Retinyl Palmitate Concentrations Research Articles

Exposure to Polybrominated Diphenyl Ethers (PBDEs): Changes in Thyroid, Vitamin A, Glutathione Homeostasis, and Oxidative Stress in American Kestrels (Falco sparverius)

Polybrominated diphenyl ethers (PBDEs), a class of additive flame retardants, are temporally increasing in wildlife tissues and capable of disrupting normal endocrine function. We determined whether in ovo and post-hatch exposure of captive American kestrels (Falco sparverius) to environmentally relevant PBDEs alter thyroid, retinol, and oxidative stress measures. Control eggs were injected with safflower oil and subsequent nestlings fed the same vehicle; dosed eggs received PBDE congeners (BDE-47, -99, -100, -153), which mainly comprise the Penta-BDE commercial mixture, dissolved in safflower oil at concentrations (1500 ng/g total [Sigma] PBDEs) approximating those in Great Lakes gull eggs. Nestlings hatching from dosed eggs were orally exposed for 29 days to variable SigmaPBDE concentrations that are similar to levels reported in tissues of Great Lakes trout (100 ng/g). Treatment kestrels had lower plasma thyroxine (T(4)), plasma retinol, and hepatic retinol and retinyl palmitate concentrations, but unaltered triiodothyronine (T(3)) concentrations and thyroid glandular structure. BDE-47, -100, and -99 were negatively associated with plasma T(4), plasma retinol (BDE-100, -99) and hepatic retinol (BDE-47). Despite an antioxidant-rich diet, PBDE exposure induced hepatic oxidative stress, particularly in females, with an increased hepatic GSSG:GSH ratio, a marginal increase in lipid peroxidation, and increased oxidized glutathione. Positive associations were found between concentrations of BDE-183 and thiols and, in males, between BDE-99 and reduced GSH, but a negative association occurred between BDE-99 and TBARS. Subsequently, concentrations of PBDE congeners in wild birds may alter thyroid hormone and vitamin A concentrations, glutathione metabolism and oxidative stress.

Relationship of retinoid and carotenoid metabolism with caecotrophy in rabbits.

The relationship of retinoid and carotenoid metabolism with caecotrophy was studied in adult female New Zealand White rabbits kept in individual metabolic cages. Caecotrophy was prevented by the use of plastic collars. The dry matter, crude protein, fibre, fat and ash contents of hard and soft faecal samples were determined. The retinoid (retinol and retinyl palmitate) and carotenoid (canthaxanthin, beta-carotene, beta-cryptoxanthin, lutein and zeaxanthin) levels of the blood, liver, kidney, caecal content and faeces were also measured by high-performance liquid chromatography (HPLC). The prevention of caecotrophy resulted in a significant decrease of blood retinol (P < 0.001) and retinyl palmitate (P < 0.01) concentration but it did not cause any significant change in the retinol and retinyl palmitate contents of the liver and kidney. The caecal content (25.78 +/- 6.87 microg/g) and the soft faeces (34.52 +/- 10.48 microg/g) contained the retinoids in similar amounts. Various carotenoids were found in considerable amounts in different types of faeces, while in the tissues (blood, liver and kidney) these pigments did not occur in substantial amounts. Total carotenoid concentration was similar in the caecal content (11.23 microg/g) and in the caecotroph (13.85 microg/g). On the basis of the results it can be assumed that the retinoid content of rabbit feed could be lowered in the presence of adequate caecal function and caecotrophy.

Accumulation of retinol in the liver after prolonged hyporetinolemia in the vitamin A-sufficient rat

We assessed the effects of prolonged reduction of plasma retinol concentrations (hyporetinolemia) on the distribution of tissue vitamin A (VA) and of its active compounds using a model of continuous recombinant human interleukin-6 (rhIL-6) infusion via osmotic minipumps in VA-sufficient male rats. Plasma retinol and retinol-binding protein (RBP) concentrations remained decreased and lower in rhIL-6-treated rats compared with controls from 7.5 h throughout 7 days of infusion (P < 0.001). This reduction was accompanied by a 68% increase in hepatic retinol concentration by 7 days (P < 0.05). Hepatic and renal retinyl palmitate and retinoic acid concentrations did not change, and renal megalin content remained unchanged; hepatic RBP concentrations were 41% lower in rhIL-6-treated rats compared with controls (P < 0.05). These results indicate that instead of being lost, retinol accumulated in the liver during inflammation and that hyporetinolemia was attributable to a decrease in the availability of hepatic RBP. A plausible consequence of the effect of rhIL-6-induced hyporetinolemia is that by 7 days tissues that are dependent on plasma retinol may become deprived of VA. These results have important implications in understanding the mechanism by which measles infection induces hyporetinolemia and VA deficiency of extrahepatic tissues.

205 DEVELOPMENT OF OOCYTES FROM COWS TREATED WITH RETINOL IS COMPROMISED PRIOR TO IMPLANTATION

In the cow, sheep, and gilt, a retinoid support enhances the oocyte development (reviewed by Hidalgo C et al. 2003 Reproduction 125, 409–416). The shortest effective time interval for these animals injected with retinol (ROH) is four days, but the reasons for this have not been described. Furthermore, the development after transfer of embryos derived from oocytes of donors treated with ROH is not known. The objective of the present work was to elucidate the two above questions. We analyzed the ROH and retinyl palmitate (REP) contents in plasma and follicular fluid (FF). The estrus cycles of heifers (n = 12) were synchronized with progestagen and PGF2α. Blood samples were taken at progestagen removal (Day 0). Animals were injected with 1 × 106 ROH units (n = 6) or corn oil (vehicle; n = 6) on Days 0, 1, and 4. Contents of follicles between 3 and 10 mm were aspirated on Day 4 by an ultrasound-guided procedure. All samples were analyzed for ROH and REP by HPLC. To produce embryos for transfer, follicles were aspirated from donors treated four times with 1 × 106 ROH units/week or vehicle, starting four days before the first aspiration. Both groups of cumulus-oocyte complexes (COCs) were matured in vitro with or without 5 nM 9-cis-retinoic acid (RA). The presence of RA prolonged the exposure to retinoids (cows treated with ROH) or acted as a positive control (heifers with vehicle). Oocytes were fertilized and cultured in mSOF + 5% FCS. Embryo transfer (ET) to recipients was performed with fresh (one) or vitrified (two) good-morphology Day 7 embryos, and pregnancy monitored on Days 21, 35, and 60. Data analysis was by GLM (ROH and REP concentrations) or CATMOD (pregnancy monitoring), and Duncan's test (a,bP &lt; 0.05; v,x,y,zP &lt; 0.02). Average FF volume recovered were 351 ± 127 μL (controls; range 100–700) and 393 ± 127 μL (ROH; range 80–950). Concentrations of REP were unaffected by timing, follicle or blood, and ROH treatment (data not shown). Concentrations of ROH (μg/dL) for vehicle and ROH-treated cows were (LSM ± SE) 42.0 ± 1.8vx and 42.0 ± 2.4vx (plasma-Day 0, before ROH injection), 37.3 ± 1.8v and 47.64 ± 2.4x (plasma-Day 1), 42.6 ± 1.9vx and 45.5 ± 2.3vx (plasma-Day 4), and 6.1 ± 3.0y and 16.8 ± 2.6z (FF-day 4), respectively. Cumulatively, embryos from donors receiving ROH did not exhibit pregnancy on Day 21 (0/15x; confirmed on Day 35), which differed from vehicle donors on Day 21 (44%, 8/18y) and Day 35 (33%, 6/18y), and tended to differ (P = 0.07) on Day 60 (22%, 4/18). Pregnancy rates were independent of fresh or vitrified embryos within ETs. Injected ROH elevated blood concentrations of ROH, although values became normalized on Day 4. However, a coasting period of 4 days for ROH administration seems to be justified by increased intrafollicular levels of ROH. Development into blastocysts is disrupted before implantation, showing that ROH directly affects the oocyte during its intrafollicular growth. This work was supported by MCYT-AGL-2002-0117.

Antagonism of Hypervitaminosis A-Induced Anterior Neural Tube Closure Defects with a Methyl-Donor Deficiency in Murine Whole-Embryo Culture

The interaction of a dietary excess of vitamin A (retinoid) and deficiency of methyl-donor compounds was examined in murine early-organogenesis embryonic development. Female mice were fed one of six diets from the time of vaginal plug detection until gestational d 8.0, when embryos were removed and grown in whole embryo culture for 46 h, using serum from rats fed the same diet for 36 d as the culture medium. The six diets were either methyl-donor deficient (designated -FCM: devoid of folic acid, choline and supplemental L-methionine, but having methionine as a component of the protein portion of the diet) or methyl-donor sufficient (designated +FCM: containing folic acid, choline and L-methionine supplementation), in combination with one of three concentrations of retinyl palmitate (0.016, 0.416 or 4.016 g/kg diet). The high dose of retinyl palmitate induced a failure of anterior neuropore closure and hypoplasia of the visceral arches, both of which were significantly ameliorated by simultaneous administration of the methyl-donor-deficient diet. The primary acidic retinoid detected in the rat serum was 9,13-di-cis-retinoic acid, although we hypothesize that teratogenic retinoids were formed by embryonic biotransformation of the retinyl esters to toxic metabolites. Biochemical measurements of metabolites in relevant pathways were performed. We propose that the amelioration of these malformations may be used to determine biochemical pathways critical for retinoid teratogenesis.

Uptake of vitamin A in buccal mucosal cells after topical application of retinyl palmitate: a randomised, placebo-controlled and double-blind trial

Retinoids have been reported to produce regressions in metaplastic changes of the mucosal epithelium. In order to define the role of these micronutrients in the prevention of squamous metaplasia of the oral cavity, it is necessary to measure their uptake in target tissues such as the buccal mucosal epithelium. We demonstrated in a trial that retinyl palmitate applied topically via a toothpaste is taken up by buccal mucosal cells in young healthy volunteers. In the randomised, parallel-designed, placebo-controlled and double-blind trial, forty volunteers divided in two groups cleaned their teeth either with a placebo toothpaste or a retinyl palmitate-containing toothpaste (1 mg/g) for 56 d. Buccal mucosal cells samples were taken from the healthy volunteers during the retinyl palmitate application and the following wash-out phase to determine the concentration of retinyl palmitate and retinol by HPLC. Supplementary blood samples were taken from the volunteers on days 0 and 56 to investigate changes in plasma retinyl palmitate and retinol concentrations. Results from only thirty participants (sixteen placebo and fourteen treated subjects) were used in the statistical evaluation as the remaining sample results were spoiled by a technical defect during the HPLC analysis. A significant (P<0.05) uptake of retinyl palmitate in buccal mucosal cells after 7 d and a significant (P<0.05) increase of plasma retinol after 17 d was demonstrated in our present study. The uptake of retinyl palmitate and the following hydrolysis to retinol led to an enrichment of vitamin A in buccal mucosal cells.

Determination of Retinyl Palmitate (Vitamin A) in Fortified Fluid Milk by Liquid Chromatography: Collaborative Study

A liquid chromatographic (LC) method was developed for fast and simple measurement of retinyl palmitate (vitamin A) in fortified milk. Retinyl acetate internal standard was added to a test portion of milk followed by extraction into hexane. The hexane extract was analyzed by LC using a normal-phase silica gel column equilibrated with mobile phase (conditioned hexane-isopropanol, 99.85 + 0.15, v/v) about 1 h before injections. The retinyl palmitate concentration was calculated by using a relative response factor determined with calibration standards. In the collaborative study, 11 laboratories analyzed 13 pairs of fluid milk materials in blind duplicate. Twelve of the materials were composed of skim milk (< 0.5% fat), 1% fat milk, 2% fat milk, and 1% fat chocolate milk. Each material was fortified at 3 concentrations of retinyl palmitate of approximately 581 microg/L (1000 IU/qt), 1163 microg/L (2000 IU/qt), and 2236 microg/L (4000 IU/qt). The 13th material, unfortified skim milk, served as a matrix blank. Repeatability standard deviations (RSDr) without outliers ranged from 1.5 to 5.7% and reproducibility standard deviations (RSDR) without outliers ranged from 5.0 to 22.7%. cis-Isomers co-eluted with the predominant trans-retinyl palmitate isomer and were included in the results reported by all the collaborative laboratories. Endogenous long-chain esters from milk fat were also measured with the retinyl palmitate additive. The Study Director recommends that this method for determination of retinyl palmitate in fluid milk by LC be adopted First Action.

Prospective study of antioxidant micronutrients in the blood and the risk of developing prostate cancer.

Antioxidant micronutrients may have chemopreventive effects. The authors examined the associations between prediagnostic blood levels of micronutrients and prostate cancer risk in two nested case-control studies of 9,804 and 10,456 male residents of Washington County, Maryland, who donated blood in 1974 (CLUE I) and 1989 (CLUE II), respectively. Until 1996, 182 men for whom adequate serum remained for assays in the CLUE I cohort and 142 men in the CLUE II cohort developed prostate cancer. Each case was matched with two controls by age, gender, race, and date of blood donation. In both cohorts, cases and controls had similar concentrations of alpha-carotene, beta-carotene, total carotene, beta-cryptoxanthin, lutein, lycopene, retinol, and ascorbic acid; serum alpha-tocopherol was weakly associated with prostate cancer risk. Higher retinyl palmitate concentrations were associated with a lower risk in CLUE I but not CLUE II. In CLUE I, cases had lower concentrations of gamma-tocopherol than did controls (p = 0.02), but no dose-response trend was observed. A strong inverse association between gamma-tocopherol and prostate cancer risk was observed in CLUE II. Findings do not replicate previous reports of a protective association between lycopene and prostate cancer, but they suggest potential chemopreventive effects of gamma-tocopherol on prostate cancer.

Biological effects of marine PCB contamination on black guillemot nestlings at Saglek, Labrador: liver biomarkers.

Black guillemots (Cepphus grylle) in Saglek Bay, Labrador have elevated polychlorinated biphenyl (PCB) concentrations due to marine sediment contamination around a former military site. We measured liver biomarkers and sigmaPCB concentrations in 31 nestlings from three PCB-exposure groups: Reference group (range: 15-46 ng/g liver, wet wt.), moderately exposed Islands group (24-150 ng/g), and highly exposed Beach group (170-6200 ng/g). Biomarker responses were dose-dependent and in some cases sex-dependent. Livers of female Beach nestlings were enlarged 36% relative to Reference females. In both sexes, Beach nestlings had liver ethoxyresorufin-O-deethylase (EROD) activities elevated 79% and liver retinol concentrations reduced 47%. Retinyl palmitate concentrations were reduced 50% but only among female nestlings. Island nestlings also exhibited EROD induction (57%) and reductions in retinol and retinyl palmitate concentrations (28 and 58%, respectively). Liver lipid content increased with sigmaPCBs in both sexes, and correlated with liver mass in males. Malic enzyme activity and porphyrin concentrations showed little association with sigmaPCBs. Although similar associations between liver biomarkers and organochlorine exposure in fish-eating birds are well documented, typically exposures involve multiple contaminants and there is uncertainty about specific PCB effects. Our findings indicate that liver biomarkers respond to relatively low PCB exposures (approximately 73 ng/g liver) in guillemots.

Health of Tree Swallow (Tachycineta Bicolor) Nestlings Exposed to Chlorinated Hydrocarbons in the St. Lawrence River Basin. Part I. Renal and Hepatic Vitamin A Concentrations

Sixteen-day-old tree swallows (Tachycineta bicolor), near fledging, were collected in 1999 and 2000 from nine sites representing a gradient of dioxin concentrations, within the vicinity of the St. Lawrence River in Canada and the United States, to determine if organochlorine contaminants correlated with vitamin A levels measured as retinol and retinyl palmitate. Mean concentrations of hepatic retinol ranged from 3mg /kg to 13 mg /kg, and from 0.35 mg /kg to 1.5mg /kg for renal retinol. Mean concentrations of hepatic retinyl palmitate ranged from 18 mg /kg to 146mg /kg, and 1mg /kg to 6mg/kg for renal retinyl palmitate. In 1999, molar ratio of renal retinol:retinyl palmitate was significantly and positively correlated with total polychlorinated dibenzodioxin (PCDD) concentration. Among sites, total PCDDs ranged from 5.4 ng /kg wet weight to 79.5 ng /kg wet weight in tree swallows. These results suggest that current levels of organochlorine contaminants in the St. Lawrence River and surrounding tributaries may be interacting with the vitamin A pathway. Lower circulating levels and higher tissue concentrations of retinoids may result in compromised immune function and reduced reproductive success in adult birds.

Retinoids, LRAT and REH activities in eggs of Japanese quail following maternal and in ovo exposures to 3,3',4,4'-tetrachlorobiphenyl.

Environmental monitoring for the effects of organochlorine contaminants has identified changes in egg yolk retinoid concentrations, however underlying mechanisms are unknown. Breeding female Japanese quail, Coturnix coturnix japonica, received 3 bimonthly injections of a low dose (5 microg/g) of the coplanar polychlorinated biphenyl (PCB) 3,3',4,4'-tetrachlorobiphenyl. No overt maternal toxicity or embryotoxicity was detected. For the exposed females, oocytes (yolk deposition nearly completed) and freshly laid eggs contained higher concentrations of retinol. For PCB-exposed female quail, freshly laid eggs and fertile eggs incubated for 6 days contained higher concentrations of retinyl palmitate. These results suggest that the PCB congener caused greater deposition of retinoids in yolks during late oogenesis or greater net esterification in the yolk-sac membrane. Retinyl ester hydrolase (REH) activity, assayed in the microsomal fraction of the yolk-sac membrane, was significantly inhibited in the PCB-exposed group. Therefore, the higher retinyl palmitate concentrations may be explained by less retinyl palmitate conversion to retinol. In a second experiment, fertile quail eggs were injected with 2, 10 or 20 microg of the PCB congener and incubated for 6 days. In contrast to the maternal exposure, yolk retinol decreased and retinyl ester hydrolase increased in the PCB-injected eggs. The difference in the results of adult versus egg injections may be related to several factors including adult transformation of the PCB to toxic metabolites and differences in post-transcriptional regulation of REH expression.

Effect of citric pectin on beta-carotene bioavailability in rats.

The effect of citric pectin on the bioavailability of synthetic beta-carotene was studied. Thirty Wistar rats were used, ten animals were sacrificed at the beginning of the experiment and remaining animals were divided into two groups and received the following diets for 30 days: control group (CG)--24 micrograms beta-carotene/g diet + 0% citric pectin; experimental group (EG)--24 micrograms beta-carotene/g diet + 7% citric pectin. Plasma and liver beta-carotene, vitamin A, and retinyl palmitate concentrations were determined by high-performance liquid chromatography (HPLC). Plasma retinol concentration was 1.42 +/- 0.36 mumol/L for CG and 1.10 +/- 0.24 mumol/L for EG (p = 0.1), and plasma beta-carotene concentration was 0.20 +/- 2.51 mumol/L for CG and 0.07 +/- 0.04 mumol/L for EG (p = 0.01). Only traces of retinyl palmitate were detected in CG and none in EG. Retinol did not differ significantly between groups CG and EG, while a significantly higher beta-carotene concentration was observed for CG. Liver concentrations of retinol (CG: 4.90 +/- 2.51 micrograms/g; EG: 2.68 +/- 1.12 micrograms/g), beta-carotene (CG: 0.98 +/- 0.28 microgram/g; EG: 0.11 +/- 0.06 microgram/g), and retinyl palmitate (CG: 95.47 +/- 45.13 micrograms/g, EG: 37.01 +/- 17.20 micrograms/g) differed significantly between groups (p < 0.05), with a lower concentration being observed for EG. We conclude that 7% citric pectin in the rat diet decreases the bioavailability of synthetic beta-carotene, reducing the liver reserves of vitamin A and beta-carotene.

Antioxidant systems of the developing quail embryo are compromised by mycotoxin aurofusarin

The effects of aurofusarin in the quail diet on the antioxidant systems of the developing embryo are investigated. Thirty eight 45-day-old Japanese quails ( Coturnix japonica) were divided into two groups and were fed on a corn-soya diet or the same diet supplemented with aurofusarin at the level of 26.4 mg/kg feed in the form of Fusarium graminearum culture enriched with aurofusarin. Eggs obtained after 7 weeks of feeding were incubated. Samples of quail tissues were collected at day 17 of embryonic development and from day old hatchlings. Antioxidants and malondialdehyde were analysed by HPLC-based methods. Inclusion of aurofusarin in the maternal diet was associated with decreased concentrations of α- and γ-tocopherols, α- and γ-tocotrienols, retinol, lutein and zeaxanthin in egg yolk. The vitamin E (tocopherols and tocotrienols) concentration in the liver and yolk sac membrane (YSM) of the day 17 embryos and the hatchlings from aurofusarin-fed group was significantly decreased. Alpha-tocopherol concentration was also reduced in kidney, lung, heart, muscle and brain of day-old quails. In the liver of day-old quails, concentrations of lutein, zeaxanthin, retinol, retinyl linoleate, retinyl oleate, retinyl palmitate and retinyl stearate were also reduced. As a result of these diminished antioxidant concentrations, tissue susceptibility to lipid peroxidation was significantly increased. It is suggested that a compromised antioxidant system of the egg yolk and embryonic tissues could predispose quails to increased mortality at late stages of their embryonic development.

Effects of β-carotene and vitamin A on oval cell proliferation and connexin 43 expression during hepatic differentiation in the rat

The effects of β-carotene and vitamin A administrations were evaluated in an in vivo model of hepatic cell differentiation. For this purpose, male Wistar rats received β-carotene (70 mg/kg of body weight), vitamin A (10 mg/kg of body weight) or corn oil (control group), by gavage and at every other day during the entire experimental period. After 4 consecutive weeks of treatment, the animals were submitted to the AAF/PH model of hepatic cell differentiation (6 × 20 mg of AAF [2-acetylaminofluorene]/kg of body weight and partial hepatectomy) and killed on different days following the surgery (until day 16 after hepatectomy). Liver samples were collected for determination of β-carotene, retinol and retinyl palmitate concentrations, for histopathological (hematoxilin-eosin) examination, for immunohistochemical detection of glutathione S-transferase, as well as for the evaluation of connexin 43 (a structural protein of gap junctions of oval cells) expression by northern blot analysis. Compared to controls, the oval cell proliferation peaks (observed by histopathological examination and immunohistochemistry) and connexin 43 expression peaks, were postponed to later days after hepatectomy, in a similar way in β-carotene and vitamin A treated animals. Compared to the other experimental groups, the vitamin A treated group showed an increase in connexin 43 expression. It was concluded that β-carotene and vitamin A modulated oval cell proliferation and connexin 43 expression, delaying both events. These findings suggest that β-carotene and vitamin A can modulate the hepatic differentiation process in vivo.

Postprandial chylomicron clearance rate in late teenagers with diabetes mellitus type 1.

A delayed chylomicron (CM) clearance rate, a known risk factor for atherosclerosis, has been described in adults with diabetes type 1 (DM1). We determined the CM clearance rate in late teenagers with DM1, and the relationship between CM clearance rate and elevated plasma lipid concentrations in DM1 teenagers in poor metabolic control (as characterized by HbA(1c) percentage). Plasma lipids and CM clearance were determined in nine patients with DM1 (mean age +/- SD: 17.5 +/- 0.6 y) and four healthy controls (mean age +/- SD: 20.1 +/- 0.8 y), by measuring breath (13)CO(2), plasma triglyceride, retinyl palmitate, and (13)C-labeled oleic acid concentrations, after oral administration of a fat-rich meal together with vitamin A and (13)C-oleic acid. In patients with DM1, fasting triglyceride and cholesterol concentrations were positively correlated with HbA(1c) percentage (p < 0.05). Neither in DM1 patients, nor in controls, was an elevated triglyceride concentration (above 1.7 mmol/L) found. Yet, in 22% of DM1 patients, cholesterol concentration was above 5.2 mmol/L, but not in any of the controls. CM clearance rate in DM1 patients was similar to that in controls and did not significantly correlate with HbA(1c) percentage. Fasting lipid concentrations in DM1 patients were not significantly correlated with CM clearance rate. Present data indicate that elevated lipid concentrations in late teenagers with DM1 are not attributable to a delay in CM clearance rate. A delayed CM clearance rate at late teenager age is not a risk factor contributing to the increased risk for atherosclerosis in DM1.

Acute effect of dietary stanyl ester dose on post-absorptive alpha-tocopherol, beta-carotene, retinol and retinyl palmitate concentrations.

Stanyl esters dissolved in margarine inhibit cholesterol absorption, lower sterol absorption in general, and lower serum total cholesterol, LDL-cholesterol and plant sterol levels. To find out whether stanyl esters inhibit absorption of fat-soluble vitamins and beta-carotene in acute experiments, we performed two fat-tolerance tests fortified with vitamins (retinol 0.9-3.7 mg, alpha-tocopherol 70-581 mg), beta-carotene (25-150 mg) and squalene (0.5 g) with and without 1 g of stanyl ester added to the test meal in ten healthy men. The concentrations or areas under the curves (AUC) of cholesterol, triacylglycerols, squalene and alpha-tocopherol, beta-carotene and retinyl palmitate showed typical postprandial changes in serum, chylomicrons, VLDL and VLDL infranatant (intermediate-density lipoproteins, LDL and HDL) over 24 h after the test meal without stanyl esters, and they were not affected by the addition of stanyl esters. The post-absorptive serum campesterol concentration and campesterol : cholesterol were significantly lowered at 6-9 h by stanyl ester supplementation, reflecting reduced sterol absorption efficiency. Changes in vitamin and beta-carotene AUC did not correlate with the given doses. In conclusion, the present study shows that stanyl esters dissolved in margarine do not detectably interfere in a short-term study with the absorption of alpha-tocopherol, beta-carotene or retinol measured by a 24 h oral fat-load test.

Transcutaneous Absorption of Vitamin A in Newborn Rats

Very low birth weight neonates have low tissue concentrations of vitamin A, which may contribute to the development of lung disease. These infants, however, may not receive vitamin A supplementation for several days after birth. We determined if the relatively permeable skin of a newborn could be used to administer vitamin A. 25 control rat pups were killed and lungs and livers were collected. 20 μl (1,000 IU) of retinyl palmitate were applied to the skin surface of an additional 50 two-day-old pups. At 2.5 and 5 h after application, 25 pups were killed, and lungs and livers were collected. Concentrations of retinyl palmitate and retinol were significantly higher in the lungs of pups 5 h after administration of vitamin A compared with controls. There were no differences in concentrations of retinyl palmitate or retinol in livers. We conclude that transcutaneous administration may be an effective means of delivering vitamin A to the lungs of newborn rats.

Predictors of postprandial triacylglycerol response in children: the Columbia University Biomarkers Study

Predictors of postprandial lipemia have not been explored in children. Our objective was to determine whether the postprandial triacylglycerol response is associated with low HDL-cholesterol and high fasting triacylglycerol concentrations and family history of early-onset ischemic heart disease (IHD) in children. We administered a standardized fat load (52.5 g fat/m(2)) to 60 children (mean age: 14.0 y), 20 with and 40 without a family history of early-onset IHD, and to 29 mothers, all recruited from families enrolled in the Columbia University Biomarkers Study. Plasma lipid and retinyl palmitate concentrations were measured in the fasting state and 3, 6, and 8 h after the oral fat load. In children, postprandial lipemia, as indicated by the incremental area under the triacylglycerol response curve, was associated with elevated fasting triacylglycerol concentrations (>/=1.13 mmol/L; P: < 0.01), with low fasting HDL-cholesterol concentrations (</=0.91 mmol/L; P: < 0.01), and with the combination of low HDL-cholesterol and high triacylglycerol concentrations (P: < 0.05). Family history of IHD, baseline LDL-cholesterol concentration, and apolipoprotein E genotype were not associated with the postprandial triacylglycerol or retinyl palmitate response. The mothers had fasting triacylglycerol concentrations similar to those of their children but a more prolonged response with higher triacylglycerol concentrations at 6 and 8 h (P: < 0.01 and P: < 0.05, respectively). In children, a delayed postprandial triacylglycerol response to a fat load is associated with the combination of high fasting triacylglycerol and low HDL-cholesterol concentrations. Predictors of postprandial triacylglycerol concentrations may be similar in children and adults.

Postprandial Retinyl Palmitate and Squalene Metabolism Is Age Dependent

Most of our awake time is spent in a postprandial state. It has not been investigated in detail whether the post-prandial clearance of triglyceride-rich lipoproteins is age dependent. In addition, postabsorptive squalene metabolism has not been studied in relation to age. Accordingly, we investigated postprandial lipid metabolism in six young (22-25 years of age) and eight old (78-79 years of age) healthy men by use of an oral fat load containing 345,000 IU of vitamin A and 0.5 g of squalene as postprandial markers. Postprandial samples were drawn after 3, 4, 6, 9, 12 and 24 hours after the fat load. The retinyl palmitate area under the incremental curve of the old subjects was higher in plasma than that of the young subjects (p < .01). The pattern of postprandial very low density lipoprotein squalene responses differed significantly in the old compared with the young subjects (p < .01), but the areas under the incremental curve did not differ. Postprandial retinyl palmitate and squalene concentrations correlated significantly at 3-12 hours (p < .01). These data suggest that postprandial lipoprotein metabolism measured by retinyl palmitate and squalene is retarded with increasing age.

Hepatic hyper-vitaminosis A

We report the case of a 32-year-old man with portal hypertension without cirrhosis due to chronic vitamin A intoxication. Portal hypertension revealed by oesophageal varice rupture progressively worsened and ascites occurred 5 years after the patient stopped vitamin A intake. Initially, serum retinyl palmitate concentration was increased whereas serum retinol concentration was normal. There was no hepatic fibrosis on light microscopic examination of liver biopsy specimens. Five years after the patient stopped excessive vitamin A intake, serum retinol and retinol-binding protein concentrations were below the normal range even though there was an increased hepatic retinyl ester content. This was attributed to the late development of peri-sinusoidal fibrosis. This case mainly shows the importance of retinyl ester level determination: serum retinyl palmitate should be measured immediately after intoxication and hepatic retinyl esters should be measured initially and particularly later. Indeed, later serum and hepatic retinol levels in chronic hyper-vitaminosis A may be normal and lead to under-estimation of liver vitamin A overload.