Vaccinium myrtillus is a plant that belongs to the family of Ericaceae. Is a perennial bush between 30 to 60 cms tall which grows in the mountains and forests of Europe and the north of the United States . Its branches have alternating leaves, of elliptic shape and bright green color and its flowers, which appear between April and June, are pink and heart shaped. The fruit is dark blue and is harvested from July to September. The ripeness of the fruit depends on how high the plant is growing: plants that grow in higher lands usually last more in ripping that those that grow in lower lands.

Several actives principles have been isolated from the leaves and fruits of Bilberry, including antocyanoside flavonoids (whose genies are delphinine, cyanidine , malvidine , petunidine and peonidine ), heterosides of flavonoids ( hyperosid , quercitosid ), vitamins, sugars and pectins . Besides, quecetine , catechinic tannins (up to 10% in dried fruits) iridoids and acids ( 2).

1. Grieve M. A Modern Herbal. Vol.1. New York, NY: Dover Publications; 1971: 385-386.

2. Benigni R, Capra C, Cattorini PE. Plante Medicinali - Chimica Farmacologia E Terapia . Vol.II . Milano, Italia: Inverni della Beffa ; 1962:951-958.

3 . Baj A et al. Qualitative and quantitative evaluation of Vaccinium myrtillus anthocyanins by high-resolution gas chromatography and high-performance liquid chromatography. J Chromatography 1983 ;279:365 -372.

4. Morazzoni P, Bombardelli E. Vaccinium myrtillus I. Fitoterapia 1996 ;67:3 -29.

5. Salvayre R, Braquet P, Perruchot T. Douste-Blazy L. Flavonoids and Bioflavonoids 1981. Amsterdam - Oxford - New York : Elsevier Press; 1982:437-442.

6. Kuhnau J. The flavonoids . A class of semi-essential food components. Their role in human nutrition. Wld Rev Nutr Diet 1976 ;24:117 -191.

7. Harvsteen B. Flavonoids , a class of natural products of high pharmacological potency. Biochem Pharmacol 1983 ;32:1141 -1148.

A review has been presented of the biochemistry and pharmacology of a class of natural products, the flavonoids . These substances which are widely distributed in the plant kingdom and present in considerable quantities in common food products, spices and beverages have in a concentrated form ( Propolis ) been used since ancient times by physicians and laymen to treat a great variety of human diseases but they have yet to pass the tests of modern, controlled, clinical experimentation. An attempt has been made to present the fundamental evidence from the basic biological sciences which is required to stimulate the interest of the clinicians in this new field. The few existing reports on the careful pharmacodynamic , pharmacokinetic and clinical studies which have been made have been summarized to provide a basis for a full-scale investigation of the therapeutic potential of flavonoids .

8. Gabor M. Pharmacologic effects of flavonoids on blood vessels. Angiologica 1972 ;9:355 -374.

9. Mian E, Curri SB, Lietti A, Bombardelli E. Anthocyanosides and the walls of the microvessels : further aspects of the mechanism of action of their protective effect in syndromes due to abnormal capillary fragility. Minerva Med 1977:68:3565-3581.

10. Bottecchia D, et al. Preliminary reports on the inhibitory effect of Vaccinium myrtillus anthocyanosides on platelet aggregation and clot retraction. Fitoterapia 1987 ;48:3 -8.

11. Amella M, Bronner C, Briancon F, et al. Inhibition of mast cell histamine release by flavonoids and bioflavonoids . Planta Medica 1985 ;51:16 -20.

13. Vanaclocha B, Cañigueral S. ( eds .). Fitoterapia, Vademecum de Prescripción. 4ta edición. Barcelona: Masson, 2003.

14. Caselli L. Clinical and electroretinographic study on activity of anthocyanosides . Arch Med Int 1985 ;37:29 -35.

15. Wegmann R, Maeda K, Tronch P, Bastide P. Effects of anthocyanosides on photoreceptors. Cytoenzymatic aspects. Ann Histochim 1969 ;14:237 -256.

16. Jayle GE, Aubert L. Action des glucosides d'anthocyanes sur la vision scotopique et mesopique du sujet normal. Therapie 1964:19:171-185.

17. Terrasse J, Moinade S. Premiers resultats obtenus avec un nouveau facteur vitamininique P "les anthocyanosides " extraits du Vaccinium myrtillus . Presse Med 1964 ;72:397 -400.

18. Sala D, Rolando M, Rossi PL, Pissarello L. Effect of anthocyanosides on visual performances at low illumination. Minerva Oftalmol 1979 ;21:283 -285.

19. Gloria E, Peria A. Effect of anthocyanosides on the absolute visual threshold.
Ann Ottalmol Clin Ocul 1966 ;92:595 -607.

20. Junemann G. On the effect of anthocyanosides on hemeralopia following quinine poisoning. Klin Monatsbl Augenheilkd 1967 ;151:891 -896.

21. Pautler EL, Ennis SR. The effect of diet on inherited retinal dystrophy in the rat. Curr Eye Res 1984 ;3:1221 -1224.

22. Hess H, Knapka JJ, Newsome DA, et al. Dietary prevention of cataracts in the pink-eyed RCS rat. Lag Anim Sci 1985 ;35:47 -53.

23. Bravetti G. Preventive medical treatment of senile cataract with vitamin E and anthocyanosides : clinical evaluation. Ann Ottalmol Clin Ocul 1989 ;115:109 .

24 . Scharrer A, Ober M. Anthocyanosides in the treatment of retinopathies. Klin Monatabl Augenheilkd 1981 ;178:386 -389.

25 . Chaundry PS, Cambera J, Juliana HR, Varma SD. Inhibition of human lens aldose reductase by flavonoids , sulindac and indomethacin . Biochem Pharmacol 1983 ;32:1995 -1998.

The inhibition of human lens aldose reductase by flavonoids has been studied. Quercetin , the major pentahydroxyflavone , was observed to inhibit human lens aldose reductase by 50% at a concentration of 5 X 10(-6) M. The inhibitory activity of its 3-O-glucoside was similar to that of the parent aglycon . Glycosidation with L-sugar ( quercitrin and guaijaverin ), however, improved the inhibitory activity (the IC50 values being 1 X 10(-6) M and 2.5 X 10(-6) M respectively). The improvement in inhibitory activity with glycosidation with L-sugar was also apparent from the high inhibitory activity of myricitrin as compared to myricetin , although the improvement in this case of hexahydroxy flavone glycosidation was significantly less than in the case of penthahydroxy flavone glycosidation . The structure-activity relationship observed for human lens enzyme was similar to that reported previously for rat lens enzyme. Inhibitory activity on the whole however, was lower with human lens enzyme. Some known inhibitors of cyclo-oxygenase such as indomethacin , aspirin and sulindac also inhibited human lens aldose reductase . Thus, an inhibitor of one of the enzymes may actually inhibit both and, when administered, may exert mixed physiological effects.

26. Varma SD, Mizuno A, Kinoshita JH. Diabetic cataracts and flavonoids . Science 1977 ;195:87 -89.

27. Perossini M, et al. Diabetic and hypertensive retinopathy therapy with Vaccinium myrtillus anthocyanosides : Double-blind placebo controlled clinical trial. Ann Ottalmol Clin Ocul 1987 ;113:1173 .

28. Lietti A, Cristoni A, Picci M. Studies on Vaccinium myrtillus anthocyanosides . I. Vasoprotective and anti-inflammatory activity. Arzneimittelforschung . 1976 ;26 (5):829-32.

A Vaccinium myrtillus anthocyanosides preparation (equivalent to 25% of anthocyanidins ) demonstrated significant vasoprotective and antioedema properties in exerimental animals. In rabbits, the skin capillary permeability increase, due to chloroform, was reduced both after i.p . (25--100 mg/kg) and oral administration (200--400 mg/kg) of anthocyanosides . Their activity was more lasting in comparison to rutin or mepyramine and this did not seem to be due to a specific antagonism towards inflammatory process mediators such as histamine or bradykinin . Experiments carried out in rats demonstrated that Vacinium myrtillus anthocyanosides were effective both in skin capillary permeability test as well as on vascular resistance of rats fed a P factor deficient diet. In the former test effective doses were in the range of 25--100 mg/kg (by oral route). In both the animal species investigated, anthocyanosides were two-fold more active when compared to the flavonoid rutin . Vaccinium myrtillus anthocyanosides by oral route inhibited carrageein paw oedema in rats showing a dose-response relationship. An antioedema activity was detected also after i.v . or topical application.

29. Colantuoni A, Bertuglia S, Magistretti MJ, Donato L. Effects of Vaccinium Myrtillus anthocyanosides on arterial vasomotion . Arzneimittelforschung . 1991 Sep ;41 (9):905-9.

Consiglio Nazionale delle Ricerche , Institute of Clinical Physiology , Pisa , Italy .

The effects of Vaccinium Myrtillus anthocyanosides ( Myrtocyan , VMA; CAS 84082-34-8) on arteriolar vasomotion were assessed in cheek pouch microcirculation of anesthetized hamsters and in skeletal muscle microvasculature of unanesthetized hamster skin fold window preparation. Intravenously injected VMA induced vasomotion in cheek pouch arterioles and terminal arterioles with higher frequency in smaller vessels. In the skeletal muscle arteriolar networks VMA increased vasomotion frequency and amplitude in all vessel orders. The results indicate that VMA are effective in promoting and enhancing arteriolar rhythmic diameter changes, that play a role in the redistribution of microvascular blood flow and interstitial fluid formation.

30. Pennarola R, et al. The therapeutic action of the anthocyanosides in microcirculatory changes due to adhesiveinduced polyneuritis. Gazz Med Ital 1980 ;139:485 -491.

31. Ghiringhelli C, Gregoratti L, Marastoni F. Capillarotropic activity of anthocyanosides in high doses in phlebopathic stasis. Minerva Cardioangiol 1978 ;25:255 -276

33. Bailey CJ, Day C. Traditional plant medicines as treatments for diabetes . Diabetes Care 1989 ;12:553 -564

Department of Pharmaceutical Sciences, Aston University , Birmingham , United Kingdom .

35. Bever B, Zahnd G. Plants with oral hypoglycemic action. Quart J Crude Drug Res 1979 ;17:139 -196.

36. Mills S, Bone K. Materia medica . In: Mills S, Bone K, eds. Principles and Practice of Phytotherapy . Churchill Livingstone Publishing; 2000 ;297 -302.

37. Boniface R, Robert AM. Effect of anthocyanins on human connective tissue metabolism in humans. Klinische Monatsblatter furAugenheilkunde 1996 ;209:368 -372.

Labor fur Bindegewebsbiochemie , Medizinische Fakultat , Universitat Paris, Val de Marne.

38. Detre Z, Jellinek H, Miskulin M, Robert AM. Studies on vascular permeability in hypertension: action of anthocyanosides . Clin Physiol Biochem . 1986 ;4 (2):143-9.

The initial phase of renal hypertension induced by ligature of the abdominal aorta was accompanied by a transient increase in vascular permeability. This permeability increase has not the same intensity in all parts of the organism: it is greater in the skin and in the aorta wall than in the brain vessels. Treatment of rats with a flavonoid -type drug ( anthocyanosides of Vaccinium myrtillus ) for 12 days before the induction of hypertension kept the blood-brain barrier permeability normal and limited the increase in vascular permeability in the skin and the aorta wall. As previously demonstrated, the collagens of the blood vessel walls play an important role in the control of vascular permeability. Interaction of these collagens with the drug may be partly responsible for the protection against the permeability-increasing action of hypertension observed in the treated animals.

39. Rao CN, Rao VH, Steinman B. Influence of bioflavonoids on the collagen metabolism in rats with adjuvant induced arthritis. Ital J Biochem 1981 ;30:54 -62.

In rats with adjuvant induced arthritis, the effect of CA or HR on the metabolism of collagen in skin was studied. In untreated animals with adjuvant induced arthritis, the synthesis of collagen is decreased, the catabolism of newly formed collagen is intensified and the conversion of soluble collagen to insoluble collagen is impaired. Bioflavonoids were found to increase the collagen synthesis, accelerate the conversion of soluble collagen to insoluble collagen and inhibit the catabolism of soluble collagen. The beneficial action of bioflavonoids on the metabolism of collagen in adjuvant induced arthritis was discussed.

40. Colombo D, Vescovini R. Controlled trial of anthocyanosides from Vaccinium myrtillus in primary dysmenorrhea . G Ital Ost Ginecol 1985 ;7:1033 -1038l.

41. Puilleiro G, et al. Ex vivo study of the inhibitory effects of Vaccinium myrtillus anthocyanosides on human platelet aggregation. Fitoterapia 1989 ;60:69 -75.

42. Magistretti MJ, Conti M, Cristoni A. Antiulcer activity of an anthocyanidin from Vaccinium myrtillus . Arzneimittelforschung . 1988 May ;38 (5):686-90.

Research and Development Laboratories, Inverni della Beffa S.p.A ., Milan , Italy .

The antiulcer effects of 3 ,5,7 -trihydroxy-2-(3,4-dihydroxyphenyl)-1-benzopyrylium chloride ( IdB 1027) were assessed in various experimental models. Given orally, IdB 1027 antagonized gastric ulcerations induced by pylorus ligation , stress, nonsteroidal antiinflammatory drugs, ethanol, reserpine , histamine and duodenal ulceration induced by mercaptamine ( cysteamine ). Moreover it antagonized chronic gastric ulcers induced by acetic acid. Given intraperitoneally , it was more potent than after oral administration. IdB 1027 did not affect gastric secretion in pylorus- ligated rats and increased gastric mucus in normal animals both in the absence and in the presence of indometacin treatment. Tolerability was very good. These results indicate that IdB 1027 possesses a promising antiulcer activity, probably by potentiating the defensive barriers of the gastrointestinal mucosa.

43. Laplaud PM, Lelubre A, Chapman MJ. Antioxidant action of Vaccinium myrtillus extract on human low density lipoproteins in vitro: initial observations. Fundam Clin Pharmacol . 1997 ;11 (1):35-40.

Institut National de la Sante et de la Recherche Medicale ( Inserm ), Unite 321 Lipoproteines et Atherogenese , Hopital de la Pitie , Paris, France .

Oxidative modifications of low density lipoproteins (LDL) are now recognised as one of the major processes in atherogenesis . Various drugs, as well as a number of natural products, have been proposed to inhibit such processes. Among the naturally-occurring constituents of plants which appear to possess antioxidant activity are polyphenolic compounds such as flavonoids . The aqueous extract of Vaccinium myrtillus is rich in such molecules. In this report, we describe the in vitro antioxidative potential of this extract on human LDL. The copper-induced oxidative modification of these lipoproteins was assessed using 1) measurement of oxidative resistance as determined by the lag-phase preceding conjugated diene formation; 2) quantification of the amount of lipoperoxides and thiobarbituric acid-reactive substances generated, and measurement of the modification in the net negative electrical charge of the lipoproteins, over a 7-hour time course experiment. Trace amounts of V myrtillus extract (15 to 20 micrograms/ mL ) induce statistically significant changes in the oxidation behaviour of LDL, which include 1) prolongation of the lag-phase of conjugated diene production (P < 0.01); 2) reduction in the formation of lipoperoxides and of thiobarbituric acid-reactive substances up to 7 hours and especially between 1 and 5 hours (P < 0.01); and 3) inhibition of modification in the net negative charge of LDL. These results demonstrate that V myrtillus extract exerts potent protective action on LDL particles during in vitro copper-mediated oxidation. Calculation of IC50 values indicates that, on a molar basis, this extract may indeed be more potent than either ascorbic acid or butylated hydroxytoluene in the protection of LDL particles from oxidative stress.

44. Eandi M. Post-marketing investigation on Tegens " preparation with respect to side effects. 1987. Cited in Morazzoni P, Bombardelli E. Vaccinium myrtillus I. Fitoterapia 1996 ;67:3 -29

45. Politzer M. Experiences in the medical treatment of progressive myopia . Klin Monatsbl Augenheilkd . 1977 Oct ;171 (4):616-9.

In order to judge the effect of anthocyanosides and vitamine E ( Difrarel E) on refraction, visual acuity and eye- fundus , we treated 36 patients with this speciality in progressive myopia. After an observation period of 14.5 months an average increase of myopia by 0.53 dpt per eye was demonstrated. The final examination of 29 patients showed a stabilization of the fundus -alterations, as well as a stable, or an improved visual acuity respectively. In 7 patients a moderate deterioration of the partial or overall medical findings occured . Our observations allow the conclusion that Difrarel E achieves therapeutically valuable results in the treatment of progressive myopia.

46. Bertuglia S, Malandrino S, Colantuoni A. Effect of Vaccinium myrtillus anthocyanosides on ischaemia reperfusion injury in hamster cheek pouch microcirculation. Pharmacol Res. 1995 Mar-Apr ;31 (3-4):183-7.

CNR Institute of Clinical Physiology, Pisa , Italy .

The effects of Vaccinium myrtillus anthocyanosides (VMA) on ischaemia reperfusion injury were investigated in the hamster cheek pouch microcirculation. Ischaemia was induced by clamping the cheek pouch for 30 min followed by 30 min of reperfusion. The microvasculature was visualized by a fluorescence technique. VMA [10 mg ( 100 g body weight)-1] were orally administered for 2 and 4 weeks. The number of adhering leukocytes to venular vessel walls, the perfused capillary length, the increase in permeability, the arteriolar diameter changes were determined. Ischaemia and reperfusion were associated with increased number of leukocytes sticking to venules , decreased number of perfused capillaries, and increased permeability. VMA decreased the number of leukocytes sticking to the venular wall and preserved the capillary perfusion; the increase in permeability was significantly reduced after reperfusion. VMA saved the arteriolar tone and induced the appearance of rhythmic diameter changes of arterioles. These results demonstrate the ability of Vaccinium myrtillus anthocyanosides to reduce microvascular impairments due to ischaemia reperfusion injury, with preservation of endothelium, attenuation of leukocyte adhesion and improvement of capillary perfusion.

48: Sparrow JR, Vollmer- Snarr HR, Zhou J, Jang YP, Jockusch S, Itagaki Y, Nakanishi K. A2E-epoxides damage DNA in retinal pigment epithelial cells. Vitamin E and other antioxidants inhibit A2E-epoxide formation. J Biol Chem. 2003 May 16 ;278 (20):18207-13.

Department of Ophthalmology and Chemistry, Columbia University , New York , USA .

The autofluorescent pigments that accumulate in retinal pigment epithelial cells with aging and in some retinal disorders have been implicated in the etiology of macular degeneration. The major constituent is the fluorophore A2E, a pyridinium bisretinoid . Light-exposed A2E-laden retinal pigment epithelium exhibits a propensity for apoptosis with light in the blue region of the spectrum being most damaging. Efforts to understand the events precipitating the death of the cells have revealed that during irradiation (430 nm), A2E self-generates singlet oxygen with the singlet oxygen in turn reacting with A2E to generate epoxides at carbon-carbon double bonds. Here we demonstrate that A2E-epoxides, independent of singlet oxygen, exhibit reactivity toward DNA with oxidative base changes being at least one of these lesions. Mass spectrometry revealed that the antioxidants vitamins E and C, butylated hydroxytoluene , resveratrol , a trolox analogue (PNU-83836-E), and bilberry extract reduce A2E-epoxidation, whereas single cell gel electrophoresis and cell viability studies revealed a corresponding reduction in the incidence of DNA damage and cell death. Vitamin E, a lipophilic antioxidant, produced a more pronounced decrease in A2E-epoxidation than vitamin C, and treatment with both vitamins simultaneously did not confer additional benefit. Studies in which singlet oxygen was generated by endoperoxide in the presence of A2E revealed that vitamin E, butylated hydroxytoluene , resveratrol , the trolox analogue, and bilberry reduced A2E-epoxidation by quenching singlet oxygen. Conversely, vitamin C and ginkgolide B were not efficient quenchers of singlet oxygen under these conditions.

49: Hou DX. Potential mechanisms of cancer chemoprevention by anthocyanins. Curr Mol Med. 2003 Mar ;3 (2):149-59.

Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University , Korimoto , Kagoshima , Japan .

Anthocyanins are the chemical components that give the intense color to many fruits and vegetables, such as blueberries, red cabbages and purple sweet potatoes. Epidemiological investigations have indicated that the moderate consumption of anthocyanin products such as red wine or bilberry extract is associated with a lower risk of cardiovascular disease and improvement of visual functions. Recently, there is increasing interesting in the pharmaceutical function of anthocyanins . This review summarizes current knowledge on the various molecular evidences of cancer chemoprevention by anthocyanins . These mechanisms can be subdivided into the following aspects: 1) the antioxidation ; 2) the molecular mechanisms involved in anticarcinogenesis ; 3) the molecular mechanisms involved in the apoptosis induction of tumor cells. Finally, the bioavailability and structure-activity relationship of anthocyanins are also summarized.

50: Erlund I, Marniemi J, Hakala P, Alfthan G, Meririnne E, Aro A. Consumption of black currants, lingonberries and bilberries increases serum quercetin concentrations. Eur J Clin Nutr . 2003 Jan ;57 (1):37-42.

Biomarker Laboratory, National Public Health Institute, Helsinki , Finland .

OBJECTIVE: To study serum quercetin concentrations of subjects consuming berries or habitual Finnish diets. DESIGN: Randomized parallel dietary intervention. SUBJECTS: Forty healthy men (age 60 y). INTERVENTION: Twenty subjects consumed 100 g/day of berries (black currants, lingonberries and bilberries) for 8 weeks. Twenty subjects consuming their habitual diets served as controls. Fasting blood samples were obtained 2 weeks prior to the study, at baseline, and at 2, 4 and 8 weeks. Intake of quercetin was assessed from 3 day food records collected at baseline and at 8 weeks. RESULTS: The serum quercetin concentrations were significantly higher in the subjects consuming berries compared to the control group (P=0.039 ANCOVA with repeated measures). During the berry consumption period the mean serum concentrations of quercetin ranged between 21.4 and 25.3 micro g/l in the berry group, which was 32-51% higher compared with the control group. According to 3 day food records, there was no difference in quercetin intake at baseline, but at 8 weeks the intake was 12.31.4 mg/day ( means.e.m .) in the berry group and 5.80.6 mg/day in the control group (P=0.001). CONCLUSIONS: The results indicate that the berries used in this study are a good source of bioavailable quercetin .

51: Roy S, Khanna S, Alessio HM, Vider J, Bagchi D, Bagchi M, Sen CK. Anti- angiogenic property of edible berries. Free Radic Res. 2002 Sep ;36 (9):1023-31.

Laboratory of Molecular Medicine, Department of Surgery, 512 Heart and Lung Research Institute, Ohio State University Medical Center, 473 W. 12th Avenue, Columbus, OH 43210, USA.

Recent studies show that edible berries may have potent chemopreventive properties. Anti- angiogenic approaches to prevent and treat cancer represent a priority area in investigative tumor biology. Vascular endothelial growth factor (VEGF) plays a crucial role for the vascularization of tumors. The vasculature in adult skin remains normally quiescent. However, skin retains the capacity for brisk initiation of angiogenesis during inflammatory skin diseases such as psoriasis and skin cancers. We sought to test the effects of multiple berry extracts on inducible VEGF expression by human HaCaT keratinocytes . Six berry extracts (wild blueberry, bilberry, cranberry, elderberry, raspberry seed, and strawberry) and a grape seed proanthocyanidin extract (GSPE) were studied. The extracts and uptake of their constituents by HaCaT were studied using a multi-channel HPLC- CoulArray approach. Antioxidant activity of the extracts was determined by ORAC. Cranberry, elderberry and raspberry seed samples were observed to possess comparable ORAC values. The antioxidant capacity of these samples was significantly lower than that of the other samples studied. The ORAC values of strawberry powder and GSPE were higher than cranberry, elderberry or raspberry seed but significantly lower than the other samples studied. Wild bilberry and blueberry extracts possessed the highest ORAC values. Each of the berry samples studied significantly inhibited both H2O2 as well as TNF alpha induced VEGF expression by the human keratinocytes . This effect was not shared by other antioxidants such as alpha- tocopherol or GSPE but was commonly shared by pure flavonoids . Matrigel assay using human dermal microvascular endothelial cells showed that edible berries impair angiogenesis.

52: Seeram NP, Momin RA, Nair MG, Bourquin LD. Cyclooxygenase inhibitory and antioxidant cyanidin glycosides in cherries and berries. Phytomedicine . 2001 Sep ;8 (5):362-9.

Department of Horticulture and National Food Safety and Toxicology Center , Michigan State University, East Lansing 48824 , USA .

Anthocyanins from tart cherries, Prunus cerasus L. ( Rosaceae ) cv . Balaton and Montmorency; sweet cherries, Prunus avium L. ( Rosaceae ); bilberries, Vaccinum myrtillus L. ( Ericaceae ); blackberries, Rubus sp. ( Rosaceae ); blueberries var. Jersey, Vaccinium corymbosum L. ( Ericaceae ); cranberries var. Early Black, Vaccinium macrocarpon Ait . ( Ericaceae ); elderberries, Sambucus canadensis ( Caprifoliaceae ); raspberries, Rubus idaeus ( Rosaceae ); and strawberries var. Honeoye, Fragaria x ananassa Duch . ( Rosaceae ), were investigated for cyclooxygenase inhibitory and antioxidant activities. The presence and levels of cyanidin-3-glucosylrutinoside 1 and cyanidin-3-rutinoside 2 were determined in the fruits using HPLC. The antioxidant activity of anthocyanins from cherries was comparable to the commercial antioxidants, tert-butylhydroquinone , butylated hydroxytoluene and butylated hydroxyanisole , and superior to vitamin E, at a test concentration of 125 microg /ml. Anthocyanins from raspberries and sweet cherries demonstrated 45% and 47% cyclooxygenase -I and cyclooxygenase -II inhibitory activities, respectively, when assayed at 125 microg /ml. The cyclooxygenase inhibitory activities of anthocyanins from these fruits were comparable to those of ibuprofen and naproxen at 10 microM concentrations. Anthocyanins 1 and 2 are present in both cherries and raspberry. The yields of pure anthocyanins 1 and 2 in 100 g Balaton and Montmorency tart cherries, sweet cherries and raspberries were 21, 16.5; 11, 5; 4.95, 21; and 4.65, 13.5 mg, respectively. Fresh blackberries and strawberries contained only anthocyanin 2 in yields of 24 and 22.5 mg/100 g, respectively. Anthocyanins 1 and 2 were not found in bilberries, blueberries, cranberries or elderberries.

53: Nyman NA, Kumpulainen JT. Determination of anthocyanidins in berries and red wine by high-performance liquid chromatography. J Agric Food Chem. 2001 Sep ;49 (9):4183-7.

Food Research, L-Building, MTT Agrifood Research Finland , FIN-31600 Jokioinen , Finland .

A high-performance liquid chromatographic (HPLC) method for the determination of anthocyanidins from berries and red wine is described. Delphinidin , cyanidin , petunidin , pelargonidin , peonidin , and malvidin contents of bilberry ( Vaccinium myrtillus ), black currant ( Ribes nigrum ), strawberry ( Fragaria ananassa cv . Jonsok ), and a Cabernet sauvignon ( Vitis vinifera ) red wine were determined. The aglycon forms of the anthocyanins present in the samples were revealed by acid hydrolysis. A reversed phase analytical column was employed to separate the anthocyanidins before identification by diode array detection. The suitability of the method was tested by determining the recovery (95-102% as aglycons and 69-104% from glycosides) for each anthocyanidin . Method repeatability was tested by charting the total aglycon content of two samples over a period of 14 analyses and determining the coefficients of variation (1.41% for bilberry and 2.56% for in-house reference material). The method developed proved thus to be effective for reliable determination of anthocyanidins from freeze-dried berry samples and red wine. The total anthocyanidin content of the tested samples was as follows: in-house reference material, 447 8 mg/100 g; strawberry, 23.8 0.4 mg/100 g; black currant, 135 3 mg/100 g; bilberry, 360 3 mg/100 g; and Cabernet sauvignon red wine, 26.1 0.1 mg/100 mL .

54: Kahkonen MP, Hopia AI, Heinonen M. Berry phenolics and their antioxidant activity. J Agric Food Chem. 2001 Aug ;49 (8):4076-82.

Department of Applied Chemistry and Microbiology, Division of Food Chemistry, P.O. Box 27 , FIN-00014 University of Helsinki , Finland .

Phenolic profiles of a total of 26 berry samples, together with 2 apple samples, were analyzed without hydrolysis of glycosides with HPLC. The phenolic contents among different berry genera varied considerably. Anthocyanins were the main phenolic constituents in bilberry, bog-whortleberry, and cranberry, but in cowberries, belonging also to the family Ericaceae genus Vaccinium , flavanols and procyanidins predominated. In the family Rosaceae genus Rubus (cloudberry and red raspberry), the main phenolics found were ellagitannins , and in genus Fragaria (strawberry), ellagitannins were the second largest group after anthocyanins . However, phenolic acids were dominant in rowanberries (genus Sorbus ) and anthocyanins in chokeberry (genus Aronia ). In the family Grossulariaceae genus Ribes (currants and gooseberry), anthocyanins predominated, as well as in crowberries (family Empetraceae genus Empetrum ). In apples, hydroxycinnamic acids were the main phenolic subgroup. Extraction methods for berries and apples were studied to produce phenolic extracts with high antioxidant activity. Evaluation of antioxidant activity was performed by autoxidazing methyl linoleate (40 degrees C, in the dark). The extraction method affected remarkably both the phenolic composition and the antioxidant activity, but with statistical analysis the observed activity could not be well explained with the contents of individual phenolic subgroups.

55: Dugo P, Mondello L, Errante G, Zappia G, Dugo G. Identification of anthocyanins in berries by narrow-bore high-performance liquid chromatography with electrospray ionization detection. J Agric Food Chem. 2001 Aug ;49 (8):3987-92.

Dipartimento di Chimica Organica e Biologica Facolta di Scienze MM.FF.NN., Universita di Messina, Salita Sperone , 98166 - Messina , Italy .

Qualitative determination of anthocyanins in extracts of red fruits by narrow-bore HPLC/ESI-MS was carried out. This method was used to investigate anthocyanin contents of black bilberry ( Vaccinium myrtillus L.), blackberry ( Rubus sp.), and mulberry ( Morus nigra ). An ultraviolet diode array and a mass spectrometer with ESI source were used for detection. Anthocyanin identifications were made by using retention time data and UV- vis and mass spectra and comparing them with those of commercially available standard compounds. The method allowed the identification of fourteen anthocyanins in black bilberry extract, six anthocyanins in blackberry extract, and five anthocyanins in mulberry extract.

56: Head KA. Natural therapies for ocular disorders, part two: cataracts and glaucoma. Altern Med Rev. 2001 Apr ;6 (2):141-66.

Thorne Research, Inc., P.O. Box 25, Dover, ID 83825 ,US A.

Pathophysiological mechanisms of cataract formation include deficient glutathione levels contributing to a faulty antioxidant defense system within the lens of the eye. Nutrients to increase glutathione levels and activity include lipoic acid, vitamins E and C, and selenium. Cataract patients also tend to be deficient in vitamin A and the carotenes, lutein and zeaxanthin . The B vitamin riboflavin appears to play an essential role as a precursor to flavin adenine dinucleotide (FAD), a co-factor for glutathione reductase activity. Other nutrients and botanicals, which may benefit cataract patients or help prevent cataracts, include pantethine , folic acid, melatonin, and bilberry. Diabetic cataracts are caused by an elevation of polyols within the lens of the eye catalyzed by the enzyme aldose reductase . Flavonoids , particularly quercetin and its derivatives, are potent inhibitors of aldose reductase . Glaucoma is characterized by increased intraocular pressure (IOP) in some but not all cases. Some patients with glaucoma have normal IOP but poor circulation, resulting in damage to the optic nerve. Faulty glycosaminoglycan (GAG) synthesis or breakdown in the trabecular meshwork associated with aqueous outflow has also been implicated. Similar to patients with cataracts, those with glaucoma typically have compromised antioxidant defense systems as well. Nutrients that can impact GAGs such as vitamin C and glucosamine sulfate may hold promise for glaucoma treatment. Vitamin C in high doses has been found to lower IOP via its osmotic effect. Other nutrients holding some potential benefit for glaucoma include lipoic acid, vitamin B12, magnesium, and melatonin. Botanicals may offer some therapeutic potential. Ginkgo biloba increases circulation to the optic nerve; forskolin (an extract from Coleus forskohlii ) has been used successfully as a topical agent to lower IOP; and intramuscular injections of Salvia miltiorrhiza have shown benefit in improving visual acuity and peripheral vision in people with glaucoma.

57: Marniemi J, Hakala P, Maki J, Ahotupa M. Partial resistance of low density lipoprotein to oxidation in vivo after increased intake of berries. Nutr Metab Cardiovasc Dis. 2000 Dec ;10 (6):331-7.

Research and Development Centre, Social Insurance Institution, University of Turku , Finland .

BACKGROUND AND AIM: The health-promoting effects of fruit- and vegetable-based diets are known to be associated with their antioxidative components. We found in our preliminary in vitro laboratory tests that extracts of many common Finnish edible berries are potent scavengers of peroxyl radicals and inhibitors of lipid peroxidation . We therefore designed the current study to evaluate both the long-term (8 weeks) and short-term (5 hours) effects of increased intake of three berries on antioxidant potential and lipid peroxidation . METHODS AND RESULTS: Healthy 60-year-old men were randomized to berry, supplement and control groups (20 men in each group). The berry group ate, in addition to their normal diet, a 100 g portion of deep-frozen berries (bilberries, lingonberries , or black currants) daily for 8 weeks. The other groups ingested daily 100 mg of alpha- tocopherol and 500 mg of ascorbic acid (supplement group) or 500 mg of calcium gluconate (control group). In the short-term experiment 6 men ate 80 g of each of the three berries in one go. Serum ascorbate concentrations increased significantly in both the berry and the supplement group. Serum alpha- tocopherol levels and the antioxidant potential (TRAP) in low density lipoprotein (LDL) increased in the supplement group only. In the berry group, slightly lowered LDL diene conjugation (p = 0.074) and slightly increased total serum TRAP (p = 0.084) values were observed. No changes were found in these measures in the supplement or the control group. In the short-term experiment, LDL TRAP showed a small increase (about 10%, p = 0.039) during five hours after the intake of 240 g berries. CONCLUSIONS: The effects of consumption of berries on antioxidant potential and diene conjugation in LDL particles in vivo appear to be small.

58: Cohen- Boulakia F, Valensi PE, Boulahdour H, Lestrade R, Dufour-Lamartinie JF, Hort-Legrand C, Behar A. In vivo sequential study of skeletal muscle capillary permeability in diabetic rats: effect of anthocyanosides . Metabolism. 2000 Jul ;49 (7):880-5.

Laboratory of Biophysics, Paris 6 University, France .

Alterations in the capillary filtration of macromolecules are well documented in diabetic patients and experimental diabetes. Various flavonoids including anthocyanosides and ginkgo biloba extracts have been shown to be effective against experimentally induced capillary hyperfiltration . The aim of the present study was to test the effects of anthocyanosides on capillary filtration in diabetic rats. For this purpose, we have validated the use of our previously described in vivo method for measurement of the capillary filtration of albumin (CFA) in rats. Male Wistar rats with streptozotocin (STZ)-induced diabetes were randomized in 3 groups to receive either ginkgo biloba (group A), Vaccinium myrtillus (group B), or no treatment (group C). The isotopic test of CFA consisted of intravenously injecting 99mtechnetium-labeled albumin, inducing venous compression on a hindquarter, and measuring radioactivity externally on the limb before, during, and after removal of venous compression. After removal of the tourniquet, the radioactivity curve decreased. Interstitial albumin retention (AR) and the ratio of the amplitudes of the low- and high-frequency peaks (LF/HF ratio), an index of lymphatic function obtained by the fast Fourier transform of the last part of the radioactivity curve, were calculated. In STZ-treated animals, the isotopic test was performed at a mean age of 97 days (time 1) and after 6 weeks (time 2) and 12 weeks (time 3) of treatment, ie , 6 and 12 weeks after time 1. At time 1, AR was significantly higher in the 3 diabetic groups than in the control rats, without a significant difference between these groups. In group B, AR decreased significantly (P = .015) at times 2 and 3. In group C, AR increased significantly (P < .0005) from time 1 to time 3. In group A, AR increased slightly (NS) between time 1 and time 3. In groups A and C, the LF/HF ratio significantly increased with time (P < .0005) and the levels at time 3 were significantly higher versus control rats (P < .0001). In group B, the LF/HF ratio remained unchanged from time 1 to time 3 and similar to the values found in the control rats. In conclusion, these data show that (1) this new in vivo noninvasive method can be used to study CFA in skeletal muscle in diabetic rats, (2) it is reproducible and may be repeated over several months to evaluate spontaneous microcirculatory changes, and (3) anthocyanosides appear to be effective in preventing the increase in CFA and the failure of lymphatic uptake of interstitial albumin in diabetic animals.

59: Cignarella A, Nastasi M, Cavalli E, Puglisi L. Novel lipid-lowering properties of Vaccinium myrtillus L. leaves, a traditional antidiabetic treatment, in several models of rat dyslipidaemia : a comparison with ciprofibrate . Thromb Res. 1996 Dec 1 ;84 (5):311-22.

Institute of Pharmacological Sciences, University of Milano , Italy .

Vaccinium myrtillus L. (blueberry) leaf infusions are traditionally used as a folk medicine treatment of diabetes. To further define this therapeutical action, a dried hydroalcoholic extract of the leaf was administered orally to streptozotocin -diabetic rats for 4 days. Plasma glucose levels were consistently found to drop by about 26% at two different stages of diabetes. Unexpectedly, plasma triglyceride (TG) were also decreased by 39% following treatment. Subsequent to the latter observation, possible lipid-lowering properties of the extract were investigated on other models of hyperlipidaemia and ciprofibrate , a well-established hypolipidaemic drug, was used as a reference compound. Both drug reduced TG levels of rats on hyperlipidaemic diet in a dose-dependent fashion. When administered at single doses over the same experimental period, blueberry and ciprofibrate were effective in lowering TG concentrations in ethanol-treated normolipidaemic animals and in genetically hyperlipidaemic Yoshida rats. Unlike ciprofibrate , however, blueberry failed to prevent the rise in plasma TG elicited by fructose and did not affect free fatty acid levels in any of the above experimental conditions. In rats treated with Triton WR-1339, blueberry feeding induced an hypolipidaemic activity one hour after injection but proved to be ineffective at later time points, thus suggesting that its hypolipidaemic action may reflect improved TG-rich lipoprotein catabolism. In addition, ciprofibrate and the extract were tested for antithrombotic activity using a collagen-triggered model of venous thrombosis in diabetic and Yoshida rats. Only ciprofibrate , however, significantly reduced thrombus formation in diabetics, possibly because of its effects on free fatty acid metabolism, whereas no effect was observed in Yoshida rats. In conclusion, the present findings indicate that active consituent (s) of Vaccinium myrtillus L. leaves may prove potentially useful for treatment of dyslipidaemiae associated with impaired TG-rich lipoprotein clearance.