Antioxidant Flavonoids: Structure, Function and Clinical Usage

Alan L. Miller, ND Abstract

Flavonoids occur in most plant species, and account for a significant percentage of the chemical constituents of some; e.g. dried green tea leaves contain approximately 30% flavonoids by weight. Flavonoids have been shown to have antibacterial, anti-inflammatory, antiallergic, antimutagenic, antiviral, antineoplastic, anti-thrombotic, and vasodilatory activity. The potent antioxidant activity of flavonoidstheir ability to scavenge hydroxyl radicals, superoxide anions, and lipid peroxy radicalsmay be the most important function of flavonoids, and underlies many of the above actions in the body. Oxidative damage is implicated in most disease processes, and epidemiological, clinical, and laboratory research on flavonoids and other antioxidants suggest their use in the prevention and treatment of a number of these. Catechin and its derivatives, oligomeric proanthocyanidins, quercetin and quercetin chalcone, Ginkgo flavone glycosides, silymarin, and others can be utilized in preventative and treatment protocols for cardiovascular disease, cancer, inflammatory conditions, asthma, periodontal disease, liver disease, cataracts and macular degeneration.
Source: Alternative Medicine Review 1996 (Jul); 1 (2): 103111 http://www.chiro.org/nutrition/ABSTRACTS/Antioxidant_Flavonoids.shtml

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Pure dietary flavonoids quercetin and ()epicatechin augment nitric oxide products and reduce endothelin-1 acutely in healthy men
Wai Mun Loke, Jonathan M Hodgson, Julie M Proudfoot, Allan J McKinley, Ian B Puddey and Kevin D Croft Dietary flavonoids may improve endothelial function and ultimately lead to beneficial cardiovascular effects. The objective was to assess whether pure dietary flavonoids can modulate nitric oxide and endothelin-1 production and thereby improve endothelial function. A randomized, placebo-controlled, crossover trial in 12 healthy men was conducted to compare the acute effects of the oral administration of 200 mg quercetin, ()-epicatechin, or epigallocatechin gallate on nitric oxide, endothelin-1, and oxidative stress after nitric oxide production was assessed via the measurement of plasma Snitrosothiols and plasma and urinary nitrite and nitrate concentrations. The effects on oxidative stress were assessed by measuring plasma and urinary F2-isoprostanes. Plasma and urinary concentrations of quercetin, ()-epicatechin, and epigallocatechin gallate were measured to establish the absorption of these flavonoids. Relative to water (control), quercetin and ()-epicatechin resulted in a significant increase in plasma Snitrosothiols, plasma nitrite, and urinary nitrate concentrations (P < 0.05), but not in plasma nitrate or urinary nitrite. Epigallocatechin gallate did not alter any of the measures of nitric oxide production. Quercetin and ()-epicatechin resulted in a significant reduction in plasma endothelin-1 concentration (P < 0.05), but only quercetin significantly decreased the urinary endothelin-1 concentration. None of the 3 treatments significantly changed plasma or urinary F2-isoprostane concentrations. Significant increases in the circulating concentrations of the 3 flavonoids were observed (P < 0.05) after the corresponding treatment. Dietary flavonoids, such as quercetin and ()-epicatechin, can augment nitric oxide status and reduce endothelin-1 concentrations and may thereby improve endothelial function.

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Flavonoids From Argentine Tagetes (Asteraceae) With Antimicrobial Activity

Mara L. Tereschuk, Mario D. Baigor, Lucia I. C. de Figueroa, Lidia R. Abdala
ABSTRACT The flavonoids, constituting one of the most numerous and widespread groups of natural plant constituents, are important to humans not only because they contribute to plant colors but also because many members are physiologically active. These lowmolecular-weight substances, found in all vascular plants, are phenylbenzopyrones. Over 4000 structures have been identified in plant sources, and they are categorized into several groups Primarily recognized as pigments responsible for the autumnal burst of hues and the many shades of yellow, orange, and red in flowers and food, the flavonoids are found in fruits, vegetables, nuts, seeds, stems, flowers, and leaves as well as tea and wine and are important constituents of the human diet . They are prominent components of citrus fruits and other food sources. Flavonols (quercetin, myricetin, and kaempferol) and flavones (apigenin and luteolin) are the most common phenolics in plantbased foods. Quercetin is also a predominant component of onions, apples, and berries. Such flavanones as naringin are typically present in citrus fruit, and flavanols, particularly catechin, are present as catechin gallate in such beverages as green or black tea and wine. Some major sources of flavonoids are outlined in Table 1 (2). The daily intake of flavonoids in humans has been estimated to be approx 25 mg/d, a quantity that could provide pharmacologically significant concentrations in body fluids and tissues, assuming good absorption from the gastrointestinal tract (3).

Book Title: Public Health Microbiology: Methods and Protocols Series: Methods in Molecular Biology | Volume: 268 | Pub. Date: May-20-2004 | Page Range: 317-330 | DOI: 10.1385/1-59259-766-1:317

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Flavonoid-Rich Dark Chocolate Improves Endothelial Function and Increases Plasma Epicatechin Concentrations in Healthy Adults
Mary B. Engler, PhD, Marguerite M. Engler, PhD, Chung Y. Chen, PhD, Mary J. Malloy, MD, Amanda Browne, BS, Elisa Y. Chiu, BS, MS, Ho-Kyung Kwak, PhD, Paul Milbury, MS, Steven M. Paul, PhD, Jeffrey Blumberg, PhD, FACN and Michele L. Mietus-Snyder, MD Dark chocolate derived from the plant (Theobroma cacao) is a rich source of flavonoids. Cardioprotective effects including antioxidant properties, inhibition of platelet activity, and activation of endothelial nitric oxide synthase have been ascribed to the cocoa flavonoids. To investigate the effects of flavonoid-rich dark chocolate on endothelial function, measures of oxidative stress, blood lipids, and blood pressure in healthy adult subjects. The study was a randomized, double-blind, placebo-controlled design conducted over a 2 week period in 21 healthy adult subjects. Subjects were randomly assigned to daily intake of high-flavonoid (213 mg procyanidins, 46 mg epicatechin) or low-flavonoid dark chocolate bars (46 g, 1.6 oz). High-flavonoid chocolate consumption improved endothelium-dependent flow-mediated dilation (FMD) of the brachial artery (mean change = 1.3 0.7%) as compared to low-flavonoid chocolate consumption (mean change = 0.96 0.5%) (p = 0.024). No significant differences were noted in the resistance to LDL oxidation, total antioxidant capacity, 8-isoprostanes, blood pressure, lipid parameters, body weight or body mass index (BMI) between the two groups. Plasma epicatechin concentrations were markedly increased at 2 weeks in the highflavonoid group (204.4 18.5 nmol/L, p 0.001) but not in the low-flavonoid group (17.5 9 nmol/L, p = 0.99). Flavonoid-rich dark chocolate improves endothelial function and is associated with an increase in plasma epicatechin concentrations in healthy adults. No changes in oxidative stress measures, lipid profiles, blood pressure, body weight or BMI were seen.

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Flavonoids activate pregnane X receptor-mediated CYP3A4 gene expression by inhibiting cyclindependent kinases in HepG2 liver carcinoma cells
Hanqing Dong, Wenwei Lin, Jing Wu, Taosheng Chen

The expression of the drug-metabolizing enzyme cytochrome P450 3A4 (CYP3A4) is regulated by the pregnane X receptor (PXR), which is modulated by numerous signaling pathways, including the cyclin-dependent kinase (Cdk) pathway. Flavonoids, commonly consumed by humans as dietary constituents, have been shown to modulate various signaling pathways (e.g., inhibiting Cdks). Flavonoids have also been shown to induce CYPs expression, but the underlying mechanism of action is unknown. Here, we report the mechanism responsible for flavonoid-mediated PXR activation and CYP expression. In a cell-based screen designed to identify compounds that activate PXR-mediated CYP3A4 gene expression in HepG2 human carcinoma cells, we identified several flavonoids, such as luteolin and apigenin, as PXR activators. The flavonoids did not directly bind to PXR, suggesting that an alternative mechanism may be responsible for flavonoid-mediated PXR activation. Consistent with the Cdk5-inhibitory effect of flavonoids, Cdk5 and p35 (a non-cyclin regulatory subunit required to activate Cdk5) were expressed in HepG2. The activation of Cdk5 attenuated PXR-mediated CYP3A4 expression whereas its downregulation enhanced it. The Cdk5-mediated downregulation of CYP3A4 promoter activity was restored by flavonoids, suggesting that flavonoids activate PXR by inactivating Cdk5. In vitro kinase assays showed that Cdk5 directly phosphorylates PXR. The Cdk kinase profiling assay showed that apigenin inhibits multiple Cdks, suggesting that several Cdks may be involved in activation of PXR by flavonoids. Our results for the first time link the stimulatory effect of flavonoids on CYP expression to their inhibitory effect on Cdks, through a PXR-mediated mechanism. These results may have important implications on the pharmacokinetics of drugs coadministered with herbal remedy and herbal-drug interactions. BMC Biochemistry 2010, 11:23doi:10.1186/1471-2091-11-23 Published: 16 June 2010

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Antioxidant activity of dietary fruits, vegetables, and commercial frozen fruit pulps.
Hassimotto NM, Genovese MI, Lajolo FM. ABSTRACT Fruits, vegetables, and commercial frozen pulps (FP) consumed in the Brazilian diet were analyzed for antioxidant activities using two different methods, one that determines the inhibition of copper-induced peroxidation of liposome and another based on the inhibition of the co-oxidation of linoleic acid and beta-carotene. The anthocyanin-rich samples showed the highest, concentration-dependent, antioxidant activities in both systems. In the liposome system, at both 10 and 50 microM gallic acid equivalent (GAE) addition levels, the neutral and acidic flavonoids of red cabbage, red lettuce, black bean, mulberry, Gala apple peel, jambolao, acai FP, mulberry FP, and the acidic flavonoids of acerola FP showed the highest antioxidant activities (>85% inhibition). In the beta-carotene bleaching system, the samples cited above plus red guava gave inhibition values >70%. On the other hand, some samples showed prooxidant activity in the liposome system coincident with a low antioxidant activity in the beta-carotene system. There was no relationship between total phenolics content, vitamin C, and antioxidant activity, suggesting that the antioxidant activity is a result of a combination of different compounds having synergic and antagonistic effects. PMID: 15826041 PubMed

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Mammalian Cells: Implications for Inflammation, Heart Disease, and Cancer

Elliott Middleton, Jr., Chithan Kandaswami and Theoharis C. Theoharides ABSTRACT Flavonoids are nearly ubiquitous in plants and are recognized as the pigments responsible for the colors of leaves, especially in autumn. They are rich in seeds, citrus fruits, olive oil, tea, and red wine. They are low molecular weight compounds composed of a three-ring structure with various substitutions. This basic structure is shared by tocopherols (vitamin E). Flavonoids can be subdivided according to the presence of an oxy group at position 4, a double bond between carbon atoms 2 and 3, or a hydroxyl group in position 3 of the C (middle) ring. These characteristics appear to also be required for best activity, especially antioxidant and antiproliferative, in the systems studied. The particular hydroxylation pattern of the B ring of the flavonoles increases their activities, especially in inhibition of mast cell secretion. Certain plants and spices containing flavonoids have been used for thousands of years in traditional Eastern medicine. In spite of the voluminous literature available, however, Western medicine has not yet used flavonoids therapeutically, even though their safety record is exceptional. Suggestions are made where such possibilities may be worth pursuing.

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Flavonoid intake and coronary mortality in Finland: a cohort study

Paul Knekt, Ritva Jarvinen, Antti Reunanen, , Jouni Maatela ABSTRACT

Objective: To study the association between dietary intake of flavonoids and subsequent coronary mortality. Design: A cohort study based on data collected at the Finnish mobile clinic health examination survey from 1967-72 and followed up until 1992. Settings: 30 communities from different parts of Finland. Subjects: 5133 Finnish men and women aged 30-69 years and free from heart disease at baseline. Main outcome measure: Dietary intake of flavonoids, total mortality, and coronary mortality. Results: In women a significant inverse gradient was observed between dietary intake of flavonoids and total and coronary mortality. The relative risks between highest and lowest quarters of flavonoid intake adjusted for age, smoking, serum cholesterol concentration, blood pressure, and body mass index were 0.69 (95% confidence interval 0.53 to 0.90) and 0.54 (0.33 to 0.87) for total and coronary mortality, respectively. The corresponding values for men were 0.76 (0.63 to 0.93) and 0.78 (0.56 to 1.08), respectively. Adjustment for intake of antioxidant vitamins and fatty acids weakened the associations for women; the relative risks for coronary heart disease were 0.73 (0.41 to 1.32) and 0.67 (0.44 to 1.00) in women and men, respectively. Intakes of onions and apples, the main dietary sources of flavonoids, presented similar associations. The relative risks for coronary mortality between highest and lowest quarters of apple intake were 0.57 (0.36 to 0.91) and 0.81 (0.61 to 1.09) for women and men, respectively. The corresponding values for onions were 0.50 (0.30 to 0.82) and 0.74 (0.53 to 1.02), respectively. Conclusion: The results suggest that people with very low intakes of flavonoids have higher risks of coronary disease.

Dietary Flavonoids, Antioxidant Vitamins, and Incidence of Stroke

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Sirving O. Keli, MD, PhD; Michael G. L. Hertog, PhD; Edith J. M. Feskens, PhD; Daan Kromhout, PhD, MPH Background Epidemiological studies suggested that consumption of fruit and vegetables may protect against stroke. The hypothesis that dietary antioxidant vitamins and flavonoids account for this observation is investigated in a prospective study. Methods A cohort of 552 men aged 50 to 69 years was examined in 1970 and followed up for 15 years. Mean nutrient and food intake was calculated from crosscheck dietary histories taken in 1960, 1965, and 1970. The association between antioxidants, selected foods, and stroke incidence was assessed by Cox proportional hazards regression analysis. Adjustment was made for confounding by age, systolic blood pressure, serum cholesterol, cigarette smoking, energy intake, and consumption of fish and alcohol. Results Forty-two cases of first fatal or nonfatal stroke were documented. Dietary flavonoids (mainly quercetin) were inversely associated with stroke incidence after adjustment for potential confounders, including antioxidant vitamins. The relative risk (RR) of the highest vs the lowest quartile of flavonoid intake ( 28.6 mg/d vs <18.3 mg/d) was 0.27 (95% confidence interval [CI], 0.11 to 0.70). A lower stroke risk was also observed for the highest quartile of -carotene intake (RR, 0.54; 95% CI, 0.22 to 1.33). The intake of vitamin C and vitamin E was not associated with stroke risk. Black tea contributed about 70% to flavonoid intake. The RR for a daily consumption of 4.7 cups or more of tea vs less than 2.6 cups of tea was 0.31 (95% CI, 0.12 to 0.84). Conclusions The habitual intake of flavonoids and their major source (tea) may protect against stroke.
(Arch Intern Med. 1996;154:637-642)

Effect of Acute and Chronic Tea Consumption on Platelet Aggregation in Patients With Coronary Artery Disease
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Stephen J. Duffy; Joseph A. Vita; Monika Holbrook; Peter L. Swerdloff; John F. Keaney, Jr ABSTRACT Epidemiological studies suggest that tea consumption is associated with a decreased risk of cardiovascular events, but the mechanisms of benefit remain undefined. Platelet aggregation is a precipitating event in cardiovascular disease, and tea contains antioxidant flavonoids that are known to decrease platelet aggregation in vitro. To test the effect of tea consumption on platelet aggregation, we randomized 49 patients with coronary artery disease to either 450 mL of black tea or water consumed initially, followed by 900 mL of tea or water daily for 4 weeks in a crossover design. Ex vivo platelet aggregation in platelet-rich plasma was assessed in response to ADP and thrombin receptoractivating peptide at baseline and 2 hours and 4 weeks after beverage consumption. We observed dose-dependent platelet aggregation in response to each agonist, and neither relation was altered by acute or chronic tea consumption. Plasma flavonoids increased with acute and chronic tea consumption, indicating adequate absorption of tea flavonoids. In conclusion, these results demonstrate that acute and chronic black tea consumption does not affect ex vivo platelet aggregation in patients with coronary artery disease. These findings suggest that an effect of tea flavonoids on platelet aggregation is unlikely to be the explanation for the reduction in risk of cardiovascular events noted in epidemiological studies.

A Review of Potential Health Benefits of Flavonoids

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Jay M. Patel

ABSTRACT
Flavonoids occur naturally in fruit, vegetables, and beverages such as tea and wine and over 4000 structurally unique flavonoids have been identified in plant sources. Plants and spices containing flavonoids have historically been used in traditional eastern medicine. Research in the field of flavonoids has increased since the discovery of the French paradox i.e. the low cardiovascular mortality rate observed in Mediterranean populations in association with red wine consumption and a high saturated fat intake. The aim of this review is to obtain a further understanding on the reported beneficial health effects of these substances and discuss the action of flavonoids as antioxidants. The major actions of flavonoids are those against cardiovascular diseases, ulcers, viruses, inflammation, osteoporosis, diarrhea and arthritis. Brief description about the disease causing effect of free radicals is given and ways by which flavonoids neutralize free radicals has also been mentioned.

Flavonoids Affect Actin Functions in Cytoplasm and Nucleus

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Markus Bhl, Simon Tietze, Andrea Sokoll, Sineej Madathil, Frank Pfennig, Joannis Apostolakis, Karim Fahmy and Herwig O. Gutzeit

Abstract
Based on the identification of actin as a target protein for the flavonol quercetin, the binding affinities of quercetin and structurally related flavonoids were determined by flavonoid-dependent quenching of tryptophan fluorescence from actin. Irrespective of differences in the hydroxyl pattern, similar Kd values in the 20M range were observed for six flavonoids encompassing members of the flavonol, isoflavone, flavanone, and flavane group. The potential biological relevance of the flavonoid/actin interaction in the cytoplasm and the nucleus was addressed using an actin polymerization and a transcription assay, respectively. In contrast to the similar binding affinities, the flavonoids exert distinct and partially opposing biological effects: although flavonols inhibit actin functions, the structurally related flavane epigallocatechin promotes actin activity in both test systems. Infrared spectroscopic evidence reveals flavonoid-specific conformational changes in actin which may mediate the different biological effects. Docking studies provide models of flavonoid binding to the known small moleculebinding sites in actin. Among these, the mostly hydrophobic tetramethylrhodaminebinding site is a prime candidate for flavonoid binding and rationalizes the high efficiency of quenching of the two closely located fluorescent tryptophans. The experimental and theoretical data consistently indicate the importance of hydrophobic, rather than H-bond-mediated, actin-flavonoid interactions. Depending on the rigidity of the flavonoid structures, different functionally relevant conformational changes are evoked through an induced fit.

Preparative Isolation of Flavonoids from Mulberry (Morus Alba L.) Leaves by Macroscopous Adsorption
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DONGDONG JIA, SHUFEN LI and ZHIPENG GU

ABSTRACT
The flavonoids in mulberry leaves have good antioxidant activities. In this study, ultrasound-assisted extraction was used to obtain the crude flavonoids, and the macroporous resin adsorption technology was further employed to purify the flavonoids. The nonpolar resin labeled as H-103 was selected from eight kinds of macroporous resins as the adsorbent. Static and dynamic adsorptiondesorption were tested to optimize the operation parameters. The experimental results show that the purity of the total flavonoids can be 36.3%, with a recovery of 83.4% under the optimum conditions. The flavonoids of rutin and isoquercitrin in the extract were detected by high-performance liquid chromatography, and their recoveries were above 80%. The devised method of isolation of the flavonoids from mulberry leaves may be used to develop a new process for medicinal purpose.

Growth, weaning performance and blood indicators of humoral immunity in Holstein calves fed supplemental flavonoids
S. M. J. Yaghoubi , G. R. Ghorbani , H. R. Rahmani and A. Nikkhah Copyright Journal compilation 2008 Blackwell Publishing Ltd

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ABSTRACT
The primary objective was to test the hypothesis that flavonoids mediate immune response and affect calf performance. Twenty Holstein calves [7 2 days age; 41.4 0.7 kg body weight (BW)] were randomly assigned to four treatments of (i) no; (ii) low (7.3 105 g/kg BW); (iii) medium (7.3 104 g/kg BW); and (iv) high (3.6 103 g/kg BW) doses of flavonoids intake in a completely randomized design. Calves received the treatments as a tablet until weaning or a daily intake of 680 g starter. After weaning, calves received no supplemental flavonoids and monitored until 120 days of age. The flavonoids were extracted from propolis. Treatments did not affect body length, wither height and the severity of scours. At week 5 of age, BW was higher when calves fed the high compared to the low dose of flavonoids. At week 6, calves fed the high dose of flavonoids had higher BW than those fed no or low doses of flavonoids. The serum immunoglobulin G (IgG) concentrations remained lower at the first 3 weeks of the experiment when calves received the low but not the high doses of flavonoids. At week 4, both medium and low doses of flavonoids moderated serum IgG. At week 8, the medium and high but not the low doses of flavonoids lowered serum IgG. At week 6, calves fed high and medium flavonoids doses had lower blood immunoglobulin M (IgM) than control calves. Results suggest that flavonoids affect the humoral immune response and can improve growth in young calves. This response depended on calf age. Future studies are needed to further evaluate the premise that dietary forages or the main source of flavonoids are helpful for a less stressful weaning in the modern calf raising.

Dietary Flavonoids and the Risk of Colorectal Cancer

Evropi Theodoratou, Janet Kyle, Roseanne Cetnarskyj, Susan M. Farrington, Albert Tenesa, Rebecca Barnetson, Mary Porteous, Malcolm Dunlop and Harry Campbell

ABSTRACT
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In vitro and in vivo laboratory data point to chemoprotective effects of flavonoids on colorectal cancer. However, there has been limited epidemiologic research on the dietary intake of flavonoids and risk of colorectal cancer. Recent expansions of dietary databases to include flavonoid data now make such studies feasible. Association between the six main classes of flavonoids and the risk of colorectal cancer was examined using data from a national prospective case-control study in Scotland, including 1,456 incident cases and 1,456 population-based controls matched on age, sex, and residence area. Dietary, including flavonoid data, were obtained from a validated, self-administered food frequency questionnaire. Risk of colorectal cancer was estimated using conditional logistic regression models in the whole sample and stratified by sex, smoking status, and cancer site and adjusted for established and putative risk factors. After energy adjustment, reductions in colorectal cancer risk associated with the highest quartiles of intake (versus the lowest quartile) were 27% for flavonols [odds ratio (OR), 0.73; Ptrend = 0.012], 32% for quercetin (OR, 0.68; Ptrend = 0.001), 32% for catechin (OR, 0.68; Ptrend < 0.0005); 26% for epicatechin (OR, 0.74; Ptrend = 0.019), and 22% for procyanidins (OR, 0.78; Ptrend = 0.031). The significant dose-dependent reductions in colorectal cancer risk that were associated with increased consumption of flavonols, quercetin, catechin, and epicatechin remained robust after controlling for overall fruit and vegetable consumption or for other flavonoid intake. The risk reductions were greater among nonsmokers, but no interaction beyond a multiplicative effect was present. Sex-specific or cancer-type differences were not observed. No risk reductions were associated with intake of flavones (Ptrend = 0.64), flavonones (Ptrend = 0.22), and phytoestrogens (Ptrend = 0.26). This was the first of several a priori hypotheses to be tested in this large study and showed strong and linear inverse associations of flavonoids with colorectal cancer risk. (Cancer Epidemiol Biomarkers Prev 2007;16(4):68493)

Flavonoids: a review of probable mechanisms of action and potential applications

Robert J Nijveldt, Els van Nood, Danny EC van Hoorn, Petra G Boelens, Klaske van Norren and Paul AM van Leeuwen
ABSTRACT

The aim of this review, a summary of the putative biological actions of flavonoids, was to obtain a further understanding of the reported beneficial health effects of these substances. Flavonoids occur naturally in fruit, vegetables, and beverages such as tea
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and wine. Research in the field of flavonoids has increased since the discovery of the French paradox,ie, the low cardiovascular mortality rate observed inMediterranean populations in association with red wine consumptionand a high saturated fat intake. Several other potentialbeneficial properties of flavonoids have since been ascertained.We review the different groups of known flavonoids, the probablemechanisms by which they act, and the potential clinicalapplications of these fascinating natural substances American Journal of Clinical Nutrition, Vol. 74, No. 4, 418-425, October 2001 2001 American Society for Clinical Nutrition

Flavonoid intake and cardiovascular disease mortality: a prospective study in postmenopausal women
Pamela J Mink, Carolyn G Scrafford, Leila M Barraj, Lisa Harnack, ChingPing Hong, Jennifer A Nettleton and David R Jacobs, Jr Background: Dietary flavonoids may have beneficial cardiovascular effects in human populations, but epidemiologic study results have not been conclusive.
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Objective: We used flavonoid food composition data from 3 recently available US Department of Agriculture databases to improve estimates of dietary flavonoid intake and to evaluate the association between flavonoid intake and cardiovascular disease (CVD) mortality. Design: Study participants were 34 489 postmenopausal women in the Iowa Women's Health Study who were free of CVD and had complete food-frequency questionnaire information at baseline. Intakes of total flavonoids and 7 subclasses were categorized into quintiles, and food sources were grouped into frequency categories. Proportional hazards rate ratios (RR) were computed for CVD, coronary heart disease (CHD), stroke, and total mortality after 16 y of follow-up. Results: After multivariate adjustment, significant inverse associations were observed between anthocyanidins and CHD, CVD, and total mortality [RR (95% CI) for any versus no intake: 0.88 (0.78, 0.99), 0.91 (0.83, 0.99), and 0.90 (0.86, 0.95)]; between flavanones and CHD [RR for highest quintile versus lowest: 0.78 (0.65, 0.94)]; and between flavones and total mortality [RR for highest quintile versus lowest: 0.88 (0.82, 0.96)]. No association was found between flavonoid intake and stroke mortality. Individual flavonoid-rich foods associated with significant mortality reduction included bran (added to foods; associated with stroke and CVD); apples or pears or both and red wine (associated with CHD and CVD); grapefruit (associated with CHD); strawberries (associated with CVD); and chocolate (associated with CVD). Conclusion: Dietary intakes of flavanones, anthocyanidins, and certain foods rich in flavonoids were associated with reduced risk of death due to CHD, CVD, and all causes. American Journal of Clinical Nutrition, Vol. 85, No. 3, 895-909, March 2007 2007 American Society for Nutrition

Flavonoid intake and risk of chronic diseases

Paul Knekt, Jorma Kumpulainen, Ritva Jrvinen, Harri Rissanen, Markku Helivaara, Antti Reunanen, Timo Hakulinen and Arpo Aromaa ABSTRACT Background: Flavonoids are effective antioxidants and may protect against several chronic diseases.

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Objective: The association between flavonoid intake and risk of several chronic diseases was studied. Design: The total dietary intakes of 10 054 men and women during the year preceding the baseline examination were determined with a dietary history method. Flavonoid intakes were estimated, mainly on the basis of the flavonoid concentrations in Finnish foods. The incident cases of the diseases considered were identified from different national public health registers. Results: Persons with higher quercetin intakes had lower mortality from ischemic heart disease. The relative risk (RR) between the highest and lowest quartiles was 0.79 (95% CI: 0.63, 0.99: P for trend = 0.02). The incidence of cerebrovascular disease was lower at higher kaempferol (0.70; 0.56, 0.86; P = 0.003), naringenin (0.79; 0.64, 0.98; P = 0.06), and hesperetin (0.80; 0.64, 0.99; P = 0.008) intakes. Men with higher quercetin intakes had a lower lung cancer incidence (0.42; 0.25, 0.72; P = 0.001), and men with higher myricetin intakes had a lower prostate cancer risk (0.43; 0.22, 0.86; P = 0.002). Asthma incidence was lower at higher quercetin (0.76; 0.56, 1.01; P = 0.005), naringenin (0.69; 0.50, 0.94; P = 0.06), and hesperetin (0.64; 0.46, 0.88; P = 0.03) intakes. A trend toward a reduction in risk of type 2 diabetes was associated with higher quercetin (0.81; 0.64, 1.02; P = 0.07) and myricetin (0.79; 0.62, 1.00; P = 0.07) intakes. Conclusion: The risk of some chronic diseases may be lower at higher dietary flavonoid intakes. American Journal of Clinical Nutrition, Vol. 76, No. 3, 560-568, September 2002 2002 American Society for Clinical Nutrition

Flavonoid intake and the risk of cardiovascular disease in women

Howard D Sesso, J Michael Gaziano, Simin Liu and Julie E Buring ABSTRACT Background: Despite emerging evidence of the role of flavonoids in cardiovascular disease (CVD) prevention, the association remains unclear.

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Objective: We examined whether flavonoids and selected flavonols and flavones or their food sources are associated with CVD risk. Design: Women (n = 38 445) free of CVD and cancer participated in a prospective study with a mean follow-up of 6.9 y. On the basis of a food-frequency questionnaire, total flavonoids and selected flavonols and flavones were categorized into quintiles, and food sources were categorized into 4 groups. Relative risks were computed for important vascular events (519 events; excluding revascularizations) and CVD (729 events), including myocardial infarction, stroke, revascularization, and CVD death. Results: The mean flavonoid intake was 24.6 18.5 mg/d, primarily as quercetin (70.2%). For both CVD and important vascular events, no significant linear trend was observed across quintiles of flavonoid intake (P = 0.63 and 0.80, respectively). No individual flavonol or flavone was associated with CVD. Broccoli and apple consumption were associated with nonsignificant reductions in CVD risk: 2530% and 1322%, respectively. A small proportion of women (n = 1185) consuming 4 cups (946 mL) tea/d had a reduction in the risk of important vascular events but with a nonsignificant linear trend (P = 0.07). Conclusions: Flavonoid intake was not strongly associated with a reduced risk of CVD. The nonsignificant inverse associations for broccoli, apples, and tea with CVD were not mediated by flavonoids and warrant further study. American Journal of Clinical Nutrition, Vol. 77, No. 6, 1400-1408, June 2003 2003 American Society for Clinical Nutrition

Comparison of the antioxidant effects of Concord grape juice flavonoids -tocopherol on markers of oxidative stress in healthy adults
Dawn J OByrne, Sridevi Devaraj, Scott M Grundy and Ishwarlal Jialal ABSTRACT

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Background: Concord grape juice (CGJ) is a rich source of flavonoids, which have greater antioxidant efficacy in vitro than does -tocopherol; however, the efficacies of flavonoids and -tocopherol in vivo have not been compared. Objective: We compared the in vivo antioxidant efficacy of CGJ with that of tocopherol in healthy adults. Design: Subjects were randomly assigned to receive either 400 IU RRR- -tocopherol/d (n = 17) or 10 mL CGJ kg-1 d-1 (n = 15) for 2 wk. Serum oxygen radical absorbance capacity, plasma protein carbonyls, urinary F2-isoprostanes, and resistance of LDL to ex vivo oxidation were measured before and after supplementation as markers of antioxidant status and oxidative stress. Results: After supplementation, plasma -tocopherol increased 92% in subjects who received -tocopherol (P < 0.001); plasma total and conjugated phenols increased 17% (P < 0.01) and 22% (P < 0.001), respectively, in subjects who received CGJ. There was a significant change in plasma triacylglycerols in both groups, but the concentrations were within the normal range. CGJ supplementation was associated with significantly higher triacylglycerols than was -tocopherol supplementation. Both supplementation regimens significantly increased serum oxygen radical absorbance capacity (P < 0.001) and LDL lag time (P < 0.001) and significantly decreased the LDL oxidation rate (P < 0.01), with no significant difference in effectiveness. Protein carbonyl concentrations in native plasma decreased 20% after CGJ supplementation, which was a significantly different response than that after -tocopherol supplementation (P < 0.05). Conclusions: In healthy adults, 10 mL CGJ kg-1 d-1 increased serum antioxidant capacity and protected LDL against oxidation to an extent similar to that obtained with 400 IU -tocopherol/d but decreased native plasma protein oxidation significantly more than did -tocopherol. CGJ flavonoids are potent antioxidants that may protect against oxidative stress and reduce the risk of free radical damage and chronic diseases. American Journal of Clinical Nutrition, Vol. 76, No. 6, 1367-1374, December 2002 2002 American Society for Clinical Nutrition

The flavonoids quercetin and catechin synergistically inhibit platelet function by antagonizing the intracellular production of hydrogen peroxide
Pasquale Pignatelli, Fabio M Pulcinelli, Andrea Celestini, Luisa Lenti, Andrea Ghiselli, Pier Paolo Gazzaniga and Francesco Violi ABSTRACT
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Background: Epidemiologic studies have shown an inverse relation between moderate consumption of red wine and cardiovascular disease. Studies have shown that red wine and its component flavonoids inhibit in vivo platelet activation, but the underlying mechanism has not yet been identified. Objective: Because we showed previously that collagen-induced platelet aggregation is associated with a burst of hydrogen peroxide, which in turn contributes to stimulating the phospholipase C pathway, the aim of this study was to investigate whether flavonoids synergize in inhibiting platelet function and interfere with platelet function by virtue of their antioxidant effect. Design: We tested the effect of 2 flavonoids, quercetin and catechin, on collageninduced platelet aggregation and hydrogen peroxide and on platelet adhesion to collagen. Results: Catechin (50100 mol/L) and quercetin (1020 mol/L) inhibited collageninduced platelet aggregation and platelet adhesion to collagen. The combination of 25 mol catechin/L and 5 mol quercetin/L, neither of which had any effect on platelet function when used alone, significantly inhibited collagen-induced platelet aggregation and platelet adhesion to collagen. Such a combination strongly inhibited collageninduced hydrogen peroxide production, calcium mobilization, and 1,3,4-inositol triphosphate formation. Conclusions: These data indicate that flavonoids inhibit platelet function by blunting hydrogen peroxide production and, in turn, phospholipase C activation and suggest that the synergism among flavonoids could contribute to an understanding of the relation between the moderate consumption of red wine and the decreased risk of cardiovascular disease. American Journal of Clinical Nutrition, Vol. 72, No. 5, 1150-1155, November 2000 2000 American Society for Clinical Nutrition

Inverse association of tea and flavonoid intakes with incident myocardial infarction: the Rotterdam Study
Johanna M Geleijnse, Lenore J Launer, Deirdre AM van der Kuip, Albert Hofman and Jacqueline CM Witteman ABSTRACT Background: Dietary flavonoids may protect against cardiovascular disease, but evidence is still conflicting. Tea is the major source of flavonoids in Western populations.
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Objective: The association of tea and flavonoid intake with incident myocardial infarction was examined in the general Dutch population. Design: A longitudinal analysis was performed with the use of data from the Rotterdam Studya population-based study of men and women aged 55 y. Diet was assessed at baseline (19901993) with a validated semiquantitative food-frequency questionnaire. The analysis included 4807 subjects with no history of myocardial infarction, who were followed until 31 December 1997. Data were analyzed in a Cox regression model, with adjustment for age, sex, body mass index, smoking status, pack-years of cigarette smoking, education level, and daily intakes of alcohol, coffee, polyunsaturated fat, saturated fat, fiber, vitamin E, and total energy. Results: During 5.6 y of follow-up, a total of 146 first myocardial infarctions occurred, 30 of which were fatal. The relative risk (RR) of incident myocardial infarction was lower in tea drinkers with a daily intake >375 mL (RR: 0.57; 95% CI: 0.33, 0.98) than in nontea drinkers. The inverse association with tea drinking was stronger for fatal events (0.30; 0.09, 0.94) than for nonfatal events (0.68; 0.37, 1.26). The intake of dietary flavonoids (quercetin + kaempferol + myricetin) was significantly inversely associated only with fatal myocardial infarction (0.35; 0.13, 0.98) in upper compared with lower tertiles of intake. American Journal of Clinical Nutrition, Vol. 75, No. 5, 880-886, May 2002 2002 American Society for Clinical Nutrition

Flavonoids, flavonoid-rich foods, and cardiovascular risk: a meta-analysis of randomized controlled trials
Lee Hooper, Paul A Kroon, Eric B Rimm, Jeffrey S Cohn, Ian Harvey, Kathryn A Le Cornu, Jonathan J Ryder, Wendy L Hall and Aedn Cassidy ABSTRACT Background:The beneficial effects of flavonoid consumption on cardiovascular risk are supported by mechanistic and epidemiologic evidence. Objective:We aimed to systematically review the effectiveness of different flavonoid subclasses and flavonoid-rich food sources on cardiovascular disease (CVD) and risk factorsie, lipoproteins, blood pressure, and flow-mediated dilatation (FMD).
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Design:Methods included a structured search strategy on MEDLINE, EMBASE, and Cochrane databases; formal inclusion or exclusion, data extraction, and validity assessment; and meta-analysis. Results:One hundred thirty-three trials were included. No randomized controlled trial studied effects on CVD morbidity or mortality. Significant heterogeneity confirmed differential effects between flavonoid subclasses and foods. Chocolate increased FMD after acute (3.99%; 95% CI: 2.86, 5.12; 6 studies) and chronic (1.45%; 0.62, 2.28; 2 studies) intake and reduced systolic (5.88 mm Hg; 9.55, 2.21; 5 studies) and diastolic (3.30 mm Hg; 5.77, 0.83; 4 studies) blood pressure. Soy protein isolate (but not other soy products or components) significantly reduced diastolic blood pressure ( 1.99 mm Hg; 2.86, 1.12; 9 studies) and LDL cholesterol (0.19 mmol/L; 0.24, 0.14; 39 studies). Acute black tea consumption increased systolic (5.69 mm Hg; 1.52, 9.86; 4 studies) and diastolic (2.56 mm Hg; 1.03, 4.10; 4 studies) blood pressure. Green tea reduced LDL (0.23 mmol/L; 0.34, 0.12; 4 studies). For many of the other flavonoids, there was insufficient evidence to draw conclusions about efficacy. Conclusions:To date, the effects of flavonoids from soy and cocoa have been the main focus of attention. Future studies should focus on other commonly consumed subclasses (eg, anthocyanins and flavanones), examine dose-response effects, and be of long enough duration to allow assessment of clinically relevant endpoints. American Journal of Clinical Nutrition, Vol. 88, No. 1, 38-50, July 2008 2008 American Society for Nutrition

Dietary intake of selected flavonols, flavones, and flavonoid-rich foods and risk of cancer in middleaged and older women
Lu Wang, I-Min Lee, Shumin M Zhang, Jeffrey B Blumberg, Julie E Buring and Howard D Sesso Abstarct Background: Flavonoids may protect against cancer development through several biological mechanisms. However, epidemiologic studies on dietary flavonoids and cancer risk have yielded inconsistent results. Objective: We prospectively investigated the association between the intake of selected flavonoids and flavonoid-rich foods and risk of cancers in the Women's Health Study.

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Design: A total of 3234 incident cancer cases were identified during 11.5 y of follow-up among 38,408 women aged 45 y. Intake of individual flavonols (quercetin, kaempferol, and myricetin) and flavones (apigenin and luteolin) was assessed from food-frequency questionnaires. Cox regression models were used to estimate the relative risk (RR) of total and site-specific cancer across increasing intakes of total and individual selected flavonoids and flavonoid-rich foods (tea, apple, broccoli, onion, and tofu). Results: The multivariate RRs of total cancer across increasing quintiles of total quantified flavonoid intake were 1.00, 1.00, 0.93, 0.94, and 0.97 (P for trend = 0.72). For site-specific cancers, the multivariate RRs in the highest quintile of total quantified flavonoid intake compared with the lowest quintile were 1.03 for breast cancer, 1.01 for colorectal cancer, 1.03 for lung cancer, 1.15 for endometrial cancer, and 1.09 for ovarian cancer (all P > 0.05). The associations for the individual flavonoid intakes were similar to those for the total intake. There was also no significant association between intake of flavonoid-rich foods and the incidence of total and site-specific cancers. Conclusion: Our results do not support a major role of 5 common flavonols and flavones or selected flavonoid-rich foods in cancer prevention.
American Journal of Clinical Nutrition, doi:10.3945/ajcn.2008.26913 Vol. 89, No. 3, 905-912, March 2009 2009 American Society for Clinical Nutrition

Dietary flavonoid intake and non-Hodgkin lymphoma risk

Cara L Frankenfeld, James R Cerhan, Wendy Cozen, Scott Davis, Maryjean Schenk, Lindsay M Morton, Patricia Hartge and Mary H Ward ABSTRACT Background: The role of dietary factors in non-Hodgkin lymphoma (NHL) risk is not yet well understood. Dietary flavonoids are polyphenolic compounds proposed to be anticarcinogenic. Flavonoids are well-characterized antioxidants and metal chelators, and certain flavonoids exhibit antiproliferative and antiestrogenic effects. Objective: We aimed to evaluate the hypothesis that higher flavonoid intake is associated with lower NHL risk.
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Design: During 19982000, we identified incident NHL cases aged 2074 y from 4 US Surveillance, Epidemiology, and End Results cancer registries. Controls without history of NHL were selected by random-digit dialing or from Medicare files and frequencymatched to cases by age, center, race, and sex. Using 3 recently developed US Department of Agriculture nutrient-specific databases, flavonoid intake was estimated from participant responses to a 117-item food-frequency questionnaire (n = 466 cases and 390 controls). NHL risk in relation to flavonoid intake in quartiles was evaluated after adjustment for age, sex, registry, education, NHL family history, and energy intake. Results: Higher total flavonoid intake was significantly associated with lower risk of NHL (P for trend < 0.01): a 47% lower risk in the highest quartile of intake than in the lowest (95% CI: 31%, 73%). Higher intakes of flavonols, epicatechins, anthocyanidins, and proanthocyanidins were each significantly associated with decreased NHL risk. Similar patterns of risk were observed for the major NHL subtypesdiffuse large B-cell lymphoma (n = 167) and follicular lymphoma (n = 146). Conclusion: A higher intake of flavonoids, dietary components with several putative anticarcinogenic activities, may be associated with lower NHL risk.
American Journal of Clinical Nutrition, Vol. 87, No. 5, 1439-1445, May 2008 2008 American Society for Nutrition

Health promotion by flavonoids, tocopherols, tocotrienols, and other phenols: direct or indirect effects? Antioxidant or not?
Barry Halliwell, Joseph Rafter and Andrew Jenner

ABSTRACT
Foods and beverages rich in phenolic compounds, especially flavonoids, have often been associated with decreased risk of developing several diseases. However, it remains unclear whether this protective effect is attributable to the phenols or to other agents in the diet. Alleged health-promoting effects of flavonoids are usually attributed to their powerful antioxidant activities, but evidence for in vivo antioxidant effects of flavonoids is confusing and equivocal. This may be because maximal plasma concentrations, even after extensive flavonoid intake, may be low (insufficient to exert significant systemic
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antioxidant effects) and because flavonoid metabolites tend to have decreased antioxidant activity. Reports of substantial increases in plasma total antioxidant activity after flavonoid intake must be interpreted with caution; findings may be attributable to changes in urate concentrations. However, phenols might exert direct effects within the gastrointestinal tract, because of the high concentrations present. These effects could include binding of prooxidant iron, scavenging of reactive nitrogen, chlorine, and oxygen species, and perhaps inhibition of cyclooxygenases and lipoxygenases. Our measurements of flavonoids and other phenols in human fecal water are consistent with this concept. We argue that tocopherols and tocotrienols may also exert direct beneficial effects in the gastrointestinal tract and that their return to the gastrointestinal tract by the liver through the bile may be physiologically advantageous. American Journal of Clinical Nutrition, Vol. 81, No. 1, 268S-276S, January 2005 2005 American Society for Clinical Nutrition

Consumption of flavonoids in onions and black tea: lack of effect on F2-isoprostanes and autoantibodies to oxidized LDL in healthy humans
James D O'Reilly, Anthony I Mallet, Gareth T McAnlis, Ian S Young, Barry Halliwell, Tom AB Sanders and Helen Wiseman Abstact Background: Oxidative damage to lipids in vivo may be involved in the development of atherosclerosis and cancer. Onions and black tea are foods rich in flavonoids, predominantly the flavonoid quercetin, which is a potent in vitro inhibitor of membrane lipid peroxidation and LDL oxidation. Objective: Our objective was to investigate the effects of consuming a high-flavonoid (HF) diet enriched with onions and black tea on indexes of oxidative damage in vivo compared with a low-flavonoid (LF) diet.

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Design: Thirty-two healthy humans were studied in a randomized crossover design. Indexes of oxidative damage used were plasma F2-isoprostanes (a biomarker of lipid peroxidation in vivo) and the titer of antibodies to malondialdehyde (MDA)-modified LDL. Results: There were no significant differences in the intake of macronutrients or assessed micronutrients, plasma F2-isoprostane concentrations, and MDA-LDL autoantibody titer between the HF and LF dietary treatments. In the men, however, plasma concentrations of the F2-isoprostane 8-epi-prostaglandin F2 were slightly higher after the HF treatment phase than after the LF treatment [0.31 0.029 nmol/L (111 10.4 ng/L) compared with 0.26 0.022 nmol/L (92 7.8 ng/L); P = 0.041]. In all subjects, plasma quercetin concentrations were significantly higher after the HF treatment phase than after the LF treatment: 221.6 37.4 nmol/L compared with less than the limit of detection of 66.2 nmol/L. Conclusion: Flavonoid consumption in onions and tea had no significant effect on plasma F2-isoprostane concentrations and MDA-LDL autoantibody titer in this study and thus does not seem to inhibit lipid peroxidation in humans. American Journal of Clinical Nutrition, Vol. 73, No. 6, 1040-1044, June 2001 2001 American Society for Clinical Nutrition

Effects of flavonoids and vitamin C on oxidative DNA damage to human lymphocytes

M Noroozi, WJ Angerson and ME Lean This study assessed the antioxidant potencies of several widespread dietary flavonoids across a range of concentrations and compared with vitamin C as a positive control. The antioxidant effects of pretreatment with flavonoids and vitamin C, at standardized concentrations (7.6, 23.2, 93, and 279.4 micromol/L), on oxygen radical- generated DNA damage from hydrogen peroxide (100 micromol/L) in human lymphocytes were examined by using the single-cell gel electrophoresis assay (comet assay). Pretreatment with all flavonoids and vitamin C produced dose-dependent reductions in oxidative DNA damage. At a concentration of 279 micromol/L, they were ranked in decreasing order of potency as follows: luteolin (9% of damage from unopposed hydrogen peroxide), myricetin (10%), quercetin (22%), kaempferol (32%), quercitrin (quercetin-3-Lrhamnoside) (45%), apigenin (59%), quercetin- 3-glucoside (62%), rutin (quercetin-3beta-D-rutinoside) (82%), and vitamin C (78%). The protective effect of vitamin C against DNA damage at this concentration was significantly less than that of all the flavonoids except apigenin, quercetin-3-glucoside, and rutin. The ranking was similar
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with estimated ED50 (concentration to produce 50% protection) values. The protective effect of quercetin and vitamin C at a concentration of 23.2 micromol/L was found to be additive (quercetin: 71% of maximal DNA damage from unopposed hydrogen peroxide; vitamin C: 83%; both in combination: 62%). These data suggest that the free flavonoids are more protective than the conjugated flavonoids (eg, quercetin compared with its conjugate quercetin-3-glucoside, P < 0.001). Data are also consistent with the hypothesis that antioxidant activity of free flavonoids is related to the number and position of hydroxyl groups. American Journal of Clinical Nutrition, Vol 67, 1210-1218, Copyright 1998 by The American Society for Clinical Nutrition, Inc

Cocoa polyphenols and inflammatory mediators

Helmut Sies, Tankred Schewe, Christian Heiss and Malte Kelm

ABSTRACT
Cocoa products are sources of flavan-3-ols, which have attracted interest regarding cardiovascular health. This review provides a survey of our research on the effects of cocoa polyphenols on leukotriene and nitric oxide (NO) metabolism and on myeloperoxidase-induced modification of LDL. Because intake of flavonoid-rich chocolate by human subjects was reported to decrease the plasma concentrations of proinflammatory cysteinyl leukotrienes, we assessed whether cocoa polyphenols inhibited human 5-lipoxygenase, the key enzyme of leukotriene synthesis. ()Epicatechin and other cocoa flavan-3-ols proved to be inhibitory at the enzyme level. This action may confer antileukotriene action in vivo. In a double-blind crossover study, 20 individuals at risk for cardiovascular diseases received cocoa beverages with high or low contents of flavan-3-ols. NO-dependent, flow-mediated dilation of the brachial artery and concentrations of nitroso compounds in plasma were measured, and it was shown that ingestion of the high-flavanol coca drink but not the low-flavanol cocoa drink
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significantly increased plasma concentrations of nitroso compounds and flow-mediated dilation of the brachial artery. Therefore, ingested flavonoids may reverse endothelial dysfunction through enhancement of NO bioactivity. Oxidative modification of LDL appears to be crucial for atherogenesis, and one of the mediators is the proinflammatory proatherogenic enzyme myeloperoxidase. Micromolar concentrations of ()-epicatechin or other flavonoids were found to suppress lipid peroxidation in LDL induced by myeloperoxidase in the presence of physiologically relevant concentrations of nitrite, an NO metabolite. Adverse effects of NO metabolites, such as nitrite and peroxynitrite, were thus attenuated.
American Journal of Clinical Nutrition, Vol. 81, No. 1, 304S-312S, January 2005 2005 American Society for Clinical Nutrition

The transparent testa4 Mutation Prevents Flavonoid Synthesis and Alters Auxin Transport and the Response of Arabidopsis Roots to Gravity and Light
Charles S. Buer and Gloria K. Muday

ABSTRACT
We examined whether flavonoids act as endogenous auxin transport regulators during gravity vector and light intensity changes in Arabidopsis thaliana roots. Flavonoid deficient transparent testa4 [tt4(2YY6)] seedlings had elevated root basipetal auxin transport compared with the wild type, consistent with the absence of a negative auxin transport regulator. The tt4(2YY6) roots had delayed gravitropism that was chemically complemented with a flavonoid intermediate. Flavonoid accumulation was found in wild-type columella cells, the site of gravity perception, and in epidermal and cortical cells, the site of differential growth, but flavonoid accumulation was absent in tt4(2YY6) roots. Flavonoid accumulation was higher in gravity-stimulated root tips as compared with vertical controls, with maximum differences coinciding with the timing of gravitropic bending, and was located in epidermal cells. Exogenous indole-3-acetic acid (IAA) also elevated flavonoid accumulation, suggesting that flavonoid changes in response to gravity might be partly as a result of changing IAA distribution. Acropetal
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IAA transport was also elevated in roots of tt4(2YY6). Flavonoid synthesis was repressed in the dark, as were differences in root acropetal transport in tt4(2YY6). These results are consistent with light- and gravity-induced flavonoid stimulation that alters auxin transport in roots and dependent physiological processes, including gravitropic bending and root development. The Plant Cell 16:1191-1205 (2004) 2004 American Society of Plant Biologists

Dietary intake of flavonoids and asthma in adults

V. Garcia, I. C. W. Arts, J. A. C. Sterne, R. L. Thompson and S. O. Shaheen

ABSTRACT
Epidemiological studies have suggested that a high consumption of apples may protect against asthma and chronic obstructive pulmonary disease. This effect has been attributed to their high flavonoid content, but few studies have investigated the relationship between flavonoid intake and obstructive lung disease directly. In a population-based, case-control study of 1,471 adults aged 1650 yrs in London (UK), the present study examined whether dietary intake of catechins, flavonols and flavones was negatively associated with asthma, asthma severity and chronic sputum production. Asthma was defined by positive responses to a standard screening questionnaire in 1996 and information about usual diet was obtained by a food frequency questionnaire in 1997. After controlling for potential confounders, dietary intake of these three flavonoid subclasses was not significantly associated with asthma, (odds ratio per quintile (95% confidence interval) = 0.94 (0.861.02); 1.00 (0.921.09); 0.98 (0.88 1.08) for flavones, flavonols and total catechins, respectively) nor was it associated with asthma severity, or chronic sputum production.
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In conclusion, no evidence was found for a protective effect of three major subclasses of dietary flavonoids on asthma. It is possible that other flavonoids or polyphenols present in apples may explain the protective effect of apples on obstructive lung disease. Eur Respir J 2005; 26:449-452 Copyright ERS Journals Ltd 2005 doi: 10.1183/09031936.05.00142104

Select Flavonoids and Whole Juice From Purple Grapes Inhibit Platelet Function and Enhance Nitric Oxide Release
Jane E. Freedman, MD; Crawford Parker, III, MD; Liqing Li, MS; Jacob A. Perlman; Balz Frei, PhD; Vadim Ivanov, PhD; Leslie R. Deak, BS; Mark D. Iafrati, MD; John D. Folts, PhD

ABSTRACT
BackgroundModerate red wine consumption is inversely associated with coronary ischemia, and both red wine and purple grape juice (PGJ) contain flavonoids with antioxidant and antiplatelet properties believed to be protective against cardiovascular events. Acute cardiac events are also associated with decreased platelet-derived nitric oxide (NO) release. In this study, the effects of PGJ and PGJ-derived flavonoids on platelet function and platelet NO production were determined. Methods and ResultsIncubation of platelets with dilute PGJ led to inhibition of aggregation, enhanced release of platelet-derived NO, and decreased superoxide production. To confirm the in vivo relevance of these findings, 20 healthy subjects consumed 7 mL kg-1 d-1 of PGJ for 14 days. Platelet aggregation was inhibited after PGJ supplementation, platelet-derived NO production increased from 3.51.2 to 6.01.5 pmol/108 platelets, and superoxide release decreased from 29.55.0 to 19.23.1 arbitrary units (P<0.007 and P<0.05, respectively). -Tocopherol levels increased significantly after PGJ consumption (from 15.60.7 to 17.60.9 mol/L; P<0.009), and
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the plasma proteinindependent antioxidant activity increased by 50.0% (P<0.05). Last, incubation of platelets with select flavonoid fractions isolated from PGJ consistently attenuated superoxide levels but had variable effects on whole-blood aggregation, platelet aggregation, and NO release. ConclusionsBoth in vitro incubation and oral supplementation with PGJ decrease platelet aggregation, increase platelet-derived NO release, and decrease superoxide production. These findings may be a result of antioxidant-sparing and/or direct effects of select flavonoids found in PGJ. The suppression of platelet-mediated thrombosis represents a potential mechanism for the beneficial effects of purple grape products, independent of alcohol consumption, in cardiovascular disease. (Circulation. 2001;103:2792.) 2001 American Heart Association, Inc.

Common dietary flavonoids inhibit the growth of the intraerythrocytic malaria parasite
Adele M Lehane and Kevin J Saliba
BMC Research Notes 2008, 1:26doi:10.1186/1756-0500-1-26 Published:18 June 2008

ABSTRACT
Flavonoids are abundant plant phenolic compounds. More than 6000 have been identified to date, and some have been shown to possess antiparasitic activity. Here we investigate the effects of a range of common dietary flavonoids on the growth of two strains of the human malaria parasite Plasmodium falciparum.A chloroquine-sensitive (3D7) and a chloroquine-resistant (7G8) strain of P. falciparum were tested for in vitro susceptibility to a range of individual dietary flavonoids and flavonoid combinations. Parasite susceptibility was measured in 96-well plates over 96 h using a previously described [3H]hypoxanthine incorporation assay. Of the eleven flavonoids tested, eight showed antiplasmodial activity against the 3D7 strain (with IC50 values between 11 and 66 M), and all showed activity against the 7G8 strain (with IC50 values between 12 and 76 M). The most active compound against both strains was luteolin, with IC50 values of 11 1 M and 12 1 M for 3D7 and 7G8, respectively. Luteolin was found to prevent the progression of parasite growth beyond the young trophozoite stage, and did not affect parasite susceptibility to the antimalarial drugs chloroquine or artemisinin. Combining low concentrations of flavonoids was found to produce an apparent additive antiplasmodial effect. Certain common dietary flavonoids inhibit the intraerythrocytic
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growth of the 3D7 and 7G8 strains of P. falciparum. Flavonoid combinations warrant further investigation as antiplasmodial agents.

Identification of the Flavonoid Hydroxylases from Grapevine and Their Regulation during Fruit Development
Jochen Bogs, Ali Ebadi, Debra McDavid and Simon P. Robinson ABSTRACT Flavonoids are important secondary metabolites in many fruits, and their hydroxylation pattern determines their color, stability, and antioxidant capacity. Hydroxylation of the B-ring of flavonoids is catalyzed by flavonoid 3'-hydroxylase (F3'H) and flavonoid 3',5'hydroxylase (F3'5'H), and may also require cytochrome b5. We report the identification of genes encoding F3'H, F3'5'H, and a putative cytochrome b5 from grapevine (Vitis vinifera L. cv Shiraz) and their transcriptional regulation in fruit. Functionality of the genes VvF3'H and VvF3'5'H1 was demonstrated by ectopic expression in petunia (Petunia hybrida), which altered flower color and flavonoid composition as expected. VvF3'H was expressed in grapes before flowering, when 3'-hydroxylated flavonols are made, and all three genes were expressed after flowering, when proanthocyanidins (PAs) are synthesized. In berry skin, expression of all three genes was low at the onset of ripening (vraison) but increased after vraison concomitant with the accumulation of 3'- and 3',5'-hydroxylated anthocyanins. VvF3'H and VvCytoB5 were expressed in seeds but not VvF3'5'H1, consistent with the accumulation of 3'-hydroxylated PAs in this tissue. VvCytoB5 expression was correlated with expression of both VvF3'H and VvF3'5'H1 in the different grape tissues. In contrast to red grapes, where VvF3'H, VvF3'5'H1, and VvCytoB5 were highly expressed during ripening, the expression of VvF3'5'H1 and VvCytoB5 in white grapes during ripening was extremely low, suggesting a difference in transcriptional regulation. Our results show that temporal and tissuespecific expression of VvF3'H, VvF3'5'H1, and VvCytoB5 in grapes is coordinated with
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the accumulation of the respective hydroxylated flavonols and PAs, as well as anthocyanins. Understanding the regulation of flavonoid hydroxylases could be used to modify flavonoid composition of fruits.
Plant Physiology 140:279-291 (2006) 2006 American Society of Plant Biologists

Flavonoid-membrane interactions: possible consequences for biological effects of some polyphenolic compounds
Andrzej B Hendrich

Flavonoids are found ubiquitously in higher plants and constitute an important component of the majority of peoples' daily diets. The biological activities of flavonoids cover a very broad spectrum, from anticancer and antibacterial activities through to inhibition of bone resorption. In the present paper, the interactions between flavonoids and lipid bilayers as well as biological membranes and their components are reviewed, with special emphasis on the structure-activity relationships and mechanisms underlying the biological activity of flavonoids.
Acta Pharmacologica Sinica (2006) 27, 2740; doi:10.1111/j.17457254.2006.00238.x

http://www.nature.com/aps/journal/v27/n1/abs/aps20062a.html

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Bimodal action of the flavonoid quercetin on basophil function: an investigation of the putative biochemical targets
Salvatore Chirumbolo, Marta Marzotto, Anita Conforti, Antonio Vella, Riccardo Ortolani and Paolo Bellavit
ABSTRACT

Flavonoids, a large group of polyphenolic metabolites derived from plants have received a great deal of attention over the last several decades for their properties in inflammation and allergy. Quercetin, the most abundant of plant flavonoids, exerts a modulatory action at nanomolar concentrations on human basophils. As this mechanism needs to be elucidated, in this study we focused the possible signal transduction pathways which may be affected by this compound. Methods: K2-EDTA derived leukocyte buffy coats enriched in basophil granulocytes were treated with different concentrations of quercetin and triggered with anti-IgE, fMLP, the calcium ionophore A23187 and the phorbol ester PMA in different experimental conditions. Basophils were captured in a flow cytometry analysis as CD123bright/HLADRnon expressing cells and fluorescence values of the activation markers CD63-FITC or
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CD203c-PE were used to produce dose response curves. The same population was assayed for histamine release. Quercetin inhibited the expression of CD63 and CD203c and the histamine release in basophils activated with anti-IgE or with the ionophore: the IC50 in the antiIgE model was higher than in the ionophore model and the effects were more pronounced for CD63 than for CD203c. Nanomolar concentrations of quercetin were able to prime both markers expression and histamine release in the fMLP activation model while no effect of quercetin was observed when basophils were activated with PMA. The specific phosphoinositide-3 kinase (PI3K) inhibitor wortmannin exhibited the same behavior of quercetin in anti-IgE and fMLP activation, thus suggesting a role for PI3K involvement in the priming mechanism. These results rule out a possible role of protein kinase C in the complex response of basophil to quercetin, while indirectly suggest PI3K as the major intracellular target of this compound also in human basophils. Clinical and Molecular Allergy 2010, 8:13doi:10.1186/1476-7961-8-13

Published:17 September 2010

http://www.clinicalmolecularallergy.com/content/8/1/13/abstract

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