Lipoprotein carotenoid profiles and the susceptibility of low density lipoprotein to oxidative modification in healthy elderly volunteers

OBJECTIVES: To determine antioxidant levels in plasma, low density lipoprotein (LDL) and high density lipoprotein (HDL) before and after supplementation with a carotene mixture or lycopene; to examine the interrelationships between carotenoids and tocopherols in plasma, LDL and HDL under normal dietary conditions and after supplementation with carotene or lycopene; and to investigate whether supplementation with a carotene mixture or lycopene could enhance the ability of LDL to withstand oxidative stress in vitro, in a group of healthy elderly people aged > or =65 y. DESIGN: Randomized placebo controlled double blind study. SETTING: Free living urban adults in Ireland. Subjects: Fifty-one volunteers aged > or =65 y. INTERVENTIONS: Volunteers were each provided with capsules providing either 13.3 mg lycopene, or 11.9 mg carotene or placebo for 12 weeks. RESULTS: Both absolute and cholesterol standardized plasma carotenoid concentrations correlated strongly with LDL and HDL concentrations of carotenoids before and after supplementation with carotene or lycopene. Supplementation with a carotene mixture or lycopene had no effect on oxidative modification of LDL in vitro despite significant increases in plasma and LDL concentrations of lycopene, alpha-carotene and beta-carotene. CONCLUSIONS: The results of this study suggest that, in unsupplemented individuals, plasma can act as a biomarker of carotenoid and gamma-tocopherol concentrations in both LDL and HDL. Supplementation with carotenes or lycopene do not reduce or delay oxidation of LDL. These results support the assumption that carotenoids, such as beta-carotene and lycopene, may show protective effects because they are good markers of fruit and vegetable intake.     

Modification of lymphocyte DNA damage by carotenoid supplementation in postmenopausal women

BACKGROUND: Oxidative stress has been implicated in the pathogenesis of chronic diseases related to aging such as cancer and cardiovascular disease. Carotenoids could be a part of a protective strategy to minimize oxidative damage in vulnerable populations, such as the elderly. OBJECTIVE: Our aim was to determine the protective effect of carotenoids against DNA damage. DESIGN: A randomized, double-blind, placebo-controlled intervention study was conducted. Thirty-seven healthy, nonsmoking postmenopausal women aged 50-70 y were randomly assigned to 1 of 5 groups and were instructed to consume a daily dose of mixed carotenoids (beta-carotene, lutein, and lycopene; 4 mg each), 12 mg of a single carotenoid (beta-carotene, lutein, or lycopene), or placebo for 56 d. Plasma carotenoid concentrations were analyzed by using HPLC, and lymphocyte DNA damage was measured by using a single-cell gel electrophoresis (comet) assay. RESULTS: At day 57, all carotenoid-supplemented groups showed significantly lower endogenous DNA damage than at baseline (P < 0.01), whereas the placebo group did not show any significant change. Significantly less (P < 0.05) endogenous DNA damage was found as early as day 15 in the mixed carotenoid (P < 0.01) and beta-carotene (P < 0.05) groups. CONCLUSIONS: The results indicate that carotenoid supplementation decreases DNA damage and that a combination of carotenoids (4 mg each of lutein, beta-carotene, and lycopene), an intake that can be achieved by diet, or a larger dose (12 mg) of individual carotenoids exerts protection against DNA damage.     

Airway and circulating levels of carotenoids in asthma and healthy controls

BACKGROUND: Elevated oxidative stress and impaired antioxidant defences are increasingly recognised features of asthma. Carotenoids are potent dietary antioxidants that may protect against asthma by reducing oxidative damage. OBJECTIVES: This study aimed firstly, to characterise circulating and airway levels of carotenoids in asthma compared to healthy controls, in relation to dietary intake. Secondly, the study aimed to test whether airway lycopene defences can be improved using oral supplements. METHODS: Induced sputum and peripheral blood samples were collected from subjects with asthma (n = 15) and healthy controls (n = 16). Dietary carotenoid intakes were estimated using the 24-hour recall method and analysed using a modified version of the Foodworks 210 Nutrient Calculation Software. Another group of healthy controls (n = 9) were supplemented with 20 mg/day lycopene for 4 weeks. Carotenoids (beta-carotene, lycopene, alpha-carotene, beta-cryptoxanthin, lutein/zeaxanthin) were measured by HPLC. RESULTS: Despite similar dietary intake, whole blood levels of total carotenoids, lycopene, lutein, beta-cryptoxanthin, alpha-carotene and beta-carotene were significantly lower in asthma than controls. However, there were no differences in plasma or sputum carotenoid levels. Induced sputum carotenoid levels were significantly lower than plasma and whole blood levels, but correlated strongly with plasma levels (r = 0.798, p < 0.001). Although there were no overall increases in either plasma or sputum lycopene levels following supplementation, changes in airway lycopene levels correlated with changes in plasma levels (r = 0.908, p < 0.002). CONCLUSIONS: Whole blood, but not plasma or sputum, carotenoid levels are deficient in asthma. Plasma carotenoid levels reflect airway carotenoid levels and when plasma levels are improved using oral supplements this is reflected in the airways.     

Effect of beta-carotene supplementation and lactation on carotenoid metabolism and mitogenic T lymphocyte proliferation

BACKGROUND: Information is lacking regarding the effects of beta-carotene supplementation, early lactation, or both on circulating carotenoid concentrations and T lymphocyte proliferation. OBJECTIVES: This study investigated the effects of short-term beta-carotene supplementation (30 mg/d for 28 d) during early lactation (days 4-32 postpartum) on circulating carotenoid concentrations and on the T lymphocyte proliferative response to phytohemagglutinin. DESIGN: Subjects aged 19-39 y were paired [lactating (4 d postpartum) and nonlactating (never pregnant, healthy women)] and randomly assigned to receive either beta-carotene or a placebo. During the study, subjects provided eight 24-h food records for analysis with the NUTRITIONIST IV and US Department of Agriculture carotenoid databases. Nonfasting blood samples were collected at baseline and at 28 d. Plasma analysis included quantification of alpha-carotene, beta-carotene, lutein, lycopene, retinol, and alpha-tocopherol, complete differential blood cell counts, and lymphocyte proliferative activity. RESULTS: beta-Carotene supplementation increased beta-carotene (P < 0.001) and alpha-carotene (P < 0.05) concentrations but did not affect lycopene concentrations significantly. Supplemented women showed significant decreases in plasma lutein (P < 0.03), as did lactating subjects (P < 0.02). Neither lactation nor beta-carotene supplementation affected the T lymphocyte proliferative response to phytohemagglutinin. CONCLUSIONS: Our results suggest that beta-carotene supplementation as well as some events related to parturition, initiation of lactation, or both alter circulating concentrations of lutein. beta-Carotene supplementation does not enhance T lymphocyte immune competence in healthy women.     

Effect of beta-carotene supplementation on the concentrations and distribution of carotenoids,vitamin E,vitamin A,and cholesterol in plasma lipoprotein and non-lipoprotein fractions in healthy older women.

We studied the effect of beta-carotene supplementation on the concentrations and distribution in plasma lipoprotein and non-lipoprotein fractions of carotenoids, alpha-tocopherol, retinol, and cholesterol.

Ten women ingested either 90 mg of beta-carotene or placebo daily for 3 weeks while residing in their homes and eating their usual meals. Carotenoids (beta-carotene, lycopene, lutein/zeaxanthin), retinol, alpha-tocopherol, and cholesterol were measured in plasma lipoprotein and non-lipoprotein fractions before and after treatment.

In the beta-carotene-supplemented group, total plasma beta-carotene increased 14-fold from 0.48 +/? 0.13 to 6.83 +/? 2.12 mumol/L (p = 0.04). Although the greatest increase in beta-carotene was in low-density-lipoproteins (LDL), the magnitude of increase was similar in LDL, high-density-lipoproteins (HDL), and very-low-density-lipoproteins (VLDL). Thus, the relative distribution of beta-carotene in lipoproteins was unchanged: approximately 71% was in LDL, approximately 15% in HDL and approximately 12% in VLDL, before and after beta-carotene supplementation. There were no changes in amounts and distribution in lipoproteins of the other carotenoids, alpha-tocopherol, and cholesterol. There was no change in the amount of retinol in lipoprotein-deficient plasma. There were no changes in total plasma triglycerides. Significant positive correlations were found between LDL- or VLDL-cholesterol and alpha-tocopherol in LDL or VLDL, respectively; between LDL- or VLDL-cholesterol and lutein/zeaxanthin in LDL or VLDL, respectively; and between HDL-cholesterol and beta-carotene in HDL.

beta-Carotene supplementation (90 mg/day for 3 weeks) in healthy older women results in an enrichment of all plasma lipoprotein fractions with beta-carotene, but does not alter the relative distribution of beta-carotene in lipoproteins. beta-Carotene supplementation has no effect on the amounts and relative distribution of lycopene, lutein/zeaxanthin, and alpha-tocopherol in lipoproteins, or of retinol in the non-lipoprotein fraction of plasma. Short-term beta-carotene supplementation has no effect on the concentrations of plasma total triglycerides, total cholesterol, HDL-, LDL-, and VLDL-cholesterol.     

Effects of tomato juice consumption on plasma and lipoprotein carotenoid concentrations and the susceptibility of low density lipoprotein to oxidative modification

Effects of tomato juice supplementation on the carotenoid concentration in lipoprotein fractions and the oxidative susceptibility of LDL were investigated in 31 healthy Japanese female students. These subjects were randomized to one of three treatment groups; Control, Low and High. The Control, Low and High groups consumed 480 g of a control drink, 160 g of tomato juice plus 320 g of the control drink, and 480 g of tomato juice, providing 0, 15 and 45 mg of lycopene, respectively, for one menstrual cycle. The ingestion of tomato juice, rich in lycopene but having little beta-carotene, increased both lycopene and beta-carotene. Sixty-nine percent of lycopene in plasma was distributed in the LDL fraction and 24% in the HDL fraction. In the Low group, the lycopene concentration increased 160% each in the VLDL+IDL, LDL and HDL fractions (p<0.01). In the High group, the lycopene concentration increased 270% each in the VLDL+IDL and LDL fractions, and 330% in the HDL fraction (p<0.01). Beta-carotene also increased 120% and 180% in LDL fractions of the Low and the High groups, respectively. Despite these carotenoid increases in LDL, the lag time before oxidation was not prolonged as compared with that of the Control group. The propagation rate decreased significantly after consumption in the High group. Multiple regression analysis showed a positive correlation between lag time changes and changes in the alpha-tocopherol concentration per triglyceride in LDL, and a negative correlation between propagation rate changes and changes in the lycopene concentration per phospholipid in LDL. These data suggest that alpha-tocopherol is a major determinant in protecting LDL from oxidation, while lycopene from tomato juice supplementaion may contribute to protect phospholipid in LDI, from oxidation. Thus, oral intake of lycopene might be beneficial for ameliorating atherosclerosis.     

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.     

Carotenoids normally present in serum inhibit proliferation and induce differentiation of a human monocyte/macrophage cell line (U937)

Carotenoids, plant pigments with potent antioxidant activity, are implicated in chronic disease protection. They are absorbed from the diet and transported by plasma lipoproteins. Monocytes, as circulating blood cells, are exposed to carotenoid-rich lipoproteins. Such exposure may lead to enrichment with carotenoids and may affect the functions of monocyte-derived macrophages. This study explored the effect of cellular enrichment in vitro with beta-carotene, lycopene, or lutein on monocyte/macrophage function, using U937 cells as a model. Cell proliferation, production of reactive oxygen species, and cell-substrate adhesion were examined. Maximal carotenoid levels in medium supplemented with preenriched human serum were 2-8 mumol/L; incubation for 1-6 d resulted in 0.2-1.1 nmol carotenoid/mg cell protein (0.25-1 nmol/10(6) cells), approximately 10-fold more than that reported in normal tissue in vivo but within the range that might be anticipated with dietary supplementation. beta-Carotene, lycopene, and lutein markedly inhibited the proliferation of U937 cells, to an extent similar to or greater than that due to phorbol myristic acetate, a known differentiation/activation agent. Lycopene, but not beta-carotene or lutein, caused a significant increase in reactive oxygen species, indicating the induction of cell differentiation. Adhesion and LDL oxidation were unaffected. Thus, cellular carotenoids inhibit proliferation, and for lycopene at least, this may involve cell differentiation. The effectiveness of lycopene, a nonprovitamin A carotenoid, is consistent with a vitamin A-independent pathway modulating cell function.     

Supplementation with beta-carotene or a similar amount of mixed carotenoids protects humans from UV-induced erythema

Carotenoids are useful oral sun protectants, and supplementation with high doses of beta-carotene protects against UV-induced erythema formation. We compared the erythema-protective effect of beta-carotene (24 mg/d from an algal source) to that of 24 mg/d of a carotenoid mix consisting of the three main dietary carotenoids, beta-carotene, lutein and lycopene (8 mg/d each). In a placebo-controlled, parallel study design, volunteers with skin type II (n = 12 in each group) received beta-carotene, the carotenoid mix or placebo for 12 wk. Carotenoid levels in serum and skin (palm of the hand), as well as erythema intensity before and 24 h after irradiation with a solar light simulator were measured at baseline and after 6 and 12 wk of treatment. Serum beta-carotene concentration increased three- to fourfold (P < 0.001) in the beta-carotene group, whereas in the mixed carotenoid group, the serum concentration of each of the three carotenoids increased one- to threefold (P < 0.001). No changes occurred in the control group. The intake of either beta-carotene or a mixture of carotenoids similarly increased total carotenoids in skin from wk 0 to wk 12. No changes in total carotenoids in skin occurred in the control group. The intensity of erythema 24 h after irradiation was diminished in both groups that received carotenoids and was significantly lower than baseline after 12 wk of supplementation. Long-term supplementation for 12 wk with 24 mg/d of a carotenoid mix supplying similar amounts of beta-carotene, lutein and lycopene ameliorates UV-induced erythema in humans; the effect is comparable to daily treatment with 24 mg of beta-carotene alone.     

Circulating levels of retinol, tocopherol and carotenoid in Nepali pregnant and postpartum women following long-term beta-carotene and vitamin A supplementation

OBJECTIVE: To characterize circulating carotenoid and tocopherol levels in Nepali women during pregnancy and post-partum and to determine the effects of beta-carotene and vitamin A supplementation on their concentration in serum. DESIGN: Randomized community supplementation trial. SETTING: The study was carried out from 1994 to 1997 in the Southern, rural plains District of Sarlahi, Nepal. SUBJECTS: A total of 1431 married women had an ascertained pregnancy, of whom 1186 (83%) provided an analyzable serum sample during pregnancy; 1098 (77%) provided an analyzable 3-4 months post-partum serum sample. INTERVENTIONS: Women received a weekly dose of vitamin A (7000 microg RE), beta-carotene (42 mg) or placebo before, during and after pregnancy. Serum was analyzed for retinol, alpha-tocopherol, gamma-tocopherol, beta-carotene, alpha-carotene, lycopene, lutein + zeaxanthin, and beta-cryptoxanthin concentrations during mid-pregnancy and at approximately 3 months post-partum. RESULTS: Compared to placebo, serum retinol, beta-carotene, gamma-tocopherol, beta-cryptoxanthin and lutein + zeaxanthin concentrations were higher among beta-carotene recipients during pregnancy and, except for beta-cryptoxanthin, at postpartum. In the vitamin A group, serum retinol and beta-cryptoxanthin were higher during pregnancy, and retinol and gamma-tocopherol higher at postpartum. Lutein + zeaxanthin was the dominant carotenoid, regardless of treatment group, followed by serum beta-carotene. Serum lycopene level was lowest, and very low compared to the US population. Serum retinol was higher, and carotenoid and alpha-tocopherol lower, at postpartum than during pregnancy in all groups. CONCLUSIONS: Pregnant and lactating Nepali women have lower serum carotenoid and tocopherol levels than well-nourished populations. beta-carotene supplementation appeared to increase levels of tocopherol and other carotenoids in this population.     

Changes in serum retinol,alpha-tocopherol,vitamin C,carotenoids, xinc and selenium after micronutrient supplementation during alcohol rehabilitation

OBJECTIVE: To test the effect of a 21-day supplementation with moderate doses of antioxidant nutrients on biochemical indicators of vitamin, carotenoid and trace element levels in alcohol-dependent patients during a program of alcohol rehabilitation. DESIGN: A randomized double-blind trial was performed comparing two groups receiving daily either a combination of micronutrients (beta-carotene: 6 mg, vitamin C: 120 mg, vitamin E: 30 mg, zinc: 20 mg, selenium: 100 micro g) or a placebo. SUBJECTS: 106 alcohol-dependent patients 20 to 60 years of age without severe liver disease, hospitalized for a 21-day rehabilitation program. Measure of Outcome: Vitamin C, retinol, alpha-tocopherol, zeaxanthin/lutein, beta-cryptoxanthin, lycopene, alpha- and beta-carotene, zinc and selenium were measured in serum, initially and after supplementation. RESULTS: (1) In the placebo group, after 21 days of rehabilitation, serum concentrations of vitamin C and all five carotenoids significantly increased, whereas retinol and alpha-tocopherol concentrations decreased; zinc and selenium levels were unaffected. (2) At the end of the hospital stay, serum indicators were significantly improved in the supplement group as compared to the placebo group for vitamin C, alpha-tocopherol, beta-carotene, zinc and selenium; conversely, lycopene changes were higher in the placebo group than in supplement group. (3) Of the serum antioxidants measured at entrance, only vitamin C was significantly depleted in heavy smokers, and, after the supplementation period, vitamin C was efficiently repleted in this later group. CONCLUSION: Our results indicate that a short-term supplementation with physiological doses of antioxidant vitamins, carotenoids and trace elements during alcohol rehabilitation clearly improves micronutrient status indicators. Heavy smokers in particular seem to respond to vitamin C supplementation.     

Effect of tomato product consumption on the plasma status of antioxidant microconstituents and on the plasma total antioxidant capacity in healthy subjects

OBJECTIVES: to identify the plasma antioxidant microconstituents mainly affected by tomato product consumption, to check whether tomato product consumption can affect antioxidant status, and to identify tomato-product antioxidant-microconstituents mainly involved in the effect of these products on oxidative stress. DESIGN: Medium-term dietary supplementation study. SETTING: Human Nutrition Laboratory, Clermont-Ferrand, France. SUBJECTS: Twenty healthy young (20 < years < 40), non obese (18 < BMI (kg/m2) < 25), females were recruited by advertisement. All of them completed the study. INTERVENTION: The usual diet of the subjects was supplemented for three weeks with 96 g/day tomato puree. The volunteers then avoided tomato-product-rich foods for a subsequent three-week period. Measures of Outcome: Fasting blood samples were collected the day before supplementation, the day after the supplementation period, and the day after the depletion period. The status of several antioxidant microconstituents (plasma microconstituent concentrations), and the antioxidant status (plasma total antioxidant capacity) were assessed. RESULTS: Supplementation with tomato puree significantly increased plasma lycopene, beta-carotene and lutein. Conversely it did not significantly affect plasma vitamin C and E, plasma antioxidant trace metals (Cu, Zn and Se), and plasma total antioxidant capacity. Avoidance of tomato-product-rich foods for three weeks significantly (p < 0.05) decreased plasma lycopene, beta-carotene, lutein and vitamin C, as well as plasma total antioxidant capacity. Plasma total antioxidant capacity, as measured by chemiluminescence, was positively related (p < 0.05) to the status of lycopene, vitamin C and beta-carotene. CONCLUSIONS: Tomato product consumption can affect not only the lycopene status, but also that of other antioxidant microconstituents (beta-carotene and lutein). Lycopene, but also beta-carotene, are apparently the main tomato microconstituents responsible for the effect of tomato products on antioxidant status.     

Bioavailability of lutein from vegetables is 5 times higher than that of beta-carotene.

BACKGROUND: To gain more insight into the relation between vegetable consumption and the risk of chronic diseases, it is important to determine the bioavailability of carotenoids from vegetables and the effect of vegetable consumption on selected biomarkers of chronic diseases. OBJECTIVE: To assess the bioavailability of beta-carotene and lutein from vegetables and the effect of increased vegetable consumption on the ex vivo oxidizability of LDL. DESIGN: Over 4 wk, 22 healthy adult subjects consumed a high-vegetable diet (490 g/d), 22 consumed a low-vegetable diet (130 g/d), and 10 consumed a low-vegetable diet supplemented with pure beta-carotene (6 mg/d) and lutein (9 mg/d). RESULTS: Plasma concentrations of vitamin C and carotenoids (ie, alpha-carotene, beta-carotene, lutein, zeaxanthin, and beta-cryptoxanthin) were significantly higher after the high-vegetable diet than after the low-vegetable diet. In addition to an increase in plasma beta-carotene and lutein, the pure carotenoid-supplemented diet induced a significant decrease in plasma lycopene concentration of -0.11 micromol/L (95% CI: -0.21, -0.0061). The responses of plasma beta-carotene and lutein to the high-vegetable diet were 14% and 67%, respectively, of those to the pure carotenoid- supplemented diet. Conversion of beta-carotene to retinol may have attenuated its plasma response compared with that of lutein. There was no significant effect on the resistance of LDL to oxidation ex vivo. CONCLUSIONS: Increased vegetable consumption enhances plasma vitamin C and carotenoid concentrations, but not resistance of LDL to oxidation. The relative bioavailability of lutein from vegetables is higher than that of beta-carotene.     

Carotenoid composition of human milk during the first month postpartum and the response to beta-carotene supplementation

BACKGROUND: Information is lacking regarding normal changes in milk carotenoid concentrations in healthy, well-nourished women during the first month of lactation. OBJECTIVES: This study investigated milk carotenoid concentrations during days 4-32 postpartum and assessed the effects of maternal beta-carotene supplementation. DESIGN: Subjects (n = 21; aged 19-39 y) were randomly assigned to receive beta-carotene (30 mg/d) or placebo from days 4 to 32 postpartum. Each subject provided 8 diet records and 8 milk samples during the study. Diet records were analyzed for energy, macronutrients, vitamins A and E, and carotenoids. Milk samples were analyzed with HPLC for concentrations of carotenoids, retinol, and alpha-tocopherol. Data were analyzed by using repeated-measures analysis and orthogonal contrasts. RESULTS: No significant differences in average dietary intakes, body mass index, age, or parity were found between groups at baseline or after supplementation. Milk carotenoid concentrations decreased over time (P < 0.01), as did retinol and alpha-tocopherol concentrations (P < 0.003). Concentrations of most carotenoids decreased to those reported for mature milk by day 32 postpartum. Milk lutein concentrations remained elevated throughout the study compared with values reported for mature milk, whereas plasma lutein concentrations decreased significantly over time. beta-carotene supplementation did not significantly change the milk concentrations of beta-carotene, the other carotenoids, retinol, or alpha-tocopherol. CONCLUSIONS: The lack of increase in milk beta-carotene despite supplementation suggests that transitional milk may be already nearly saturated with beta-carotene. The elevated milk lutein concentration and simultaneous decrease in plasma lutein suggest that lutein metabolism may be altered during early lactation.     

Dietary supplementation with a natural carotenoid mixture decreases oxidative stress

OBJECTIVE: To determine whether dietary supplementation with a natural carotenoid mixture counteracts the enhancement of oxidative stress induced by consumption of fish oil. DESIGN: A randomised double-blind crossover dietary intervention. SETTING: Hugh Sinclair Unit of Human Nutrition, School of Food Biosciences, The University of Reading, Whiteknights PO Box 226, Reading RG6 6AP, UK. SUBJECTS AND INTERVENTION: A total of 32 free-living healthy nonsmoking volunteers were recruited by posters and e-mails in The University of Reading. One volunteer withdrew during the study. The volunteers consumed a daily supplement comprising capsules containing fish oil (4 x 1 g) or fish oil (4 x 1 g) containing a natural carotenoid mixture (4 x 7.6 mg) for 3 weeks in a randomised crossover design separated by a 12 week washout phase. The carotenoid mixture provided a daily intake of beta-carotene (6.0 mg), alpha-carotene (1.4 mg), lycopene (4.5 mg), bixin (11.7 mg), lutein (4.4 mg) and paprika carotenoids (2.2 mg). Blood and urine samples were collected on days 0 and 21 of each dietary period. RESULTS: The carotenoid mixture reduced the fall in ex vivo oxidative stability of low-density lipoprotein (LDL) induced by the fish oil (P=0.045) and it reduced the extent of DNA damage assessed by the concentration of 8-hydroxy-2'-deoxyguanosine in urine (P=0.005). There was no effect on the oxidative stability of plasma ex vivo assessed by the oxygen radical absorbance capacity test. beta-Carotene, alpha-carotene, lycopene and lutein were increased in the plasma of subjects consuming the carotenoid mixture. Plasma triglyceride levels were reduced significantly more than the reduction for the fish oil control (P=0.035), but total cholesterol, HDL and LDL levels were not significantly changed by the consumption of the carotenoid mixture. CONCLUSIONS: Consumption of the natural carotenoid mixture lowered the increase in oxidative stress induced by the fish oil as assessed by ex vivo oxidative stability of LDL and DNA degradation product in urine. The carotenoid mixture also enhanced the plasma triglyceride-lowering effect of the fish oil. SPONSORSHIP:: The study was supported by funding from the Greek Studentship Foundation and from Unilever Bestfoods plc. Carotenoids were contributed by Overseal Foods plc.     

Airway and Circulating Levels of Carotenoids in Asthma and Healthy Controls

Background: Elevated oxidative stress and impaired antioxidant defences are increasingly recognised features of asthma. Carotenoids are potent dietary antioxidants that may protect against asthma by reducing oxidative damage.

Objectives: This study aimed firstly, to characterise circulating and airway levels of carotenoids in asthma compared to healthy controls, in relation to dietary intake. Secondly, the study aimed to test whether airway lycopene defences can be improved using oral supplements.

Methods: Induced sputum and peripheral blood samples were collected from subjects with asthma (n = 15) and healthy controls (n = 16). Dietary carotenoid intakes were estimated using the 24-hour recall method and analysed using a modified version of the Foodworks 210 Nutrient Calculation Software. Another group of healthy controls (n = 9) were supplemented with 20 mg/day lycopene for 4 weeks. Carotenoids (β-carotene, lycopene, α-carotene, β-cryptoxanthin, lutein/zeaxanthin) were measured by HPLC.

Results: Despite similar dietary intake, whole blood levels of total carotenoids, lycopene, lutein, β-cryptoxanthin, α-carotene and β-carotene were significantly lower in asthma than controls. However, there were no differences in plasma or sputum carotenoid levels. Induced sputum carotenoid levels were significantly lower than plasma and whole blood levels, but correlated strongly with plasma levels (r = 0.798, p < 0.001). Although there were no overall increases in either plasma or sputum lycopene levels following supplementation, changes in airway lycopene levels correlated with changes in plasma levels (r = 0.908, p < 0.002).

Conclusions: Whole blood, but not plasma or sputum, carotenoid levels are deficient in asthma. Plasma carotenoid levels reflect airway carotenoid levels and when plasma levels are improved using oral supplements this is reflected in the airways.     

Daily intake of a formulated tomato drink affects carotenoid plasma and lymphocyte concentrations and improves cellular antioxidant protection

The salutary characteristics of the tomato are normally related to its content of carotenoids, especially lycopene, and other antioxidants. Our purpose was to verify whether the daily intake of a beverage prototype called Lyc-o-Mato((R)) containing a natural tomato extract (Lyc-o-Mato((R)) oleoresin 6 %) was able to modify plasma and lymphocyte carotenoid concentrations, particularly those of lycopene, phytoene, phytofluene and beta-carotene, and to evaluate whether this intake was sufficient to improve protection against DNA damage in lymphocytes. In a double-blind, cross-over study, twenty-six healthy subjects consumed 250 ml of the drink daily, providing about 6 mg lycopene, 4 mg phytoene, 3 mg phytofluene, 1 mg beta-carotene and 1.8 mg alpha-tocopherol, or a placebo drink. Treatments were separated by a wash-out period. Plasma and lymphocyte carotenoid and alpha-tocopherol concentrations were determined by HPLC, and DNA damage by the comet assay. After 26 d of consumption of the drink, plasma carotenoid levels increased significantly: concentrations of lycopene were 1.7-fold higher (P<0.0001); of phytofluene were 1.6-fold higher (P<0.0001); of phytoene were doubled (P<0.0005); of beta-carotene were 1.3-fold higher (P<0.05). Lymphocyte carotenoid concentrations also increased significantly: that of lycopene doubled (P<0.001); that of phytofluene was 1.8-fold higher (P<0.005); that of phytoene was 2.6-fold higher (P<0.005); that of beta-carotene was 1.5-fold higher (P<0.01). In contrast, the alpha-tocopherol concentration remained nearly constant. The intake of the tomato drink significantly reduced (by about 42 %) DNA damage (P<0.0001) in lymphocytes subjected to oxidative stress. In conclusion, the present study supports the fact that a low intake of carotenoids from tomato products improves cell antioxidant protection.     

Dietary fiber reduces the antioxidative effect of a carotenoid and α-tocopherol mixture on LDL oxidation ex vivo in humans

BACKGROUND: Antioxidant concentrations in low density lipoproteins (LDL) are an important determinant for their susceptibility to oxidation and can be modulated by dietary intake. AIM OF THE STUDY: In the present study, the influence of dietary fiber on the antioxidant enrichment and the oxidation resistance of LDL after antioxidant supplementation is investigated. METHOD: An antioxidant supplement consisting of beta-carotene, lycopene, lutein, canthaxanthin and alpha-tocopherol was given to six young women together with a standard meal. Using a cross-over study design, each subject received the standard meal without additional dietary fiber and enriched with pectin, guar, or cellulose in a random order. To determine the resistance of LDL against copper ion-induced oxidation, the formation of conjugated dienes was measured. RESULTS: Eight, 10, and 24 hours after antioxidant supplementation the isolated LDL revealed significantly (p < 0.05) increased antioxidant concentrations; addition of pectin, guar, or cellulose to the meal depressed this increase. Concomitantly, the observed increase in the resistance of LDL against oxidation (measured as lag phase) was lower with dietary fiber supplementation than that found without. On average, pectin, guar, and cellulose reduced the increase of the lag phase (measured without addition of dietary fiber) by 38%, 22%, and 18%, respectively. CONCLUSIONS: These results indicate that dietary fiber supplementation decreases the antioxidative effect of a supplement consisting of carotenoids and alpha-tocopherol in LDL, an effect that is likely to be mediated by a reduced bioavailability of these antioxidants in the gut.     

DNA damage and susceptibility to oxidative damage in lymphocytes: effects of carotenoids in vitro and in vivo

Reports on the effects of carotenoids are conflicting. The present paper examines similarities and differences from contiguous studies in vitro and in vivo. Single-cell gel electrophoresis was used to measure the frequency of single-strand breaks (SSB) in the cell line MOLT-17 (as a model system) and human peripheral blood lymphocytes (PBL). MOLT-17 cells were supplemented with beta-carotene, lutein or lycopene at a range of concentrations (0.00-8.00 micromol/l) using a liposome delivery method. Uptake was dose-dependent. beta-Carotene concentration in the media had no effect on SSB in control cells, but incubation with lycopene or lutein (>2.00 micromol/l) increased the numbers of SSB in control cells. MOLT-17 DNA was less susceptible to oxidative damage (100 micromol H2O2/l, 5 min, 4 degrees C) following incubation with carotenoids between 0.50 and 1.00 micromol/l; at >1.00 micromol/l the effects were ambiguous. Apparently healthy male volunteers supplemented their habitual diets with lutein, beta-carotene or lycopene (natural isolate capsules, 15 mg/d, 4 weeks) in three independent studies, raising plasma concentrations to different extents. Lycopene and lutein had no effect on SSB in control PBL or following oxidative challenge. However, increased plasma beta-carotene was associated with more SSB in control cells whilst PBL DNA resistance to oxidative damage ex vivo was unaffected. These results suggest that the carotenoids are capable of exerting two overlapping but distinct effects: antioxidant protection by scavenging DNA-damaging free radicals and modulation of DNA repair mechanisms.     

No antioxidant effect of combined HRT on LDL oxidizability and oxidative stress biomarkers in treated post-menopausal women

OBJECTIVE: To compare oxidative stress and LDL oxidizability in postmenopausal women with and without HRT. METHODS: In a cross sectional study, two groups of women, with or without combined per os HRT (1.5-2 mg estrogen associated with 10 mg dydrogesteron), were age and duration of menopause matched. Women were recruited after medical examination at LBSO (Oxidative Stress Laboratory), Joseph Fourier University, Grenoble, and Department of Gynecology, Grenoble University Hospital, France. Main outcome measures included determination of lipid profile and oxidative stress biomarkers (TBARS, LDL oxidizability, autoantibodies against oxidized-LDL). Measurement of circulating levels of vitamin C, E, beta-carotene, lycopene and total antioxidant plasma capacity. RESULTS: HRT led to decreased plasma total and LDL cholesterol (p < 0.05), but did not affect oxidizability and oxidation of LDL. Circulating levels of antioxidant vitamins (beta-carotene, vitamin C, vitamin E/triglycerides) and total antioxidant capacity of plasma and lipid peroxidation, assessed by plasma TBARs, were not different from controls in postmenopausal women receiving HRT. CONCLUSION: This study suggests that even if combined HRT modifies the blood lipid profile, it does not appear to influence oxidative status.