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Broccoli sprouts are considered functional food as they are naturally enriched in glucoraphanin (GR) that is the biological precursor of the anticancer compound sulforaphane (SFN). Due to its health promoting value, also broccoli sprout juice is becoming very popular. The present study aimed to quantitatively assess the conversion of GR to its hydrolysis products, SFN and SFN-nitrile, during the juice preparation process. We demonstrated that SFN plus SFN-nitrile yield from glucoraphanin is quite low (≈25%) and that some SFN is lost during the juice preparation partially due to the spontaneous conversion to sulforaphane-amine or conjugation to GSH and proteins naturally present in the juice. Our results demonstrate that the detection of the sole SFN free form does not provide reliable information about the real concentration of this functional compound in the juice.

Epidemiological studies show an inverse association between Brassica consumption and chronic diseases. Phytochemicals are thought to be beneficial for human health and therefore responsible for this protective effect. Increasing their levels into Brassica food is considered an expedient nutritional strategy that can be achieved through the manipulation of growth conditions by elicitors. In this work we systematically evaluated the influence of treatment with different elicitors (sucrose, mannitol, NaCl, 1-aminocyclopropane-L-carboxylic acid, salicylic acid and methyl jasmonate) on the phytochemical composition of broccoli sprouts. The content of total and single glucosinolates, total phenolic compounds, total flavonoids, total anthocyanins, vitamin C and E and β-carotene was assessed. The exposure to different elicitors produced concentration- and elicitor-dependent specific changes in the content of all the phytochemicals considered. Sucrose, identified as the most effective elicitor by principal component analysis, induced a significant increase of total and specific glucosinolates, vitamin C, total anthocyanins and polyphenols. Sucrose is likely to represent an effective tool to increase the nutritional value of broccoli sprouts.

Apples are one of the most commonly consumed fruits and their high polyphenol content is considered one of the most important determinants of their health-promoting activities. Here we studied the nutrikinetics of apple polyphenols by UHPLC-HRMS metabolite fingerprinting, comparing bioavailability when consumed in a natural or a polyphenol-enriched cloudy apple juice. Twelve men and women participated in an acute single blind controlled crossover study in which they consumed 250 mL of cloudy apple juice (CAJ), Crispy Pink apple variety, or 250 mL of the same juice enriched with 750 mg of an apple polyphenol extract (PAJ). Plasma and whole blood were collected at time 0, 1, 2, 3 and 5 h. Urine was collected at time 0 and 0-2, 2-5, 5-8, and 8-24 h after juice consumption. Faecal samples were collected from each individual during the study for 16S rRNA gene profiling. As many as 110 metabolites were significantly elevated following intake of polyphenol enriched cloudy apple juice, with large inter-individual variations. The comparison of the average area under the curve of circulating metabolites in plasma and in urine of volunteers consuming either the CAJ or the PAJ demonstrated a stable metabotype, suggesting that an increase in polyphenol concentration in fruit does not limit their bioavailability upon ingestion. Faecal bacteria were correlated with specific microbial catabolites derived from apple polyphenols. Human metabolism of apple polyphenols is a co-metabolic process between human encoded activities and those of our resident microbiota. Here we have identified specific blood and urine metabolic biomarkers of apple polyphenol intake and identified putative associations with specific genera of faecal bacteria, associations which now need confirmation in specifically designed mechanistic studies.

The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower) is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response after light exposure. Broccoli seeds were germinated and grown hydroponically for five days in total darkness or with a light/dark photoperiod (16 h light/8 h dark cycle). We used an ultra-performance liquid-chromatography system coupled to an ion-mobility, time-of-flight mass spectrometer to profile the large array of metabolites present in the sprouts. Differences at the metabolite level between groups were analyzed using multivariate statistical analyses, including principal component analysis and correlation analysis. Altered metabolites were identified by searching publicly available and in-house databases. Metabolite pathway analyses were used to support the identification of subtle but significant changes among groups of related metabolites that may have gone unnoticed with conventional approaches. Besides the chlorophyll pathway, light exposure activated the biosynthesis and metabolism of sterol lipids, prenol lipids, and polyunsaturated lipids, which are essential for the photosynthetic machinery. Our results also revealed that light exposure increased the levels of polyketides, including flavonoids, and oxylipins, which play essential roles in the plant’s developmental processes and defense mechanism against herbivores. This study highlights the significant contribution of light exposure to the ultimate metabolic phenotype, which might affect the cellular physiology and nutritional value of broccoli sprouts. Furthermore, this study highlights the potential of an unbiased omics approach for the comprehensive study of the metabolism.

Benefits to health from a high consumption of fruits and vegetables are well established and have been attributed to bioactive secondary metabolites present in edible plants. However, the effects of specific health-related phytochemicals within a complex food matrix are difficult to assess. In an attempt to address this problem, we have used elicitation to improve the nutraceutical content of seedlings of Brassica oleracea grown under controlled conditions. Analysis, by LC-MS, of the glucosinolate, isothiocyanate and phenolic compound content of juices obtained from sprouts indicated that elicitation induces an enrichment of several phenolics, particularly of the anthocyanin fraction. To test the biological activity of basal and enriched juices we took advantage of a recently developed in vitro model of inflamed human intestinal epithelium. Both sprouts’ juices protected intestinal barrier integrity in Caco-2 cells exposed to tumor necrosis factor α under marginal zinc deprivation, with the enriched juice showing higher protection. Multivariate regression analysis indicated that the extent of rescue from stress-induced epithelial dysfunction correlated with the composition in bioactive molecules of the juices and, in particular, with a group of phenolic compounds, including several anthocyanins, quercetin-3-Glc, cryptochlorogenic, neochlorogenic and cinnamic acids.

Glucosinolates are an important class of secondary plant metabolites, possessing health-promoting properties. Young broccoli plants are a very good source of glucosinolates with concentrations several times greater than in mature plants. The aim of our study was to develop a liquid chromatography–mass spectrometry and liquid chromatography/tandem mass spectrometry qualitative and quantitative method for the measure of glucosinolates in broccoli sprouts. The described method provides high sensitivity and specificity, allowing a rapid and simultaneous determination of 14 glucosinolates. The proposed method has been validated for eight glucosinolates: glucobrassicin, glucoraphanin, glucoiberin, glucoerucin, progoitrin, gluconapin, sinigrin and glucocheirolin. The linear range was 1–150 µg ml−1, the intra-day and inter-day precision values are within 6% and 8% at the lower limit of quantification, while the overall recovery of the eight glucosinolates was 99 ± 9%. This validated method was used successfully for analysis of glucosinolates content of broccoli sprouts grown in different conditions. Copyright © 2012 John Wiley & Sons, Ltd.

Coffee consumption has been associated with a lower risk of type 2 diabetes. This association does not depend on race, gender, geographic distribution of the study populations, or the type of coffee consumed (i.e., caffeinated or decaffeinated). This review discusses the strength of this relationship, examines the possibility that the pattern of coffee consumption could influence the association, and evaluates the possible relationship between coffee consumption and other risk factors associated with diabetes. Particular attention is paid to the identification, on the basis of the scientific evidence, of the possible mechanisms by which coffee components might affect diabetes development, especially in light of the paradoxical effect of caffeine on glucose metabolism. In addition to the role of coffee in reducing the risk of developing type 2 diabetes, the possible role of coffee in the course of the illness is explored. Finally, the possibility that coffee can also affect the risk of other forms of diabetes (e.g., type 1 diabetes and gestational diabetes) is examined.

Moderate wine consumption has been shown to lower cardiovascular risk when part of a healthy life style. Red wine compounds, especially polyphenols, might play a role in preventing the development and progression of atherosclerosis, through different modalities of action, including inhibition of lipid peroxidation, metal chelation, free radical scavenging, inhibition of platelet aggregation, anti-inflammatory and estrogenic activity, improvement of endothelial function, lowering of blood pressure and modulation of lipoprotein metabolism. The attenuation of the postprandial oxidative stress could be another mechanism involved in protection by wine phenols, as the absorption of prooxidant/oxidized species with a meal can induce physiological events, such as the formation of mildly oxidized lipoprotein or endothelial dysfunction and inflammatory responses, all linked to the development of cardiovascular disease. As regards oxidizable/oxidized dietary fats, the typical Western diet contains substantial quanti...

The antioxidant activity of eight synthetic 4-methylcoumarins was systematically studied. The antioxidant capacity was measured using: (i) a competition kinetic test, to measure the relative capacity to quench peroxyl radical; (ii) the in vitro oxidative modification of human low-density lipoprotein, initiated by AAPH or catalyzed by copper. In both models, the ortho-OH substitutes were found to be better antioxidant than the meta one. The most efficient antioxidant was the 7,8-dihydroxy-4-methylcoumarin and the corresponding diacetoxy-substituted was unexpectedly a good antioxidant. Finally, the presence of an ethoxycarbonylethyl substituent at the C-3 position increased the antioxidant capacity of both 7,8-dihydroxy-4-methylcoumarin and 7,8-diacetoxy-4-methylcoumarin.

Despite the vast literature describing the biological effects of phenolic compounds, rather scarce data are available on their absorption from diet in humans. The present study focused on the absorption in humans of phenolic acids from white wine, particularly hydroxycinnamic acids and their esters with tartaric acid. The results obtained indicate that, following a single wine drink, hydroxycinnamic acids from white wine are absorbed from the gastrointestinal tract and circulate in the blood after being largely metabolized to the form of glucuronide and sulfate conjugates. Unmodified tartaric acid esters of hydroxycinnamic acids from wine are present in human plasma at low levels, if any. Wine hydroxycinnamic acids, although present in wine as conjugated forms, are still bioavailable to humans.

Epidemiological studies suggest that high polyphenols intake from diet is associated with reduced risk for cardiovascular diseases. Platelet aggregation is a crucial mechanism in the pathogenesis and clinical expression of coronary acute syndrome, and there is extensive evidence that antiplatelet therapy reduces cardiovascular disease risk. In this review, the available literature on the effect of polyphenols supplementation on platelet aggregation in humans or animal models has been critically analyzed, taking into consideration the different experimental protocols employed. In some studies, polyphenols supplementation did not show any effect on platelet aggregation. However, in the most of the studies, polyphenols supplementation, either as purified compounds or food extracts, showed some inhibitory effects, both in humans and in animal models. The extent of the inhibition varies in a wide range, depending on the experimental conditions used. The observed inhibitory effect of polyphenols on platelet aggregation might explain, at least in part, the epidemiological data on beneficial effect of dietary polyphenols on cardiovascular disease risk and suggests a role for polyphenols in helping to prevent cardiovascular diseases.

In spite of the wide literature describing the biological effects of phenolic compounds, scarce data are available on their absorption from diet. In the present work, we studied the absorption in humans of phenolic acids from beer, a common beverage rich in different phenolic acids with related chemical structures. Beer was analyzed for free and total (free+bound) phenolic acids. Ferulic, caffeic and sinapic acids were present in beer mainly as bound forms, while 4-hydroxyphenylacetic acid and p-coumaric acid were present mainly as free forms. Vanillic acid was present equally in the free and bound forms. Plasma samples were collected before and 30 and 60 min after beer administration and analyzed for free and conjugated phenolic acid content. A significant two- to fourfold increase in plasma levels of phenolic acids was detected with peak concentrations at 30 min after beer ingestion. 4-Hydroxyphenylacetic acid was present in plasma mainly as nonconjugated forms while p-coumaric acid was present equally as nonconjugated and conjugated forms. Ferulic, vanillic and caffeic acids were present in plasma predominantly as conjugated forms, with a slight prevalence of sulfates with respect to glucuronates. Our results indicate that phenolic acids from beer are absorbed from the gastrointestinal tract and are present in blood after being largely metabolized to the form of glucuronide and sulfate conjugates. The extent of conjugation is related to the chemical structure of phenolic acids: the monohydroxy derivatives showing the lowest conjugation degree and the dihydroxy derivatives showing the highest one.

The experimental conditions commonly used to detect bound phenolic acids by alkaline hydrolysis result in loss of several phenolic acids, particularly dihydroxy-derivatives (caffeic acid, dihydrocaffeic acid, homogentisic acid). In this study we show that the addition of ascorbic acid, a strong antioxidant, and ethylenediaminetetraacetic acid, a metal chelator, totally prevent the loss of phenolic acids during alkaline hydrolysis. In these conditions, a complete recovery of caffeic acid following hydrolysis of chlorogenic acid (5′-caffeoylquinic acid, an ester of caffeic acid with quinic acid) was found. This procedure has been successfully applied to quantitatively detect bound phenolic acids in coffee brew and apple.

Coffee and tea are widely consumed beverages, but only tea has been studied for its antioxidant capacity (AC) in vivo. The aim of this study was to compare the capacities of coffee and tea to affect plasma redox homeostasis in humans. The AC of plasma before and after supplementation with 200 mL of beverages (0, 1, and 2 h) was measured by the TRAP and crocin tests. The crocin test detected an increase in plasma AC only in subjects supplemented with coffee (+7% at peak time), whereas the TRAP method showed an increase in plasma AC after consumption of both coffee and tea (+6 and +4%, respectively, at peak time). Both beverages induced a significant increase in plasma uric acid (+5 and +7%, respectively). Uric acid strongly affects the results obtained by the TRAP test and does not affect those obtained by the crocin test. We can thus argue that uric acid is the main component responsible for the plasma AC increase after tea drinking, whereas molecules other than uric acid (probably phenolic compounds) are likely to be responsible for the increase in plasma AC after coffee drinking.

Despite extensive literature describing the biological effects of polyphenols, little is known about their absorption from diet, one major unresolved point consisting of the absorption of the bound forms of polyphenols. In this view, in the present work we studied the absorption in humans of phenolic acids from coffee, a common beverage particularly rich in bound phenolic acids, such as caffeic acid, ferulic acid, and p-coumaric acid. Coffee brew was analyzed for free and total (free + bound) phenolic acids. Chlorogenic acid (5'-caffeoylquinic acid), a bound form of caffeic acid, was present in coffee at high levels, while free phenolic acids were undetectable. After alkaline hydrolysis, which released bound phenolic acids, ferulic acid, p-coumaric acid, and high levels of caffeic acid were detected. Plasma samples were collected before and 1 and 2 h after coffee administration and analyzed for free and total phenolic acid content. Two different procedures were applied to release bound phenolic acids in plasma: beta-glucuronidase treatment and alkaline hydrolysis. Coffee administration resulted in increased total plasma caffeic acid concentration, with an absorption peak at 1 h. Caffeic acid was the only phenolic acid found in plasma samples after coffee administration, while chlorogenic acid was undetectable. Most of caffeic acid was present in plasma in bound form, mainly in the glucuronate/sulfate forms. Due to the absence of free caffeic acid in coffee, plasma caffeic acid is likely to be derived from hydrolysis of chlorogenic acid in the gastrointestinal tract.

The aim of the present study was to verify the extent of oxidative stress induced by a meal at plasma and LDL level, and to investigate the capacity of red wine to counteract this action. In two different sessions, six healthy men ate the same test meal consisting of "Milanese" meat and fried potatoes. The meal was taken either with 400 ml red wine or with an isocaloric hydroalcoholic solution. Oxidative stress at plasma level was estimated through the measure of ascorbic acid, alpha-tocopherol, protein SH groups, uric acid, and antioxidant capacity, measured before and 1 and 3 h after the meal. The change in the resistance of LDL to oxidative modification was taken as an index of exposure to pro-oxidants. The susceptibility to Cu(II)-catalyzed oxidation of baseline and postprandial LDL was measured as conjugated dienes formation, tryptophan residues, and relative electrophoretic mobility. The experimental meal taken with wine provoked a significant increase in the total plasma antioxidant capacity and in the plasma concentration of alpha-tocopherol and SH groups. Postprandial LDL was more susceptible to metal-catalyzed oxidation than the homologous baseline LDL after the ethanol meal. On the contrary, postprandial LDL obtained after the wine meal was as resistant or more resistant to lipid peroxidation than fasting LDL.

The concentration of phenols (total, tannins, non-tannin flavonoids, non-tannin non-flavonoids), selected minerals (selenium, zinc and copper) and dextrins were measured in wort and beer produced in Italy. Samples were withdrawn at different phases of beer brewing: wort (step 1), beer at the end of maturation (step 2), filtered beer (step 3), beer after bottling and pasteurization (step 4). On the same samples, the total antioxidant capacity and the inhibitory effect on low density lipoprotein (LDL) oxidative modification were tested. Total antioxidant capacity was measured by monitoring the loss of fluorescence of a probe compound. During beer brewing the phenolic concentration decreased by 28% corresponding to a parallel decrease of the antioxidant activity (−29%). This decay was explained by the loss of tannins and non-tannic flavonoids during the technological process. Wort was more efficient than beer in inhibiting LDL oxidation at step 4, both by metal catalyst or peroxyl radicals. However, the better protection provided by both wort and beer on metal-catalyzed LDL oxidation indicates a prevalent chelating action. The data suggest that beer in the diet can supply molecules with antioxidant activity which could play a role in antioxidant activityin vivo.

The relationship between chronic moderate beer consumption and oxidative stress was studied in rats. Animals were fed three different isocaloric diets for six weeks: a beer-containing diet (30% w/w), an ethanol-supplemented diet (1.1 g/100 g, the same as in the beer diet) and an alcohol-free basal diet. At the end of the feeding period, rats were analyzed for plasma and liver oxidative status. Some livers were isolated and exposed to ischemia-reperfusion to assess the additional oxidative stress determined by reperfusion. No significant differences in plasma antioxidant status were found among the three dietary groups. Lipoproteins from the beer group, however, showed a greater propensity to resist lipid peroxidation. Ischemia caused a decrease in liver energy and antioxidant status in all groups. Nevertheless, ATP was lower in the livers of rats exposed to the ethanol diet. During reperfusion, lipoperoxidation increased significantly in all groups. However, livers obtained from ethanol-treated rats showed the higher formation of lipoperoxides. In conclusion, a moderate consumption of beer in a well-balanced diet did not appear to cause oxidative stress in rats; moreover, probably through its minor components, beer could attenuate the oxidative action of ethanol by itself.

Glucosinolates are a class of secondary plant metabolites particularly occurring in Cruciferae with potential health-promoting properties, as their hydrolysis products, isothiocyanates, possess chemopreventive and antioxidant activities. In the present study, we systematically studied the in vitro redox behaviour of 15 glucosinolates, by using a range of analytical methods measuring different activities: (i) radical scavenging activity toward peroxyl and toward ABTS radical (chain-breaking activity); (ii) capacity in modulating the in vitro resistance of human low-density lipoprotein (LDL) catalysed by copper (chelating and chain-breaking activity). Data obtained from different assays were compared and analysed by principal component analysis (PCA). PCA allowed us to identify a big cluster of glucosinolates (10 out 15 tested) that do not possess any antioxidant capacity; while, the other five glucosinolates showed moderate and specific antioxidant capacity. Notably, sinalbin and gluconasturtiin were highly active in scavenging ABTS radical and in protecting LDL from copper-catalysed oxidation, respectively. The overall results of this study indicate that just few glucosinolates can act as antioxidants.

Background. Increases in blood lipids have been observed in humans when coffee is brewed by the boiling method. The purpose of this study was to evaluate if giving up Italian coffee might reduce blood cholesterol levels. Methods. Eighty-four normolipidaemic young adult males, after a 3-week baseline (BL), were randomly assigned to three different regimens of coffee consumption : espresso (E), mocha (M), and no coffee, but tea (T). The average coffee consumption during intervention (I) was 3.1 ± 1.2 and 2.8 ± 1.1 cups per day for espresso and mocha group respectively (espresso : 25-35 ml/cup ; mocha : 40-50 ml/cup). Total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides were measured eight times during the study. Dietary pattern, alcohol consumption, smoking habits, drug use, and anthropometric data were also recorded. Results. The changes observed in serum cholesterol concentration between baseline and intervention were not statistically different in all groups. The changes were 0.0 mmol/l (T), +0.10 mmol/l (E) and +0.05 mmol/l (M) for total serum cholesterol ; 0 mmol/l (T), -0.02 mmol/l (E) and -0.03 mmol/l (M) for HDL-C ; -0.13 mmol/l (T), +0.02 mmol/l (E) and -0.05 mmol/l (M) for LDL-C. Serum triglycerides showed a significant increase during intervention (P < 0.01 by ANOVA) in all groups with a change of 0.18 mmol/l, 0.18 mmol/l and 0.22 mmol/l, for tea, espresso and mocha group respectively. Conclusion. The results indicate that coffee brewed in the Italian way does not alter blood levels of total cholesterol, HDL-cholesterol and LDL-cholesterol, since no significant differences were observed in these blood parameters after a 6-week break from coffee consumption.

The antioxidant activity of the major phenols derived from hydroxycinnamic acid (caffeic, ferulic, and p-coumaric acids) on in vitro LDL oxidation was screened, using Cu2+ as catalyst. The presence of the second phenolic hydroxy group enhanced the inhibitory effect of these compounds. In fact, at 5 microM concentration, only caffeic acid completely protected LDL from modification as measured as conjugated dienes formation and apo B-100 fragmentation, also preserving alpha-tocopherol. The effect of caffeic acid in inhibiting LDL oxidative modification induced by three different oxidant systems was tested. Using both Cu2+ and 2,2'-azobis (2-amidinopropane)-hydrochloride (AAPH), the inhibitory effect of caffeic acid was dose-dependent. Yet, the better protection was achieved in the metal-ion dependent system. Also the murine macrophages-mediated LDL oxidation was efficiently inhibited by 5 microM caffeic acid. UV-VIS spectra of caffeic acid incubated with cupric ions show the formation of a caffeic acid:copper complex, responsible for a transient chelating activity. This mechanism, coupled with its free radical scavenging property, accounts for the higher inhibitory activity exhibited by caffeic in Cu(2+)-catalyzed reaction.

Incorporation of long-chain polyunsaturated fatty acids (PUFA) in low density lipoprotein (LDL) might induce a decrease in its resistance against oxidative modification because of their high degree of unsaturation. To investigate the in vivo and in vitro influence of dietary ω --3 fatty acids on plasma and LDL susceptibility to oxidative modification, a diet containing 15% wt/wt fish oil was fed to rats together with diets containing 15% wt/wt soybean oil or coconut oil for 6 weeks. The plasma lipid concentration was significantly lower after fish oil feeding compared with the two control diets. Fish oil fed rats exhibited significantly lower total (peroxyl) radical-trapping antioxidant activity (TRAP) than both soybean oil and coconut oil fed animals ( P ω --3 fatty acids induce a decrease in the plasma antioxidant potential and an increase in the ex vivo susceptibility of LDL to oxidative modification.

Moderate wine consumption has been shown to lower cardiovascular risk. One of the mechanisms could involve the control of postprandial hyperlipaemia, a well-defined risk factor for atherosclerosis, reasonably by reducing the absorption of lipid oxidised species from the meal. The objective of the present study was to investigate whether wine consumption with the meal is able to reduce the postprandial increase in plasma lipid hydroperoxides and cholesterol oxidation products, in human subjects. In two different study sessions, twelve healthy volunteers consumed the same test meal rich in oxidised and oxidisable lipids (a double cheeseburger), with 300 ml of water (control) or with 300 ml of red wine (wine). The postprandial plasma concentration of cholesterol oxidation products was measured by GC–MS. The control meal induced a significant increase in the plasma concentration of lipid hydroperoxides and of two cholesterol oxidation products, 7-β-hydroxycholesterol and 7-ketocholesterol. The postprandial increase in lipid hydroperoxides and cholesterol oxidation products was fully prevented by wine when consumed with the meal. In conclusion, the present study provides evidence that consumption of wine with the meal could prevent the postprandial increase in plasma cholesterol oxidation products.

The effect of varying unsaturation degree of dietary lipid on the oxidative response of rat liver microsomes was studied. Three groups of growing male rats were maintained for 6 weeks on 15% fat diets containing either soybean oil, olive oil, or coconut oil, with the same level of vitamin E. After 6 weeks, microsomal malondialdehyde, vitamin E, and fatty acid composition were measured in liver microsomes. The relative abundance of saturated and unsaturated fatty acids in the microsomes reflected the composition of the dietary lipid. When dietary requirement for vitamin E was satisfied, the increased polyunsaturated fatty acid intake from vegetable oils did not enhance lipid peroxidation in physiological conditions, as demonstrated by similar malondialdehyde concentrations found in the three groups. However, the somewhat lower vitamin E content measured in soybean oil-fed rats confirms an enhanced requirement for dietary antioxidant caused by the increased intake of polyunsaturated fatty acids. The susceptibility of liver microsomes to lipid peroxidation stimulated by the ADP/iron/ascorbate system was also studied. Membranes of soybean oil-fed rats exhibited the highest peroxidation rate, as shown by oxygen consumption and malondialdehyde and 4-hydroxy-2,3-trans-neonenal production, because of the lower concentration of vitamin E and of the higher content of polyunsaturated fatty acids. Microsomes of olive oil- and coconut oil-fed rats showed highest protection against lipid peroxidation.

The effect of microwave, boiling and pressure cooking on total antioxidant capacity and total phenolics content of seven vegetables were studied. Total phenolics in raw and cooked vegetables were determined by the Folin–Ciocalteu method, whereas the total antioxidant capacity of vegetables extracts was evaluated using the Crocin method. After boiling, four out of seven vegetables (cauliflower, peas, spinach and Swiss chard) showed a significant decrease in their total phenolic content (P

Several lines of evidence support a pro-atherogenic role for oxidized LDL. The most persuasive evidence, however, is that antioxidant feeding can retard the progression of atherosclerosis in animal models. This review focuses on the effect of antioxidants (probucol, a-tocopherol, ascorbate, (i-carot

In spite of the wide literature describing the biological effects of phenolic compounds, scarce data are available on their absorption from diet and metabolism in humans. Recently, a renewal interest has been focused on beer, a common beverage with low ethanol content and rich in phenolic compounds, particularly caffeic, vanillic, p-coumaric, sinapic, syringic and ferulic acids. In this study, the absorption in humans of phenolic acids from beer was studied. Firstly, a recently developed hydrolytic procedure was applied to quantitatively measure total (free plus bound) phenolic acids in beer. Secondly, we measured plasma concentration of phenolic acids in samples collected before and 30 and 60 min after beer administration in healthy humans. The results obtained show that the most of phenolic acids are present in beer as bound forms and only a small portion can be detected as free forms. Moreover, phenolic acids from beer are absorbed from the gastrointestinal tract and are present in blood after being largely metabolized to the form of glucuronide and sulfate conjugates. The extent of conjugation is related to the chemical structure of phenolic acids, the monohydroxy derivatives showing the lowest conjugation degree and the dihydroxy derivatives showing the highest one.

Epidemiological studies indicate a J-shaped relationship linking coffee consumption and cardiovascular risk, suggesting that moderate coffee consumption can be beneficial. Platelet aggregation is of critical importance in thrombotic events, and platelets play a major role in the aetiology of several CVD. The aim of this study was to evaluate the effect of coffee drinking on platelet aggregationex vivo, using caffeine as control. A crossover study was performed on ten healthy subjects. In two different sessions, subjects drank 200 ml coffee, containing 180 mg caffeine, or a capsule of caffeine (180 mg) with 200 ml water. Platelets were separated from plasma at baseline and 30 and 60 min after coffee drinking. Platelet aggregation was induced with three different agonists: collagen, arachidonic acid and ADP. Coffee drinking inhibited collagen (P P sem0·1) to 2·4 (sem0·6) ng/mg (P

Evidence is accumulating that postprandial phenomena play a role in atherogenesis. Dietary lipid hydroperoxides that escape from the gastrointestinal barrier can be incorporated into plasma lipoproteins, leading to a modified form of LDL (LDL minus). The present human study was designed to investigate the effect of selenium supplementation on the formation of LDL minus in the postprandial phase. Fourteen healthy subjects ate the same test meal, high in lipid hydroperoxides, at baseline and after 10-day selenium supplementation (110 μg/day). Plasma selenium, LDL minus, LDL resistance to oxidative modification, plasma antioxidants (ascorbic acid, GSH and GPx activity) and MDA were measured in preprandial (time 0) and postprandial (3 h) phases. Supplementation did not induce changes in the concentration of selenium in fasting plasma, but, at the same time, it induced a significant decrease in preprandial plasma GPx activity and inhibited the meal-induced increase in GPx activity. Selenium supplementation fully prevented the meal-induced increase in both LDL minus level and LDL susceptibility to oxidation. This study demonstrated the efficacy of selenium in preventing postprandial oxidative stress. The results, obtained on subjects adequately supplied with selenium, suggest that a non-limiting selenium availability counteracts the postprandial formation of the atherogenic form of LDL and provide a rationale for the epidemiological evidence of the inverse correlation between selenium intake and the incidence of chronic and degenerative diseases.

Several food items of plant origin, and in particular red wine, have been reported to protect from cardiovascular disease (CVD) development, thanks to their polyphenol components. Polyphenols undergo complex metabolic transformation during digestion and intestinal absorption. Here we report a novel model to study the effects of complex food matrices, applied to red wine, on gene expression in cultured primary human endothelial cells that takes into account the polyphenol metabolic transformation. Red wine was administered to human volunteers acting as ‘bio-reactors’. Serum (RWS) obtained after 40 min was utilized to enrich endothelial cell culture media. The expression of specific genes involved in cell adhesion (vascular cell adhesion molecule (VCAM), intercellular adhesion molecule (ICAM) and monocytes chemoattractant protein (MCP-1)) and fibrinolysis (tissue-plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2)) was considered as a molecular marker of cell function and related to the effects of RWS. The gene expression profile determined by RWS incubation was significantly different from that observed after the addition of red wine. Data obtained by this approach indicate the importance of taking into account the complex metabolic transformation of polyphenols that occurs during absorption when studying their effect on human health.

Postprandial hyperlipemia is a well-defined risk factor for atherosclerosis. A reasonable contributing mechanism could involve the postprandial increase of plasma lipid hydroperoxides (LPO) affecting the oxidant/antioxidant balance and increasing the susceptibility of LDL to oxidation. Wine has been shown to prevent both these events. The present study was designed to investigate the effect of supplementing a meal with grape seed proanthocyanidins (the main phenolic antioxidant of red wine) on plasma postprandial oxidative stress. In two different sessions, 8 healthy volunteers consumed the same test meal rich in oxidized and oxidizable lipids without (control) or with 300 mg of a proanthocyanidin-rich grape seeds extract (GSE). Lipid hydroperoxide concentration, antioxidant status, and LDL resistance to oxidative modification were measured in postprandial plasma. The content of LPO in chylomicrons was 1.5-fold higher after the control meal than after the GSE-supplemented meal. Plasma LPO increased only after consumption of the control meal. The plasma antioxidant capacity increased in the postprandial phase only following the GSE supplemented meal. LDL isolated 3 h after the control meal tended to be more susceptible to oxidative modification (but the difference did not reach statistical significance). An opposite trend was observed following the GSE supplemented meal. In conclusion, the supplementation of a meal with GSE minimizes the postprandial oxidative stress by decreasing the oxidants and increasing the antioxidant levels in plasma, and, as a consequence, enhancing the resistance to oxidative modification of LDL.

The measure of antioxidant capacity (AC) considers the cumulative action of all the antioxidants present in plasma and body fluids, thus providing an integrated parameter rather than the simple sum of measurable antioxidants. The capacity of known and unknown antioxidants and their synergistic interaction is therefore assessed, thus giving an insight into the delicate balance in vivo between oxidants and antioxidants. Measuring plasma AC may help in the evaluation of physiological, environmental, and nutritional factors of the redox status in humans. Determining plasma AC may help to identify conditions affecting oxidative status in vivo (e.g., exposure to reactive oxygen species and antioxidant supplementation). Moreover, changes in the plasma AC after supplementation with galenic antioxidants or with antioxidant-rich foods may provide information on the absorption and bioavailability of nutritional compounds. Consequently, this review discusses the rationale, interpretation, confounding factors, measurement limits, and human applications of the measure of plasma AC.

Caffeic acid (CA) is a common constituent of human diet while pine bark extract (PBE) is utilized either as nutritional supplement or as phytochemical remedy for different diseases. CA and PBE, are reported as efficient antioxidants and more recently have been described to modulate cellular response to oxidative challenge and to possess many other biological activities, i.e. anti-inflammatory, antimutagenic, antitumoral effects. In order to investigate in depth the mechanism of action of these polyphenols, the effects of CA and PBE on the activity of some protein kinases involved in the regulation of fundamental cellular processes were studied in vitro: phosphorylase kinase (PhK), protein kinase A (PKA), protein kinase C (PKC). PBE at the concentration of 20 microg/ml (corresponding to 69 microM catechin equivalents) inhibited PKA, PhK and PKC by about 90, 59, 57%, respectively, while 100 microM CA inhibited by 37, 52 and 54%, respectively. Considerable inhibitions have been still observed at even lower concentrations of CA and PBE. For PhK and PKA, the inhibition follows a non-competitive mechanism. CA also inhibits PKC activity in a partially purified cellular extract. The results suggest a possible involvement of CA and PBE in modulation of cellular functions.

The positive association of a moderate intake of alcoholic beverages with a low risk for cardiovascular disease, in addition to ethanol itself, may be linked to their polyphenol content. This article describes the effect of acute ingestion of beer, dealcoholized beer, and ethanol (4.5% v/v) on the total plasma antioxidant status of subjects, and the change in the high performance liquid chromatography profile of some selected phenolic acids (caffeic, sinapic, syringic, and vanillic acids) in 14 healthy humans. Plasma was collected at various times: before (T0), 1 hour after (T1), and 2 hours after (T2) drinking. The study is part of a larger research planned to identify both the impact of brewing on minor components potentially present in beer and their metabolic fate in humans. Beer was able to induce a significant (P < 0.05) increase in plasma antioxidant capacity at T1 (mean +/- SD: T0 1,353 +/- 320 microM; T1 1,578 +/- 282 microM), returning close to basal values at T2. All phenolic acids measured in plasma tended to increase after beer intake (20% at T1, 40% at T2). Syringic and sinapic acid reached statistical significance (P < 0.05 by one-way analysis of variance-Fisher's test) at T1 and T2, respectively. Plasma metabolic parameters (glucose, total cholesterol, triglycerides, and uric acid) and plasma antioxidants (alpha-tocopherol and glutathione) remained unchanged. Ethanol removal impaired the absorption of phenolic acids, which did not change over the time of the experiment, accounting for the low (and not statistically significant) increase in plasma antioxidant capacity after dealcoholized beer drinking. Ethanol alone did not affect plasma antioxidant capacity or any of the antioxidant and metabolic parameters measured.

The antioxidant activity of four derivatives of benzoic acid was systematically compared with the activity of the four homologous derivatives of cinnamic acid. The couples of compounds differed for the kind of aromatic substitution (p-hydroxy, p-hydroxymethoxy, p-hydroxydimethoxy, dihydroxy). The antioxidant activity was measured using (i) a competition kinetic test, to measure the relative capacity to quench peroxyl radical and (ii) the in vitro oxidative modification of human low-density lipoprotein (LDL), initiated by 2,2'-azobis(amidinopropane) dihydrochloride or catalyzed by Cu(II). In both models, cinnamic acids were more efficient than their benzoic counterparts. As for the influence of the aromatic substitution, in the kinetic test the antioxidant activity increased in the sequence p-hydroxy < p-hydroxymethoxy < dihydroxy < p-hydroxydimethoxy. In contrast, in the LDL system, the dihydroxy acids had an antioxidant capacity equal to or higher than that of the p-hydroxydimethoxy acids.

Heme-peroxidases, such as horseradish peroxidase (HRP), are among the most popular catalysts of low density lipoprotein (LDL) peroxidation. In this model system, a suitable oxidant such as H2O2 is required to generate the hypervalent iron species able to initiate the peroxidative chain. However, we observed that traces of hydroperoxides present in a fresh solution of linoleic acid can promote lipid peroxidation and apo B oxidation, substituting H2O2. Spectral analysis of HRP showed that an hypervalent iron is generated in the presence of H2O2 and peroxidizing linoleic acid. Accordingly, careful reduction of the traces of linoleic acid lipid hydroperoxide prevented formation of the ferryl species in HRP and lipid peroxidation. However, when LDL was oxidized in the presence of HRP, the ferryl form of HRP was not detectable, suggesting a Fenton-like reaction as an alternative mechanism. This was supported by the observation that carbon monoxide, a ligand for the ferrous HRP, completely inhibited peroxidation of LDL. These results are in agreement with previous studies showing that myoglobin ferryl species is not produced in the presence of phospholipid hydroperoxides, and emphasize the relevance of a Fenton-like chemistry in peroxidation of LDL and indirectly, the role of pre-existing lipid hydroperoxides.

Nonvitamin phenolic compounds are ubiquitous in food plants and therefore potentially present in human plasma in a diet-dependent concentration. The aim of this study was to evaluate the ability of caffeic acid, a phenolic acid with antioxidant activity, to affect cellular response in U937 human monocytic cells to t-butyl hydroperoxide-induced oxidative stress. In our experimental conditions caffeic acid was incorporated into cells without any cytotoxic effect. Caffeic acid-treated cells showed an increased resistance to oxidative challenge, as revealed by an higher percent of survival and the maintenance of an higher proliferative capacity in respect to control cells. This effect seems to be due to the ability of caffeic acid to reduce glutathione depletion and to inhibit lipid peroxidation during tBOOH treatment. It can be concluded that caffeic acid exerts an antioxidant action inside the cell, responsible for the observed modulation of the cellular response to oxidative challenge. Due to its presence in the diet, therefore, caffeic acid may play a role in the modulation of oxidative processes in vivo.

Dietary supplementation of caffeic acid (0.2 and 0.8% w/w) in rats resulted in a statistically significant increase of α-tocopherol both in plasma and lipoprotein. While caffeic acid was not detectable in plasma under fasting conditions, in postprandial plasma it was present at micromole concentrations, doubling plasma total antioxidant capacity. Lipoproteins from caffeic acid-fed rats were more resistant than control to Cu 2+ -catalyzed oxidation, despite the lack of incorporation of caffeic acid in the particles. No significant effects on plasma and liver copper concentration, nor the increase in liver of Mn-superoxide dismutase reported in copper deficiency, were detected. These results demonstrate the physiological relevance of caffeic acid and its antioxidant action in vivo, through both a direct contribution to the antioxidant defense system and a sparing effect on α-tocopherol.

The purpose of this study was to determine the most potent activator of the respiratory burst of polymorphonuclear leukocytes (PMN) with respect to the oxidative modification of low density lipoprotein (LDL). Phorbol-12-myristate-13-acetate (PMA), n-formyl-methionyl-leucylphenylalanine (nFMLP), lipopolysaccharide (LPS), and opsonized zymosan (OZ) were tested. The generation of reactive oxygen species by PMN was assayed as superoxide anion production. Oxidative modification of LDL was monitored by thiobarbituric acid reactive substance (TBARS) activity, by conjugated dienes formation at 234nm and by electrophoretic mobility on agarose gel. PMA was the most potent activator of PMN, inducing a 6-fold increase in the superoxide anion production, followed by OZ (3-fold increase). PMA activation also induced the greatest modification of LDL by PMN: 700% increase of conjugated dienes formation, 222% increase of TBARS, and 70% increase in the electrophoretic mobility. The indices of oxidative modification significantly correlated with the superoxide anion generated by different activators. Also, LDL oxidation by PMN was inhibited by superoxide dismutase but not by catalase, methionine, or hydroxyl radical scavengers. Our data indicate that PMNs activated by PMA produce a mildly oxidized form of LDL by a mechanism that appears to involve the superoxide anion.

The foods which contribute most to the intake of selected vitamins and minerals were evaluated for the elderly in Italy and compared to food sources of nutrients in the US. A sample of 945 Italians, aged > or = 60, from 14 centres throughout Italy, each completed weighed 7-day food records during the period 1983-1987. These data were used to quantify the contribution of individual foods to the total intake of and variability in intake of vitamin A, thiamin, riboflavin, vitamin C, calcium, and iron. In addition, those foods contributing most to micronutrient intake for the Italians were compared to those foods contributing most for the elderly in the US as recorded by the NHANES II Survey (1976-1980). Overall, there was a preponderance of vegetable rather than animal sources, and of less processed food sources in Italy. In Italy, wine was an important source of several nutrients, including iron and riboflavin. These data are useful for characterizing diet in a well-nourished population with marked differences in diet from the US. They may also form the basis for a quantitative food frequency questionnaire for epidemiological studies in Italy.

A study was carried out on 48 healthy middle-age men and women habitually subsisting on a "Mediterranean type" diet in a rural area of southern Italy. Their freely chosen natural diet was modified for a period of 42 days by partially substituting animal fats for olive oil. Currently available foods were used, and the subjects maintained their habitual lifestyle. Dietary fat content changed from 33 to 37\% of total energy and the polyunsaturated to saturated fatty acid ratio changed from 0.48 to 0.22. The base-line serum total cholesterol of men increased during the dietary intervention period from 214 +/- 30 mg/dl (mean and SD) to 245 +/- 33 mg (+15\%). Low-density lipoprotein cholesterol increased 19\%, while high-density lipoprotein cholesterol remained unmodified. Women, while exhibiting a similar trend in serum total cholesterol (+16\%), showed also a 19\% increase in their high-density lipoprotein cholesterol (p less than 0.001). Apoprotein B increased in parallel with low-density lipoprotein cholesterol in both sexes. The results of the study confirm the impact of the dietary factor on blood lipids. They also provide additional evidence on the response of high-density lipoprotein cholesterol to diet in free-living populations.