Processing tangerine tomatoes: effects on lycopene-isomer concentrations and profile.

Because lycopene is a powerful biological antioxidant, its delivery to humans is of major concern. cis-Lycopene isomers are more bioavailable than the all-trans isomers and thus more efficiently absorbed. Tangerine tomatoes, whose lycopene isomeric content is almost all tetra-cis, provide a useful food source for comparing cis- and trans-isomer absorption. Tangerine tomatoes were processed into sauce in the Univ. of California, Davis Pilot Plant for subsequent use in a human feeding study described in another publication. Samples were taken at several stages during processing and carotenoids extracted and analyzed for carotenoid-isomer profiles and concentrations. Analyses showed that total lycopene concentration decreased considerably during the 1st step of processing, which included heating and juicing operations. Processing resulted in a large decrease in tetra-cis lycopene concentration accompanied by increases in trans- and other cis-lycopene isomers.

Serum carotenoid depletion follows first-order kinetics in healthy adult women fed naturally low carotenoid diets.

Dietary intakes of carotenoids are highly variable in human populations as are serum carotenoid concentrations. However, there are few controlled data relating carotenoid intake to concentration. Most of the data that are available are from measurements of the absorption and decay of large pharmacologic doses of carotenoids, and are therefore of unknown physiologic relevance. Our objective was to determine the half-life (t(1/2)) of the most abundant carotenoids in blood serum from healthy adult women living under controlled conditions. As part of two carotenoid isotopic studies, we measured serum concentrations of beta-carotene, alpha-carotene, lutein, zeaxanthin, beta-cryptoxanthin and lycopene in 19 healthy young adult women that were fed controlled low carotenoid diets for approximately 10 wk. All other nutrients (vitamins A, E and C) were provided at 100-150% of the 1989 U.S. recommended dietary allowance levels. Exercise and activities were controlled throughout the studies to simulate usual activity patterns. Carotenoid concentrations were measured by reversed-phase HPLC. Serum carotenoid concentration decreases during depletion followed first-order kinetics. The half-lives determined in decreasing order were as follows: lutein (76 d) > alpha-carotene (45 d) = beta-cryptoxanthin (39 d) = zeaxanthin (38 d) = beta-carotene (37 d) > lycopene (26 d). Half-lives were unrelated to physical or demographic characteristics such as body mass, body fat, racial background or age in these relatively homogeneous groups. Carotenoids decreased by similar first-order mechanisms, although the rates differed for individual carotenoids.

Variability of the conversion of beta-carotene to vitamin A in women measured by using a double-tracer study design.

BACKGROUND: Blood beta-carotene and vitamin A responses to oral beta-carotene are variable in humans. Some individuals are characterized as responders and others as low- or nonresponders. A better understanding of the conditions that produce the variability is important to help design public health programs that ensure vitamin A sufficiency. OBJECTIVE: Our objective was to assess variability in absorption and conversion of beta-carotene to vitamin A in vivo in humans by using a novel double-tracer ¿hexadeuterated (D(6)) beta-carotene and D(6) retinyl acetate approach. DESIGN: Eleven healthy women were housed at the US Department of Agriculture Western Human Nutrition Research Center metabolic unit for 44 d, where they consumed diets adequate in vitamins and minerals except for carotenoids. After an adaptation period, the women were given 30 micromol D(6) retinyl acetate orally, followed 1 wk later with 37 micromol D(6) beta-carotene (approximately equimolar doses). Time-dependent plasma concentration curves were determined for D(6) retinol, D(6) beta-carotene, and trideuterated (D(3)) retinol (derived from D(6) beta-carotene). RESULTS: Mean (+/-SE) absorption of D(6) beta-carotene was 3.3 +/- 1.3% for all subjects. The mean conversion ratio was 0.81 +/- 0.34 mol D(3) retinol to 1 mol D(6) beta-carotene for all subjects. However, only 6 of the 11 subjects had plasma D(6) beta-carotene and D(3) retinol concentrations that we could measure. The mean absorption of D(6) beta-carotene in these 6 subjects was 6.1 +/- 0.02% and their conversion ratio was 1.47 +/- 0.49 mol D(3) retinol to 1 mol D(6) beta-carotene. The remaining 5 subjects were low responders with

Effect of oral beta-carotene supplementation on plasma human immunodeficiency virus (HIV) RNA levels and CD4+ cell counts in HIV-infected patients.

We conducted a pilot, open-label study to assess the effect of short-term beta-carotene administration (180 mg/d with meals for 4 weeks) on the plasma human immunodeficiency virus (HIV) RNA levels and CD4+ lymphocyte counts in 21 HIV-infected patients. We found that plasma HIV RNA levels and CD4+ lymphocyte counts did not change following this short course of beta-carotene supplementation. Patients with lower serum concentrations of beta-carotene before supplementation were no more likely to have an increase in their CD4+ lymphocyte count or plasma HIV RNA copy number than were those with higher concentrations. No correlation was found between pre- or postsupplementation beta-carotene or vitamin A concentrations and pre- or postsupplementation CD4+ lymphocyte counts or plasma HIV RNA titers. This study provides no support for beta-carotene supplementation for HIV-infected subjects with normal baseline serum levels of beta-carotene and vitamin A.

Compartmental models of vitamin A and beta-carotene metabolism in women.

We have developed compartmental models of vitamin A and beta-carotene (beta C) metabolism in women living under controlled conditions on diets with known concentrations of vitamins and carotenoids. Fourteen healthy adult women were given either retinyl-d4 acetate, or beta C-d8 before breakfast. Natural and stable-isotopes of retinol and beta C were collected in serum for up to 95 days or 20 days, respectively. Stable isotopes were separated from other components and measured by GC-MS or HPLC-UV. Preformed retinyl-d4 acetate metabolism in all women tested can be accurately described by a simple four-compartment model. However, the model did not fit one women initially, when she had marginal vitamin A status. We tested the hypothesis that dietary changes of beta C intake have important roles on the kinetics of vitamin A metabolism. Dietary changes of beta C intake did not influence the turnover rate of retinol in any compartment. However, it did result in changes in steady-state masses and residence times of retinol in several compartments. A working compartmental model for beta C metabolism was developed. The kinetics of retinol-d4 formed from beta C is more complicated than the pre-formed retinol-d4. Results suggest that beta C-d8 readily converts into retinol-d4 with high inter-individual variability.

Estimating the concentration of beta-carotene required for maximal protection of low-density lipoproteins in women.

The reportedly inconsistent antioxidant protective effect of beta-carotene on plasma LDL may depend on LDL's beta-carotene concentration. We measured carbonyl production by CuSO4-challenged LDL from nine healthy women living at the US Department of Agriculture-Western Human Nutrition Research Center and consuming a natural food diet that provided only 0.14 micromol beta-carotene/d for 120 d. During the first 60 d, four women received a placebo and the remaining five women received too small a supplement (0.93 micromol beta-carotene/d) to increase plasma or LDL beta-carotene; therefore, the data for all nine women during this time were pooled. From days 61 to 120, all subjects received the small supplement. From days 101 to 120 they all received an additional, larger, mixed carotenoid supplement (6.16 micromol beta-carotene/d). Plasma beta-carotene dropped from 0.76 +/- 0.21 micromol/L (x +/- SEM) on day 2 to 0.33 +/- 0.08 on day 60 (P = 0.035) and rose to 1.73 +/- 0.18 (P = 0.001) on day 120. LDL beta-carotene dropped from 1.67 +/- 0.53 micromol/g LDL protein on day 2 to 1.27 +/- 0.28 micromol/g LDL protein on day 60 (P = 0.650) and rose to 10.04 +/- 1.07 micromol/g LDL protein (P = 0.001) on day 120. Plasma lycopene dropped from 0.20 micromol/L on day 2 to 0.02 micromol/L on day 60 and did not increase by day 120. Carbonyl production rose from 24 +/- 6 micromol/g LDL protein on day 2 to 42 +/- 4 micromol/g LDL protein (P = 0.001) on day 60 and dropped to 6 +/- 1 micromol/g LDL protein (P = 0.001) on day 120. LDL seemed fully protected with 9.7 +/- 2.5 micromol beta-carotene/g LDL protein, or 2.3 +/- 1.8 micromol beta-carotene/L plasma.

The effect of a low carotenoid diet on malondialdehyde-thiobarbituric acid (MDA-TBA) concentrations in women: a placebo-controlled double-blind study.

OBJECTIVE: The purpose of the study was to evaluate the effect of a low carotenoid diet (83 micrograms Beta-carotene) on malondialdehyde-thiobarbituric acid (MDA-TBA) concentrations of nine pre-menopausal women. METHODS: Subjects lived on the metabolic research unit of the Western Human Nutrition Research Center (WHNRC), where diet, exercise and other activities were controlled. Five subjects (Group C, control group) consumed a low carotenoid diet and received an additional 0.5 mg/day of Beta-carotene while four subjects (Group P, placebo group) received only the low carotenoid diet during days 1 to 60 (period 1). All subjects received 0.5 mg/day of Beta-carotene during days 60 to 100 (period 2), plus three capsules/day mixed carotenoid supplement (Neo-Life Company) during study days 100 to 120. Changes in MDA-TBA concentrations were analyzed during the study periods and between the groups. RESULTS: At the start of the study (day 1), no significant difference in the MDA-TBA concentration was observed between the control (Group C) and the placebo (Group P) subjects. During period 1 (days 2 to 60), when Group P subjects consumed the low carotenoid diet without supplementation, the MDA-TBA values for Group P rose markedly and were significantly (p < 0.05) higher than the MDA-TBA values for Group C subjects who were receiving carotenoid supplementation. During period 2 (days 60 to 100) when both groups received carotenoid supplementation, the MDA-TBA values of Group P subjects were significantly (p < 0.05) reduced to the point where they were similar to the MDA-TBA values for Group C subjects. CONCLUSIONS: These findings provide evidence to support the beneficial effects of carotenoids in preventing lipid peroxidation in the cells. Further studies are needed to identify the exact mechanism by which carotenoids prevent lipid peroxidation and the amount needed for normal activity.

Modulated mitogenic proliferative responsiveness of lymphocytes in whole-blood cultures after a low-carotene diet and mixed-carotenoid supplementation in women.

To determine the effects of dietary carotenes on the mitogenic proliferative responsiveness of blood lymphocytes in vitro, nine premenopausal women were fed a low-carotene diet for 120 d. Low-dose beta-carotene (0.5 mg/d) was given to five subjects on days 1-60, while four received a placebo. All subjects received a low-dose beta-carotene (0.5 mg/d) supplement on days 61-120, plus a carotenoid complex on days 101-120. The mean (+/-SEM) serum beta-carotene concentration for the combined beta-carotene supplemented and placebo subjects (n = 9) was not significantly reduced from that on day 1 (1.27 +/- 0.24 mumol/L) on days 60 (0.66 +/- 0.14 mumol/L) and 100 (0.91 +/- 0.38 mumol/L), but on day 120 (3.39 +/- 0.44 mumol/L) it was increased above that on days 1, 60, and 100. Maximum mitogenic proliferative responsiveness of blood lymphocytes in vitro to optimal dose phytohemagglutinin (PHA) was reduced on days 60 (P = 0.025) and 100 (P < 0.0001), but corrected itself on day 120 to a value above those on day 1 (P = 0.04), day 60 (P = 0.0001), and day 100 (P < 0.0001). Present findings show that a diet low in carotene had a suppressive effect on the maximum mitogenic proliferative responsiveness of blood lymphocytes in vitro, which was not corrected with low-dose beta-carotene supplementation but was with a carotenoid complex from vegetables rich in carotenoids.

Supercritical fluid extraction and reversed-phase liquid chromatography methods for vitamin A and beta-carotene heterogeneous distribution of vitamin A in the liver.

We developed supercritical fluid procedures for extracting vitamin A and beta-carotene from vitamin supplements and calf liver tissue. The SF extracts could be injected onto an HPLC column without further pretreatment. Samples were analysed by RP-HPLC using diode array detection or by spectrophotometry. Recoveries were very good. SF extracts from a vitamin preparation of uniform composition had an R.S.D. of 4%. Extracts from calf liver supplements were predictably more heterogeneous. The SF extraction method is less labor intensive then traditional liquid-liquid procedures for extracting vitamin A and carotenoids from tissues.

Nutrient density estimates from an average of food frequency and food records correlate well with serum concentration of vitamins E and the carotenoids in free-living adults.

Dietary intakes are usually estimated by either a food frequency test, or by food records. We hypothesized that dietary intake estimates for fat soluble vitamins might be more accurate if information from both food frequency tests and food records were used. We estimated dietary intakes in 10 healthy adults by 4 food frequency questionnaires and ten 3-day food records collected over a year. Serum antioxidant nutrient concentrations (vitamins A, E, and the carotenoids) were measured by HPLC throughout the year. Few changes in intake occurred over the year. Estimates of nutrient densities correlated more often than estimates of nutrient intakes to serum nutrient concentrations. Nutrient density estimates derived from the average of food frequency and food record estimates were significantly correlated with serum nutrient concentrations more often than dietary estimates derived from either food frequency or food records alone. We suggest that nutrient density estimates derived from a combination of food frequency and food records may be useful for studies of free-living individuals.

Blood antioxidants changes in young women following beta-carotene depletion and repletion.

OBJECTIVE: This study was undertaken to investigate the relationship between beta-carotene intake and biochemical indices of antioxidant status in the blood of nine premenopausal women ages 18 to 42. METHODS: Nine healthy adult women were fed a low beta-carotene diet for 68 days. They were repleted with the same diet supplemented with beta-carotene (15 mg beta-carotene) for 28 days. During the last week of the study, they received an additional mixed carotenoid supplement. Indices of blood antioxidant status were measured on days 1, 29, 36, 43, 50, 64, 71, 92, and 99. RESULTS: We found significant increases of erythrocyte conjugated dienes between the 71st and 99th day of the study; increases of glutathione (GSH) peroxidase (GP) on day 43 and day 92 compared to a decrease on day 29; and decreases of GSH reductase throughout the treatment period. Erythrocyte catalase activities seemed to parallel GP activities. Erythrocyte oxidized glutathione (GSSG) levels were depressed both after beta-carotene depletion and repletion. beta-Carotene depletion/repletion had no effect on plasma vitamin E or GSH levels. Platelet GSH levels were depressed after beta-carotene depletion followed by elevated GSH levels after beta-carotene repletion. CONCLUSION: A diet low in beta-carotene and adequate in all other nutrients, including vitamin A, resulted in altered erythrocyte and platelet antioxidant indices; however, it had little impact on plasma GSH or vitamin E levels in young healthy women. Our results are consistent with the suggestion that carotenes may be important in the prevention of oxidative damage.

Oxidative damage and defense.

Increased production of reactive oxygen species is a feature of most, if not all, human disease, including cardiovascular disease and cancer. Dietary antioxidants may be especially important in protecting against human diseases associated with free radical damage to cellular DNA, lipids, and proteins. Ascorbic acid is an effective water-soluble antioxidant, and epidemiologic studies suggest that increased ascorbate nutriture is associated with reduced risk of some degenerative diseases, especially cancer and eye cataracts. Population studies have also shown that high vitamin E intakes are associated with decreased risk of coronary heart disease, possibly as a result of inhibition of atherogenic forms of oxidized low-density lipoprotein. Recent data suggest that beta-carotene provides protection against lipid peroxidation in humans, as well as provitamin A activity. Yet, present data are not sufficient to quantitate micronutrient requirements needed to protect against oxidative damage. The antioxidant roles of many food constituents, such as polyphenols, have not been clarified. Most antioxidants can act as prooxidants under certain conditions, and more research is needed to determine the occurrence and importance of this in vivo. The few controlled intervention trials carried out so far have shown mixed results as to the potential of antioxidant supplements for reducing the incidence of chronic diseases. Definitive recommendations on antioxidant intakes for disease prevention must await evidence from controlled studies and intervention trials, some currently in progress. Overall, the present data suggest that protection against oxidative damage and related disease is best served by the variety of antioxidant substances found in fruit and vegetables.

A genetic model for absent chylomicron formation: mice producing apolipoprotein B in the liver, but not in the intestine.

The formation of chylomicrons by the intestine is important for the absorption of dietary fats and fat-soluble vitamins (e.g., retinol, alpha-tocopherol). Apo B plays an essential structural role in the formation of chylomicrons in the intestine as well as the VLDL in the liver. We have developed genetically modified mice that express apo B in the liver but not in the intestine. By electron microscopy, the enterocytes of these mice lacked nascent chylomicrons in the endoplasmic reticulum and Golgi apparatus. Because these mice could not form chylomicrons, the intestinal villus enterocytes were massively engorged with fat, which was contained in cytosolic lipid droplets. These mice absorbed D-xylose normally, but there was virtually no absorption of retinol palmitate or cholesterol. The levels of alpha-tocopherol in the plasma were extremely low. Of note, the absence of chylomicron synthesis in the intestine did not appear to have a significant effect on the plasma levels of the apo B-containing lipoproteins produced by the liver. The mice lacking intestinal apo B expression represent the first genetic model of defective absorption of fats and fat-soluble vitamins and provide a useful animal model for studying nutrition and lipoprotein metabolism.

Effect of a low beta-carotene diet on the immune functions of adult women.

We examined the effect of beta-carotene depletion and repletion on the immune status of nine healthy women who lived in the metabolic suite for 100 d. For the first 4 d all women were fed a basal diet supplemented with 1.5 mg beta-carotene/d (baseline). During the next 68 d, the basal diet without beta-carotene supplementation was fed to all subjects (depletion), and during the last 28 d the diet of each women was supplemented with 15.0 mg beta-carotene/d (repletion). Neither beta-carotene depletion nor repletion significantly (P < or = 0.05) altered proliferation of peripheral blood mononuclear cells cultured with phytohemagglutinin or concanavalin A, in vitro production of soluble interleukin 2 receptor, or the concentration of circulating lymphocytes and their subsets. Thus, in healthy adults consuming adequate vitamin A, beta-carotene depletion had no adverse effect on the indexes tested, nor was there any beneficial effect of modest beta-carotene supplementation.

Effects of a carotene-deficient diet on measures of oxidative susceptibility and superoxide dismutase activity in adult women.

The effect of consuming a low carotene diet (approximately 60 micrograms carotene/day) on oxidative susceptibility and superoxide dismutase (SOD) activity in women living in a metabolic research unit was evaluated. The diet had sufficient vitamins A, E, and C. The women ate the diet supplemented with 1500 micrograms/day beta-carotene for 4 days (baseline), then the unsupplemented diet for 68 days (depletion), followed by the diet supplemented with > 15,000 micrograms/day carotene for 28 days (repletion). Production of hexanal, pentanal, and pentane by copper-oxidized plasma low density lipoproteins from carotene-depleted women was greater than their production of these compounds when repleted with carotene. Erythrocyte SOD activity was depressed in carotene-depleted women; it recovered with repletion. Thiobarbituric acid reactive substances in plasma of carotene-depleted women were elevated and diminished with repletion. Dietary carotene seems to be needed, not only as a precursor of vitamin A, but also to inhibit oxidative damage and decrease oxidation susceptibility.