced changes in bcl-Xl expression in vivo. Copyright 1999 Academic Press.


ar74
Vopr Med Khim 1995 Mar-Apr;41(2):20-3
"Effect of natural complexes of biologically active substances on liver regeneration in alcohol poisoning."
Kushnerova NF, Fomenko SE, Polozhentseva MI, Bulanov AE,. 
Hepatoprotective effect of natural substances obtained from extracts of grape combs, leaves of green and black tea were studied in liver tissue of ethanol-consuming rats by means of evaluation of the neutral lipid fractions and phospholipids as well as by measurement of glucose and nicotinamide coenzymes NAD+ and NADP in blood. In all the animal groups treated with these vegetable extracts content of total phospholipids, decreased after the ethanol treatment, was increased, while fraction composition of phospholipids and of neutral lipids was normalized; in blood content of glucose, NAD+ and NADP approached to control values. These results suggest that the vegetable extracts studied exhibited the hepatoprotective effect in alcohol intoxication.


ar77
Biol Pharm Bull 1995 Oct;18(10):1347-51
"Grape extract inhibits lipid peroxidation of human low density lipoprotein."
Lanningham-Foster L, Chen C, Chance DS, Loo G
Some epidemiological data have linked dietary polyphenols with lower risk of coronary heart disease. Polyphenols might impair lipoprotein oxidation which is believed to be an important step in initiating atherogenesis. The purpose of this study was to determine if grape extract known to contain polyphenolic substances can block copper-induced oxidative modification of human low density lipoprotein (LDL).LDL oxidation was monitored spectrophotometrically by measurement of change in absorbance at 234 nm. Incubation of LDL (0.05 mg protein/ml) with 1.66 microM cupric chloride produced a lag phase of 130 min before onset of the propagation phase where polyunsaturated fatty acids undergo conversion to conjugated lipid hydroperoxides. However, in the presence of grape extract at a final concentration equal to an 8000-fold dilution, the lag phase was extended to 185 min. A 4000-fold and 2000-fold dilution of grape extract produced lag phases of 250 and 465 min, respectively. LDL oxidation was essentially blocked for at least 10 h with a 1000-fold dilution of grape extract. In other experiments, incubation of LDL (0.2 mg protein/ml) with 5 microM cupric chloride for 1-4 h increased both thiobarbituric acid-reactive substances and electrophoretic mobility of LDL on agarose gel. In addition, there was loss of immunoreactivity of LDL with a murine monoclonal antibody against human apolipoprotein B-100. However, these oxidative changes to LDL by copper were prevented when diluted grape extract was present during incubation. It is concluded that grape extract contains antioxidants in the form of polyphenols with the capacity to inhibit oxidative modification of LDL.

(Use of these supplements should not take the place of treatment by a Doctor. They are not a replacement for antibiotics, and should be used under the supervision of a Doctor.

Information on this page is not intended to replace the care of a Doctor and should only be considered to be the opinion of the authors.)

Peleliu, Tribute to a Marine
(For You, Dad. Now I understand.)



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