Ex Vivo Antioxidant Capacities of Fruit and Vegetable Juices. Potential In Vivo Extrapolation.

BACKGROUND: In support of claims that their products have antioxidant properties, the food industry and dietary supplement manufacturers rely solely on the in vitro determination of the ORAC (oxygen radical antioxidant capacity) value, despite its acknowledged lack of any in vivo relevance. It thus appears necessary to use tests exploiting biological materials (blood, white blood cells) capable of producing physiological free radicals, in order to evaluate more adequately the antioxidant capacities of foods such as fruit and vegetable juices. MATERIALS: Two approaches to assessing the antioxidant capacities of 21 commercial fruit and vegetable juices were compared: the ORAC assay and the "PMA-whole blood assay," which uses whole blood stimulated by phorbol myristate acetate to produce the superoxide anion. We described in another paper the total polyphenol contents (TPCs) and individual phenolic compound contents of all the juices were investigated. RESULTS: Ranking of the juices from highest to lowest antioxidant capacity differed considerably according to the test used, so there was no correlation (r = 0.33, p = 0.13) between the two assays when considering all juices. Although the results of the ORAC assay correlated positively with TPC (r = 0.50, p = 0.02), a much stronger correlation (r = 0.70, p = 0.004) emerged between TPC and % superoxide anion inhibition. In the PMA-whole blood assay, peonidin-3-O-glucoside, epigallocatechin gallate, catechin, and quercetin present in juices were found to inhibit superoxide anion production at concentrations below 1 µM, with a strong positive correlation. CONCLUSIONS: Associated with the determination of total and individual phenolic compounds contained in fruit and vegetable juices, the PMA-whole blood assay appears better than the ORAC assay for evaluating juice antioxidant capacity.

Compared Phenolic Compound Contents of 22 Commercial Fruit and Vegetable Juices: Relationship to ex-vivo Vascular Reactivity and Potential in vivo Projection.

The real impact of polyphenol-rich vegetable and fruit juice intake on cardiovascular health remains a matter of controversy. In the present study, rat aorta segments immersed in an organ bath (OB) were used to explore whether the total polyphenol content and/or individual phenolic compound contents of 22 commercial vegetable (n = 3) and fruit juices [(citrus (n = 5), berries (n = 10), apple (n = 2), pineapple (n = 2)] might be associated with vascular tone. Red juices (particularly blackcurrant) and lemon juice caused the most marked vasorelaxation, its amplitude being endothelium dependent or not according to the volume ratio of juice to initial OB solution Vjuice/VOBS). At volume ratios 5% and 10%, both the juice and OB total polyphenol for all juices and total anthocyanin contents for berry juices significantly correlated with aorta vasorelaxation intensity. This was not the case for total or individual flavonols (except kaempferol) or for total or individual flavanols (except epigallocatechin gallate). If one relates our measured concentrations of individual phenolic compounds in OB to what is known about their physiological concentrations, and given our evidenced correlations between compound concentrations and vasorelaxation intensity, kaempferol, epigallocatechin gallate and peonidin-3-O-glucoside seem to emerge as the interesting phenolic compounds likely to be responsible for the potent vasorelaxation observed with fruit juices, and more particularly blackcurrant ones. Clinical investigation is required, however, to confirm our observations.

PAOT-Liquid® Technology: An Easy Electrochemical Method for Evaluating Antioxidant Capacity of Wines.

Polyphenol compounds present in high quantity in wines are well-known to have potent cardio-protective properties through several biological mechanisms including antioxidant activity [1]. A large number of methods have been developed for evaluating the antioxidant capacity of food matrices. Most of them have, however, the disadvantage of being time consuming and require specific analytical protocols and devices. In the present study, we present the electrochemical PAOT (Pouvoir Antioxydant Total)-Liquid® Technology which can be easily used by winemakers for evaluating antioxidant activity of wine during all steps of making process. The methodology is based on the measurement of electric potential variation resulting from chemical reactions between wine polyphenols and a free radical mediator M• as source of oxidants. Total antioxidant activity as estimated by the PAOT-Liquid® activity was 6.8 fold higher in red wines (n = 14) when compared to rosé (n = 3) and white (n = 3) wines bought in a commercial market. Moreover, PAOT-Liquid® activity was highly correlated with total polyphenols content (TPC) of all wines (r = 0.9540, p < 0.0001) and the classical DPPH (2,2-diphenyl-1-picryhydrazyl) assay which is often used for evaluating antioxidant capacity of food matrices (r = 0.9102, p < 0.0001).

Validation of a food frequency questionnaire assessing dietary polyphenol exposure using the method of triads.

When conducting research on polyphenols and their effects on health, it is of primary importance to use standardised and validated dietary assessment tools. This paper aims at assessing the validity of a food frequency questionnaire (FFQ) for quantifying dietary polyphenol exposure among healthy adults using the method of triads. Fifty-three healthy adults, aged 20-60, were included in the study. Total dietary polyphenol intake (TDP) estimated by the FFQ was compared with TDP measured by a 3-day food record (FR) and with urinary excretion levels of total polyphenols (TUP). Pearson correlations were calculated between methods. Validity coefficients (VC) were estimated between the three measurements and the 'unknown' true intake. There was a strong correlation between both dietary methods (r = 0.70, p < 0.0001). A moderate but significant association was observed between FFQ-derived TDP and TUP (r = 0.32, p = 0.020). The method of triads yielded a VC for the FFQ of 0.63 (95%CI: 0.41-0.84), indicating a strong relationship between FFQ-derived TDP and the true polyphenol intake. This study shows that the FFQ is an adequate tool not only for measuring dietary polyphenol exposure in nutrition epidemiological studies but also for guiding clinicians in dietary advice and counselling.

The potency of commercial blackcurrant juices to induce relaxation in porcine coronary artery rings is not correlated to their antioxidant capacity but to their anthocyanin content.

OBJECTIVE: Polyphenol-rich products such as fruit juices have been found to have strong antioxidant capacities and to induce potent endothelium-dependent relaxation. We evaluated whether the commercial blackcurrant juices induced endothelium-dependent relaxation of isolated coronary arteries can be related to their antioxidant capacity and/or phenolic content. METHODS: Six different commercial blackcurrant juices were selected. Their main phenolic compounds were measured by ultra-performance liquid chromatography and antioxidant capacity was evaluated by spectrometric methods. Vascular reactivity studies with these juices were done using isolated porcine coronary arteries. RESULTS: The six different commercial blackcurrant juices induced relaxation ranging from 21% to 100% at the concentration of 0.5% volume per volume (v/v). The relaxation induced at 0.5% v/v was not correlated to their antioxidant capacity measured by either oxygen radical antioxidant capacity or DPPH (2,2-diphenyl-1-picrylhydrazyl) assays and also not to the ascorbic acid, total polyphenols, total flavanols, and total phenolic acid contents. In contrast, the amplitude of the relaxation was correlated to the total anthocyanins content and the individual anthocyanin concentration. CONCLUSIONS: Correlations between relaxation amplitude and total anthocyanin or individual anthocyanin contents are of interest for the development of functional blackcurrant beverages with the potential to promote vascular protection.

Polyphenol-Rich Blackcurrant Juice Prevents Endothelial Dysfunction in the Mesenteric Artery of Cirrhotic Rats with Portal Hypertension: Role of Oxidative Stress and the Angiotensin System.

Chronic liver diseases with portal hypertension are characterized by a progressive vasodilatation, endothelial dysfunction, and NADPH oxidase-derived vascular oxidative stress, which have been suggested to involve the angiotensin system. This study evaluated the possibility that oral intake of polyphenol-rich blackcurrant juice (PRBJ), a rich natural source of antioxidants, prevents endothelial dysfunction in a rat model of cirrhosis induced by chronic bile duct ligation (CBDL), and, if so, determined the underlying mechanism. Male Wistar rats received either control drinking water or water containing 60 mg/kg gallic acid equivalents of PRBJ for 3 weeks before undergoing surgery with CBDL or sham surgery. After 4 weeks, vascular reactivity was assessed in mesenteric artery rings using organ chambers. Both the acetylcholine-induced nitric oxide (NO)- and endothelium-dependent hyperpolarization (EDH)-mediated relaxations in mesenteric artery rings were significantly reduced in CBDL rats compared to sham rats. An increased level of oxidative stress and expression of NADPH oxidase subunits, COX-2, NOS, and of the vascular angiotensin system are observed in arterial sections in the CBDL group. Chronic intake of PRBJ prevented the CBDL-induced impaired EDH-mediated relaxation, oxidative stress, and expression of the different target proteins in the arterial wall. In addition, PRBJ prevented the CBDL-induced increase in the plasma level of proinflammatory cytokines (interleukin [IL]-1α, monocyte chemotactic protein 1, and tumor necrosis factor α) and the decrease of the anti-inflammatory cytokine, IL-4. Altogether, these observations indicate that regular ingestion of PRBJ prevents the CBDL-induced endothelial dysfunction in the mesenteric artery most likely by normalizing the level of vascular oxidative stress and the angiotensin system.

Bilberry extract (Antho 50) selectively induces redox-sensitive caspase 3-related apoptosis in chronic lymphocytic leukemia cells by targeting the Bcl-2/Bad pathway.

Defect in apoptosis has been implicated as a major cause of resistance to chemotherapy observed in B cell chronic lymphocytic leukaemia (B CLL). This study evaluated the pro-apoptotic effect of an anthocyanin-rich dietary bilberry extract (Antho 50) on B CLL cells from 30 patients and on peripheral blood mononuclear cells (PBMCs) from healthy subjects, and determined the underlying mechanism. Antho 50 induced concentration- and time-dependent pro-apoptotic effects in B CLL cells but little or no effect in PBMCs. Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect. Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2. Antho 50 significantly induced PEG-catalase-sensitive formation of reactive oxygen species in B CLL cells. PEG-catalase prevented the Antho 50-induced induction of apoptosis and related signaling. The present findings indicate that Antho 50 exhibits strong pro-apoptotic activity through redox-sensitive caspase 3 activation-related mechanism in B CLL cells involving dysregulation of the Bad/Bcl-2 pathway. This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin. They further suggest that Antho 50 has chemotherapeutic potential by targeting selectively B CLL cells.

Antioxidant potential of different plum cultivars during storage.

Plums, the most commonly consumed fruits from Romania, are fruits rich in bioactive compounds, such as antioxidants. This research work was carried out to investigate the antioxidant potential of twelve plum cultivars, fresh and stored during 10days at 4°C by using different methods (DPPH, ORAC and erythrocyte resistance to haemolysis). The contents of total phenolic compounds and total anthocyanins were also determined by specific spectrometric methods. Significant differences between fresh and stored plum cultivars (p<0.05) were found. Storage at 4°C resulted in an increase in antioxidant potential and anthocyanins content of the autumn plum varieties. Autumn plum varieties also showed a higher antioxidant capacity than summer varieties, as assessed by the ORAC and the haemolysis resistance assays. Our results suggest that, even after storage, plums could be a good source of antioxidants, which may provide health-promoting effects for humans.

Cultivars, culture conditions, and harvest time influence phenolic and ascorbic acid contents and antioxidant capacity of strawberry (Fragaria x ananassa).

UNLABELLED: Strawberries are a good source of micronutrients, especially antioxidant phenolics. More information is needed to better exploit the health-promoting effect of such fruits. Several studies focused on the effects of genotype, cultural practices, and seasonal variation on the antioxidant potential of strawberries, but often yielding contradictory results and/or focusing on only 1 source of variability. In the present study, we showed that total phenolic compounds, ascorbic acid, and antioxidant capacity strongly differed between genotypes of strawberry. The precise results revealed the importance of genetic background for the antioxidant capacity and for the content of total phenolics (with up to 3.3-fold variations). Other parameters may also influence the antioxidant capacity in strawberry such as harvest time, culture conditions, and environmental factors. Moreover, in this study, the harvesting time (at the same ripening stage) appeared to be very important, more important than genotype. Variations of the antioxidant capacity of up to 4.1-folds were observed following the harvesting time of "Elsanta" cultivar. PRACTICAL APPLICATION: This article compares the antioxidant capacity and the content in ascorbic acid and phenolic compounds of strawberries of different varieties and of fruits harvested from April to December at the same ripening stage. The importance of strawberry antioxidant capacity resides in its benefits for human health.

Influence of cultivar, harvest time, storage conditions, and peeling on the antioxidant capacity and phenolic and ascorbic acid contents of apples and pears.

Apple and pear fruits are important sources of secondary plant metabolites and one of the major sources of dietary phenolics consumed all year round. The aim of this work was to identify the main variables influencing phenolic content and antioxidant capacity in apples. Higher phenolic and antioxidant contents were observed in some varieties (such as the Delbar Estival apple and Durondeau pear). Storage conditions were important. Our results also showed that fruits should be consumed rapidly after purchase and with their peel. After one week of domestic storage, the ascorbic acid content was found to decrease by 75%. Peeling led to a more than 25% decrease in total phenolics and ascorbic acid. The harvest time (at normal ripeness) had only a limited impact, but significant year-to-year variations were observed. In conclusion, well-chosen and well-stored apples and pears may contribute to an antioxidant-rich diet if consumed rapidly and with their peel.

Adventitious rooting is enhanced by methyl jasmonate in tobacco thin cell layers.

Adventitious roots (ARs) are induced by auxins. Jasmonic acid (JA) and methyl jasmonate (MeJA) are also plant growth regulators with many effects on development, but their role on ARs needs investigation. To this aim, we analyzed AR formation in tobacco thin cell layers (TCLs) cultured with 0.01-10 microM MeJA, either under root-inductive conditions, i.e., on medium containing 10 microM indole-3-butyric acid (IBA) and 0.1 microM kinetin, or without hormones. The explants were excised from the cultivars Samsun, Xanthii and Petite Havana, and from genotypes with altered AR-forming ability in response to auxin, namely the non-rooting rac mutant and the over-rooting Agrobacterium rhizogenes rolB transgenic line. Results show that NtRNR1 (G1/S) and Ntcyc29 (G2/M) gene activity, cell proliferation and meristemoid formation were stimulated in hormone-cultured TCLs by submicromolar MeJA concentrations. The meristemoids developed either into ARs and xylogenic nodules, or into xylogenic nodules only (rac TCLs). MeJA-induced meristemoid over-production characterized rolB TCLs. No rooting or xylogenesis occurred under hormone-free conditions, independently of MeJA and genotype. Endogenous JA progressively (days 1-4) increased in hormone-cultured TCLs in the absence of MeJA. JA levels were enhanced by 0.1 microM MeJA, on both days 1 and 4. Endogenous IBA was the only auxin detected, both in the free form and as IBA-glucose. Free IBA increased up to day 2, remaining constant thereafter (day 4). Its level was enhanced by 0.1 microM MeJA only on day 1, while IBA conjugation was not affected by MeJA. Taken together, these results show that an interplay between jasmonates and auxins regulates AR formation and xylogenesis in tobacco TCLs.

Evolution of antioxidant capacity during storage of selected fruits and vegetables.

Interest in the consumption of fresh fruits and vegetables is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. Among all of the selected fruits and vegetables, strawberries and black grapes have relatively high antioxidant capacities associated with high contents of total phenolic compounds, ascorbic acid, and flavonols. More interesting, the results of this study indicated that in most fruits and vegetables storage did not affect negatively the antioxidant capacity. Better, in some cases, an increase of the antioxidant capacity was observed in the days following their purchase, accompanied by an increase in phenolic compounds. In general, fruits and vegetables visually spoil before any significant antioxidant capacity loss occurs except in banana and broccoli. When ascorbic acid or flavonoids (aglycons of flavonols and anthocyanins) were concerned, the conclusions were similar. Their content was generally stable during storage.

Antioxidant capacity of black currant varies with organ, season, and cultivar.

Small berries such as black currant constitute one of the important sources of potential health-promoting phytochemicals because these fruits are rich sources of compounds with high antioxidant properties. In this work, antioxidant capacities of different parts (buds, leaves, fruits) of various black currant cultivars were compared throughout the growing season with the aim to prepare extracts with high antioxidant capacity. Buds (opened, at the end of March) and leaves (in June) had a higher content in phenolics and antioxidants than fully ripened berries (in July) and the best yield (per branch) was obtained with the leaves collected in June due to their higher biomass. The differences observed among the eight cultivars tested were small. Concerning flavonols, quercetin was dominant in all organs and cultivars, myricetin varied widely among the cultivars, and kampferol was very low.

Cuttings of the non-rooting rac tobacco mutant overaccumulate phenolic compounds.

The auxin and phenolic contents, as well as phenylalanine ammonia-lyase (PAL) activity, were determined in in vitro cultured shoots of the recalcitrant-to-root rac mutant of tobacco, and compared with wild-type shoots. The mutant and wild-type shoots showed similar auxin changes during the culture cycle, but with higher contents for the mutant. A transient peak of auxin (corresponding to the achievement of the rooting inductive phase) occurred at day 14 in both types of shoots, but earlier in the basal parts of the wild-type stems. The rac shoots contained more phenolics, corresponding with an increased PAL activity. The most abundant phenolic compound found in the two types of tobacco was chlorogenic acid, which was more abundant in the rac shoots. Rutin was also detected at a higher concentration in the mutant shoots. Basal parts of wild-type shoots treated with 10-3 M chlorogenic acid reacted by accumulating auxins and, unlike untreated controls, did not form adventitious roots. The relationships between these biochemical analyses in relation to the growth limitation of the rac mutant, and the inhibition of its root development, are discussed.