Potential of golden potatoes to improve vitamin A and vitamin E status in developing countries.

Potato (Solanum tuberosum L.) is the third most widely consumed plant food by humans. Its tubers are rich in starch and vitamin C, but have low or null levels of essential nutrients such as provitamin A and vitamin E. Transformation of potato with a bacterial mini-pathway for ß-carotene in a tuber-specific manner results in a "golden" potato (GP) tuber phenotype resulting from accumulation of provitamin A carotenoids (α- and ß-carotene) and xanthophylls. Here, we investigated the bioaccessibility of carotenoids and vitamin E as α-tocopherol (αTC) in boiled wild type and golden tubers using in vitro digestion. Golden tubers contained up to 91 µg provitamin A carotenes (PAC)/g D, increased levels of xanthophylls, phytoene and phytofluene, as well as up to 78 µg vitamin E/g DW. Cubes from wild type and GP tubers were boiled and subjected to simulated digestion to estimate bioaccessibility of carotenoids and αTC. Retention in boiled GPs exceeded 80% for ß-carotene (ßC), α-carotene (αC), lutein, phytoene ± and αTC, but less than 50% for phytofluene. The efficiency of partitioning of total ßC, αC, E-lutein, phytoene, phytofluene and αTC in the mixed micelle fraction during small intestinal digestion was influenced by genotype, tuber content and hydrophobicity. Apical uptake of the compounds that partitioned in mixed micelles by monolayers of human intestinal Caco-2 cells during incubation for 4h was 14-20% for provitamin A and xanthophylls, 43-45% for phytoene, 23-27% for phytofluene, and 53% for αTC. These results suggest that a 150 g serving of boiled golden potatoes has the potential to contribute 42% and 23% of the daily requirement of retinol activity equivalents (RAE), as well as 34 and 17% of the daily vitamin E requirement for children and women of reproductive age, respectively.

Bioaccessibility and intestinal cell uptake of astaxanthin from salmon and commercial supplements.

Although the keto-carotenoid astaxanthin (Ast) is not typically present in human plasma due to its relative scarcity in the typical diet, global consumption of salmon, the primary source of Ast in food, and Ast supplements continues to increase. The first objective of the present study was to investigate the bioaccessibility of Ast from uncooked and cooked fillets of wild and aquacultured salmon, Ast-supplements and krill oil, during simulated gastric and small intestinal digestion. Uptake of E-Ast from micelles generated during digestion of wild salmon by monolayers of Caco-2 was also monitored. Both wild and aquacultured salmon flesh contained E-Ast and Z-isomers of unesterified Ast, whereas Ast esters were the predominant form of the carotenoid in commercial supplements and krill oil. Flesh from wild salmon contained approximately 10 times more Ast than aquacultured salmon. Common styles of cooking flesh from wild and aquacultured salmon decreased Ast content by 48-57% and 35-47%, respectively. Ast in salmon flesh, supplements and krill oil was relatively stable (>80% recovery) during in vitro digestion. The efficiency of transfer of Ast into mixed micelles during digestion of uncooked wild salmon was 43%, but only 12% for uncooked acquacultured salmon. Cooking wild salmon significantly decreased Ast bioaccessibility. The relative bioaccessibility of Ast (41-67%) after digestion of oil vehicle in commercial supplements was inversely proportional to carotenoid content (3-10mg/capsule), whereas bioaccessibility of endogenous Ast in phospholipid-rich krill oil supplement was 68%. >95% of Ast in mixed micelles generated during digestion of supplements and krill oil was unesterified. Caco-2 intestinal cells accumulated 11-14% of E-Ast delivered in mixed micelles generated from digested wild salmon. Apical uptake and basolateral secretion of E-Ast by Caco-2 cells grown on inserts were greater after digestion of Ast-enriched krill oil compared to uncooked wild salmon. These data suggest that the bioacessibility of Ast in wild salmon and soft-gel capsules is greater than that in aquacultured salmon, and that uptake and basolateral secretion of the carotenoid by enterocyte-like cells is enhanced by the digestion products of phospholipid-rich krill oil.

α-Tocopherol bioavailability is lower in adults with metabolic syndrome regardless of dairy fat co-ingestion: a randomized, double-blind, crossover trial.

BACKGROUND: Increasing dietary fat intake is expected to improve α-tocopherol bioavailability, which could be beneficial for improving α-tocopherol status, especially in cohorts at high cardiometabolic risk who fail to meet dietary α-tocopherol requirements. OBJECTIVE: Our objective was to assess dose-dependent effects of dairy fat and metabolic syndrome (MetS) health status on α-tocopherol pharmacokinetics in plasma and lipoproteins. DESIGN: A randomized, crossover, double-blind study was conducted in healthy and MetS adults (n = 10/group) who ingested encapsulated hexadeuterium-labeled (d6)-RRR-α-tocopherol (15 mg) with 240 mL nonfat (0.2 g fat), reduced-fat (4.8 g fat), or whole (7.9 g fat) milk before blood collection at regular intervals for 72 h. RESULTS: Compared with healthy participants, those with MetS had lower (P < 0.05) baseline plasma α-tocopherol (µmol/mmol lipid) and greater oxidized low-density lipoprotein (LDL), interleukin (IL)-6, IL-10, and C-reactive protein. Regardless of health status, d6-α-tocopherol bioavailability was unaffected by increasing amounts of dairy fat provided by milk beverages, but MetS participants had lower estimated d6-α-tocopherol absorption (±SEM) than did healthy participants (26.1% ± 1.0% compared with 29.5% ± 1.1%). They also had lower plasma d6-α-tocopherol AUC from 0 to 72 h, as well as maximal concentrations (Cmax: 2.04 ± 0.14 compared with 2.73 ± 0.18 µmol/L) and slower rates of plasma disappearance but similar times to Cmax. MetS participants had lower d6-α-tocopherol AUC from t = 0-12 h (AUC0- t final) in lipoprotein fractions [chylomicron, very-low-density lipoprotein (VLDL), LDL, high-density lipoprotein]. Percentages of d6-α-tocopherol AUC0- t final in both the chylomicron (r = -0.46 to -0.52) and VLDL (r = -0.49 to -0.68) fractions were inversely correlated with oxidized LDL, IL-10, IL-6, and C-reactive protein. CONCLUSIONS: At dietary intakes equivalent to the Recommended Dietary Allowance, α-tocopherol bioavailability is unaffected by dairy fat quantity but is lower in MetS adults, potentially because of greater inflammation and oxidative stress that limits small intestinal α-tocopherol absorption and/or impairs hepatic α-tocopherol trafficking. These findings support higher dietary α-tocopherol requirements for MetS adults. This trial was registered at www.clinicaltrials.gov as NCT01787591.

Intestinal microbial dysbiosis and colonic epithelial cell hyperproliferation by dietary α-mangostin is independent of mouse strain.

Beverages and supplements prepared from mangosteen fruit are claimed to support gut health and immunity, despite the absence of supporting evidence from clinical trials. We recently reported that α-mangostin (α-MG), the most abundant xanthone in mangosteen fruit, altered the intestinal microbiome, promoted dysbiosis, and exacerbated colitis in C57BL/6J mice. The objective of this study was to determine whether induction of dysbiosis by dietary α-MG is limited to the C57BL/6J strain or represents a more generic response to chronic intake of the xanthone on the gut microbiota of mice. C3H, Balb/c, Nude FoxN1nu, and C57BL/6J mice, each demonstrating unique microbiomes, were fed standard diet or diet containing 0.1% α-MG for four weeks. Dietary α-MG significantly altered the cecal and colonic microbiota in all four strains of mice, promoting a reduction in generally assumed beneficial bacterial groups while increasing the abundance of pathogenic bacteria. Consumption of α-MG was associated with reduced abundance of Firmicutes and increased abundance of Proteobacteria. The abundance of Lachnospiraceae, Ruminococcaceae, and Lactobacillaceae was reduced in α-MG-fed mice, while that of Enterobacteriaceae and Enterococcaceae was increased. Dietary α-MG also was associated with increased proliferation of colonic epithelial cells, infiltration of immune cells, infiltration of immune cells and increased fluid content in stool. These results suggest that ingestion of pharmacologic doses of xanthones in mangosteen-containing supplements may adversely alter the gut microbiota and should be used with caution.

Impact of genotype and cooking style on the content, retention, and bioacessibility of ß-carotene in biofortified cassava (Manihot esculenta Crantz) conventionally bred in Brazil.

Biofortification is a strategy for decreasing micronutrient deficiencies in vulnerable populations by increasing nutrient density in staple food crops. Roots from five varieties of cassava biofortified with ß-carotene (ßC), three parental accessions, and one variety of commonly consumed white cassava from Brazil were investigated. Roots from biofortified varieties contained up to 23-fold higher ßC than white cassava, and the additional complement of ßC was primarily the all-trans isomer. At least 68% of ßC per gram fresh weight was retained after boiling or boiling and briefly frying. Micellarization of ßC during simulated digestion of fried root exceeded that of boiled root. Apical uptake of all-trans-ßC from mixed micelles by Caco-2 cells was affected by an interaction between variety and cooking style. These results suggest that Brazilian cassava biofortified with ßC has the potential to reduce vitamin A deficiency without requiring major changes in local and ethnic styles of home cooking.

Unsaturated fatty acids promote bioaccessibility and basolateral secretion of carotenoids and α-tocopherol by Caco-2 cells.

Bioavailability of carotenoids and tocopherols from foods is determined by the efficiency of transfer from food/meal to mixed micelles during digestion, incorporation into chylomicrons for trans-epithelial transport to lymphatic/blood system, and distribution to target tissues. Fats and oils are important factors for facilitating the absorption of lipophilic compounds. However, dietary fats and oils are composed of various types of saturated and unsaturated fatty acids which may differentially impact the bioavailability of carotenoids and tocopherols from foods. We have investigated the effects of several common commercial lipids on bioavailability using an in vitro digestion model and Caco-2 human intestinal cells. Meals consisted of mixed salad vegetables containing a single test lipid. Micellarization and cellular uptake of ß-carotene (ßC) and lycopene (LYC) during small intestinal digestion was increased by lipids rich in unsaturated fatty acids: soybean oil > olive > canola > butter. In contrast, type of lipid minimally affected the bioaccessibility of lutein (LUT) and zeaxanthin (ZEA). To examine the influence of type of dietary triglyceride on uptake and basolateral secretion of carotenoids, Caco-2 cells grown on Transwell membranes were incubated with micellar mixtures of fatty acids (1.0 mM) mimicking the types and ratio of saturated to unsaturated (mono- + poly-unsaturated) fatty acids (FA) present in butter (70 : 30), olive oil (7 : 93) and soybean oil (11 : 89). Cells were exposed to micelles containing ßC, LUT, α-tocopherol (α-TC) and a mixture of test fatty acids. Uptake and basolateral secretion of ßC, LUT and α-TC were greater in cells pre-treated with mixtures enriched in unsaturated compared to saturated FA and these effects were mediated by increased assembly and secretion of chylomicrons. These results suggest that dietary fats/oils rich in unsaturated fatty acids promote carotenoid and α-TC bioavailability by enhancing their micellarization during digestion and intestinal transport.

Dietary α-mangostin, a xanthone from mangosteen fruit, exacerbates experimental colitis and promotes dysbiosis in mice.

SCOPE: Ulcerative colitis (UC) is a chronic inflammatory disease of the colon. α-Mangostin (α-MG), the most abundant xanthone in mangosteen fruit, exerts anti-inflammatory and antibacterial activities in vitro. We evaluated the impact of dietary α-MG on murine experimental colitis and on the gut microbiota of healthy mice. METHODS AND RESULTS: Colitis was induced in C57BL/6J mice by administration of dextran sulfate sodium (DSS). Mice were fed control diet or diet with α-MG (0.1%). α-MG exacerbated the pathology of DSS-induced colitis. Mice fed diet with α-MG had greater colonic inflammation and injury, as well as greater infiltration of CD3(+) and F4/80(+) cells, and colonic myeloperoxidase, than controls. Serum levels of granulocyte colony-stimulating factor, IL-6, and serum amyloid A were also greater in α-MG-fed animals than in controls. The colonic and cecal microbiota of healthy mice fed α-MG but no DSS shifted to an increased abundance of Proteobacteria and decreased abundance of Firmicutes and Bacteroidetes, a profile similar to that found in human UC. CONCLUSION: α-MG exacerbated colonic pathology during DSS-induced colitis. These effects may be associated with an induction of intestinal dysbiosis by α-MG. Our results suggest that the use of α-MG-containing supplements by patients with UC may have unintentional risk.

Biological activities and bioavailability of mangosteen xanthones: a critical review of the current evidence.

Mangosteen (Garcinia mangostana L.) is a tropical tree native to Southeast Asia that produces a fruit whose pericarp contains a family of tricyclic isoprenylated polyphenols referred to as xanthones. Numerous in vitro studies have shown that these xanthones possess anti-oxidant, anti-proliferative, pro-apoptotic, anti-inflammatory and anti-carcinogenic activities. Aggressive marketing of such health promoting benefits has resulted in mangosteen's classification as a "superfruit". This has led to sales of mangosteen containing beverages in USA alone exceeding $200 million in 2008 despite very limited animal and human studies. This review will (a) critically address recent reports of in vivo studies on the bioavailability and metabolism of mangosteen xanthones, (b) update the in vitro and in vivo data on anti-cancer and anti-inflammatory activities of mangosteen xanthones, and (c) suggest needed areas of inquiry regarding the absorption, metabolism and efficacy of mangosteen xanthones.

Carotenoid bioavailability from raw vegetables and a moderate amount of oil in human subjects is greatest when the majority of daily vegetables are consumed at one meal.

While the impact of food composition and processing on carotenoid bioavailability has been the subject of several investigations, the effect of meal patterning remains unknown. The aim of this pilot study was to assess the impact of select consumption patterns on the bioavailability of carotenoids from vegetables. On three randomized testing days, subjects consumed raw salad vegetables and 8 g canola oil over a two meal period in three meal patterns. Meal patterns included consumption of 100% of vegetables and oil in the first meal and 0% in the second, 75% in the first meal and 25% in the second, and 50% in the first meal and 50% in the second. Additional protein-rich "chef's salad" ingredients were distributed equally between meals. We hypothesized that carotenoid absorption would be highest when 50% of vegetables and oil were consumed at each meal and lowest when 100% were consumed at once. Blood was collected 0 to 12 hours postprandially and triacylglycerol-rich lipoprotein fractions (TRL) were isolated by ultracentrifugation. TRL carotenoid concentrations were analyzed by high performance liquid chromatography-diode array detector. Considering all carotenoids, absorption expressed as area under the curve was greatest when ≥75% of vegetables were consumed in a single meal (P < .05). Absorption of carotenes also followed this trend (P < .05 for α- and ß-carotene). For xanthophylls, consuming all vegetables in one meal increased absorption compared to intake of 50% at each meal (P < .05). These data suggest that carotenoid absorption may be the greatest when daily recommended vegetables are consumed in one meal compared to smaller doses over multiple meals.

Anti-tumorigenicity of dietary α-mangostin in an HT-29 colon cell xenograft model and the tissue distribution of xanthones and their phase II metabolites.

SCOPE: This study investigated the in vivo and in vitro activity of α-mangostin (α-MG), the most abundant xanthone in mangosteen pericarp, on HT-29 cell tumorigenicity, proliferation, and several markers of tumor cell activity, as well as the profile and amounts of xanthones in serum, tumor, liver, and feces. METHODS AND RESULTS: Balb/c nu/nu mice were fed either control diet or diet containing 900 mg α-MG/kg. After 1 week of acclimation to diet, mice were injected subcutaneously with HT-29 cells and fed the same diets ad libitum for an additional 2 or 4 weeks. After 2 and 4 weeks, tumor mass and the concentrations of BcL-2 and ß-catenin in tumors of mice fed diet with α-MG were significantly less than in mice fed control diet. Xanthones and their metabolites were identified in serum, tumor, liver, and feces. In vitro treatment of HT-29 cells with α-MG also inhibited cell proliferation and decreased expression of BcL-2 and ß-catenin. CONCLUSION: Our data demonstrate that the anti-neoplastic effect of dietary α-MG is associated with the presence of xanthones in the tumor tissue. Further investigation of the impact of beverages and food products containing xanthones on the prevention of colon cancer or as complementary therapy is merited.

Isoflavone retention during processing, bioaccessibility, and transport by Caco-2 cells: effects of source and amount of fat in a soy soft pretzel.

The impact of source and amount of lipid used to prepare a soy soft pretzel on the bioaccessibility and transport of isoflavones was investigated using the coupled in vitro digestion/Caco-2 human cell model. Pretzels were prepared without or with 2.9 or 6.0% exogenous lipid from either shortening, canola oil, ground almond, or ground hazelnut. The isoflavone backbone structure was stable during pretzel production, although there was partial conversion from malonyl and acetyl glucosides to simple glucosides and aglycones. Endogenous ß-glucosidase activity in ground almond facilitated partial conversion of simple glucosides to aglycones during proofing, resulting in a slight decrease in bioaccessibility of isoflavones as compared with other sources of lipid. Amount and source of lipid did not affect bioaccessibility or uptake and metabolism of isoflavones by Caco-2 cells, although transport across the monolayer was greater with the lesser amount of shortening. These results suggest that the isoflavone structure, but not source or amount of lipid in a soy pretzel, may affect bioavailability of isoflavones.

Susceptibility of anthocyanins to ex vivo degradation in human saliva.

Some fruits and their anthocyanin-rich extracts have been reported to exhibit chemopreventive activity in the oral cavity. Insights regarding oral metabolism of anthocyanins remain limited. Anthocyanin-rich extracts from blueberry, chokeberry, black raspberry, red grape, and strawberry were incubated ex vivo with human saliva from 14 healthy subjects. All anthocyanins were partially degraded in saliva. Degradation of chokeberry anthocyanins in saliva was temperature dependent and decreased by heating saliva to 80 °C and after removal of cells. Glycosides of delphinidin and petunidin were more susceptible to degradation than those of cyanidin, pelargonidin, peonidin and malvidin in both intact and artificial saliva. Stability of di- and tri-saccharide conjugates of anthocyanidins slightly, but significantly, exceeded that of monosaccharide compounds. Ex vivo degradation of anthocyanins in saliva was significantly decreased after oral rinsing with antibacterial chlorhexidine. These results suggest that anthocyanin degradation in the mouth is structure-dependent and largely mediated by oral microbiota.

Meal triacylglycerol profile modulates postprandial absorption of carotenoids in humans.

SCOPE: Dietary lipids are considered to be primary potentiators of carotenoid absorption, yet the amount and source required to optimize bioavailability has not been systematically evaluated. The objective of this study was to examine the impact of both amount and source of triacylglycerols on postprandial absorption of carotenoids from vegetable salads. METHODS AND RESULTS: Healthy subjects (n = 29) were randomized using a Latin square design (3 × 3) and consumed three identical salads with 3, 8, or 20 g of canola oil, soybean oil, or butter. Blood was collected from 0-10 h and triacylglycerol-rich fractions (TRLs) were isolated by ultracentrifugation. Carotenoid contents of TRL fractions were analyzed by HPLC-DAD. Considering all lipid sources, 20 g of lipid promoted higher absorption compared to 3 and 8 g for all carotenoid species (p < 0.05), except for α-carotene (p = 0.07). The source of lipid had less impact on the absorption of carotenoids than amount of lipid. Pooling results from all lipid amounts, monounsaturated fatty acid rich canola oil trended toward enhancing absorption of lutein and α-carotene compared to saturated fatty acid rich butter (p = 0.06 and p = 0.08, respectively). CONCLUSION: While both amount and source of co-consumed lipid affect carotenoid bioavailability from vegetables, amount appears to exert a stronger effect.

Estrogen status alters tissue distribution and metabolism of selenium in female rats.

A reported association between estrogen and selenium status may be important in the regulation of selenium metabolism. In this study, the effect of estrogen status on the metabolism of orally administered (75)Se-selenite and tissue selenium status was investigated. Female Sprague-Dawley rats were bilaterally ovariectomized at 7 weeks of age and implanted with either a placebo pellet (OVX) or pellet containing estradiol (OVX+E2), or were sham operated (Sham). At 12 weeks of age, 60 µCi of (75)Se as selenite was orally administered to OVX and OVX+E2 rats. Blood and organs were collected 1, 3, 6 and 24 h after dosing. Estrogen status was associated with time-dependent differences in distribution of (75)Se in plasma, red blood cell (RBC), liver, heart, kidney, spleen, brain and thymus and incorporation of (75)Se into plasma selenoprotein P (Sepp1) and glutathione peroxidase (GPx). Estrogen treatment also significantly increased selenium concentration and GPx activity in plasma, liver and brain, selenium concentration in RBC and hepatic Sepp1 and GPx1 messenger RNA. These results suggest that estrogen status affects tissue distribution of selenium by modulating Sepp1, as this protein plays a central role in selenium transport.

Anti-inflammatory activities of extracts of Thai spices and herbs with lipopolysaccharide-activated RAW 264.7 murine macrophages.

Nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) play important roles in inflammatory processes. This study examined whether 13 spices/herbs commonly used in Thai dishes modulate the production of NO and TNF-alpha by the RAW 264.7 mouse macrophage cell line pretreated with plant extracts (1-100 microg/mL) prior to activation by bacterial lipopolysaccharide (LPS). Tested plant tissues were extracted with ethanol with the exception of roselle, which was extracted with 70% acetone. Eight of the 13 plant extracts inhibited NO and TNF-alpha production in a dose-dependent manner without exerting cytotoxicity. Extract from Limnophila aromatica (Kyeng) was the most robust suppressor of NO production, followed by dill, kaffer lime, chili, Teaw, mint, sweet basil, and pea eggplant, respectively (range of 50% inhibitory concentration [IC(50)] = 11.4-74.6 microg/mL). Kyeng also exhibited the greatest inhibition of TNF-alpha production (IC(50) = 10.5 microg/mL). IC(50) values for NO and TNF-alpha production in LPS-activated RAW 264.7 cells for these extracts were highly correlated (r = 0.772, P = .025). These results suggest that extracts from some spices/herbs in the habitual Thai diet possess anti-inflammatory activity. Moreover, the results support the use of NO production in LPS-activated RAW 264.7 cells as a rapid and cost-effective tool for screening the anti-inflammatory activity of extracts of spices/herbs.

Sweet potato beta-carotene bioefficacy is enhanced by dietary fat and not reduced by soluble fiber intake in Mongolian gerbils.

Orange-fleshed sweet potato (OFSP) is an important source of beta-carotene (betaC). Provitamin A bioefficacy from plant foods is influenced by dietary fat and fiber. We fed 3% OFSP powder diets with varying amounts of fat and soluble fiber to vitamin A (VA)-depleted Mongolian gerbils (n = 85) for 3 wk (8 groups, n = 10/group; control, n = 9) following a baseline kill (n = 6). OFSP diets differing in fat (3, 6, and 12%) contained 0.24% soluble fiber. Two additional 3% OFSP diets contained 6% fat and 3 or 9% white-fleshed sweet potato (WFSP) powder with soluble fiber contents of 0.42 and 0.80%, respectively. Control, VA-, and betaC-supplemented groups were included. Simulated digestion experiments compared the bioaccessibility of betaC from boiled vs. oil stir-fried OFSP. All OFSP diets maintained VA status and 12% fat and WFSP-added diets improved VA status above baseline (P < 0.05). Bioefficacy, as bioconversion factors, in gerbils fed 12% fat (3.5 +/- 1.4 microg betaC:1 microg VA) was improved over the 3% fat and betaC groups (6.5 +/- 3.7 and 6.7 +/- 3.7 microg betaC:1 microg VA, respectively) (P < 0.05) but did not differ from WFSP-added groups or the 6% fat group with no WFSP. Stir-frying doubled the efficiency of betaC incorporation into micelles during small intestinal digestion in support of the stimulatory effect of dietary fat on bioefficacy in vivo. Soluble fiber intake derived from WFSP did not influence bioefficacy. Replacing WFSP with OFSP will affect VA status if adopted by target groups.

Bioaccessibility of pro-vitamin A carotenoids is minimally affected by non pro-vitamin a xanthophylls in maize (Zea mays sp.).

The absorption of some carotenoids has been reported to be decreased by coingestion of relatively high concentrations of other carotenoids. It is unclear if such interactions occur among carotenoids during the digestion of plant foods. Current varieties of maize contain limited amounts of the pro-vitamin A (pro-VA) carotenoids beta-carotene (BC) and beta-cryptoxanthin (BCX) and relatively higher levels of their oxygenated metabolites lutein (LUT) and zeaxanthin (ZEA). Here, we examined if LUT and ZEA attenuate the bioaccessibility of pro-VA carotenoids at amounts and ratios present in maize. BC incorporation into bile salt mixed micelles during chemical preparation and during simulated small intestinal digestion of carotenoid-enriched oil was slightly increased when the concentration of LUT was sixfold or more greater than BC. Likewise, the efficiency of BC micellarization was slightly increased during simulated small intestinal digestion of white maize porridge supplemented with oil containing ninefold molar excess of LUT to BC. Mean efficiencies of micellarization of BC, BCX, LUT, and ZEA were 16.7, 27.7, 30.3, and 27.9%, respectively, and independent of the ratio of LUT plus ZEA to pro-VA carotenoids during simulated digestion of maize porridge prepared from flours containing 0.4-11.3 microg/g endogenous pro-VA carotenoids. LUT attenuated uptake of BC by differentiated cultures of Caco-2 human cells from medium-containing micelles in a dose-dependent manner with inhibition reaching 35% when the molar ratio of LUT to BC was 13. Taken together, these results suggest that the bioaccessibility of pro-VA carotenoids in maize is likely to be minimally affected by the relative levels of xanthophylls lacking pro-VA activity present in cultivars of maize.

Micellarization and intestinal cell uptake of beta-carotene and lutein from drumstick (Moringa oleifera) leaves.

The leaves and pods of the drumstick tree are used as food and medicine in some Asian and African countries. Although relatively high concentrations of beta-carotene and lutein have been reported in the leaves, the bioavailability of these carotenoids from this source is unknown. We have analyzed the digestive stability and bioaccessibility of carotenoids in fresh and lyophilized drumstick leaves using the coupled in vitro digestion/Caco-2 cell model. Beta-carotene and lutein were stable during simulated gastric and small intestinal digestion. The efficiency of micellarization of lutein during the small intestinal phase of digestion exceeded that of beta-carotene. Addition of peanut oil (5% vol/wt) to the test food increased micellarization of both carotenoids, and particularly beta-carotene. Caco-2 cells accumulated beta-carotene and lutein from micelles generated during digestion of drumstick leaves in a time- and concentration-dependent manner. The relatively high bioaccessibility of beta-carotene and lutein from drumstick leaves ingested with oil supports the potential use of this plant food for improving vitamin A nutrition and perhaps delaying the onset of some degenerative diseases such as cataracts.

Supplementation of test meals with fat-free phytosterol products can reduce cholesterol micellarization during simulated digestion and cholesterol accumulation by Caco-2 cells.

Phytosterols have been shown to reduce cholesterol absorption in humans. Supplementing phytosterols in fat-free formulations, however, has yielded controversial results. In the present study, we investigated the effect of supplementing test meals with different fat-free phytosterol products on cholesterol incorporation into mixed micelles during simulated digestion and accumulation of micellar cholesterol by Caco-2 cells: control orange juice (OJ), orange juice supplemented with either multivitamin/multimineral tablets (MVT), multivitamin/multimineral tablets containing phytosterols (MVT+P), and phytosterol powder (PP). These combinations were added to Ensure-based test meals and spiked with cholesterol of natural isotopic composition or 13C2-cholesterol to differentiate external from endogenous cholesterol. After simulated gastric/small intestinal digestion, micelle fractions were analyzed for cholesterol enzymatically (n = 6-20/product) and by high-performance liquid chromatography-tandem mass spectrometry (n = 12/product) and added to Caco-2 cells to determine the accumulation of 13C2-cholesterol (n = 10-24/product). As compared to OJ, PP and MVT+P significantly decreased cholesterol micellarization (determined enzymatically) by 70 +/- 39 (mean +/- SD) and 70 +/- 39%, respectively (P < 0.001, Bonferroni). The stable isotope experiments revealed that both PP and MVT+P reduced cholesterol micellarization [by 25 +/- 12 (P = 0.055) and 21 +/- 8% (P = 0.020), respectively, Fisher's protected LSD test] and Caco-2 cell accumulation (by 28 +/- 8 and 10 +/- 8%, respectively; P < 0.010, Bonferroni). OJ+P did not inhibit micellarization or accumulation of cholesterol by Caco-2 cells. This study shows that fat-free phytosterol-containing products can significantly inhibit cholesterol micellarization and Caco-2 cell bioaccessibility, albeit to different extents depending on individual formulations. This is most likely explained by inhibition of cholesterol micellarization.

Assessment of lutein bioavailability from meals and a supplement using simulated digestion and caco-2 human intestinal cells.

Lutein and zeaxanthin are selectively accumulated in the lens and macular region of the retina. It was suggested that these xanthophylls protect ocular tissues from free-radical damage that can cause cataracts and age-related macular degeneration. Insights regarding the absorption of dietary xanthophylls for delivery to ocular tissues are limited. Our primary objective was to examine factors affecting the transfer of lutein from foods to absorptive intestinal epithelial cells during digestion. Lutein and other carotenoids present in spinach purée and lutein from a commercial supplement were relatively stable during in vitro digestion. Micellarization of lutein and zeaxanthin during the small intestinal phase of digestion exceeded that of beta-carotene and was greater for xanthophylls in oil-based supplements than in spinach. Apical uptake of lutein from micelles by Caco-2 human intestinal cells was linear for at least 8 h, and accumulation from synthetic micelles exceeded that from micelles generated during simulated digestion. Stimulation of chylomicron synthesis resulted in the secretion of 7.6 +/- 0.1% of cellular lutein into the triglyceride-rich fraction in the basolateral chamber. These data support the use of simulated digestion and the Caco-2 cell model as effective tools for identifying factors affecting absorption of dietary carotenoids.