Early molecular oxidative stress biomarkers of ischemic penumbra in acute stroke.

OBJECTIVES: To assess whether plasma biomarkers of oxidative stress predict diffusion-perfusion mismatch in patients with acute ischemic stroke (AIS). METHODS: We measured plasma levels of oxidative stress biomarkers such as F2-isoprostanes (F2-isoPs), total and perchloric acid Oxygen Radical Absorbance Capacity (ORACTOT and ORACPCA), urinary levels of 8-oxo-7,8-dihydro-2'-deoxyguoanosine, and inflammatory and tissue-damage biomarkers (high-sensitivity C-reactive protein, matrix metalloproteinase-2 and -9) in a prospective study of patients with AIS presenting within 9 hours of symptom onset. Diffusion-weighted (DWI) and perfusion-weighted (PWI) MRI sequences were analyzed with a semiautomated volumetric method. Mismatch was defined as baseline mean transit time volume minus DWI volume. A percent mismatch cutoff of >20% was considered clinically significant. A stricter definition of mismatch was also used. Mismatch salvage was the region free of overlap by final infarction. RESULTS: Mismatch >20% was present in 153 of 216 (70.8%) patients (mean [±SD] age 69.2 ± 14.3 years, 41.2% women). Patients with mismatch >20% were more likely to have higher baseline plasma levels of ORACPCA (p = 0.020) and F2-isoPs (p = 0.145). Multivariate binary logistic regression demonstrated that lnF2-isoP (odds ratio [OR] 2.44, 95% confidence interval [CI] 1.19-4.98, p = 0.014) and lnORACPCA (OR 4.18, 95% CI 1.41-12.41, p = 0.010) were independent predictors of >20% PWI-DWI mismatch and the stricter mismatch definition, respectively. lnORACTOT significantly predicted mismatch salvage volume (>20% mismatch p = 0.010, stricter mismatch definition p = 0.003). CONCLUSIONS: Elevated hyperacute plasma levels of F2-isoP and ORAC are associated with radiographic evidence of mismatch and mismatch salvage in patients with AIS. If validated, these findings may add to our understanding of the role of oxidative stress in cerebral tissue fate during acute ischemia.

Polyphenols in Almond Skins after Blanching Modulate Plasma Biomarkers of Oxidative Stress in Healthy Humans.

Almond skins are a waste byproduct of blanched almond production. Polyphenols extracted from almond skins possess antioxidant activities in vitro and in vivo. Thus, we examined the pharmacokinetic profile of almond skin polyphenols (ASP) and their effect on measures of oxidative stress. In a randomized crossover trial, seven adults consumed two acute ASP doses (225 mg (low, L) or 450 mg (high, H) total phenols) in skim milk or milk alone. Plasma flavonoids, glutathione peroxidase (GPx), glutathione (GSH), oxidized GSH (GSSG), and resistance of low- density lipoprotein (LDL) to oxidation were measured over 10 h. The H dose increased catechin and naringenin in plasma, with maximum concentrations of 44.3 and 19.3 ng/mL, respectively. The GSH/GSSG ratio at 3 h after the H doses was 212% of the baseline value, as compared to 82% after milk (p = 0.003). Both ASP doses upregulated GPx activity by 26-35% from the baseline at 15, 30, 45, and 120 min after consumption. The in vitro addition of α-tocopherol extended the lag time of LDL oxidation at 3 h after L and H consumption by 144.7% and 165.2% of that at 0 h compared to no change after milk (p ≤ 0.05). In conclusion, ASP are bioavailable and modulate GSH status, GPx activity, and the resistance of LDL to oxidation.

Oxidative Stress Biomarkers of Brain Damage: Hyperacute Plasma F2-Isoprostane Predicts Infarct Growth in Stroke.

BACKGROUND AND PURPOSE: Oxidative stress is an early response to cerebral ischemia and is likely to play an important role in the pathogenesis of cerebral ischemic injury. We sought to evaluate whether hyperacute plasma concentrations of biomarkers of oxidative stress, inflammation, and tissue damage predict infarct growth (IG). METHODS: We prospectively measured plasma F2-isoprostane (F2-isoP), urinary 8-oxo-7,8-dihydro-2'-deoxyguoanosine, plasma oxygen radical absorbance capacity assay, high sensitivity C reactive protein, and matrix metalloproteinase 2 and 9 in consecutive patients with acute ischemic stroke presenting within 9 hours of symptom onset. Patients with baseline diffusion-weighted magnetic resonance imaging and follow-up diffusion-weighted imaging or computed tomographic scan were included to evaluate the final infarct volume. Baseline diffusion-weighted imaging volume and final infarct volume were analyzed using semiautomated volumetric method. IG volume was defined as the difference between final infarct volume and baseline diffusion-weighted imaging volume. RESULTS: A total of 220 acute ischemic stroke subjects were included in the final analysis. One hundred seventy of these had IG. Baseline F2-isoP significantly correlated with IG volume (Spearman ρ=0.20; P=0.005) and final infarct volume (Spearman ρ=0.19; P=0.009). In a multivariate binary logistic regression model, baseline F2-isoP emerged as an independent predictor of the occurrence of IG (odds ratio, 2.57; 95% confidence interval, 1.37-4.83; P=0.007). In a multivariate linear regression model, baseline F2-isoP was independently associated with IG volume (B, 0.38; 95% confidence interval, 0.04-0.72; P=0.03). CONCLUSIONS: Elevated hyperacute plasma F2-isoP concentrations independently predict the occurrence of IG and IG volume in patients with acute ischemic stroke. If validated in future studies, measuring plasma F2-isoP might be helpful in the acute setting to stratify patients with acute ischemic stroke for relative severity of ischemic injury and expected progression.

Effect of almond consumption on vascular function in patients with coronary artery disease: a randomized, controlled, cross-over trial.

OBJECTIVE: Almonds reduce cardiovascular disease risk via cholesterol reduction, anti-inflammation, glucoregulation, and antioxidation. The objective of this randomized, controlled, cross-over trial was to determine whether the addition of 85 g almonds daily to a National Cholesterol Education Program (NCEP) Step 1 diet (ALM) for 6 weeks would improve vascular function and inflammation in patients with coronary artery disease (CAD). RESEARCH DESIGN AND METHODS: A randomized, controlled, crossover trial was conducted in Boston, MA to test whether as compared to a control NCEP Step 1 diet absent nuts (CON), incorporation of almonds (85 g/day) into the CON diet (ALM) would improve vascular function and inflammation. The study duration was 22 weeks including a 6-weeks run-in period, two 6-weeks intervention phases, and a 4-weeks washout period between the intervention phases. A total of 45 CAD patients (27 F/18 M, 45-77 y, BMI = 20-41 kg/m(2)) completed the study. Drug therapies used by patients were stable throughout the duration of the trial. RESULTS: The addition of almonds to the CON diet increased plasma α-tocopherol status by a mean of 5.8%, reflecting patient compliance (P ≤0.05). However, the ALM diet did not alter vascular function assessed by measures of flow-mediated dilation, peripheral arterial tonometry, and pulse wave velocity. Further, the ALM diet did not significantly modify the serum lipid profile, blood pressure, C-reactive protein, tumor necrosis factor-α or E-selectin. The ALM diet tended to decrease vascular cell adhesion molecule-1 by 5.3% (P = 0.064) and increase urinary nitric oxide by 17.5% (P = 0.112). The ALM intervention improved the overall quality of the diet by increasing calcium, magnesium, choline, and fiber intakes above the Estimated Average Requirement (EAR) or Recommended Dietary Allowance (RDA). CONCLUSIONS: Thus, the addition of almonds to a NECP Step 1 diet did not significantly impact vascular function, lipid profile or systematic inflammation in CAD patients receiving good medical care and polypharmacy therapies but did improve diet quality without any untoward effect. TRIAL REGISTRATION: The trial was registered with the ClinicalTrials.Gov with the identifier: NCT00782015.

White matter hyperintensity volume correlates with matrix metalloproteinase-2 in acute ischemic stroke.

BACKGROUND: White matter hyperintensity (WMH), a common radiographic finding associated with stroke risk and outcome, has been linked to matrix metalloproteinase (MMP) activity and increased levels of oxidative stress in nonstroke populations. We sought to determine whether WMH severity is associated with plasma levels of MMPs and oxidative stress (F2-isoprostane) in subjects with acute ischemic stroke (AIS). METHODS: We measured plasma biomarker levels at baseline and 48 hours in consecutive AIS subjects. White matter hyperintensity volume (WMHv) was quantified on admission magnetic resonance imaging using a validated semiautomated protocol, and Spearman correlation coefficients were derived for all measured biomarkers. RESULTS: We enrolled 405 AIS subjects (mean age 70±15 years; 58% male; median WMHv 3.4 cm3, interquartile range 1.4-9.5). WMHv and age were strongly correlated (ρ=.57, P<.0001). WMHv and MMP-2 levels were correlated at baseline (ρ=.23, P<.0001) and at 48 hours poststroke (ρ=.19, P=.002). In multivariate analysis, 48-hour MMP-2 levels were independently associated with WMHv (ß=.12, P=.04). MMP-9 and F2-isioprostane levels did not correlate with WMHv. CONCLUSIONS: In AIS patients, MMP-2 levels are associated with the pre-existing burden of WMH. If validated, these findings may further elucidate the role of MMP-2 in pathophysiology of chronic cerebrovascular injury, such as WMH, and in brain susceptibility to acute ischemia.

Flavonoid intake and eye health.

In the modern era of evidence-based scientific medicine, there is little recognition of centuries of shaman observational evidence. Yet it is extremely difficult to conduct long duration controlled studies of large populations. The controversy surrounding the issue of flavonoid bioactivity and alleged benefits for eye health is also plagued by natural product industry marketing efforts that rely on small, often poorly designed studies. Ample laboratory evidence exists from in vitro and in vivo studies that provide plausible mechanistic evidence for flavonoid interactions relevant to visual function. Lacking are large randomized double-blind placebo-controlled studies in older subjects who have early signs of vision impairment. These studies could link flavonoid intake and bioavailability to efficacy in prevention of age related vision disorders that develop over decades. Support for clinical trials remains to be found before a full "recommendation" can be made regarding the value of diets high in flavonoids for eye health.

Effects of cranberry juice consumption on vascular function in patients with coronary artery disease.

BACKGROUND: Cranberry juice contains polyphenolic compounds that could improve endothelial function and reduce cardiovascular disease risk. OBJECTIVE: The objective was to examine the effects of cranberry juice on vascular function in subjects with coronary artery disease. DESIGN: We completed an acute pilot study with no placebo (n = 15) and a chronic placebo-controlled crossover study (n = 44) that examined the effects of cranberry juice on vascular function in subjects with coronary artery disease. RESULTS: In the chronic crossover study, subjects with coronary heart disease consumed a research preparation of double-strength cranberry juice (54% juice, 835 mg total polyphenols, and 94 mg anthocyanins) or a matched placebo beverage (480 mL/d) for 4 wk each with a 2-wk rest period between beverages. Beverage order was randomly assigned, and participants refrained from consuming other flavonoid-containing beverages during the study. Vascular function was measured before and after each beverage, with follow-up testing ≥12 h after consumption of the last beverage. Mean (±SD) carotid-femoral pulse wave velocity, a measure of central aortic stiffness, decreased after cranberry juice (8.3 ± 2.3 to 7.8 ± 2.2 m/s) in contrast with an increase after placebo (8.0 ± 2.0 to 8.4 ± 2.8 m/s) (P = 0.003). Brachial artery flow-mediated dilation, digital pulse amplitude tonometry, blood pressure, and carotid-radial pulse wave velocity did not change. In the uncontrolled pilot study, we observed improved brachial artery flow-mediated dilation (7.7 ± 2.9% to 8.7 ± 3.1%, P = 0.01) and digital pulse amplitude tonometry ratio (0.10 ± 0.12 to 0.23 ± 0.16, P = 0.001) 4 h after consumption of a single 480-mL portion of cranberry juice. CONCLUSIONS: Chronic cranberry juice consumption reduced carotid femoral pulse wave velocity-a clinically relevant measure of arterial stiffness. The uncontrolled pilot study suggested an acute benefit; however, no chronic effect on measures of endothelial vasodilator function was found. This trial was registered at clinicaltrials.gov as NCT00553904.

Anthocyanins are bioavailable in humans following an acute dose of cranberry juice.

Research suggests that anthocyanins from berry fruit may affect a variety of physiological responses, including endothelial function, but little information is available regarding the pharmacokinetics of these flavonoids in humans. To determine the pharmacokinetics of cranberry anthocyanins, a study was undertaken in 15 participants (age: 62 +/- 8 y) with coronary artery disease. Blood and urine samples were collected between baseline (0 h) and 4 h after consumption of 480 mL cranberry juice (54% juice; 835 mg total polyphenols; 94.47 mg anthocyanins). Marked inter-individual differences in plasma anthocyanin pharmacokinetics were observed with maximum anthocyanin concentrations detected between 1 and 3 h. Cranberry anthocyanins were bioavailable but with notable differences in the maximum concentration and area under the curve(0-4h) between individual participants. The pattern of anthocyanin glucosides observed in plasma and urine generally reflected the relative concentration determined in the juice. Plasma concentrations of the individual anthocyanins ranged between 0.56 and 4.64 nmol/L. Total recovery of urinary anthocyanin was 0.79 +/- 0.90% of the dose delivered. These data are in agreement with the pharmacokinetics of anthocyanins from other foods suggesting that cranberry anthocyanins are poorly absorbed and rapidly removed from plasma. Observed concentrations of plasma anthocyanins appear insufficient to alter radical load or redox potential but may be adequate to affect signal transduction and/or gene expression.

Xenobiotic metabolism and berry flavonoid transport across the blood-brain barrier.

A compelling body of literature suggests berry phytochemicals play beneficial roles in reversing age-related cognitive impairment and protect against neurodegenerative disorders. Anthocyanins are bioactive phytochemicals in berries suspected to be responsible for some of these neuroprotective effects. The plausible mechanisms of anthocyanin bioactivity in brain tissue are dependent on their bioavailability to the brain. Pigs were fed 2% whole freeze-dried, powdered blueberry in the diet for 8 weeks. Anthocyanin and anthocyanin glucuronides were measured in the cortex, cerebellum, and midbrain and diencephalon by LC-MS/MS. Anthocyanins and their glucuronides were found in the range of femtomoles per gram of fresh weight of tissue at 18 h postprandial, after anthocyanins had been removed from the blood by xenobiotic metabolism. Xenobiotic metabolism, anthocyanin interaction, and transporter barriers to brain bioavailability are briefly discussed. The plausible mechanism of neuroprotective action of anthocyanins may be via modulation of signal transduction processes and/or gene expression in brain tissue rather than by direct antioxidant radical quenching.

Dietary blueberry attenuates whole-body insulin resistance in high fat-fed mice by reducing adipocyte death and its inflammatory sequelae.

Adipose tissue (AT) inflammation promotes insulin resistance (IR) and other obesity complications. AT inflammation and IR are associated with oxidative stress, adipocyte death, and the scavenging of dead adipocytes by proinflammatory CD11c+ AT macrophages (ATMPhi). We tested the hypothesis that supplementation of an obesitogenic (high-fat) diet with whole blueberry (BB) powder protects against AT inflammation and IR. Male C57Bl/6j mice were maintained for 8 wk on 1 of 3 diets: low-fat (10% of energy) diet (LFD), high-fat (60% of energy) diet (HFD) or the HFD containing 4% (wt:wt) whole BB powder (1:1 Vaccinium ashei and V. corymbosum) (HFD+B). BB supplementation (2.7% of total energy) did not affect HFD-associated alterations in energy intake, metabolic rate, body weight, or adiposity. We observed an emerging pattern of gene expression in AT of HFD mice indicating a shift toward global upregulation of inflammatory genes (tumor necrosis factor-alpha, interleukin-6, monocyte chemoattractant protein 1, inducible nitric oxide synthase), increased M1-polarized ATMPhi (CD11c+), and increased oxidative stress (reduced glutathione peroxidase 3). This shift was attenuated or nonexistent in HFD+B-fed mice. Furthermore, mice fed the HFD+B were protected from IR and hyperglycemia coincident with reductions in adipocyte death. Salutary effects of BB on adipocyte physiology and ATMPhi gene expression may reflect the ability of BB anthocyanins to alter mitogen-activated protein kinase and nuclear factor-kappaB stress signaling pathways, which regulate cell fate and inflammatory genes. These results suggest that cytoprotective and antiinflammatory actions of dietary BB can provide metabolic benefits to combat obesity-associated pathology.

Identification of anthocyanins in the liver, eye, and brain of blueberry-fed pigs.

Dietary intervention with anthocyanins may confer benefits in brain function, including vision. Research to date indicates that animals have only a limited capacity to absorb anthocyanins, compared to other types of flavonoids. Pigs, which are a suitable model for human digestive absorption, were used to examine the deposition of anthocyanins in tissues including the liver, eye, and brain tissue. Pigs were fed diets supplemented with 0, 1, 2, or 4% w/w blueberries ( Vaccinium corymbosum L. 'Jersey') for 4 weeks. Prior to euthanasia, pigs were fasted for 18-21 h. Although no anthocyanins were detected in the plasma or urine of the fasted animals, intact anthocyanins were detected in all tissues where they were sought. LC-MS/MS results are presented for the relative concentration of 11 intact anthocyanins in the liver, eye, cortex, and cerebellum. The results suggest that anthocyanins can accumulate in tissues, including tissues beyond the blood-brain barrier.

Antioxidant activity and metabolite profile of quercetin in vitamin-E-depleted rats.

Dietary antioxidants interact in a dynamic fashion, including recycling and sparing one another, to decrease oxidative stress. Limited information is available regarding the interrelationships in vivo between quercetin and vitamin E. We investigated the antioxidant activity and metabolism of quercetin (Q) in 65 F-344 rats (n=13 per group) randomly assigned to the following vitamin E (VE)-replete and -deficient diets: (a) VE replete (30 mg alpha-tocopherol acetate/kg diet) control ad libitum (C-AL), (b) VE replete pair fed (C-PF), (c) VE replete+5.0 g Q/kg diet (R-VE+5Q), (d) VE deplete (<1 mg/kg total tocopherols)+5.0 g Q/kg diet (D-VE+5Q) and (e) D-VE. After 12 weeks, blood and tissue were collected for measurement of plasma vitamin E, quercetin and its metabolites, serum pyruvate kinase (PK), plasma protein carbonyls, malondialdehyde (MDA) and oxygen radical absorbance capacity. D-VE diets decreased serum alpha-tocopherol and increased PK activity in a time-dependent manner. The D-VE diet increased plasma protein carbonyls but did not affect MDA. Dietary quercetin supplementation increased quercetin and its metabolites in plasma and liver but did not affect D-VE-induced changes in plasma alpha-tocopherol, PK or protein carbonyls. Plasma isorhamnetin and its disposition in muscle were enhanced by the D-VE diet, as compared to the R-VE diet. Conversely, tamarixetin disposition in muscle was decreased by the D-VE diet. Thus, quercetin did not slow vitamin E decline in vivo; neither did it provide antioxidant activity in vitamin-E-depleted rats. However, vitamin E status appears to enhance the distribution of isorhamnetin into the circulation and its disposition in muscle.

Avenanthramides are bioavailable and have antioxidant activity in humans after acute consumption of an enriched mixture from oats.

The consumption of polyphenols is associated with a decreased risk of cardiovascular disease. Avenanthramides (AV), alkaloids occurring only in oats, may have anti-atherosclerotic activity, but there is no information concerning their bioavailability and bioactivity in humans. We characterized the pharmacokinetics and antioxidant action of avenanthramide A, B, and C in healthy older adults in a randomized, placebo-controlled, 3-way crossover trial with 1-wk washout periods. Six free-living subjects (3 mol/L, 3 F; 60.8 +/- 3.6 y) consumed 360 mL skim milk alone (placebo) or containing 0.5 or 1 g avenanthramide-enriched mixture (AEM) extracted from oats. Plasma samples were collected over a 10-h period. Concentrations of AV-A, AV-B, and AV-C in the AEM were 154, 109, and 111 micromol/g, respectively. Maximum plasma concentrations of AV (free + conjugated) after consumption of 0.5 and 1 g AEM were 112.9 and 374.6 nmol/L for AV-A, 13.2 and 96.0 nmol/L for AV-B, and 41.4 and 89.0 nmol/L for AV-C, respectively. Times to reach the C(max) for both doses were 2.30, 1.75, and 2.15 h for AV-A, AV-B, and AV-C and half times for elimination were 1.75, 3.75, and 3.00 h, respectively. The elimination kinetics of plasma AV appeared to follow first-order kinetics. The bioavailability of AV-A was 4-fold larger than that of AV-B at the 0.5 g AEM dose. After consumption of 1 g AEM, plasma reduced glutathione was elevated by 21% at 15 min (P < or = 0.005) and by 14% at 10 h (P < or = 0.05). Thus, oat AV are bioavailable and increase antioxidant capacity in healthy older adults.

Effect of almond skin polyphenolics and quercetin on human LDL and apolipoprotein B-100 oxidation and conformation.

Almond skin polyphenolics (ASP) and vitamin C (VC) or E (VE) inhibit the Cu(2+)-induced generation of conjugated dienes in human low-density lipoprotein (LDL) in a synergistic manner. However, the mechanism(s) by which this synergy occurs is unknown. As modification of apolipoprotein (apo) B-100 is an early, critical step in LDL oxidation, we examined the effects of combining ASP or quercetin and antioxidant vitamins on the oxidation of this moiety as well as on the alteration of LDL conformation and electronegativity (LDL-). In a dose-dependent manner, ASP (0.12-2.0 micromol/L gallic acid equivalents) decreased tryptophan (Trp) oxidation by 6.7-75.7%, increased the generalized polarity (Gp) of LDL by 21.0-81.5% at 90 min and reduced the ratio of LDL- to total LDL (tLDL) by 38.2-83.8% at 5 h. The actions of ASP on these parameters were generally additive to those of VC and VE. However, a 10-25% synergy of ASP plus VC in protecting apo B-100 Trp against oxidation may result from their synergistic interaction in prolonging the lag time to oxidation. ASP and VE acted in synergy to reduce LDL-/tLDL by 24-43%. Quercetin's actions were similar to ASP, though more effective at inhibiting Trp oxidation. Thus, ASP and quercetin reduce the oxidative modification of apo B-100 and stabilize LDL conformation in a dose-dependent manner, acting in an additive or synergistic fashion with VC and VE.

Bilberry (Vaccinium myrtillus) anthocyanins modulate heme oxygenase-1 and glutathione S-transferase-pi expression in ARPE-19 cells.

PURPOSE: To determine whether anthocyanin-enriched bilberry extracts modulate pre- or posttranslational levels of oxidative stress defense enzymes heme-oxygenase (HO)-1 and glutathione S-transferase-pi (GST-pi) in cultured human retinal pigment epithelial (RPE) cells. METHODS: Confluent ARPE-19 cells were preincubated with anthocyanin and nonanthocyanin phenolic fractions of a 25% enriched extract of bilberry (10(-6)-1.0 mg/mL) and, after phenolic removal, cells were oxidatively challenged with H(2)O(2). The concentration of intracellular glutathione was measured by HPLC and free radical production determined by the dichlorofluorescin diacetate assay. HO-1 and GST-pi protein and mRNA levels were determined by Western blot and RT-PCR, respectively. RESULTS: Preincubation with bilberry extract ameliorated the intracellular increase of H(2)O(2)-induced free radicals in RPE, though H(2)O(2) cytotoxicity was not affected. By 4 hours, the extract had upregulated HO-1 and GST-pi protein by 2.8- and 2.5-fold, respectively, and mRNA by 5.5- and 7.1-fold, respectively, in a dose-dependent manner. Anthocyanin and nonanthocyanin phenolic fractions contributed similarly to mRNA upregulation. CONCLUSIONS: Anthocyanins and other phenolics from bilberry upregulate the oxidative stress defense enzymes HO-1 and GST-pi in RPE, suggesting that they stimulate signal transduction pathways influencing genes controlled by the antioxidant response element.

Determination of flavonoids and phenolics and their distribution in almonds.

Limited information is available concerning the qualitative and quantitative composition of polyphenolic compounds, especially flavonoids, in almonds. We determined total phenols, flavonoids, and phenolic acids in California almond (Prunus dulcis) skins and kernels among the principal almond varieties (Butte, Carmel, Fritz, Mission, Monterey, Nonpareil, Padre, and Price) with high-performance liquid chromatography (HPLC)/electrochemical detection and UV detection. Liquid chromatography/tandem mass spectrometry under identical HPLC conditions was utilized to verify identities of the predominant flavonoids and phenolic acids. Total phenols ranged from 127 (Fritz) to 241 (Padre) mg gallic acid equivalents/100 g of fresh weight. The analyses were compiled to produce a data set of 18 flavonoids and three phenolic acids. The predominant flavonoids were isorhamnetin-3-O-rutinoside and isorhamnetin-3-O-glucoside (in combination), catechin, kaempferol-3-O-rutinoside, epicatechin, quercetin-3-O-galactoside, and isorhamnetin-3-O-galactoside at 16.81, 1.93, 1.17, 0.85, 0.83, and 0.50 mg/100 g of fresh weight almonds, respectively. Using the existing approach of calculating only the aglycone form of flavonoids for use in the U.S. Department of Agriculture nutrient database, whole almonds would provide the most prevalent aglycones of isorhamnetin at 11.70 (3.32), kaempferol at 0.60 (0.17), catechin at 1.93 (0.55), quercetin at 0.72 (0.20), and epicatechin at 0.85 (0.24) mg/100 g of fresh weight (mg/oz serving), respectively. These data can lead to a better understanding of the mechanisms of action underlying the relationship between almond consumption and health-related outcomes and provide values for whole and blanched almonds suitable for inclusion in nutrient databases.

Rat gastrointestinal tissues metabolize quercetin.

Quercetin and quercetin glycosides from food or dietary supplements appear in body tissues almost exclusively as glucuronated, sulfated, and methylated quercetin conjugates, suggesting that the in vivo bioactivity of quercetin may be due to its metabolites. In this study, pre- and postabsorptive metabolism of orally ingested quercetin was examined by comparing the metabolite pattern in gastrointestinal (GI) tissues, contents, and internal tissues. F344 rats (n = 6) were fed for 6 wk a diet containing 0.45% quercetin and the metabolite patterns were determined in the tissues and contents of stomach, small intestine, cecum, and colon and in liver, kidney, and plasma using LC-MS/MS. GI contents contained predominantly unmetabolized quercetin at 94-100%, whereas quercetin in GI tissues was present as 11 different sulfated, glucuronated, and methylated metabolites at 32% in stomach, 88% in small intestine, 27% in cecum, and 46% in colon. Quercetin was further metabolized postabsorption and found in liver, kidney, and plasma almost exclusively as sulfated methyl-quercetin glucuronide. The unique pattern of quercetin metabolites in each GI tissue indicates extensive biotransformation before absorption and distribution in rats.

Flavonols, flavones, flavanones, and human health: epidemiological evidence.

Polyphenolic flavonoids are among a wide variety of phytochemicals present in the human diet. Basic research, animal model, and human studies suggest flavonoid intake may reduce the risk of several age-related chronic diseases. The vast number of flavonoids and mixtures of their subclasses, including flavonols, flavones, and flavanones, and the variety of agricultural practices that affect their concentration in foods have presented a challenge to the development of adequate food composition databases for these compounds. Nonetheless, dietary assessments have been applied to cohort and case-control epidemiological studies, and several reveal an inverse association with risk of some forms of cancer, cardiovascular disease, and other chronic conditions. Those observational studies that have examined these relationships with regard to flavonols, flavones, and flavanones are reviewed. The requirement for caution in interpreting these studies is discussed with regard to the limited information available on the bioavailability and biotransformation of these flavonoids. As the totality of the available evidence on these flavonoids suggests a role in the prevention of cancer and cardiovascular disease, further research is warranted, particularly in controlled clinical trials.

Antioxidant status and biomarkers of oxidative stress in dogs with congestive heart failure.

Alterations in antioxidant status and oxidative stress have been documented in dogs with dilated cardiomyopathy (DCM). The purpose of this study was to more broadly assess this relationship in dogs with congestive heart failure (CHF). Malondialdehyde (MDA), 8-F(2alpha)-isoprostane, protein carbonyls, reduced (GSH) and oxidized (GSSG) glutathione, vitamins A, C, and E, and oxygen radical absorbance capacity (ORAC) were measured from a single venous blood sample from dogs with CHF secondary to DCM or chronic valvular disease (CVD) and in healthy controls. Nineteen dogs with CHF (14 CVD and 5 DCM) and 12 healthy controls were enrolled in the study. Concentrations of 8-F(2alpha)-isoprostane (CHF: 44.6 pg/mL [range, 27.1-98.0 pg/mL], controls: 25.3 pg/ mL [range, 11.1-80.4 pg/mL]) but not MDA (CHF: 4.11 microM [range, 1.89-6.39 microM], controls: 3.88 microM [range, 2.14-4.72 microM]) or protein carbonyls (0.69 nmol/mg protein [range, 0.37-1.67 nmol/mg protein], controls: 0.80 nmol/mg protein [range, 0.40-1.14 nmol/mg protein]) were significantly higher in the dogs with CHF than in the controls. Vitamin E concentration (CHF: 2,215 microg/ dL [range, 916-3,499 microg/dL], controls: 2,820 microg/dL [range, 1,738-3,775 microg/dL]) and GSH:GSSG (CHF: 12.0 [range, 3.69-30.1], controls: 22.7 [range, 12.5-227]) were significantly lower, whereas ORAC (CHF: 824 micromol Trolox equivalent/L [range, 304-984], controls: 497 micromol Trolox equivalent/L [range, 258-759]) and vitamin C (CHF: 0.90 mg/dL [range, 0.55-2.02 mg/dL], controls: 0.72 mg/dL [range, 0.43-0.85 mg/dL]) concentrations were higher in dogs with CHF than in controls. Vitamin A concentrations were not different between dogs with CHF and controls. No differences in any of the parameters were detected between dogs with DCM versus those with CVD. Some antioxidant defenses are decreased in dogs with CHF, and some biomarkers of oxidative stress are increased in dogs with CHF. The effect of dietary interventions to correct this imbalance in antioxidant defenses warrants further study.

Flavonoids from almond skins are bioavailable and act synergistically with vitamins C and E to enhance hamster and human LDL resistance to oxidation.

Consumption of tree nuts such as almonds has been associated with a reduced risk of coronary heart disease. Flavonoids, found predominantly in the skin of almonds, may contribute to their putative health benefit, but their bioactivity and bioavailability have not previously been studied. Almond skin flavonoids (ASF) were extracted with HCl:H2O:methanol (1:19:80) and their content of catechins and flavonols identified by HPLC with electrochemical detection. ASF bioactivity was assessed in vitro by their capacity to increase the resistance of human LDL to oxidation induced by 10 micromol/L Cu2+. ASF from 0.18 to 1.44 mumol gallic acid equivalent (GAE)/L increased the lag time to LDL oxidation in a dose-dependent manner (P < or = 0.0001). Combining ASF with vitamin E or ascorbic acid extended the lag time >200% of the expected additive value (P < or = 0.05). The bioavailability and in vivo antioxidant activity of 40 micromol ASF were examined in BioF1B hamsters. Peak plasma concentrations of catechin, epicatechin, and flavonols (quercetin, kaempferol, and isorhamnetin) occurred at 60, 120, and 180 min, respectively. The concentration of isorhamnetin was significantly elevated in liver at 180 min. Absorbed ASF enhanced the ex vivo resistance of hamster LDL collected at 60 min to oxidation by 18.0% (P = 0.028), and the in vitro addition of 5.5 micromol/L vitamin E synergistically extended the lag time of the 60-min sample by 52.5% (P < or = 0.05). Thus, ASF possess antioxidant capacity in vitro; they are bioavailable and act in synergy with vitamins C and E to protect LDL against oxidation in hamsters.