Discovery and biological relevance of 3,4-didehydroretinol (vitamin A2) in small indigenous fish species and its potential as a dietary source for addressing vitamin A deficiency.

Discovered in the late 1920s, 3,4-didehydroretinol (DROL, vitamin A2) plays a significant biological role in freshwater fish. The functions of this vitamin have been investigated but to a far lesser extent than those of retinol (ROL, vitamin A1). A recent study indicating all-trans DROL has 119-127% vitamin A biological activity compared to that of all-trans ROL suggests the significance of DROL for addressing vitamin A deficiency (VAD) in comparison to ROL may be currently overlooked. Freshwater fish such as small indigenous fish species (SIS), with high DROL content can be a promising dietary source for reducing VAD in areas where SIS are readily available and consumed. In this paper, the discovery and biological relevance of DROL are reviewed and furthermore, the vast potential of production and consumption of DROL-rich SIS in food-based strategies to combat VAD in Bangladesh and other developing countries with high prevalence of VAD is highlighted.

Consumption of canned citrus fruit meals increases human plasma ß-cryptoxanthin concentration, whereas lycopene and ß-carotene concentrations did not change in healthy adults.

Several studies suggest that ß-cryptoxanthin has a greater plasma response from its common food sources than other carotenoids such as ß-carotene and lycopene. The hypothesis of this study is that changes in plasma ß-cryptoxanthin concentrations will be greater than changes in plasma ß-carotene or lycopene concentrations even if these carotenoids are fed in a similar food matrix, such as citrus fruit. We tested this hypothesis by measuring changes in plasma concentrations of ß-cryptoxanthin, lycopene, and ß-carotene after feeding measured amounts of canned tangerines and pink grapefruit to healthy nonsmoking adult humans. Volunteers served as their own controls and received both citrus fruit treatments randomly. In the first study, 8 subjects ate single meals of 234-304g of tangerines or 60-540g of pink grapefruit. The second study compared changes in plasma carotenoid concentration caused by feeding 234g of tangerines or 540g of pink grapefruit to 11 subjects. Blood was collected 5 times within 24hours after each citrus meal. Carotenoid concentrations were analyzed by reversed-phase high-performance liquid chromatography. Plasma ß-cryptoxanthin concentrations increased within 5hours and then stabilized, remaining high throughout the 24hours measured. Plasma concentrations of lycopene and ß-carotene did not change. These results show that ß-cryptoxanthin concentrations increased after a citrus fruit meal, but lycopene and ß-carotene concentrations did not change after a similar citrus fruit meal. These results support our hypothesis that changes in plasma ß-cryptoxanthin are greater than changes in plasma lycopene or ß-carotene, even when these carotenoids are fed in a similar food matrix.

Absorption, metabolism, and functions of ß-cryptoxanthin.

ß-Cryptoxanthin, a carotenoid found in fruits and vegetables such as tangerines, red peppers, and pumpkin, has several functions important for human health. Most evidence from observational, in vitro, animal model, and human studies suggests that ß-cryptoxanthin has relatively high bioavailability from its common food sources, to the extent that some ß-cryptoxanthin-rich foods might be equivalent to ß-carotene-rich foods as sources of retinol. ß-Cryptoxanthin is an antioxidant in vitro and appears to be associated with decreased risk of some cancers and degenerative diseases. In addition, many in vitro, animal model, and human studies suggest that ß-cryptoxanthin-rich foods may have an anabolic effect on bone and, thus, may help delay osteoporosis.

Red palm oil-supplemented and biofortified cassava gari increase the carotenoid and retinyl palmitate concentrations of triacylglycerol-rich plasma in women.

Boiled biofortified cassava containing ß-carotene can increase retinyl palmitate in triacylglycerol-rich plasma. Thus, it might alleviate vitamin A deficiency. Cassava requires extensive preparation to decrease its level of cyanogenic glucosides, which can be fatal. Garification is a popular method of preparing cassava that removes cyanogen glucosides. Our objective was to compare the effectiveness of biofortified gari to gari prepared with red palm oil. The study was a randomized crossover trial in 8 American women. Three gari preparations separated by 2-week washout periods were consumed. Treatments (containing 200-225.9 g gari) were as follows: biofortified gari (containing 1 mg ß-carotene), red palm oil-fortified gari (1 mg ß-carotene), and unfortified gari with a 0.3-mg retinyl palmitate reference dose. Blood was collected 6 times from -0.5 to 9.5 hours after ingestion. Triacylglycerol-rich plasma was separated by ultracentrifugation and analyzed by high-performance liquid chromatography (HPLC) with diode array detection. Area under the curve for ß-carotene, α-carotene, and retinyl palmitate increased after the fortified meals were fed (P < .05), although the retinyl palmitate increase induced by the red palm oil treatment was greater than that induced by the biofortified treatment (P < .05). Vitamin A conversion was 2.4 ± 0.3 and 4.2 ± 1.5 µg pro-vitamin A carotenoid/1 µg retinol (means ± SEM) for red palm oil and biofortified gari, respectively. These results show that both treatments increased ß-carotene, α-carotene, and retinyl palmitate in triacylglycerol-rich plasma concentrations in healthy well-nourished adult women, supporting our hypothesis that both interventions could support efforts to alleviate vitamin A deficiency.

Beta-cryptoxanthin as a source of vitamin A.

Beta-cryptoxanthin is a common carotenoid that is found in fruit, and in human blood and tissues. Foods that are rich in beta-cryptoxanthin include tangerines, persimmons and oranges. Beta-cryptoxanthin has several functions that are important for human health, including roles in antioxidant defense and cell-to-cell communication. Most importantly, beta-cryptoxanthin is a precursor of vitamin A, which is an essential nutrient needed for eyesight, growth, development and immune response. We evaluate the evidence for beta-cryptoxanthin as a vitamin A-forming carotenoid in this paper. Observational, in vitro, animal model and human studies suggest that beta-cryptoxanthin has greater bioavailability from its common food sources than do alpha- and beta-carotene from theirs. Although beta-cryptoxanthin appears to be a poorer substrate for beta-carotene 15,15' oxygenase than is beta-carotene, animal model and human studies suggest that the comparatively high bioavailability of beta-cryptoxanthin from foods makes beta-cryptoxanthin-rich foods equivalent to beta-carotene-rich foods as sources of vitamin A. These results mean that beta-cryptoxanthin-rich foods are probably better sources of vitamin A, and more important for human health in general, than previously assumed.

Bioavailability of iron, zinc, and provitamin A carotenoids in biofortified staple crops.

International research efforts, including those funded by HarvestPlus, a Challenge Program of the Consultative Group on International Agricultural Research (CGIAR), are focusing on conventional plant breeding to biofortify staple crops such as maize, rice, cassava, beans, wheat, sweet potatoes, and pearl millet to increase the concentrations of micronutrients that are commonly deficient in specific population groups of developing countries. The bioavailability of micronutrients in unfortified staple crops in developing regions is typically low, which raises questions about the efficacy of these crops to improve population micronutrient status. This review of recent studies of biofortified crops aims to assess the micronutrient bioavailability of biofortified staple crops in order to derive lessons that may help direct plant breeding and to infer the potential efficacy of food-based nutrition interventions. Although reducing the amounts of antinutrients and the conduction of food processing generally increases the bioavailability of micronutrients, antinutrients still possess important benefits, and food processing results in micronutrient loss. In general, biofortified foods with relatively higher micronutrient density have higher total absorption rates than nonbiofortified varieties. Thus, evidence supports the focus on efforts to breed plants with increased micronutrient concentrations in order to decrease the influence of inhibitors and to offset losses from processing.

Assessment of tissue distribution and concentration of ß-cryptoxanthin in response to varying amounts of dietary ß-cryptoxanthin in the Mongolian gerbil.

There is a general lack of knowledge regarding the absorption and tissue storage of the provitamin A carotenoid ß-cryptoxanthin. The present study investigated the whole-body tissue distribution of ß-cryptoxanthin in an appropriate small animal model, the Mongolian gerbil (Meriones unguiculatus), for human provitamin A carotenoid metabolism. After 5 d of carotenoid depletion, five gerbils were euthanised for baseline measurements. The remaining gerbils were placed in three weight-matched treatment groups (n 8). All the groups received 20 µg/d of ß-cryptoxanthin from tangerine concentrate, while the second and third groups received an additional 20 and 40 µg/d of pure ß-cryptoxanthin (CX40 and CX60), respectively, for 21 d. During the last 2 d of the study, urine and faecal samples of two gerbils from each treatment group were collected. ß-Cryptoxanthin was detected in the whole blood, and in twelve of the fourteen tissues analysed. Most tissues resembled the liver, in which the concentrations of ß-cryptoxanthin were significantly higher in the CX60 (17·8 (sem 0·7) µg/organ; P= 0·004) and CX40 (16·2 (sem 0·9) µg/organ; P= 0·006) groups than in the CX20 group (13·3 (sem 0·4) µg/organ). However, in intestinal tissues, the concentrations of ß-cryptoxanthin increased only in the CX60 group. Despite elevated vitamin A concentrations in tissues at baseline due to pre-study diets containing high levels of vitamin A, ß-cryptoxanthin maintained those vitamin A stores. These results indicate that ß-cryptoxanthin is stored in many tissues, potentially suggesting that its functions are widespread.

Gastric digestion of raw and roasted almonds in vivo.

Almonds are an important dietary source of lipids, protein, and α-tocopherol. It has been demonstrated that the physical form of almond kernels influences their digestion and absorption, but the role of thermal processes on the digestion of almonds has received little attention. The objectives of this study were to examine the gastric emptying and nutrient composition of gastric chyme from pigs (used as a model for the adult human) fed a single meal of either raw or roasted almonds over a 12-h postprandial period (72 pigs total, 6 pigs at each diet-time combination). Concentrations of glucose, triacylglycerols, and α-tocopherol in peripheral plasma during the 12-h postprandial period were determined. For dry matter and lipid, the gastric emptying profile was not different between raw and roasted almonds. Roasting almonds also did not influence gastric pH, or plasma glucose or triacylglycerols levels. In contrast, the gastric emptying of protein was more rapid for raw almonds compared to roasted almonds (P < 0.01) and intragastric protein content exhibited segregation (P < 0.001) throughout the stomach, with raw almonds having a higher level of segregation compared to roasted almonds. Postprandial plasma α-tocopherol levels were, on average 33% greater (P < 0.001) after consumption of raw almonds, most likely as a result of the higher concentration of α-tocopherol in raw almonds compared to roasted almonds. Roasting of almonds did not influence the overall gastric emptying process, but did lead to differences in the distribution of protein in the stomach and to the gastric emptying of protein.

The effects of daily consumption of ß-cryptoxanthin-rich tangerines and ß-carotene-rich sweet potatoes on vitamin A and carotenoid concentrations in plasma and breast milk of Bangladeshi women with low vitamin A status in a randomized controlled trial.

BACKGROUND: The potential of ß-cryptoxanthin (CX)-rich foods to form vitamin A (VA) in humans in not well understood. OBJECTIVE: We measured the effects of consuming CX- and ß-carotene (BC)-rich foods on plasma and breast milk VA and carotenoids in lactating women with low VA status. DESIGN: Participants were randomly assigned to 4 groups (n = 34, 34, 34, and 33, respectively) receiving orange-fleshed sweet potatoes (OFSPs) (12 mg BC/d), tangerines (5.3 mg CX/d), white-fleshed sweet potatoes (WFSPs) with a VA supplement (0.5 mg/d), or WFSPs 2 times/d, 6 d/wk for 3 wk. All except the VA group received placebo capsules identical in appearance to VA supplements. Changes in plasma and breast milk VA, BC, and CX were measured. RESULTS: Plasma retinol increased in the VA group. Plasma BC in the OFSP group and CX in the tangerine group increased 250% and 830%, respectively; apparent relative absorption in the CX group, considering the amounts consumed, was 4 times that in the BC group. Mean (±SEM) changes in milk VA in the OFSP (0.028 ± 0.074 µmol/L) and tangerine (0.067 ± 0.091 µmol/L) groups did not differ from those in the control (-0.077 ± 0.068 µmol/L) or VA (0.277 ± 0.094 µmol/L) group. Milk CX increased in the tangerine group. CONCLUSIONS: VA capsules increased plasma and milk VA concentrations. The greater change in CX concentrations in the tangerine group than in BC concentrations in the OFSP group suggests that CX in tangerines was better absorbed, but both foods failed to increase milk VA concentrations. This trial was registered at clinicaltrials.gov as NCT01420406.

Biofortified cassava increases ß-carotene and vitamin A concentrations in the TAG-rich plasma layer of American women.

Biofortification of cassava with the provitamin A carotenoid ß-carotene is a potential mechanism for alleviating vitamin A deficiency. Cassava is a staple food in the African diet, but data regarding the human bioavailability of ß-carotene from this food are scarce. The objective of the present study was to evaluate provitamin A-enhanced cassava as a source of ß-carotene and vitamin A for healthy adult women. The study was a randomised, cross-over trial of ten American women. The subjects consumed three different porridges separated by 2 week washout periods. Treatment meals (containing 100 g cassava) included: biofortified cassava (2 mg ß-carotene) porridge with added oil (15 ml peanut or rapeseed oil, 20 g total fat); biofortified cassava porridge without added oil (6 g total fat); unfortified white cassava porridge with a 0·3 mg retinyl palmitate reference dose and added oil (20 g total fat). Blood was collected six times from - 0·5 to 9·5 h post-feeding. TAG-rich lipoprotein (TRL) plasma was separated by ultracentrifugation and analysed using HPLC with coulometric array electrochemical detection. The AUC for retinyl palmitate increased after the biofortified cassava meals were fed (P< 0·05). Vitamin A conversion was 4·2 (sd 3·1) and 4·5 (sd 3·1) µg ß-carotene:1 µg retinol, with and without added oil, respectively. These results show that biofortified cassava increases ß-carotene and retinyl palmitate TRL plasma concentrations in healthy well-nourished adult women, suggesting that it is a viable intervention food for preventing vitamin A deficiency.

Modelling potential ß-carotene intake and cyanide exposure from consumption of biofortified cassava.

Vitamin A (VA) deficiency causes disability and mortality. Cassava can be crossbred to improve its ß-carotene (BC) content; typical white cassava contains negligible amounts of BC. However, cassava contains cyanide and its continued consumption may lead to chronic disability. Our objective was to estimate the risk-benefit of consuming BC-enhanced cassava to increase VA intake. A total of ten American women were fed white and BC-enhanced cassava. BC and cyanide data from the feeding study were combined with African cassava consumption data to model the potential daily BC, VA and cyanide intakes of African women. If BC-enhanced cassava replaced white cassava in the diets, it could theoretically meet recommended VA intakes for the following percentages of individuals from six African countries that consume cassava as a staple crop: Angola (95 %), Central African Republic (95 %), Congo (about 100 %), Ghana (99 %), Mozambique (99 %) and Nigeria (92 %). Cyanide intake after minimal preparation of cassava could be thirteen to thirty-two times the reference dose (RfD), a toxicological exposure reference, but could be completely removed by extensive soaking. This study demonstrates that consumption of BC-enhanced cassava, processed to maintain BC and remove cyanide, theoretically increases VA intakes for African populations and other areas of the world where cassava is a staple crop.

Release and bioaccessibility of ß-carotene from fortified almond butter during in vitro digestion.

The objective of this study was to determine the release and bioaccessibility of ß-carotene from fortified almond butter using in vitro digestion models. Two types of fortifiers were investigated: ß-carotene oil (oil) and whey protein isolate (WPI)-alginate-chitosan capsules containing ß-carotene oil (capsule). Shaking water bath and Human Gastric Simulator (HGS) digestion models assessed the impact of gastric peristalsis on the release of ß-carotene. Bioaccessibility of ß-carotene was measured as percent recovered from the micelle fraction. There was greater release of ß-carotene from oil fortified almond butter in the HGS model (87.1%) due to peristalsis than the shaking water bath model (51.0%). More ß-carotene was released from capsule fortified almond butter during intestinal digestion. However, more ß-carotene was recovered from the micelle fraction of oil fortified almond butter. These results suggest that a WPI-alginate-chitosan capsule coating may inhibit the bioaccessibility of ß-carotene from fortified almond butter.

ß-Cryptoxanthin- and α-carotene-rich foods have greater apparent bioavailability than ß-carotene-rich foods in Western diets.

ß-Carotene (BC), ß-cryptoxanthin (CX) and α-carotene (AC) are common carotenoids that form retinol. The amount of retinol (vitamin A) formed from carotenoid-rich foods should depend chiefly on the bioavailability (absorption and circulation time in the body) of carotenoids from their major food sources and the selectivity and reactivity of carotene cleavage enzymes towards them. The objective of the present study was to estimate the apparent bioavailability of the major sources of provitamin A (AC, BC and CX) from the diet by comparing the concentrations of these carotenoids in blood to their dietary intakes. Dietary intakes were estimated by FFQ (three studies in this laboratory, n 86; apparent bioavailability calculated for six other studies, n 5738) or by food record (two studies in our laboratory, n 59; apparent bioavailability calculated for two other studies, n 54). Carotenoid concentrations were measured by reversed-phase HPLC. Apparent bioavailability was calculated as the ratio of concentration in the blood to carotenoid intake. Then apparent bioavailabilities for AC and CX were compared to BC. Eating comparable amounts of AC-, CX- and BC-rich foods resulted in 53 % greater AC (99 % CI 23, 83) and 725 % greater CX (99 % CI 535, 915) concentrations in the blood. This suggests that the apparent bioavailability of CX from typical diets is greater than that of BC. Thus, CX-rich foods might be better sources of vitamin A than expected.

A minute dose of 14C-{beta}-carotene is absorbed and converted to retinoids in humans.

Our objective was to quantify the absorption and conversion to retinoids of a 1.01-nmol, 3.7-kBq oral dose of (14)C-beta-carotene in 8 healthy adults. The approach was to quantify, using AMS, the elimination of (14)C in feces for up to 16 d after dosing and in urine for up to 30 d after dosing. The levels of total (14)C in undiluted serial plasma samples were measured for up to 166 d after dosing. Also, the levels of (14)C in the retinyl ester (RE), retinol (ROH), and beta-carotene fractions that were isolated from undiluted plasma using HPLC were measured. The apparent digestibility of the (14)C was 53 +/- 13% (mean +/- SD), based on the mass balance data, and was generally consistent with the area under the curve for zero to infinite period of (14)C that was eliminated in the feces collections made up to 7.5 d after dosing. Metabolic fecal elimination, calculated as the slope per day (% (14)C-dose/collection from d 7.5 to the final day), was only 0.05 +/- 0.02%. The portion of the (14)C dose eliminated via urine was variable (6.5 +/- 5.2%). Participants [except participant 6 (P6)] had a distinct plasma peak of (14)C at 0.25 d post-dose, preceded by a shoulder at approximately 0.1 d, and followed by a broad (14)C peak that became indistinguishable from baseline at approximately 40 d. Plasma (14)C-RE accounted for most of the absorbed (14)C early after dosing and P1 had the longest delay in the first appearance of (14)C-RE in plasma. The data suggest that plasma RE should be considered in estimating the ROH activity equivalent of ingested beta-carotene.

Effects of chronic ethanol ingestion and folate deficiency on the activity of 10-formyltetrahydrofolate dehydrogenase in rat liver.

BACKGROUND: We recently observed that ethanol feeding impairs 10-formyltetrahydrofolate (10-FTHF) dehydrogenase (EC 1.5.1.6.) and 10-FTHF hydrolase activity in rats. In the present study, we explored the effects of folate deficiency or sufficiency combined with alcoholic intake on 10-FTHF and possible mechanisms by which chronic ethanol ingestion produces folate deficiency. METHODS: Sprague-Dawley rats were fed either folate-sufficient (FS) or folate-deficient (FD) diets; with or without ethanol (E) for four weeks. Hepatic 10-FTHF dehydrogenase and hydrolase activity, plasma folate and homocysteine were measured at baseline and after feeding experimental diets. RESULTS: Liver weight increased slightly with either folate deficiency or ethanol consumption. In rats fed the folate-sufficient diet with ethanol (FSE), plasma folate was decreased slightly (p<0.05) and plasma homocysteine elevated compared to rats fed the FS diet without ethanol. Ethanol did not affect plasma folate and plasma homocysteine in FD rats. Red-blood cell (RBC) folate was increased similarly in rats by ethanol feeding (FSE and FDE>FS and FD). Feeding folate deficient or ethanol (FSE, FD and FDE) diets depressed hepatic activities of 10-FTHF dehydrogenase, which catalyzes the oxidative deformylation of 10-FTHF to tetrahydrofolate (THF) and carbon dioxide. Rats consuming the FDE diet had the lowest enzyme activities of the experimental groups, implying that folate deficiency and ethanol consumption each affect enzyme activity. CONCLUSIONS: We confirm that ethanol decreases hepatic 10-FTHF dehydrogenase activity and show that this decrease occurs irrespective of folate status. This shows that modulation of 10-FTHF is one possible mechanism by which ethanol intake decreases folate status and affects one-carbon metabolism.

The triacylglycerol synthesis enzyme DGAT1 also catalyzes the synthesis of diacylglycerols, waxes, and retinyl esters.

The final step of triacylglycerol biosynthesis is catalyzed by acyl CoA:diacylglycerol acyltransferase (DGAT) enzymes. The two known DGATs, DGAT1 and DGAT2, are encoded by unrelated genes. Although both DGAT1 and DGAT2 knockout mice have reduced tissue triacylglycerol contents, they have disparate phenotypes, prompting us to investigate whether the two enzymes have unrecognized functional differences. We now report that DGAT1 exhibits additional acyltransferase activities in vitro, including those of acyl CoA:monoacylglycerol acyltransferase (MGAT), wax monoester and wax diester synthases, and acyl CoA:retinol acyltransferase (ARAT), which catalyze the synthesis of diacylglycerols, wax esters, and retinyl esters, respectively. These activities were demonstrated in in vitro assays with membranes from insect cells or homogenates from COS7 cells overexpressing DGAT1. Wax synthase and ARAT activities were also demonstrated in intact COS7 cells expressing DGAT1. Additionally, cells and tissues from DGAT1-deficient mice exhibited reduced ARAT activity, and the mice had increased levels of unesterified retinol in their livers on a high-retinol diet. Our findings indicate that DGAT1 can utilize a variety of acyl acceptors as substrates in vitro and suggest that these activities may be relevant to the in vivo functions of DGAT1.

Carotenoid and retinoid metabolism: insights from isotope studies.

Use of isotopes as tracers has had an important role in elucidating key features of vitamin A and retinoid metabolism in animal models and humans. Their use has shown that beta-carotene absorption is variable, and that the appearance of beta-carotene and its metabolites in the blood by time since dosing follows characteristic patterns. Retinol formed from beta-carotene shows a different pattern, as does lutein. In this article, we summarize and discuss insights and some surprises into the absorption and metabolism of vitamin A, beta-carotene, and lutein that were gained with the use of isotope tracers in humans, rats, and cells as models.

Kinetic parameters and plasma zinc concentration correlate well with net loss and gain of zinc from men.

The search for a reliable, convenient indicator of Zn status was the focus of research for several decades. Plasma Zn concentration is still the most widely used clinical measurement, despite the known problems of interpretation. More recently, researchers suggested that isotopically determined kinetic parameters, such as the exchangeable Zn pool (EZP), may more accurately and reliably reflect body Zn status. The objective of this study was to examine the relationship between net body Zn loss and gain during acute changes in dietary Zn intake with biochemical and kinetic indices of Zn status. Five men participated in an 85-d Zn depletion/repletion study. Net body Zn loss and gain were determined from the difference between dietary plus intravenously administered Zn and Zn excretion. Biochemical indicators of Zn status included plasma Zn, plasma alkaline phosphatase activity, and plasma retinol binding protein concentration. Following intravenous administration of (70)Zn or (67)Zn, a compartmental model was used to determine EZP mass, fractional Zn absorption, endogenous zinc excretion (EZE), and plasma Zn flux. The changes in total body zinc correlated best with changes in plasma Zn (r(2) = 0.826, P < 0.001), EZE (r(2) = 0.773, P < 0.001), and plasma Zn flux (r(2) = 0.766, P < 0.001). This study confirms that plasma Zn concentration is a valid indicator of whole-body Zn status in the absence of confounding factors; however, further research is needed to determine how kinetic parameters respond to conditions where plasma Zn concentration is known to be unreliable.

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

BACKGROUND: The vitamin A activity of beta-carotene is variable and surprisingly low in women. The reasons for this are not well understood. The vitamin A activity of beta-carotene in men is still uncertain. Contributions of dietary factors compared with individual traits are largely unknown. OBJECTIVE: Our objective was to measure the intrinsic variability in the vitamin A activity of beta-carotene among healthy, well-fed men living in a controlled environment. DESIGN: We used a double-tracer test-retest design. We dosed 11 healthy men orally with 30 micromol hexadeuterated (D6) retinyl acetate (all-trans-19,19,19,20,20,20-[2H6]retinyl acetate) and then with 37 micromol D6 beta-carotene (19,19,19,19',19',19'-[2H6]beta-carotene) 1 wk later. Doses were taken with breakfasts containing 16 g fat. We measured D6 retinol, D6 beta-carotene, and trideuterated (D3) retinol (derived from D6 beta-carotene) concentrations in plasma. Areas under the plasma concentration x time since dosing curves (AUCs) were determined for D6 retinol, D6 beta-carotene, and D3 retinol. RESULTS: All men had detectable D6 retinol concentrations in plasma. The mean (+/-SE) absorption of D6 beta-carotene in all subjects was 2.235 +/- 0.925%, and the mean conversion ratio was 0.0296 +/- 0.0108 mol retinol to 1 mol beta-carotene. Only 6 of 11 men had sufficient plasma concentrations of D6 beta-carotene and D3 retinol that we could measure. The mean absorption of D6 beta-carotene in these 6 subjects was 4.097 +/- 1.208%, and the mean conversion ratio was 0.0540 +/- 0.0128 mol retinol to 1 mol beta-carotene. CONCLUSION: The vitamin A activity of beta-carotene, even when measured under controlled conditions, can be surprisingly low and variable.

DGAT1 is not essential for intestinal triacylglycerol absorption or chylomicron synthesis.

Dietary triacylglycerols are a major source of energy for animals. The absorption of dietary triacylglycerols involves their hydrolysis to free fatty acids and monoacylglycerols in the intestinal lumen, the uptake of these products into enterocytes, the resynthesis of triacylgylcerols, and the incorporation of newly synthesized triacylglycerols into nascent chylomicrons for secretion. In enterocytes, the final step in triacylglycerol synthesis is believed to be catalyzed primarily through the actions of acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes. In this study, we analyzed intestinal triacylglycerol absorption and chylomicron synthesis and secretion in DGAT1-deficient (Dgat1(-/-)) mice. Surprisingly, DGAT1 was not essential for quantitative dietary triacylglycerol absorption, even in mice fed a high fat diet, or for the synthesis of chylomicrons. However, Dgat1(-/-) mice had reduced postabsorptive chylomicronemia (1 h after a high fat challenge) and accumulated neutral-lipid droplets in the cytoplasm of enterocytes when chronically fed a high fat diet. These results suggest a reduced rate of triacylglycerol absorption in Dgat1(-/-) mice. Analysis of intestine from Dgat1(-/-) mice revealed activity for two other enzymes, DGAT2 and diacylglycerol transacylase, that catalyze triacylglycerol synthesis and apparently help to compensate for the absence of DGAT1. Our findings indicate that multiple mechanisms for triacylglycerol synthesis in the intestine facilitate triacylglycerol absorption.