Effects of blueberry and cranberry juice consumption on the plasma antioxidant capacity of healthy female volunteers.

OBJECTIVE: To assess whether consumption of 500 ml of blueberry juice or cranberry juice by healthy female subjects increased plasma phenolic content and antioxidant capacity. DESIGN: Latin square arrangement to eliminate ordering effects. After an overnight fast, nine volunteers consumed 500 ml of blueberry juice, cranberry juice or a sucrose solution (control); each volunteer participated on three occasions one week apart, consuming one of the beverages each time. Blood samples were obtained by venipuncture at intervals up to four hours after consumption of the juices. Urine samples were also obtained four hours after consuming the juice. RESULTS: Consumption of cranberry juice resulted in a significant increase in the ability of plasma to reduce potassium nitrosodisulphonate and Fe(III)-2,4, 6-Tri(2-pyridyl)-s-triazine, these measures of antioxidant capacity attaining a maximum after 60-120 min. This corresponded to a 30% increase in vitamin C and a small but significant increase in total phenols in plasma. Consumption of blueberry juice had no such effects. CONCLUSION: The increase in plasma antioxidant capacity following consumption of cranberry juice could mainly be accounted for by an increase in vitamin C rather than phenolics. This also accounted for the lack of an effect of the phenolic-rich but vitamin C-low blueberry juice. SPONSORSHIP: Funded by the Scottish Executive Rural Affairs Department and the Danish Government.

Relationship among antioxidant activity, vasodilation capacity, and phenolic content of red wines.

The relationship among antioxidant activity, based on the electron-spin resonance determination of the reduction of Fremy's radical, vasodilation activity, and phenolic content was investigated in 16 red wines. The wines were selected to provide a range of origins, grape varieties, and vinification methods. Sensitive and selective HPLC methods were used for the analysis of the major phenolics in red wine: free and conjugated myricetin, quercetin, kaempferol, and isorhamnetin; (+)-catechin, (-)-epicatechin, gallic acid, p-coumaric acid, caffeic acid, caftaric acid, trans-resveratrol, cis-resveratrol, and trans-resveratrol glucoside. Total anthocyanins were measured using a colorimetric assay. The total phenolic content of the wines was determined according to the Folin-Ciocalteu colorimetric assay and also by the cumulative measurements obtained by HPLC. The 16 wines exhibited a wide range in the values of all parameters investigated. However, the total phenol contents, measured both by HPLC and colorimetrically, correlated very strongly with the antioxidant activity and vasodilation activity. In addition, the antioxidant activity was associated with gallic acid, total resveratrol, and total catechin. In contrast, only the total anthocyanins were correlated with vasodilation activity. The results demonstrate that the different phenolic profiles of wines can produce varying antioxidant and vasodilatant activities, which opens up the possibility that some red wines may provide enhanced health benefits for the consumer.

Assessment of the antioxidant potential of scotch whiskeys by electron spin resonance spectroscopy: relationship to hydroxyl-containing aromatic components.

Electron spin resonance (ESR) spectroscopy has been used to assess the antioxidant capacity of eight Scotch whiskeys by measuring the extent by which the original spirits, or pyridine solutions of their residues, reduced Fremy's radical or galvinoxyl radical. All whiskeys displayed antioxidant activity greater than that of a 0.2 mM solution of Trolox in the Fremy's assay and of a 0.1 mM solution of quercetin in the galvinoxyl assay. The relative antioxidant capacities determined according to the two assays were highly correlated and strongly related to the total phenol content as determined by using the Folin-Ciocalteu method. Activity was a consequence of maturation in oak casks with the "newmake" spirit showing no effect. Of 10 aromatic constituents analyzed, activity was most strongly correlated with ellagic acid and gallic acid in both assays. The reductive capacities of four major phenolics were determined, which, in summation, accounted for 31-53% of the total antioxidant activity of the whiskeys. There was no evidence for synergistic interaction between the phenols investigated.

Antioxidant efficacy of phytoestrogens in chemical and biological model systems.

Phytoestrogens (PEs) are diphenolic compounds from plants which can bind to estrogen receptors and have estrogen and antiestrogen effects in man and animals. Like other plant phenolics, PEs may have antioxidant properties through hydrogen/electron donation via hydroxyl groups. They might therefore act as free radical scavengers and inhibit development of coronary heart disease and cancers. The hydrogen-donating ability of a range of phytoestrogens was assessed using electron spin resonance spectroscopy, the ferric-reducing ability of plasma assay, and the Trolox equivalent antioxidant capacity. In addition, the ability of compounds to inhibit lipid peroxidation was examined in vitamin E-deficient liver microsomes. Genistein had the highest activity of the isoflavones; however, the isoflavones were relatively poor hydrogen donors compared with the other estrogenic compounds examined. Coumestrol and equol were more effective antioxidants than genistein but had relatively limited activity in comparison with Trolox. The only estrogenic compound with significant antioxidant activity was kaempferol which is better known as a dietary antioxidant than a phytoestrogen. As the concentrations of PEs used in this study exceed the estimated serum concentrations, their relatively poor antioxidant ability in vitro may indicate little significance as antioxidants in vivo.

The effect of whisky and wine consumption on total phenol content and antioxidant capacity of plasma from healthy volunteers.

OBJECTIVE: To assess whether consumption of 100 ml of whisky or red wine by healthy male subjects increased plasma total phenol content and antioxidant capacity. DESIGN: A Latin square arrangement to eliminate ordering effects whereby, after an overnight fast, nine volunteers consumed 100 ml of red wine, malt whisky or unmatured 'new make' spirit. Each volunteer participated on three occasions one week apart, consuming one of the beverages each time. Blood samples were obtained from the anticubital vein at intervals up to 4h after consumption of the beverages when a urine sample was also obtained. RESULTS: Within 30 min of consumption of the wine and whisky, there was a similar and significant increase in plasma total phenol content and antioxidant capacity as determined by the ferric reducing capacity of plasma (FRAP). No changes were observed following consumption of 'new make' spirit. CONCLUSIONS: Consumption of phenolic-containing alcoholic beverages transiently raises total phenol concentration and enhances the antioxidant capacity of plasma. This is compatible with suggestions that moderate alcohol usage and increased antioxidant intake decrease the risk of coronary heart disease.

Electron spin resonance assessment of susceptibility of roe deer (Capreolus capreolus) and red deer (Cervus elaphus) to oilseed rape (Brassica napus) poisoning.

Ex vivo studies have been carried out on roe and red deer erythrocytes using electron spin resonance spectroscopy. Free radical formation in cells challenged with the brassica-derived haemolysin, dimethyldisulphide, was measured using spin trapping techniques. Significantly greater amounts of radical were trapped in the roe deer cells which may relate to differences in the antioxidant profile of the two genotypes. Results suggest that roe deer have a greater risk of developing oilseed rape poisoning than red deer.

Adaptation of the blood antioxidant defence mechanisms of sheep with a genetic lesion resulting in low red cell glutathione concentrations.

Finnish Landrace sheep with a genetic lesion which results in restricted cysteine transport across the red cell membrane have total glutathione concentrations in their red blood cells that are approximately 40% of those in normal sheep of the same breed. However, dimethyldisulphide-challenged red blood cells from both phenotypes produce an ESR-spin adduct at similar rates. The resistance of the low glutathione phenotype red cells to oxidant challenge is reflected by increases in the activities of antioxidant enzymes. Sheep with a genotypic disorder in cysteine transport may be a suitable model for studying the genetic expression of antioxidant enzymes in response to oxidant loads.

The role of free radicals in brassica-induced anaemia of sheep: an ESR spin trapping study.

The formation of reactive free radical species in sheep erythrocytes challenged with dimethyldisulphide, a brassica-derived haemolysin, has been investigated by electron spin resonance spectroscopy using the spin trap alpha-(4-pyridyl 1-oxide)-N-tert-butylnitrone. Erythrocytes exposed to this agent undergo a burst of free radical activity as demonstrated by the appearance of a spin adduct. The results suggest that haemolytic anaemia which can occur in sheep grazing forage brassicas is a consequence of oxidative stress.

Dietary vitamin E decreases ESR signal intensity in hepatic microsomal preparations from malignant hyperthermia susceptible pigs.

On incubation with the spin trap alpha-(4-pyridyl-l-oxide)-N-tert-butylnitrone (4-POBN), a characteristic electron spin resonance (ESR) signal was produced at a greater rate in hepatic microsomal fractions from malignant hyprthermia susceptible (MHS) pigs compared with resistant (MHR) pigs. This was accompanied by increased formation of thiobarbituric acid reactive substances (TBARS). Supplementation of diets for six weeks with 235 mg alpha-tocopherol acetate/kg significantly increased microsomal vitamin E content of both pigs types. Moreover, the rate of formation of TBARS and ESR signal height of incubated microsomes from supplemented MHS pigs was decreased to that of MHR pigs. Elevated pyruvate kinase activities and TBARS concentrations in plasma of MHS pigs were also moderated by dietary vitamin E. Vitamin E supplementation may decrease the peroxidative events associated with MH.

Spin trapping of free radicals and lipid peroxidation in microsomal preparations from malignant hyperthermia susceptible pigs.

Microsomes were prepared from livers of malignant hyperthermia susceptible (MHS) or resistant (MHR) pigs. On incubation with the spin trap alpha-(4-pyridyl-l-oxide)-N-tert-butylnitrone (4-POBN), the microsomes from MHS pigs produced a characteristic electron spin resonance (ESR) signal at a greater rate than those from MHR pigs. Increased formation in the incubations of thiobarbituric acid reactive substances (TBARS) by the microsomes of the MHS pigs indicated an enhanced susceptibility to free radical-mediated lipid peroxidation. These results provide further evidence that MHS pigs have an antioxidant abnormality which may contribute to the fatal MH response. However the nature of the abnormality is unclear. The enhanced formation of unstable free radicals and indices of lipid peroxidation was not due to decreased vitamin E concentration or glutathione peroxidase activity in the microsomes. Furthermore, fatty acid profiles were similar in microsomes from MHS and MHR pigs indicating similar amounts of potential substrate for TBARS formation.

An electron spin resonance investigation of internal rust spot, a physiological disorder of the potato tuber.

ESR spectroscopy has been used to investigate the formation of paramagnetic species during the development of internal rust spot (IRS) in the potato tuber. Production of free radicals and oxidation of metal ions such as Fe(II) and Mn(II) occur when necrotic tissue is formed. However, since IRS develops in periods of calcium stress (low calcium supply), it is suggested that the principal cause of the disorder is a loss of cell membrane integrity which is brought about by a lack of calcium. Cell senescence and the formation of necrotic tissue may then result either from increased oxygen radical production within the cell or from oxidation of metal complexes in the extracellular regions of the tissue.

Spin trapping of free radicals in homogenates of heart from selenium and vitamin E deficient rats.

The formation of unstable free radical species in homogenates of heart from selenium and/or vitamin E deficient rats has been investigated by electron spin resonance spectroscopy using the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone. There was only a small amount of free radical formation by heart from control or selenium deficient animals. However, the amounts and rates of formation of free radicals were significantly higher in animals with vitamin E deficiency and higher still in those with combined selenium and vitamin E deficiency.

The influence of dietary history on the production of free radicals in rat liver microsomes.

Free radicals have been trapped in microsomal fractions from rat livers by the use of alpha-(4-pyridyl-l-oxide)-N-tert-butylnitrone (4-POBN) and detected by ESR spectroscopy. Using this technique we have investigated the effects of low dietary concentrations of vitamin E and selenium (Se) on free radical production. When the microsomal fractions were incubated with Fe, ADP and NADPH significantly greater amounts of radicals were initially trapped when rats were deficient in vitamin E (whether combined with Se deficiency or not) than from Se-deficient animals or Se- and vitamin E-supplemented controls. After prolonged incubation (ca. 30-45 mins) the free radicals trapped from the microsomes from the Se-deficient animals rose to amounts comparable to those seen in the vitamin E-deficient microsomes. Over the same period of incubation approximately half this quantity of radicals were trapped in liver microsomes from Se- and vitamin E-supplemented rats. The effects of 4-POBN on the levels of peroxidation in the various microsomal fractions were assessed by measuring the thiobarbituric acid reactive substances (TBARS). The presence of 4-POBN significantly reduced the amounts of TBARS formed on incubation and there was a clear distinction between the groups on the basis of the vitamin E status of their diets.

Oxygen-induced free radical in wheat roots.

ESR experiments have shown that free radicals are formed in wheat plant roots as a result of exposure to O2. Although the radical(s) has not been positively identified, the nature of the spectra allows simple oxygen-derived radicals, such as O2-, HO2. and OH., to be excluded as possibilities. Adsorption of Cu(II), but not Zn(II) or Cd(II) by the root results in a rapid decay of the radical signal to reach a level of 10% of its original intensity after a few minutes.

The occurrence of copper, iron, zinc and other elements and the nature of some copper and iron complexes in humic substances from municipal refuse disposed of in a landfill.

Municipal refuse was allowed to decompose in a simulated landfill for 20 months. Three different models were studied in which the refuse, in 40 m3 lots, was either compacted or mixed 2:1 with sewage sludge, the latter being studied in both uncompacted and compacted states. At 2, 6, 12 and 20 months, humic substances were extracted from samples with 0.1 M Na2P4O7 and 0.1 M NaOH, and humic acids isolated by precipitation after acidification with HCl. The humic substances were examined by electron paramagnetic resonance (EPR) to determine the free radical content and the nature of some of the metal complexes present. The principal form of copper was present in either a square planar or a tetragonally-distorted octahedral environment, probably coordinated to two nitrogen and two oxygen atoms. Fe3+ was present in at least three different environments. In one, it was probably in a complex with rhombic symmetry; another showed Fe3+ in an axially-symmetric environment, most likely as a ferric porphyrin. No distinctions could be made between the concentrations or forms of metals present in the refuse humic substances as a result of adding sewage sludge, but the additions increased the yield of humic substances, particularly in the uncompacted landfill. Humic substances in the refuse retain metal elements in complexed forms which will restrict their release from the landfill.