The vitamin A status in 11 generally healthy surgical patients was estimated by measuring the dilution of a 45-mg oral dose of tetradeuterated retinyl acetate (99% pure). After purification of retinol by high-performance liquid chromatography, the ratio of 2H4-retinol:1H-retinol in plasma was measured by gas chromatography-mass spectrometry. On the basis of the observed ratios of [2H4]retinol:[1H]retinol over 19-47 d, the total body reserves and liver concentrations of vitamin A were calculated. Liver biopsy samples taken at surgery were directly analyzed for vitamin A. The correlation coefficient between calculated and measured liver vitamin A concentrations for 10 of the subjects was 0.88, and the Spearman's rank correlation coefficient was 0.95 (p less than 0.002). Thus, total body reserves of vitamin A in humans can be estimated validly in the marginal and satisfactory ranges by a benign, relatively noninvasive procedure.

Carotenoids provide pigmentation to avian species, and also have immunomodulatory potential, although experimental results are often inconsistent. Therefore, dietary carotenoid deposition into immune tissue of growing chicks was examined in relation to their maternal carotenoid status (i.e., yolk carotenoid level). Single-comb white leghorn chicks were hatched from carotenoid-replete (C+) or carotenoid-deplete (C-) eggs. For 4 wk posthatch, chicks were fed diets whose carotenoid level ranged from 0 to 38 mg total carotenoid/kg. Carotenoid additions consisted of lutein + canthaxanthin at a ratio of 4:1. After 4 wk, the carotenoid concentration of thymus, bursa, liver, plasma and shank epithelium was measured by HPLC. Egg yolk-derived carotenoids were detectable in chicks fed 0 dietary carotenoids for 4 wk. Chicks hatched from C+ eggs had significantly greater tissue lutein, zeaxanthin and/or canthaxanthin for all tissues (P < 0.05), compared to chicks hatched from C- eggs. Only bursa carotenoids were not dependent on chick diet (P = 0.24); for all other tissues, C+ chicks incorporated dietary carotenoids in a dose-dependent manner (P < 0.01), whereas C- chicks never achieved the same level of carotenoid incorporation. This study demonstrated the importance of maternal carotenoid status on incorporation of yolk- and diet-derived tissue carotenoids in an avian model, and may explain some variability in carotenoid-based research, given that maternal carotenoid status is rarely controlled.

We present the application of a nonparametric method to perform functional principal components analysis for functional curve data that consist of measurements of a random trajectory for a sample of subjects. This design typically consists of an irregular grid of time points on which repeated measurements are taken for a number of subjects. We introduce shrinkage estimates for the functional principal component scores that serve as the random effects in the model. Scatterplot smoothing methods are used to estimate the mean function and covariance surface of this model. We propose improved estimation in the neighborhood of and at the diagonal of the covariance surface, where the measurement errors are reflected. The presence of additive measurement errors motivates shrinkage estimates for the functional principal components scores. Shrinkage estimates are developed through best linear prediction and in a generalized version, aiming at minimizing one-curve-leave-out prediction error. The estimation of individual trajectories combines data obtained from that individual as well as all other individuals. We apply our methods to new data regarding the analysis of the level of 14 C -folate in plasma as a function of time since dosing healthy adults with a small tracer dose of 14 C -folic acid. A time transformation was incorporated to handle design irregularity concerning the time points on which the measurements were taken.The proposed methodology incorporating shrinkage and data-adaptive features is seen to be well suited for describing population kinetics of 14 C -folate specific activity and random effects, and can also be applied to other functional data analysis problems.

Biological and biomedical applications of accelerator mass spectrometry (AMS) use isotope ratio mass spectrometry to quantify minute amounts of long-lived radioisotopes such as 14C. AMS target preparation involves first the oxidation of carbon (in sample of interest) to CO2 and second the reduction of CO2 to filamentous, fluffy, fuzzy, or firm graphite-like substances that coat a −400-mesh spherical iron powder (−400MSIP) catalyst. Until now, the quality of AMS targets has been variable; consequently, they often failed to produce robust ion currents that are required for reliable, accurate, precise, and high-throughput AMS for biological/biomedical applications. Therefore, we described our optimized method for reduction of CO2 to high-quality uniform AMS targets whose morphology we visualized using scanning electron microscope pictures. Key features of our optimized method were to reduce CO2 (from a sample of interest that provided 1 mg of C) using 100 ± 1.3 mg of Zn dust, 5 ± 0.4 mg of −400MSIP, and a reduction temperature of 500 °C for 3 h. The thermodynamics of our optimized method were more favorable for production of graphite-coated iron powders (GCIP) than those of previous methods. All AMS targets from our optimized method were of 100% GCIP, the graphitization yield exceeded 90%, and δ13C was −17.9 ± 0.3‰. The GCIP reliably produced strong 12C− currents and accurate and precise Fm values. The observed Fm value for oxalic acid II NIST SRM deviated from its accepted Fm value of 1.3407 by only 0.0003 ± 0.0027 (mean ± SE, n = 32), limit of detection of 14C was 0.04 amol, and limit of quantification was 0.07 amol, and a skilled analyst can prepare as many as 270 AMS targets per day. More information on the physical (hardness/color), morphological (SEMs), and structural (FT-IR, Raman, XRD spectra) characteristics of our AMS targets that determine accurate, precise, and high-hroughput AMS measurement are in the companion paper.

Lutein is an oxygenated carotenoid (xanthophyll) found in dark green leafy vegetables. High intakes of lutein may lower the risk of age-related macular degeneration. Current understanding of human lutein metabolism as it might occur in vivo is incomplete. Therefore, we conducted a feasibility study where we dosed a normal adult woman with 14C-lutein (125 nmol, 36 nCi 14C), dissolved in olive oil (0.5 g/kg body weight) and mixed in a banana shake. Blood, urine, and feces collected before the dose was administered served to establish baseline values. Thereafter, blood was collected for 63 d following the dose, while feces and urine were collected for 2 wk post-dose. The 14C contents in plasma, urine, and feces were measured by accelerator MS. The 14C first appeared in plasma 1 h after dosing and reached its highest level, approximately2.08% of dose/L plasma, at 14 h post-dose. The plasma pattern of 14C did not include a chylomicrons/VLDL (intestinal) peak like that when the same subject received 14C-beta-carotene (a previous test), suggesting that lutein was handled differently from beta-carotene by plasma lipoproteins. Lutein had an elimination half-life (t1/2) of approximately10 d. Forty-five percent of the dose of 14C was eliminated in feces and 10% in urine in the first 2 d after dosing. Quantifying human lutein metabolism is a fertile area for future research.

Across subjects, folate absorption, bile folate flux, and body folate stores were larger than prior estimates. Marrow folate uptake and pteroylpolyglutamate synthesis, recycling, and catabolism are saturable processes. Visceral pteroylpolyglutamate was an immediate precursor of plasma p-aminobenzoylglutamate. The model is a working hypothesis with derived features that are explicitly model-dependent. It successfully quantitated folate metabolism, encouraging further rigorous testing.

Isotope tracer studies, particularly radiocarbon measurements, play a key role in biological, nutritional, and environmental research. Accelerator mass spectrometry (AMS) is now the most sensitive detection method for radiocarbon, but AMS is not widely used in kinetic studies of humans. Part of the reason is the expense, but costs would decrease if AMS were used more widely. One component in the cost is sample preparation for AMS. Biological and environmental samples are commonly reduced to graphite before they are analyzed by AMS. Improvements and mechanization of this multi-step procedure is slowed by a lack of organized educational materials for AMS sample preparation that would allow new investigators to work with the technique without a substantial outlay of time and effort. We present a detailed sample preparation protocol for graphitizing biological samples for AMS and include examples of nutrition studies that have used this procedure.

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