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The purpose of this article is to provide the rationale for the use of pedicled buccal fat pad grafts as an adjunct to the reconstruction of palatal or dentoalveolar clefts in cases when healing by secondary intention may need to be considered and integrated into the initial treatment plan because of the size of the defect or qualitative or quantitative tissue constraints. Four representative cases are presented in which a pedicled buccal fat graft was adjunctively used in conjunction with pedicled mucosal flaps to gain closure of large oroantral or oronasal cleft deformity.

Human subjects (n = 24) were supplemented with 100 mg beta-carotene/d for 6 d, either as synthetic all-trans-beta-carotene or a natural beta-carotene preparation derived from the alga Dunaliella salina, which consists of a 50:50 mixture of all-trans- and 6-cis-beta-carotene. This loading dose was followed by a 23-d maintenance dose consisting of alternate-day supplementation with 50 mg all-trans-beta-carotene or either 66 or 100 mg of the natural 50:50 isomeric mixture. The loading dose resulted in significant increases in plasma concentrations of both isomers, with the all-trans-beta-carotene-supplemented group showing a 7.2- and 5.0-fold increase in the all-trans and 9-cis concentrations in plasma, respectively. The group receiving the 50:50 mixture showed a 4.0- and 3.7-fold increase in the all-trans and 9-cis concentrations in plasma, respectively, without any apparent dose-dependency. However, even with the 50:50 mixture, the 9-cis concentrations were only a small fraction of the total plasma beta-carotene. Results after an additional 23-d period of alternate-day supplementation were not significantly different from those described above for the 6-d supplementation. Increases in low-density-lipoprotein concentrations of total beta-carotene correlated strongly with the increases seen in plasma concentrations. Lipid-soluble antioxidants vitamin E and ubiquinol were unaffected by beta-carotene supplementation. However, the amount of lycopene in the low-density lipoprotein decreased during this supplementation period. A strong discrimination between these two geometric isomers of beta-carotene was demonstrated, although the tissue site of discrimination was not determined.

This study was done to examine the intestinal absorption and cleavage of 9-cis-beta-carotene in vivo. A micellar solution, containing either no addition or 10 mumol of 9-cis- or all-trans-beta-carotene, was perfused for 2 h through the upper portion of the small intestine of ferrets. The effluent of a mesenteric lymph duct cannulation was collected, as well as intestinal mucosa scrapings, a portal blood sample, and a liver biopsy, both before and after perfusion. Carotenoids and retinoids were measured by reverse-phase, high performance liquid chromatography. 9-Cis- and all-trans-beta-carotene were transported equally well into mesenteric lymph, although the intestinal concentration of the corresponding isomer was tenfold higher after perfusion of the 9-cis- isomer than after perfusion of all-trans-beta-carotene. Regardless of which isomer was used, perfusion of beta-carotene resulted in the biosynthesis of similar amounts of retinoic acid in portal blood, liver, and intestine. However, after the perfusion of all-trans-beta-carotene, all the retinoic acid formed was in the all-trans- form, whereas the perfusion of 9-cis-beta-carotene resulted in the biosynthesis of about 50% of the total retinoic acid as the 9-cis-isomer. We conclude that in the in vivo ferret model, 9-cis-beta-carotene has a good bioavailability and is a precursor of 9-cis-retinoic acid.

Background/aims: beta-Carotene and alpha-tocopherol may have either antagonistic or synergistic effects on each other's absorption and metabolism. The effects of both physiological and pharmacological concentrations of alpha-tocopherol on the absorption and metabolism of beta-carotene in ferret intestine were determined., Methods: A high concentration of beta-carotene was perfused through the upper portion of the small intestine of ferrets in vivo with varying levels of alpha-tocopherol. The effluent of a mesenteric lymph duct cannulation, the intestinal mucosal scraping, and portal vein blood were sampled and analyzed by high-performance liquid chromatography., Results: The lymphatic transport of beta-carotene was enhanced 4-fold by alpha-tocopherol at a physiological dose and 12-21-fold at a pharmacological dose. The lymphatic transport of alpha-tocopherol was linearly (r = 0.8; P < 0.05) related to the luminal alpha-tocopherol concentration even in the presence of a high concentration of beta-carotene. Furthermore, alpha-tocopherol increased the conversion of beta-carotene into retinol in the intestine in a dose-dependent manner., Conclusions: alpha-Tocopherol has a positive effect on the intestinal absorption of intact beta-carotene and may modulate the metabolic conversion of beta-carotene into retinoids.

Objective: To evaluate small-intestine mucosal integrity and permeability with advancing age as measured by the lactulose/mannitol absorption test in healthy subjects., Design: Prospective cohort study., Setting: Clinical research unit of the USDA Human Nutrition Research Center on Aging., Study Participants: Fifty-six subjects were recruited in three age groups: 20 to 39 years (n = 20), 40 to 59 years (n = 19), and > or = 60 years (n = 17). Subjects were healthy, community-dwelling volunteers., Intervention: After an 8-hour fast, all subjects ingested 10 g of lactulose and 5 g of mannitol. Urine was collected for 6 hours and analyzed for lactulose and mannitol by high performance liquid chromatography. Twenty-four-hour urinary creatinine clearances were determined., Main Outcome Measures: Percentages of lactulose and mannitol excreted, the lactulose/mannitol ratio (x100), and the 24-hour creatinine clearance., Results: With increasing age, both the percentage of lactulose excreted (P = 0.09) and the percentage of mannitol excreted (P = 0.05) tended to decrease progressively. The lactulose/mannitol ratio (x100) did not change with increasing age (P = 0.65) because both the percentages of lactulose and mannitol excreted declined. The creatinine clearance decreased markedly with advancing age (P < .001) and accounted for the decline in percentages of lactulose and mannitol excreted (P < .02)., Conclusions: There is a progressive decline in the ability to excrete lactulose and mannitol with increasing age. This is probably attributable to a decline in renal function with advancing age. However, because of parallel decreases in lactulose and mannitol excretion, the lactulose/mannitol ratio does not change. Thus small-intestine "leakiness" does not increase with aging as measured by the lactulose/mannitol absorption test.

The effect of the live bacterial yogurt cultures, namely Streptococcus thermophilus and Lactobacillus bulgaricus, and a mucosal adhering strain of Lactobacillus gasseri (ADH) on small intestinal and fecal bacterial characteristics was examined in 10 elderly subjects with atrophic gastritis and 23 elderly normal volunteers (11 received yogurt and 12 received ADH). Neither S thermophilus nor L bulgaricus was recovered from the stomach or small intestine of subjects fed yogurt or pasteurized yogurt. ADH was recovered from gastric or small intestinal aspirates in three of four subjects and in the stools of four of five subjects diagnosed with atrophic gastritis. In 11 of 12 normal subjects, ADH was isolated from stools. There was a significant reduction in fecal bacterial enzyme activity in both normal volunteers and subjects with atrophic gastritis after being fed with viable ADH. Adherent strains of bacteria such as ADH are likely to survive passage through the gastrointestinal tract and thus have greater metabolic effects.

To determine whether patients with colon cancer metabolize beta-carotene differently from benign colon polyp patients, a normal control group (n = 13) and groups of resected colon polyp patients (n = 29) or resected colon cancer patients (Dukes A and B1, n = 21) were supplemented with placebo or beta-carotene (30 mg/day) taken with their morning meals for three months. Serum samples at zero and three months of the study were analyzed blindly for retinoic acid and beta-carotene. The results showed that beta-carotene levels in the serum of colon polyp and colon cancer groups were 8- to 12-fold higher than in the untreated control or the placebo-treated groups. The benign polyp subjects (n = 17) receiving beta-carotene showed a significant rise in serum trans-retinoic acid at three months compared with Time 0. The trans-retinoic acid values from the colon cancer group receiving beta-carotene (n = 11) or placebo (n = 10) were significantly lower than the values from the beta-carotene-supplemented colon polyp group. It appears that trans-retinoic acid levels in response to beta-carotene supplementation are different between treated cancer and benign patients because of different body demands for retinoic acid.

The antioxidant effect of dietary beta-carotene supplementation on the peroxidation potential of plasma was investigated in a randomized double-blind, placebo-controlled study. Twelve healthy women (62-80 y) supplemented their usual daily diet with 90 mg of beta-carotene (n = 6) or placebo (n = 6) capsules for 3 wk. Plasma concentrations of beta-carotene, alpha- and gamma-tocopherol, ascorbate, urate, bilirubin and in vitro production of phosphatidylcholine hydroperoxides (PC-OOH) and utilization of plasma antioxidants in the presence of 50 mmol/L 2,2'-azobis (2-aminopropane) hydrochloride (AAPH), a free radical generator, at 37 degrees C were measured before and after dietary treatment. Plasma beta-carotene increased from 0.76 +/- 0.16 to 6.45 +/- 1.16 micromol/L (P < 0.05) in supplemented but not placebo-treated subjects. The plasma concentrations of other antioxidants did not change significantly in either group. beta-Carotene supplementation did not affect basal levels of plasma PC-OOH as measured by HPLC post-column chemiluminescence but did affect AAPH-induced production of PC-OOH. Before supplementation, the induction period of plasma PC-OOH production was 2.4 +/- 0.4 h, with levels reaching 5.39 +/- 1.50 micromol/L after 6 h of incubation. After supplementation, the induction period increased significantly to 4.2 +/- 0.4 h (P < 0.01), with a lower PC-OOH production of 2.16 +/- 0.90 micromol/L after 6 h (P < 0.05). In this system, plasma ascorbate concentrations were depleted first, followed by loss of bilirubin and alpha-tocopherol and then by the sequential loss of gamma-tocopherol, urate and beta-carotene. These results indicate that beta-carotene supplementation increases the plasma antioxidant capacity of older women.

The role of 9-cis-beta-carotene (9-cis-beta-C) as a potential precursor of 9-cis-retinoic acid (9-cis-RA) has been examined in human intestinal mucosa in vitro. By using HPLC, uv spectra, and chemical derivatization analysis, both 9-cis-RA and all-trans-retinoic acid (all-trans-RA) have been identified in the postnuclear fraction of human intestinal mucosa after incubation with 9-cis-beta-C at 37 degrees C. The biosynthesis of both 9-cis-RA and all-trans-RA from 9-cis-beta-C was linear with increasing concentrations of 9-cis-beta-C (2-30 microM) and was linear with respect to tissue protein concentration up to 0.75 mg/ml. Retinoic acid was not detected when a boiled incubation mixture was incubated in the presence of 9-cis-beta-C. The rate of synthesis of 9-cis- and all-trans-RA from 4 microM 9-cis-beta-C were 16 +/- 1 and 18 +/- 2 pmol/hr/mg of protein, respectively. However, when 2 microM all-trans-beta-C was added to the 4 microM 9-cis-beta-C, the rate of all-trans-RA synthesis was increased to 38 +/- 6 pmol/hr/mg of protein, whereas the rate of 9-cis-RA synthesis remained the same. These results suggest that 9-cis-RA is produced directly from 9-cis-beta-C. Furthermore, incubations of either 0.1 microM 9-cis- or all-trans-retinal under the same incubation conditions showed that 9-cis-RA could also arise through oxidative conversion of 9-cis-retinal. Although only 9-cis-RA was detected when 9-cis-RA was used as the substrate, the isomerization of the all-trans-RA to 9-cis-RA cannot be ruled out, since both all-trans-RA and trace amounts of 9-cis-RA were detected when all-trans-retinal was incubated as the substrate. These data indicate that 9-cis-beta-C can be a source of 9-cis-RA in the human. This conversion may have a significance in the anticarcinogenic action of beta-C.

Background/aims: Bacterial overgrowth of the small intestine commonly occurs in association with hypochlorhydria caused by atrophic gastritis or during treatment with omeprazole. The purpose of this study was to determine the clinical significance of bacterial overgrowth on small intestinal absorption and permeability and to evaluate the reliability of noninvasive breath tests to detect bacterial overgrowth in subjects with hypochlorhydria., Methods: Seventeen healthy, elderly subjects with atrophic gastritis or omeprazole treatment (40 mg/day) and documented bacterial overgrowth were studied., Results: There was no evidence of fat malabsorption (72-hour fecal fat) or clinically significant carbohydrate malabsorption (25 g D-xylose and fecal pH) in any subject. The ratio of lactulose to mannitol excreted was normal in both atrophic gastritis and omeprazole-treated groups. Three subjects in each group had abnormally high alpha 1-antitrypsin clearances. Lactulose (10 g) and glucose (80 g) hydrogen breath tests were only abnormal in 1 out of 17 subjects, whereas the 1 g [14C]D-xylose test was abnormal in 6 out of 17 subjects., Conclusions: Bacterial overgrowth caused by atrophic gastritis or omeprazole treatment is typically not associated with clinically significant fat or carbohydrate malabsorption. Noninvasive breath tests for bacterial overgrowth are not reliable in subjects with hypochlorhydria.

Risks for gastric cancer in relation to diet and other environmental factors are receiving renewed attention. New developments include the emerging relationship between Helicobacter pylori infection and an increased risk for gastric cancer, and positive results of chemoprevention trials in decreasing gastric cancer risk with the use of beta-carotene and vitamin E supplements. Factors that may enhance risk include consumption of nitrites, nitrates, alcohol, and highly salted, pickled, fermented, or smoked foods. Other environmental factors which may promote cancer are H. pylori infection, inappropriate food storage, metal and cement dust exposure, and cigarette smoking. High intakes of fruits and vegetables or of antioxidants, such as beta-carotene, vitamin E, and vitamin C may decrease risk.

Sunlight-induced photodegradation of retinyl esters and retinol in human skin, blood and cultured keratinocytes was investigated. Using high-performance liquid chromatography with an extraction method that avoided saponification, the analysis of human foreskin (Caucasian) showed that levels of retinyl esters and retinol were approximately 3.5 and 5.0 times higher, respectively, in the epidermis than in the dermis. Upon irradiation by sunlight, a significant reduction in epidermal retinyl esters was observed in both summer and winter. However, epidermal retinol, dermal retinol and dermal retinyl esters did not show statistically significant reductions. When serum from volunteers who had taken a large dose of retinyl palmitate to elevate serum retinyl esters was exposed to sunlight, the retinyl esters in the serum rapidly disappeared after 10 min of exposure--similar to the photodegradation seen for retinyl palmitate in an organic solvent. While retinol in an organic solvent rapidly photodegraded similar to serum retinyl palmitate, serum retinol slowly declined upon sunlight irradiation. When cultured keratinocytes that took-up 3H-retinol and thereafter contained 3H-retinyl esters and 3H-retinol were exposed to sunlight, 80% of the 3H-retinyl esters disappeared upon sunlight irradiation whereas only about 20% of the 3H-retinol did so. These results suggest that the epidermis, serum and keratinocytes selectively protect retinol from sunlight-induced photodegradation. It is most likely that serum retinol-binding protein and cellular retinol-binding protein protect retinol, a vital epithelial growth factor, from photodegradation.

The bacterial overgrowth syndrome occurs when there are alterations in intestinal anatomy, gastrointestinal motility, or a lack of gastric acid secretion. Clinically, patients present with nonspecific gastrointestinal symptoms that include abdominal pain, bloating, excessive gas production, diarrhea, weight loss, and malabsorption. The nutritional consequences of intestinal bacterial overgrowth include vitamin deficiencies, fat malabsorption, and malnutrition. The diagnosis requires a high index of clinical suspicion and can be established by specialized testing, such as the 1-gram 14C-xylose breath test. The goal of treatment is eradication of the bacterial overgrowth (usually with antibiotics) and the correction of nutritional deficiencies.

This article is an update of our 1987 literature review of vitamin requirements of elderly people (Am J Clin Nutr 1987;45:501-12). Poor dietary intake is the cause of much vitamin malnutrition in elderly people. Nevertheless, there is strong evidence that aging affects the requirement for certain vitamins. The 1989 recommended dietary allowances (RDAs) appear to be too low for elderly people for vitamin D, riboflavin, vitamin B-6, and vitamin B-12, and too high for vitamin A. For several vitamins there is enough information to establish an RDA for the category > or = 70 y.

Six groups of elderly subjects from central Guatemala were assessed for riboflavin status by using the erythrocyte glutathione reductase activity coefficient (EGRAC). The prevalence of riboflavin deficiency ranged from 50% to 76% among the free-living rural elderly subjects. Milk intake data that were collected from some of the subjects showed a significant correlation (P < 0.0001) between frequency of milk intake and riboflavin status. In a short-term riboflavin supplementation experiment in which nine riboflavin-deficient subjects were given 10 mg riboflavin/d for 3 d, all the subjects' EGRACs were normalized by the supplementation. However, they returned to a state of deficiency within 2 wk while consuming their usual diets without supplementation. It appears that the high prevalence of riboflavin deficiency in elderly Guatemalan people is caused by inadequate intake of riboflavin-rich foods such as dairy products, and that sufficient amounts of riboflavin need to be ingested regularly to maintain satisfactory riboflavin status.

When the plasma concentration of retinoic acid is increased, there is an accompanying reduction of circulating levels of retinol, suggesting that retinoic acid may have a regulatory effect on retinol metabolism in vivo. To determine which specific step(s) of retinol metabolism might be regulated by retinoic acid, retinol was incubated with ferret liver microsomes or cytosol with retinoic acid in vitro. Incubating the microsomal fraction with retinoic acid resulted in a dose-dependent (up to 0.5 mumol/L) decrease in the formation of retinal. On the contrary, no retinoic acid inhibitory effect was observed on retinal synthesis in the cytosol incubation, or in the cytosol plus microsome incubation. However, when retinoic acid was added to the cytosolic incubation mixture in the presence of the retinal oxidative inhibitor, citral, a dose-dependent inhibition of retinal synthesis was observed. Furthermore, the effect of retinoic acid on retinyl ester metabolism in ferret liver was studied by using endogenous retinyl esters of ferret liver as the substrate. When retinoic acid was added to the incubation mixture of microsomes plus cytosol, small, nonsignificant increases in retinol and retinyl esters were observed. When retinoic acid was added in the presence of citral, both the inhibition of retinol oxidation and the stimulation of retinol esterification were dose dependent up to approximately 0.3 mumol/L and then remained the same up to 1.0 mumol/L. These data strongly suggest that retinoic acid has a regulatory effect on retinol metabolism in ferret liver, which may occur via feedback inhibition of retinol oxidation and stimulation of retinol esterification.

The riboflavin requirements of two groups of riboflavin-deficient, but otherwise healthy, Guatemalan elderly persons over the age of 60 y were studied by varying the fat:carbohydrate ratio in two diets. The first group consumed a diet similar in macronutrient content to a Western-type diet with low carbohydrate and high fat; the second group consumed a typical Guatemalan diet with high carbohydrate and low fat. Energy and protein intakes of both groups were similar. Riboflavin status was monitored by weekly measurements of erythrocyte glutathione reductase activity coefficient (EGRAC) and urinary riboflavin excretion. Increasing increments of riboflavin were added to the subjects' diets until their status was normalized, as indicated by EGRAC of < 1.34 and a sharp increase in urinary riboflavin excretion. Using the EGRAC method, the mean value of riboflavin intake at which the subjects' EGRAC reached the limit of normality was 1.37 +/- 0.03 mg/d in the first phase and 1.29 +/- 0.03 mg/d in the second phase. The sharp increase in urinary excretion occurred at riboflavin intakes of 1.13 and 1.03 mg/d for Groups 1 and 2, respectively. Thus, the differences between the two groups suggest that diets with a lower fat:carbohydrate ratio can decrease the dietary need for riboflavin. The dietary requirement of riboflavin, as estimated by the more reliable urinary excretion method, was 1.1-1.3 mg/d for those consuming the Western-type diet, which is similar to values found over 40 y ago in young adults. We conclude that the dietary requirements of riboflavin in the elderly do not differ from those of young adults.

To determine whether beta-carotene (beta-C) can serve as a source of intestinally-derived retinoic acid (RA), either 15,15'-[14C]beta-C or unlabeled beta-C was perfused through 30 cm jejunal segments of ferrets in vivo. Portal vein blood was sampled periodically via an indwelling catheter. RA was identified in portal blood by comparing retention times in HPLC, by UV absorption, and by derivatization (methylation) and subsequent GC-MS analysis. The RA concentration in the portal blood increased 3-fold with perfusion of beta-C (P < 0.05), and remained at 18 nmol/L during the perfusion of beta-C. The single peak of RA in HPLC was shown to consist of four separate peaks by GC-MS, which may be cis-trans isomers of RA. The concentration of RA in portal blood returned to the initial level (5 nmol/L) after a 2 h period of intestinal perfusion with 5% dextrose. Retinyl ester concentration in portal blood did not change before or after the perfusion, whereas retinol decreased significantly during the perfusion of beta-C. This study clearly indicates that a considerable quantity and number of polar metabolites, including RA, are formed from beta-C in the ferret intestine which are transported via the portal vein to the liver.

This study examined postabsorptive plasma vitamin A after doses of retinyl palmitate in healthy men (n = 28) and women (n = 31). On consecutive days one physiologic [3000 retinol equivalents (RE)] and one pharmacologic dose (105,000 RE) were administered and blood samples collected. Plasma retinol and retinyl esters were measured by HPLC. Tolerance curves were constructed by plotting plasma retinyl ester concentration vs time. Postprandial retinyl ester response was measured as peak rise in retinyl ester concentration and area under the curve (AUC). Peak plasma retinyl ester concentration occurred earlier for females but the earlier peak was significant only for younger subjects (< or = 50 y, P < 0.02) given the low dose and older subjects (> 50 y, P < 0.02) given the high dose. Peak rise and AUC were lower in females than in males, but this difference was significant for the high dose only (P < 0.05). In the high-dose experiment, when each age group was evaluated for sex differences the peak rise was significantly greater in males than in females in the older subjects (P < 0.05). Postabsorptive plasma retinol did not change from fasting concentrations. A lower plasma response in retinyl esters in women could be due to a more efficient chylomicron-remnant clearance.

The concentrations of beta-carotene, retinol and retinyl esters in serum and selected tissues of ferrets fed diets supplemented with beta-carotene (80 micrograms/g wet diet) for 3 wk were determined. The initial concentration of serum beta-carotene was 0.011 +/- 0.006 mumol/L (mean +/- SEM); at the end of the experimental period it was 5.75 +/- 1.60 mumol/L. No significant differences in serum retinol and total retinyl esters were observed between beta-carotene-fed and control ferrets that had been fed an unsupplemented diet. The predominant retinyl esters in serum were retinyl stearate (53%) and retinyl palmitate (35%). Of the tissues analyzed after beta-carotene feeding, the liver contained the highest concentration of beta-carotene (78.8 +/- 18.8 nmol/g). Other tissues that contained beta-carotene in amounts ranging from 17 to 20 nmol/g were adrenals, small intestine, stomach and colon; lesser amounts (6.9 nmol/g) were found in kidneys. Amounts ranging from 1.2 to 2.3 nmol/g were found in muscle, bladder, adipose tissue, lungs and skin; only 0.37 and 0.34 nmol/g were present in brain and eyes, respectively. Thus, like humans, ferrets have the capacity to absorb intact beta-carotene and to store this compound in tissues, especially the liver. However, compared with humans, ferrets have elevated concentrations of retinyl esters in serum, liver and other tissues.

We investigated the time course and the reproducibility of the relative-dose-response (RDR) test for assessing vitamin A status in older adults. The maximum plasma retinol response to 480 retinol equivalents (RE) of retinyl palmitate in abnormal responses was at 6 or 7 h after dosing compared with the 5-h sampling interval recommended by others for younger adults and children. With respect to reproducibility, the diagnostic concordance of two RDR tests at 7-d intervals in 14 elders was 71%. In 29% of tests, one test was abnormal and the other normal. Linear regression of the two RDR values in these 14 subjects gave a correlation coefficient of -0.08. We conclude that the procedure for the RDR should be modified when applied to persons greater than 60 y of age, and that multiple repetitions of the test are needed to provide a stable indication of vitamin A stores in an elderly individual.

Plasma and lipoprotein concentrations of beta-carotene (BC) were measured in men for 10 d after an oral dose of BC (120 mg) (experimental subjects, n = 11) or no BC (control subjects, n = 5). Lipoproteins were separated by sequential ultracentrifugation and BC was measured by HPLC. Plasma and lipoprotein BC concentrations in control subjects were steady. In experimental subjects, plasma BC content increased by 6 h postdosing (P less than 0.015), peaked at 24 h (P less than 0.05), and returned to baseline by 7 d. Maintenance of plasma BC concentrations suggests homeostatic control. Of the 11 experimental subjects, only 4 had a plasma response. Early increases in the BC content of chylomicrons, very-low-density lipoproteins, and intermediate-density lipoproteins. Intestinal input accounts for early rises in circulating BC concentrations whereas hepatic secretion is the source of later increases. Among all of the lipoproteins, transfer of BC may occur.

The catabolism of homocysteine through cystathionine synthesis requires pyridoxal-5'-phosphate, thus the effect of vitamin B-6 deficiency on plasma homocysteine concentrations was evaluated. Total fasting plasma homocysteine concentrations were measured in 11 elderly subjects aged 64.4 +/- 1.7 y (mean +/- SE) who consumed a vitamin B-6-deficient diet for less than or equal to 20 d. Only 1 of the 11 subjects was found to have elevated homocysteine concentrations even though all subjects exhibited high urinary xanthurenic acid concentrations after a tryptophan load, a measure indicative of vitamin B-6 deficiency. In a supporting study, fasting plasma homocysteine concentrations were measured in 3- and 23-mo-old rats fed vitamin B-6-deficient diets and were compared with those of vitamin B-6-replete, pair-fed controls. There was no difference in homocysteine concentrations between deficient and pair-fed animals after 6 wk of the dietary regimen for either age group; after 9 wk a modest elevation was observed in the 3-mo-old deficient rats whereas no difference was observed for the 23-mo-old rats. It is concluded that fasting plasma homocysteine concentrations are not initially elevated in vitamin B-6 deficiency and therefore fasting plasma homocysteine concentrations are not a good indicator of vitamin B-6 status.

There are numerous reports in the literature of impaired gastrointestinal function with aging. However, most gastrointestinal functions remain relatively intact because of the large reserve capacity of the intestine, pancreas, and liver. Clinically important changes in gastrointestinal function with aging in human include decreased taste thresholds, hypochlorhydria due to atrophic gastritis, and decreased liver blood flow and size. Increased absorbability of lipids and large size molecules has been demonstrated in aging animals, but this has not been studied in humans. Nutrients with impaired gastrointestinal bioavailability in aging include dietary B-12, calcium carbonate, and ferric iron in atrophic gastritis; calcium, zinc, and possibly carbohydrate in a mixed meal. The implications of these changes for health maintenance and chronic disease in elderly people are in need of study.

We evaluated vitamin A absorption in 50 healthy adults and 26 gastrointestinal-disease patients by measuring the postabsorptive response in plasma retinyl esters after oral doses of the vitamin. On 3 consecutive days, two physiologic-dose tests of 2000-2400 retinol equivalents (RE) and one pharmacologic-dose test (84,000 RE) were administered. The physiologic doses were given as an oil-soluble or a water-miscible preparation. In gastrointestinal-disease patients the physiologic-dose test was highly correlated with the pharmacologic-dose test for the oil-soluble preparation as determined by peak rise (r = 0.50, P less than 0.05) and area under the curve (r = 0.56, P less than 0.01), suggesting that the physiologic dose is valid for investigating vitamin A absorption. Intestinal-disease or resection patients absorbed preparations poorly. Pancreatic-disease patients absorbed the oil-soluble preparation poorly. Physiologic rather than pharmacologic doses of vitamin A can be used to study vitamin A absorption.

The hypothesis that retinoic acid (RA) is produced from the excentric cleavage of beta-carotene was tested in human intestinal homogenates in vitro. Significant amounts of RA were identified by HPLC and derivatization after incubation of intestinal mucosal homogenates with retinal, beta-carotene, or beta-apocarotenals at 37 degrees C for 60 min. RA formation was inhibited, in a dose-dependent fashion, when retinal was incubated in the presence of 0.1-3.0 mM citral (3,7-dimethyl-2,6-octadienal) under identical experimental conditions. The formation of RA from both beta-carotene and beta-apocarotenals was dose and time dependent and RA was the major metabolite of both beta-apo-8'-carotenal and beta-apo-12'-carotenal after the incubation. However, citral (0.1 to 4 mM) did not inhibit the formation of beta-apocarotenals and RA from 2 microM beta-carotene (P greater than 0.05), which proves the existence of an excentric cleavage mechanism for beta-carotene conversion into retinoids. Furthermore, RA formation from both beta-apo-8'-carotenal and beta-apo-12'-carotenal in human intestinal homogenate occurred in the presence of citral, which demonstrates that RA can be produced from excentric cleavage of beta-carotene via a series of beta-apocarotenals as intermediates.

At the request of UNICEF, the authors conducted a rapid nutritional assessment of children from birth to five years of age in southern Iraq during the second half of June 1991. With support from local UNICEF staff, the nutrition team examined 680 children, measuring weight, height, and mid-upper arm circumference and observing clinical signs of anemia, vitamin A deficiency, and dehydration. These measurements and observations were performed in 14 rural, urban, and periurban settings in Basrah and Amarah governorates. Evidence of both acute and chronic malnutrition was found in large numbers of the children examined. However, the acute nutritional signs of impending famine were not evident at the time of the survey. This is true notwithstanding epidemic levels of infectious disease, market prices for basic food items three to 20 times prewar levels, and a reported crisis in mothers' ability to breastfeed. Other signs of impending famine (distress sales of family assets, disintegration of social structures, etc.) may have existed, but were not evident to the team.

Two new products from the incubation of beta-carotene with intestinal mucosa homogenates of human, monkey, ferret, and rat were isolated using high-performance liquid chromatography (HPLC). Identification by comparing retention times in HPLC, by monitoring ultraviolet/visible spectra, by reduction to corresponding alcohol, by oxime formation, and by mass spectrometry demonstrated that they are beta-apo-13-carotenone and beta-apo-14'-carotenal. These compounds were not found in incubations done without intestinal homogenates or with disulfiram as an inhibitor. Under standard incubation conditions, these products increased linearly for 60 min and up to a protein concentration of 1.5 mg/mL and increased along with increasing concentrations of beta-carotene. Therefore, they are enzymatic cleavage products from beta-carotene. The formation of the beta-apo-13-carotenone and beta-apo-14'-carotenal provides direct evidence for an enzymatic excentric cleavage mechanism.

Previous studies using spectrophotometric methods for vitamin A analysis concluded that fasting prior to blood collection is not necessary for determining vitamin A status of children or young adult subjects. We measured the effect of mixed vitamin A and carotenoid containing meals with less than 3, 50, and 100% of the recommended dietary allowance (RDA) for vitamin A on serum concentrations of retinyl esters, retinol, and carotenoids in elderly and young adults after an overnight fast. Retinyl ester concentrations rose significantly in both age groups with a numerically higher rise over baseline in the elderly subjects: 6.0 +/- 0.9 micrograms/dl for elderly (p less than 0.001), 5.0 +/- 0.5 micrograms/dl for young (p less than 0.001) at 50% RDA; 9.0 +/- 1.3 micrograms/dl for elderly (p less than 0.001) and 6.8 +/- 1.6 micrograms/dl for young (p less than 0.05) at 100% RDA. We conclude that in both young and elderly adults, but especially in the elderly, fasting conditions are necessary for the accurate assessment of vitamin A status if spectrophotometric methods are used for measuring vitamin A.

The role of bacteria in the bioavailability of protein-bound vitamin B12 was examined in eight elderly subjects who had atrophic gastritis and in eight normal controls. On separate days and in random order, vitamin B12 absorption tests were performed using either radiolabeled crystalline or protein-bound vitamin B12. At the same time, bacterial samples were collected from the upper gastrointestinal tract. The tests and gastrointestinal aspirates were performed before and during tetracycline therapy. Crystalline vitamin B12 was absorbed to the same extent in the two study groups. Atrophic gastritis subjects absorbed significantly less protein-bound vitamin B12 than normal controls (mean +/- SEM, 0.7% +/- 0.2% vs. 1.9% +/- 0.5%, respectively). However, protein-bound vitamin B12 absorption in these subjects normalized after antibiotic therapy. These results suggest that the small amounts of vitamin B12 released from the protein binders is readily absorbed (as shown in vitro) and/or metabolized by bacteria.

The effect of chronic ethanol feeding on the status of alpha- and gamma-tocopherol in plasma, liver, lung, and testes of Sprague-Dawley rats was characterized. Rats were pair-fed liquid diets containing 36% of total calories either as ethanol or isocaloric carbohydrates. After 3 weeks, ethanol ingestion resulted in a significant (p less than or equal to 0.05) increase in liver weight and induced fatty liver without affecting total body weight. Ethanol feeding did not affect the plasma concentration of alpha-tocopherol but doubled that of gamma-tocopherol. When expressed per milligram of tissue, liver alpha-tocopherol did not vary with ethanol ingestion, whereas gamma-tocopherol concentration increased 2.5 times that of control animals. However, the concentration of alpha-tocopherol expressed per milligram of total lipids was significantly (p less than or equal to 0.01) decreased in the liver with ethanol feeding. In contrast to the liver, ethanol feeding significantly increased alpha- and gamma-tocopherol levels per milligram of total lipids in the testes. The concentration of gamma-tocopherol (but not alpha-tocopherol) per milligram of lung tissue and per total lung was significantly (p less than or equal to 0.05) increased with ethanol feeding. These data indicate that chronic ethanol ingestion significantly alters the distribution of alpha-tocopherol and gamma-tocopherol in hepatic and extrahepatic tissues of the rat.

The vitamin B-6 requirements of 12 men and women over 60 y old were studied. The protocol consisted of a 5-d baseline period and four experimental periods during which the subjects successively received 0.003, 0.015, 0.0225 and 0.03375 mg of vitamin B-6/(kg body wt.d). Dietary protein was 1.2 or 0.8 g/(kg body wt.d). At 5- or 6-d intervals, xanthurenic acid (XA) after a 5-g L-tryptophan load and 4-pyridoxic acid (4-PA) in 24-h urine, erythrocyte aspartate aminotransferase activity coefficient (EAST-AC) and plasma pyridoxal-5'-phosphate (PLP) were measured. These measurements were abnormal during vitamin B-6 depletion but returned to normal during repletion. Men who ingested approximately 120 g protein/d required 1.96 +/- 0.11 mg of vitamin B-6 to normalize XA; women who ingested 78 g protein/d required 1.90 +/- 0.18 mg of vitamin B-6 to normalize XA. To attain normal levels of EAST-AC and 4-PA in men, 2.88 +/- 0.17 mg of vitamin B-6 were needed; to normalize PLP, 1.96 +/- 0.11 mg of vitamin B-6 were required. Women required 1.90 +/- 0.18 mg or more of vitamin B-6 to normalize these measurements. Vitamin B-6 requirements were not decreased in two of three subjects who ingested 54 g of protein daily. Thus, vitamin B-6 requirements of elderly men and women are about 1.96 and 1.90 mg/d, respectively.

In vitro studies suggest that the effect of fiber in inhibiting calcium absorption is pH dependent. In nine normal, elderly control subjects and eight elderly subjects with achlorhydria, 47Ca was ingested with three test meals: a low-fiber meal (0.5 g dietary fiber), a high-fiber meal (10.5 g), and a high-fiber meal with 120 mL of 0.1 mol HC/L. In control subjects calcium retention, measured in a whole-body counter, was 25.7 +/- 4.0% (mean +/- SD) with the low-fiber meal, 19.1 +/- 1.9% with the high-fiber meal (P less than 0.002 vs low fiber), and 18.9 +/- 3.3% with the high-fiber-plus-acid meal (P less than 0.002 vs low fiber, NS vs high fiber). Calcium absorption in achlorhydric subjects was not different from control subjects: 26.2 +/- 8.0% with low fiber, 19.6 +/- 4.1% with high fiber (P less than 0.04 vs low fiber), and 21.0 +/- 5.8% with high fiber plus acid (P less than 0.04 vs low fiber, NS vs high fiber). We conclude that, in humans, the reduction in calcium absorption with high fiber intake is unaffected by gastric pH.

The effect of vitamin B-6 deficiency on immune response was studied in eight healthy elderly adults. The protocol consisted of a 5-d baseline (BL) period; a vitamin B-6-depletion period of less than or equal to 20 d; three stages of vitamin B-6-repletion, each lasting 21 d; and a 4-d final phase. The amounts of vitamin B-6 ingested during the different phases of the study were 3.00, 15.00, 22.50, and 33.75 micrograms.kg body wt-1.d-1, respectively. During the final phase the subjects ingested 50 mg vitamin B-6/d. Fasting blood was collected at the end of each period. Vitamin B-6 depletion significantly decreased percentage and total number of lymphocytes, mitogenic responses of peripheral blood lymphocytes to T- and B-cell mitogens, and interleukin 2 production. These indices returned to BL values after the third vitamin B-6-repletion period, when the total vitamin B-6 intakes were 1.90 +/- 0.18 mg/d for women and 2.88 +/- 0.17 mg/d for men. Vitamin B-6 deficiency impairs in vitro indices of cell-mediated immunity in healthy elderly adults. This impairment is reversible by vitamin B-6 repletion.

The present study was designed to investigate the interaction of age and ethanol on vitamin A status in rats. Rats aged 2 and 19 mo were fed a liquid diet containing 36% of total energy as ethanol or pair-fed a diet containing isoenergetic carbohydrate in place of ethanol. After 3 wk older rats had lower serum retinol (P = 0.04) and higher vitamin A concentrations in liver (P = 0.0001), esophagus (P = 0.0001) and the proximal (P = 0.03) and distal (P = 0.0001) colon than younger animals. Hepatic microsomal cytochrome P-450, retinyl ester hydrolase (REH) and cellular retinol-binding protein (cRBP) were significantly reduced; acyl coenzyme A: retinol acyltransferase (ARAT) was increased; and alcohol (retinol) dehydrogenase (ADH) activity was unchanged with age. Ethanol ingestion increased serum retinol as well as esophageal and colonic vitamin A levels in both age groups. Hepatic cRBP decreased further in the older rats with ethanol feeding, but no change was noted in the percentage of hepatic vitamin A as retinol or retinyl esters. Ethanol ingestion decreased REH (P = 0.0001) and ARAT activities (P = 0.02) and increased cytochrome P-450 (P = 0.04) but had no effect on the activity of ADH in either age group. These data indicate that, regardless of age, chronic ethanol ingestion significantly alters the tissue distribution of vitamin A; however, ethanol reduced cRBP levels only in older rats.

Whether the conversion of beta-carotene into retinoids involves an enzymatic excentric cleavage mechanism was examined in vitro with homogenates prepared from human, monkey, ferret, and rat tissue. Using high-performance liquid chromatography, significant amounts of beta-apo-12'-, -10'-, and -8'-carotenals, retinal, and retinoic acid were found after incubation of intestinal homogenates of the four different species with beta-carotene in the presence of NAD+ and dithiothreitol. No beta-apo-carotenals or retinoids were detected in control incubations done without tissue homogenates. The production of beta-apo-carotenals was linear for 30 min and up to tissue protein concentrations of 1.5 mg/ml. The rate of formation of beta-apo-carotenals from 2 microM beta-carotene was about 7- to 14-fold higher than the rate of retinoid formation in intestinal homogenates, and the rate of beta-apo-carotenal production was fivefold greater in primate intestine vs rat or ferret intestine (P less than 0.05). The amounts of beta-apo-carotenals and retinoids formed were markedly reduced when NAD+ was replaced by NADH, or when dithiothreitol and cofactors were deleted from the incubation mixture. Both beta-apo-carotenal and retinoid production from beta-carotene were inhibited completely by adding disulfiram, an inhibitor of sulfhydryl-containing enzymes. Incubation of beta-carotene with liver, kidney, lung, and fat homogenates from each species also resulted in the appearance of beta-apo-carotenals and retinoids. The identification of three unknown compounds which might be excentric cleavage products is ongoing. These data support the existence of an excentric cleavage mechanism for beta-carotene conversion.

This paper describes relationships between reported alcohol consumption and selected sociodemographic and health variables. Two hundred and four men and 367 women aged 60-95 years were examined as part of a nutritional status survey of elderly. Fifty-three percent of men and 44% of women reported drinking at least 2 g of alcohol per week. Men were more likely to drink than women, and the level of alcohol consumption decreased with age. Drinking was positively associated with education (p less than 0.01) and negatively associated with recent medical care (p less than 0.01), history of MI (p less than 0.05), and denture use (p less than 0.05). Among drinkers, reported alcohol intake was higher for subjects less than age 70 (p less than 0.01), males (p less than 0.01), the college educated (p less than 0.01), and smokers (p less than 0.05). Level of alcohol intake was lower for those who had received medical care in the year preceding study participation (p less than 0.05). Identical results were observed for alcohol intake expressed as percent of total calories. Intake ranged from 3.8% of total calories among subjects 80+ years old to 6.2% of total calories among 60-69-year-olds.

The relative dose response (RDR) test was examined with regard to specificity and reproducibility in subjects with adequate dietary intake and normal liver reserves of vitamin A. Twelve subjects were administered an RDR test four times over 22 d, including one placebo test in which the oral dose of vitamin A was omitted. Additionally, static measures of retinol, tocopherol, retinol binding protein (RBP) and prealbumin (PAL) were taken to determine the intra- and inter-individual coefficient of variation for these blood constituents. Intra-individual coefficients of variation were as follows: retinol, 8.8%; RBP, 11.5%; PAL, 7.6%; and alpha-tocopherol, 8.8%. The mean RDR in vitamin A-replete subjects was 1.2% and ranged from approximately -25% to 11%. No differences were observed between placebo and true RDR (i.e., with vitamin A) test responses, and there was no difference among the three true RDRs over a period of 22 d. Consistent with the hypothesis upon which the RDR test is based, nascently absorbed vitamin A evidently entered a storage pool in the liver of vitamin A-replete subjects without immediate release to peripheral sites of utilization. Because the RDR test results were normal in all subjects, the procedure appears to offer high test specificity and does not falsely diagnose hypovitaminosis A. Nevertheless, the magnitude and direction of the RDR within an individual over 22 d were highly variable, and this variability may preclude the use of a single measure of the RDR to grade the relative vitamin A nutriture of an individual subject.

The relative dose response (RDR) test has been used as a functional measure of whole-body stores of vitamin A in humans. We have examined the reproducibility of the RDR procedure in a population of Guatemalan adult subjects who would be expected to show a moderate prevalence of hypovitaminosis A. Fifty-one subjects were administered a standard RDR test, and the plasma samples were analyzed for retinol, tocopherol, retinol binding protein (RBP) and prealbumin (PAL). Thirty-four of the subjects underwent repeat RDR tests 7 d later. Plasma levels in fasted subjects were as follows: retinol, 1.35 +/- 0.30 mumol/L; RBP, 37.8 +/- 7.7 mg/L; PAL, 187.0 +/- 39.0 mg/L; and tocopherol, 16.6 +/- 6.2 mumol/L. RDRs ranged from -35.2% to +63.1%, with a mean of 2.6 +/- 10.4%. Overall, we observed poor within-subject reproducibility of the RDR procedure whether expressed numerically or by diagnostic classification. Moreover, in contrast to previous studies in children, we observed fewer positive RDR tests than would be expected for the population studied. Nevertheless, the mean RDR was inversely proportional to fasting retinol levels, thus confirming the validity of the biological basis of the RDR procedure in humans. Because of high intra-individual variability with this test, investigators should be cautious when using the RDR procedure in serial studies to monitor the efficacy of therapeutic interventions or subject compliance to dietary regimens.

We investigated postprandial vitamin A metabolism by measuring retinyl ester, triglyceride, and apolipoprotein (apo)B-48 in the plasma lipoproteins of human subjects before and after fat-feeding. Following a 14-hour fast, eight healthy subjects (two men, six women, 28 to 79 years) were given a fat-rich meal (1 g fat/kg body weight) containing vitamin A (40 retinol equivalents per kilogram body weight). Blood was collected every 3 hours for 12 hours and lipoproteins were isolated by sequential ultracentrifugation. Mean plasma retinyl ester concentration peaked 6 hours after the fat-rich meal, whereas mean plasma triglyceride peaked at 3 hours. Data obtained from hourly samples in 3 subjects showed that changes in the postprandial plasma concentration of retinyl ester occurred 1 to 2 hours after changes in the plasma triglyceride concentration. In triglyceride-rich lipoproteins (TRL) of d less than 1.006 g/mL, retinyl ester similarly peaked at 6 hours, whereas triglyceride as well as apoB-48 peaked at 3 hours. Although retinyl esters were found mainly in TRL in the initial postprandial period (84%, 3 hours; 83%, 6 hours), in fasting and postprandial plasma, particularly 9 or more hours after fat-feeding, a large percentage of plasma retinyl esters were in low-density lipoproteins (LDL) (44%, fasting; 9%, 3 hours; 9%, 6 hours; 19%, 9 hours; 32%, 12 hours). A small percentage of retinyl esters were also found in postprandial high-density lipoproteins (HDL) (2% to 7%). ApoB-48 was not detected in LDL of fasting or postprandial plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

Postprandial vitamin A and intestinal lipoprotein metabolism was studied in 86 healthy men and women, aged 19-76 yr. Three independent experiments were carried out. In the first experiment, a supplement dose of vitamin A (3,000 retinol equivalents [RE]) was given without a meal to 59 subjects, aged 22-76 yr. In the second experiment, 20 RE/kg body wt was given with a fat-rich meal (1 g fat/kg body wt) to seven younger subjects (aged less than 50 yr) and seven older subjects (aged greater than or equal to 50 yr). In both experiments, postprandial plasma retinyl ester response increased significantly with advancing age (P less than 0.05). In the third experiment, retinyl ester-rich plasma was infused intravenously into nine young adult subjects (aged 18-30 yr) and nine elderly subjects (aged greater than or equal to 60 yr), and the rate of retinyl ester disappearance from plasma during the subsequent 3 h was determined. Mean (+/- SE) plasma retinyl ester residence time was 31 +/- 4 min in the young adult subjects vs. 57 +/- 8 min in the elderly subjects (P less than 0.05). These data are consistent with the concept that increased postprandial plasma retinyl ester concentrations in older subjects are due to delayed plasma clearance of retinyl esters in triglyceride-rich lipoproteins of intestinal origin.

In 1981 to 1983, the nutrition and health status of 346 Chinese immigrants in Boston, Mass, aged 60 to 96 years was surveyed and analyzed for cardiovascular risk factors. These elderly Chinese were physically active and seldom obese and consumed a high-carbohydrate (57% of total energy intake), low-fat (24% of total energy intake), low-ascorbic acid (0.62 mmol/d) diet. Current cigarette smoking was common (39%) only in men, while alcoholism was rare in both sexes. Compared with elderly whites, they had lower mean blood pressure and blood levels of total, low-density lipoprotein, and high-density lipoprotein cholesterol, apolipoproteins A-I and B, and ascorbic acid. These characteristics resemble those of the urban population in mainland China, where hemorrhagic stroke is the major cause of cardiovascular mortality.

The occurrence of 13-cis-retinoic acid as an endogenous component in human serum has been confirmed by cochromatography with standards in both normal-phase and reverse-phase high-performance liquid chromatographic (HPLC) system, by the lambda max of its UV spectrum recorded simultaneously with the HPLC run, and by chromatography of its methyl derivative. The method using solid-phase extraction followed by a gradient reverse-phase HPLC procedure with an internal standard and sensitive UV detector, provides an efficient and sensitive technique for the separation and quantification of serum 13-cis- and all-trans-retinoic acid. Serum levels of 13-cis- and all-trans-retinoic acid in 26 fasting volunteers ranged from 1.0 to 2.2 ng/ml (mean +/- SEM = 1.4 +/- 0.3 ng/ml) and from 1.1 to 1.9 ng/ml (mean +/- SEM = 1.4 +/- 0.2 ng/ml), respectively. The levels determined by a liquid-liquid double-phase extraction method were 90% higher in both 13-cis- and all- trans-retinoic acid than those from a solid-phase extraction. Human small intestine can isomerize all-trans-retinoic acid. 13-cis-Retinoic acid is the predominant cis isomer after incubation of intestinal mucosa homogenates with all-trans-retinoic acid. Moreover, the concentration of retinoic acid in serum is related to diet in that the level of total retinoic acid was 36% higher (n = 10) 2 h after a nonstandard breakfast than in fasting subjects.