The Medi-RIVAGE study: reduction of cardiovascular disease risk factors after a 3-mo intervention with a Mediterranean-type diet or a low-fat diet.

BACKGROUND: Epidemiologic studies link Mediterranean-type diets to a low incidence of cardiovascular disease; however, few dietary intervention studies have been undertaken, especially in primary prevention. OBJECTIVES: In the Mediterranean Diet, Cardiovascular Risks and Gene Polymorphisms (Medi-RIVAGE) study, the effects of a Mediterranean-type diet (Med group) or a low-fat diet (low-fat group) on risk factors were evaluated in 212 volunteers (men and women) with moderate risk factors for cardiovascular disease. DESIGN: After the 3-mo dietary intervention, changes in many risk factors were evaluated. Dietary questionnaires and plasma nutritional markers were used to test compliance. RESULTS: Although the dietary goals were only partially reached, changes in dietary habits were observed in both groups (n = 169): protein, carbohydrate, and fiber intakes increased and fat quality (decreased saturated fat and increased monounsaturated or polyunsaturated fat) improved. BMI, total and triacylglycerol-rich lipoprotein (TRL) cholesterol, triacylglycerols, TRL triacylglycerols, apolipoproteins A-I and B, insulinemia, glycemia, and the homeostasis model assessment score were significantly lower after 3 mo. The reductions in total cholesterol, triacylglycerols, and insulinemia remained significant after adjustment for BMI. There was a trend for a diet-by-time interaction for LDL cholesterol (P = 0.09). Our data predicted a 9% reduction in cardiovascular disease risk with the low-fat diet and a 15% reduction with this particular Mediterranean diet. CONCLUSION: After a 3-mo intervention, both diets significantly reduced cardiovascular disease risk factors to an overall comparable extent.

Lycopene but not lutein nor zeaxanthin decreases in serum and lipoproteins in age-related macular degeneration patients.

BACKGROUND: Epidemiological studies have established that a low serum concentration of carotenoids was associated with risk of Age-Related Macular Degeneration (ARMD). The aim of this study was to determine carotenoid levels in serum and in different lipoprotein fractions in patients diagnosed for ARMD and in matched control group. METHOD: Thirty-four ARMD patients and 21 control subjects from Brest area (France) have been included to this study. Lipoproteins have been separated from serum by gradient density ultracentrifugation. We measured concentration of carotenoids and tocopherols in serum and in different lipoprotein fractions by HPLC. RESULTS: No difference was observed between ARMD patients and control subjects in total serum carotenoids. Individual carotenoid levels showed that only lycopene was decreased significantly in serum, LDL and HDL fractions in patients (P<0.05). Concentrations in serum and lipoparticle fractions of lutein and zeaxanthin, the major pigments present in macula were not modified between both groups. CONCLUSIONS: Lycopene, as liposoluble antioxidant nutrient, is the only carotenoid altered in ARMD patients. It cannot be excluded that this effect is related to different dietary habits, but we hypothesise that lower lycopene status could result also from specific antioxidant protection of lutein and zeaxanthin by lycopene.

In vivo and in vitro evidences that carotenoids could modulate the neutrophil respiratory burst during dietary manipulation.

BACKGROUND: The primary role of polymorphonuclear neutrophils (PMNs) is to destroy pathogenic microorganisms after phagocytosis by producing reactive oxygen species (ROS) and toxic molecules. However, PMNs produce sufficient amounts of ROS during an oxidative burst to be autotoxic and detrimental to their own functions and to possibly cause DNA damage, protein and lipid oxidation and cell membrane destructuration. OBJECTIVE: The aim of this study was to investigate in vivo the role of the antioxidant capacities of carotenoids in modulating ROS content in PMNs during oxidative burst. Moreover to investigate the direct or indirect effect of carotenoids, the modification of PMN ROS content was explored after in vitro supplementation with beta-carotene or lycopene, chosen taking account of their vitamin A and no vitamin A precursor effect, respectively. DESIGN: In vivo study: Venous blood was collected from 10 healthy male volunteers and ROS production from phorbol myristate acetate (PMA)-stimulated PMNs was determined, by flow cytometry using the fluorescent dye dihydrorhodamine 123, at baseline, after 3 weeks of carotenoid depletion (carotenoid intake limited to 25% of usual intake) and after 5 weeks of carotenoid repletion (30 mg beta-carotene, 15 mg lycopene and 9 mg lutein per day). In vitro study: ROS content in PMA-stimulated PMNs isolated from carotenoid depleted subjects and controls was quantified after an in vitro enrichment with beta-carotene (1 micromol/L) or lycopene (0.3 micromol/L). RESULTS: In vivo carotenoid depletion increased PMN H2O2 content after PMA activation by 38% (p < 0.05 vs baseline),while supplementation for 5 weeks restored basal H2O2 generation (p < 0.05 vs depletion). Although H2O2 measurement in PMNs from non-depleted subjects was not affected by an in vitro supply with beta-carotene or lycopene, a significant decrease in H2O2 content by 78.9 % and 81.2%, respectively, was observed in PMNs from carotenoid depleted subjects (p < 0.01 vs depleted control subjects). CONCLUSIONS: The carotenoid ROS quenching capacities control both in vivo and in vitro the PMNs ROS generation and probably protect these cells against DNA, membrane lipid and protein damages during oxidative burst. Moreover, these effects appear independent from the metabolic conversion of carotenoids to vitamin A.

Comparison of the postprandial chylomicron carotenoid responses in young and older subjects.

BACKGROUND: The plant carotenoids may contribute to the beneficial health effect of fruits- and vegetables-rich diet. Epidemiological studies consistently associated high plasma carotenoids status with reduced age-related diseases. However, the data concerning the bioavailability of carotenoids in the elderly are scarce. OBJECTIVE: To test whether there is an age effect on carotenoid bioavailability. DESIGN: Eight young (20-35 y) and eight older (60-75 y) healthy adults ingested three different meals containing 40 g triacylglycerols (TG) and vegetable sources of carotenoids. These sources were either 188 g carrot purée which provided 30 mg betacarotene as the main carotenoid, or 61 g tomato purée providing 30mg lycopene, or 260 g cooked chopped spinach providing 30 mg lutein. TG and carotenoids were assayed in chylomicrons (CM) collected for 9 h postprandially. RESULTS: There was no major effect of age on the postprandial CM/TG response (0-9 h area under the curve (AUC)). There was no major effect of age on the postprandial CM all- trans beta-carotene, cis betacarotene, alpha-carotene, and lutein responses. Adjustment of these responses by the CM TG responses did not reveal any age effect. While there was no significant effect of age on the CM lycopene response, the CM TG-adjusted lycopene response was significantly lower (-40 %) in the older than in the younger subjects (P < 0.04). The cis-trans ratios of CM betacarotene were not significantly different between the old and the young subjects. There was no significant effect of age on the ratio of CM retinyl-palmitate to the sum of alpha-carotene and beta-carotene measured after the carrot meal. CONCLUSIONS: The bioavailability of lycopene is apparently impaired in the old,while there is no major difference in the bioavailability of beta-carotene, alpha-carotene and probably lutein. There is also no major effect of age on the cis-trans isomerization of beta-carotene during absorption, and in the intestinal conversion of provitamin A carotenoids into vitamin A.

Effect of carrot intake on cholesterol metabolism and on antioxidant status in cholesterol-fed rat.

BACKGROUND: Vegetables are major dietary sources of fibers and antioxidants such as carotenoids, polyphenols and vitamin C which contribute to explain their protective effects against cardiovascular diseases. AIM OF THE STUDY: We investigated in the rat the effects of a 3-week supplementation of the diet with carrot (15% dry matter) on lipid metabolism and antioxidant status. RESULTS: A significant decrease of cholesterol level in liver (-44%; P= 0.0007) was observed together with a reduction of the level of liver triglycerides (-40%; P= 0.0005). Fecal total steroids excretion increased by 30% upon feeding the carrot diet as compared to the control. The secretion of bile acids was maintained, whereas the cholesterol apparent absorption was reduced in rats fed carrot diet. Carrot consumption also improved the antioxidant status. It significantly decreased the urinary excretion of thiobarbituric acid reactive substances (TBARS), reduced the TBARS levels in heart, increased the vitamin E plasmatic level and tended to increase the ferric reducing ability of plasma (FRAP) as compared to the controls. The carrot diet provided carotenoid antioxidants: 5.1 mg beta-carotene, 1.6 mg alpha-carotene and 0.25mg lutein per 100 g diet. No carotenoids were found in plasma whereas the three carotenoids were detected in the plasma of the rats fed the carrot diet at 125, 41, 43 nmol/L respective concentrations. beta-Carotene was also detected in liver and heart. CONCLUSION: Carrot consumption modifies cholesterol absorption and bile acids excretion and increases antioxidant status and these effects could be interesting for cardiovascular protection.

Short-term supplementation with lutein affects biomarkers of lutein status similarly in young and elderly subjects.

There is evidence that lutein may protect against age-related macular degeneration, cataract, cancers and cardiovascular diseases, but no data have been published on the effect of age on lutein status. The purpose of this work was to determine whether there are major differences in the status of this carotenoid between young and elderly subjects. Initial lutein status and the effect of a 5-week lutein supplementation (9 mg/d) on the most common markers of lutein status were compared in 12 young (26.9+/-0.8yr) and 17 older subjects (67.3+/-1.1yr). Lutein was measured by HPLC in fasting serum, adipose tissue and buccal mucosa cells (BMC) before and after supplementation. Macular pigment optical density (MPOD), which partly depends on retina lutein concentration, was measured by reflectometry before and after supplementation. Initial lutein status was not significantly different between the two groups, irrespective of the lutein status marker. Plasma and BMC lutein concentrations significantly increased in both groups after lutein supplementation, but not MPOD or adipose tissue lutein. Plasma and BMC responses to lutein supplementation (percent variation from initial values) were not significantly different between the two groups. These results suggest that there is no major effect of age on lutein status in healthy subjects.

Carotenoids, mostly the xanthophylls, exchange between plasma lipoproteins.

Carotenoids are exclusively transported by lipoproteins; in vitro studies suggest that they might protect these particles against oxidation. Little is known about the factors that govern the distribution of these micronutrients among lipoproteins. The objective of this study was to assess whether carotenoids are exchanged between lipoproteins and what factors, if any, were involved. In the first experiment, different groups of trout were fed for five days with either a carotenoid-free diet or with diets containing 80 mg pure carotenoid per kilogram of feed. Lipoproteins were separated by ultracentrifugation and carotenoid-rich, high-density lipoproteins (HDL) were incubated for two hours at 37 degrees C with carotenoid-free, very low-density lipoproteins (VLDL), and vice versa. After incubation, lipoproteins were re-separated and carotenoids were quantified to measure the transfer. The same experiments were done in the presence of cholesteryl ester transfer protein (CETP) and lecithin cholesterol acyltransferase (LCAT) inhibitors. In a second experiment, the exchange was measured between human VLDL and HDL. In trout, incubation of carotenoid-rich HDL with carotenoid-free VLDL resulted in the appearance of carotenoids in VLDL, and inversely. The higher the hydrophobicity of a carotenoid, the lower its proportion in HDL after incubation. CETP and LCAT inhibitors significantly increased the proportion of carotenoids in HDL after incubation. Results obtained with human lipoproteins showed that the xanthophyll lutein transferred between lipoproteins, but could not show any carotenes (alpha-carotene, beta-carotene, and lycopene) transfer. We conclude that carotenoids, chiefly the xanthophylls, exchange between lipoproteins. The transfer depends on plasma factor(s) sensitive to CETP and/or LCAT inhibitors.

Feedback regulation of beta,beta-carotene 15,15'-monooxygenase by retinoic acid in rats and chickens.

beta,beta-Carotene 15,15'-monooxygenase (formerly termed beta,beta-carotene 15,15'-dioxygenase, EC 1.13.11.21) catalyzes the conversion of provitamin A carotenoids to retinal in vertebrate tissues. In the present study, we investigated whether preformed vitamin A or beta-carotene and its direct metabolites can regulate the enzyme activity in vivo. We found dose-dependent decreases in intestinal beta,beta-carotene monooxygenase activity after oral administration to rats of retinyl acetate (up to -79%), beta-carotene (up to -79%), apo-8'-carotenal (up to -56%), all-trans retinoic acid (up to -88%), and 9-cis retinoic acid (up to -67%). Liver beta,beta-carotene 15,15'-monooxygenase (betaCMOOX) activity was not affected. Apo-12'carotenal and the retinoic acid receptor (RAR) alpha antagonist Ro 41-5253 significantly increased the intestinal enzyme activity by 55 and 94%, respectively. When beta-carotene was administered to rats pretreated with the two cytochrome P(450) (CYP) inducers, pentobarbital and naphthoflavone, the intestinal betaCMOOX activity increased by 39%. In a transcriptional study in chickens, treatment with retinoic acid resulted in low expression of the intestinal betaCMOOX. Our data suggest that retinoids and carotenoids might regulate betaCMOOX expression by a transcriptional feedback mechanism via interaction with members of the RAR family.

Short-term consumption of a high-sucrose diet has a pro-oxidant effect in rats.

The underlying mechanisms for the detrimental consequences of a high-fructose diet in animal models are not clear. However, the possibility exists that fructose feeding facilitates oxidative damage. Thus, the aim of the present study was to assess, in weaning rats, the effect of a high-sucrose diet v. starch diet for 2 weeks on oxidative stress variables. Plasma lipid levels were measured and lipid peroxidation was evaluated by urine and plasma thiobarbituric acid-reactive substances (TBARS). The susceptibilities of several tissues to peroxidation were determined in tissue homogenates after in vitro lipid peroxidation. Antioxidant defence variables were evaluated by measuring plasma and heart vitamin E levels, and heart superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities. Higher plasma triacylglycerol (P<0.01) and TBARS (P<0.01) levels were found in rats fed the sucrose diet as compared with the starch-fed group, whereas plasma alpha-tocopherol levels were significantly decreased in the sucrose-fed group compared with the starch-fed group (P<0.01). Higher urine TBARS (P<0.01) were found in the sucrose-fed group compared with the starch-fed group, suggesting increased production of these substances from lipid peroxidation in vivo. Higher susceptibility to peroxidation in heart, thymus and pancreas was also found in the sucrose-fed group v. the starch-fed group. No statistical differences were observed for liver TBARS level between the two groups. Heart SOD activity was significantly decreased (P<0.001) in the sucrose-fed group compared with the starch-fed group, whereas heart vitamin E level and GPX activity were not different between the groups. However, the in vitro generation of superoxide radical in heart homogenate, measured by electron spin resonance detection and spin trapping, was not increased in the sucrose-fed group compared with starch-fed rats. Altogether, the results indicate that a short-term consumption of a high-sucrose diet negatively affects the balance of free radical production and antioxidant defence in rats, leading to increased lipid susceptibility to peroxidation.

Vegetable-borne lutein, lycopene, and beta-carotene compete for incorporation into chylomicrons, with no adverse effect on the medium-term (3-wk) plasma status of carotenoids in humans.

BACKGROUND: The results of epidemiologic studies have consistently shown associations between dietary intake or plasma carotenoid status and incidence of cancers and cardiovascular and eye diseases. OBJECTIVE: The aim was to assess whether vegetable-borne carotenoids (lycopene, lutein, and beta-carotene) compete for intestinal absorption and whether this affects the plasma status of carotenoids in the medium term (ie, after 3 wk). DESIGN: During 3-wk periods separated by 3-wk washout periods, 20 women were supplemented with either 96 g tomato purée/d (14.98 mg lycopene + 1.50 mg beta-carotene), 92 g cooked chopped spinach/d (11.93 mg lutein + 7.96 mg beta-carotene), 96 g tomato purée/d + 92 g chopped spinach/d, 96 g tomato purée/d + 2 lutein pills (12 mg lutein), or 92 g chopped spinach/d + 1 lycopene pill (15 mg lycopene). Plasma carotenoids were measured before and after each supplementation period. The subjects also participated in postprandial experiments in which they ingested meals containing double amounts of the supplements described above. Carotenoids were measured in chylomicrons to assess the interaction of carotenoids on absorption. RESULTS: Adding a second carotenoid to a meal that provided a first carotenoid diminished the chylomicron response to the first carotenoid. However, cosupplementation with a second carotenoid of a diet supplemented with a first carotenoid did not diminish the medium-term plasma response to the first carotenoid. CONCLUSION: Consumption of carotenoids from different vegetable sources does not diminish plasma carotenoid concentrations in the medium term, despite the finding in postprandial testing of competitive inhibitory interactions among different carotenoids.