RESUMEN
OBJECTIVE: To examine total energy expenditure (TEE) in relation to occupation and reported leisure time activities in free-living Chinese adults, and to determine whether measured TEE values differ from current international dietary energy recommendations. SETTING AND SUBJECTS: Seventy three weight-maintaining adults aged 35-49 y, leading unrestricted lives in urban Beijing, with a wide variety of occupations. DESIGN AND METHODS: A cross-sectional study in which TEE was determined by doubly labeled water, body composition by deuterium oxide ((2)H(2)O) dilution, resting energy expenditure (pREE) by prediction equations, and occupational and leisure time activities by questionnaire. RESULTS: For men and women respectively, TEE averaged 12.10+/-0.32 and 9.53+/-0.23 MJ/day (P<0.001), and physical activity level (PAL=TEE/pREE) was 1.77+/-0.04 and 1.66+/-0.02 (P<0.05). Fat-free mass (FFM) was the single best predictor of TEE (adjusted r(2)=0.71, P<0.001). Occupational category (light, moderate and heavy) further predicted TEE, independent of FFM (adjusted multiple r(2)=0.82, P<0.001). Both TEE adjusted for weight and PAL increased with occupational category. Measured TEE was slightly but significantly higher than the 1985 FAO/WHO/UNU estimates for women with light occupations, but did not differ from estimates for men with light occupations, or for adults with moderate or heavy occupations. CONCLUSION: Level of occupational activity, but not duration or type of leisure activity, significantly predicted TEE in free-living urban Chinese adults. Current energy requirement recommendations slightly underestimated the energy needs of women with light occupations but were accurate for men and women with moderate and heavy occupations. SPONSORSHIP: NIH grants DK53404 and F32-DK09747.
Asunto(s)
Composición Corporal/fisiología , Metabolismo Energético/fisiología , Ocupaciones/clasificación , Adulto , Agua Corporal/metabolismo , China , Estudios Transversales , Deuterio , Femenino , Humanos , Técnicas de Dilución del Indicador , Actividades Recreativas , Masculino , Persona de Mediana Edad , Necesidades Nutricionales , Encuestas y Cuestionarios , Población Urbana , AguaRESUMEN
Tangier disease (TD), caused by mutations in the gene encoding ATP-binding cassette 1 (ABCA1), is a rare genetic disorder in which homozygotes have a marked deficiency of high density lipoproteins (HDL), as well as concentrations of low density lipoproteins (LDL) that are typically 40% of normal. Although it is well known that the reduced levels of HDL in TD are due to hypercatabolism, the mechanism responsible for the low LDL levels has not been defined. Recently, it has been reported that intestinal cholesterol absorption is altered in ABCA1 deficient mice, suggesting that aberrant cholesterol metabolism may contribute to the LDL reductions in TD. In order to explore this possibility, as well as to define the role that ABCA1 plays in the metabolism of apolipoprotein (apoB)-containing lipoproteins, we determined the kinetics of apoB-100 within lipoproteins, and cholesterol absorption, biosynthesis, and turnover, in a compound heterozygote for TD. The levels of HDL cholesterol, LDL cholesterol and LDL apoB-100 in this subject were 7, 27 and 69% of normal, respectively, the latter of which was due to a two-fold increase in LDL catabolism (0.54 vs. 0.26+/-0.07 poolsday(-1)) relative to controls (n=11). NMR analysis of plasma lipoproteins revealed that 91% of the LDL cholesterol in the TD subject was contained within small, dense LDL, as compared with only 20% for controls (n=70). Cholesterol absorption was 97% of the value for controls (n=15) in the TD subject, at 45%, with cholesterol synthesis and turnover increased modestly by 17 and 25%, respectively. Our data are consistent with the concept that the reductions of LDL observed in TD are due to enhanced catabolism, secondary to changes in LDL composition and size, with neither cholesterol absorption nor metabolism significantly influenced by mutations in ABCA1.
Asunto(s)
Apolipoproteínas B/metabolismo , Colesterol/metabolismo , Enfermedad de Tangier/metabolismo , Transportador 1 de Casete de Unión a ATP , Transportadoras de Casetes de Unión a ATP/genética , Anciano , Apolipoproteína B-100 , LDL-Colesterol/metabolismo , Heterocigoto , Homocigoto , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Persona de Mediana Edad , Enfermedad de Tangier/genéticaRESUMEN
BACKGROUND: Apolipoprotein (apo) A-IV is a major component of triacylglycerol-rich lipoprotein (TRL) apolipoproteins. OBJECTIVE: We investigated the effects of dietary saturated fat and cholesterol restriction on the metabolism of TRL and plasma apo A-IV. DESIGN: We assessed TRL and plasma apo A-IV kinetics in 16 and 4 subjects, respectively, consuming an average US (baseline) diet for 6 wk and a National Cholesterol Education Program Step II diet for 24 wk, respectively. At the end of each diet period, all subjects received a primed, constant infusion of deuterated leucine for 15 h with hourly feeding. Ratios of stable-isotope tracer to tracee were measured by using gas chromatography-mass spectrometry, and kinetic data were modeled by using SAAM II. RESULTS: Mean apo A-IV concentrations during the isotope infusion period were 6.9 +/- 2.6 mg/L in TRL and 2.2 +/- 3.2 mg/L in plasma with the baseline diet; these values were 37.7% (P < 0.001) and 19.4% (P < 0.01) lower with the Step II diet. Similar changes were observed in the fasting state between the 2 diets. The mean apo A-IV secretion rate decreased significantly from baseline by 59.6% in TRLs and by 40.2% in plasma. Significant correlations were observed between TRL apo A-IV concentrations and the secretion rate (r = 0.94, P < 0.001) and between TRL apo A-IV pool size and TRL-cholesterol concentrations (r = 0.48, P < 0.01). CONCLUSIONS: Our data indicate that the National Cholesterol Education Program Step II diet significantly decreases TRL and plasma apo A-IV concentrations compared with the average US diet and that this decrease is due to a decreased secretion rate.
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Apolipoproteínas A/metabolismo , Colesterol en la Dieta/administración & dosificación , Dieta con Restricción de Grasas , Grasas de la Dieta/administración & dosificación , Triglicéridos/metabolismo , Adulto , Antioxidantes , Apolipoproteínas A/sangre , Colesterol en la Dieta/metabolismo , Grasas de la Dieta/metabolismo , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Marcaje Isotópico , Cinética , Masculino , Persona de Mediana EdadRESUMEN
In order to investigate the metabolism of apo A-IV within TRL and plasma, we assessed TRL and plasma apo A-IV kinetics in 19 and 4 subjects, respectively, consuming an average US diet for a 6-week period. At the end of this diet study, each subject received a primed-constant infusion of deuterated leucine over a 15 h time period with hourly feeding, and blood samples were drawn at 10 time points. TRL was separated by ultracentrifugation. Apo A-IV was isolated by immunoprecipitation and/or SDS-PAGE. Apo A-IV concentrations were determined by immunoelectrophoresis. Stable isotope tracer/tracee ratios were measured by gas chromatography/mass spectrometry, and the data were analyzed by multicompartmental modeling. The mean concentrations of plasma and TRL apo A-IV during the isotope infusion period were 21.0+/-3.2 and 0.66+/-0.25 mg/dl, respectively, and these values were 11.5 and 30.5% higher than those of fasting samples. The mean TRL and plasma apo A-IV residence times (RT) were 1.97+/-0.57 and 2.71+/-0.65 days, and transport rates (TR) were 0.17+/-0.19 and 3.90+/-1.24 mg/kg per day, respectively. There were significant correlations between TRL apo A-IV concentrations and TR (r(2)=0.79, P<0.001), and between TRL apo A-IV pool size and TRL cholesterol levels (r(2)=0.29, P=0.02). Our data indicated that; (1) TRL apo A-IV has a RT of 1.97 days which is similar to that earlier reported for HDL apo A-IV; (2) Apo A-IV recirculates between TRL and other slowly turning over pools; (3) the primary determinant of TRL apo A-IV levels is its TR; and (4) there is no correlation between TRL apo A-IV and apo B48 fractional catabolism in TRL.
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Apolipoproteínas A/metabolismo , Lipoproteínas/metabolismo , Adulto , Anciano , Apolipoproteínas A/sangre , Enfermedad de la Arteria Coronaria/etiología , Enfermedad de la Arteria Coronaria/metabolismo , Enfermedad de la Arteria Coronaria/prevención & control , Femenino , Humanos , Masculino , Persona de Mediana Edad , Radioinmunoensayo , Valores de Referencia , Medición de Riesgo , Sensibilidad y Especificidad , Triglicéridos/metabolismoRESUMEN
Apolipoprotein (apo) E is a multifunctional protein that can act as a ligand for lipoprotein receptors. The receptor-mediated clearance of the triglyceride-rich lipoproteins (TRL) chylomicrons and VLDL from plasma is, in part, dependent on apo E. Enrichment of VLDL with apo E is thought to enhance receptor-mediated clearance of VLDL resulting in a low rate of conversion of VLDL to LDL. However, the kinetic mechanism controlling the concentration of apo E in VLDL is not known. We conducted kinetic studies on apo E in the TRL fraction (d < 1.006 g/ml) and apo B-100 in the TRL and LDL (d = 1.019-1.063 g/ml) fractions to assess the kinetic determinants of apo E concentration in TRL and to determine the effects that TRL apo E production and clearance rates have on the production rate of LDL apo B-100. Nineteen males between the ages of 24 and 73 underwent a primed-constant infusion with deuterated leucine tracer in the constantly-fed state. Apo B-100 from TRL and LDL, and apo E from TRL were isolated and their tracer incorporation measured by gas chromatography/mass spectrometry. The residence time and production rates of each protein were determined from the kinetic data using the SAAM II modeling program. The residence time and production rate of TRL apo E were about one-half that of TRL apo B-100 (1.8 +/- 1.0 vs. 2.9 +/- 2.1 h and 14.5 +/- 11.0 vs. 27.6 +/- 17.3 mg/kg per day, respectively). The production rate of TRL apo E was weakly correlated with the production rate of TRL apo B-100 (r = 0.424, P = 0.07). Multiple regression analysis showed that the residence time of TRL apo B-100 and the relative TRL apo E production rate (relative to the TRL apo B100 production rate) were negatively associated with LDL apo B-100 production rate, accounting for 68% of its variability. We conclude that (1) the concentration of apo E in TRL is highly correlated to its production rate, suggesting that production rate regulates the TRL apo E concentration, and (2) individuals with a relatively short TRL apo B-100 residence time and those producing TRL with a relatively low apo E content have the highest LDL apo B-100 production rates.
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Apolipoproteínas B/sangre , Apolipoproteínas E/sangre , Lipoproteínas LDL/sangre , Lipoproteínas VLDL/sangre , Triglicéridos/sangre , Adulto , Anciano , Apolipoproteína B-100 , Colesterol/sangre , Deuterio/farmacocinética , Ingestión de Alimentos , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Humanos , Cinética , Leucina/farmacocinética , Lipoproteínas LDL/biosíntesis , Masculino , Persona de Mediana EdadRESUMEN
Antibacterial and inflammatory responses of neutrophils and macrophages produce hypochlorite as a major oxidant. Numerous side chains of amino acids found in extracellular proteins can be modified by hypochlorite, including His, Arg, Tyr, Lys, Trp, and Met. We studied the relative reactivity of each of these amino acid residues in short N-blocked peptides, where other residues in the peptide were highly resistant to hypochlorite attack. Hypochlorite treatment led to modified peptides in each case, which were detected by changes in retention on reversed-phase HPLC. A distinct single product, consuming two equivalents of hypochlorite per equivalent of peptide, was obtained from the Lys-containing peptides. UV spectroscopy, nuclear magnetic resonance (NMR), and electrospray/mass spectroscopy identified this product as the dichloramine at the epsilon-amino group of the Lys side chain. The dichloramine at Lys did not decompose to form a detectable amount of carbonyl reactive with dinitrophenylhydrazine. The dichloramine at Lys did however quantitatively revert back to Lys during HCl digestion of the tetrapeptide for amino acid analysis, with simultaneous modification of the adjacent Phe residue. The formation of the dichloramine at Lys was not blocked by peptides or acetylated amino acids that contained Tyr, His, or Arg. In contrast, the presence of equimolar Met-containing peptide, or N-Acetyl-Trp, both inhibited the formation of the dichloramine at Lys. Thus, Met and Trp side chains of proteins might be able to protect Lys from chloramine formation under some circumstances, but this interpretation must consider that Met and Trp are typically found in relatively inaccessible hydrophobic sites, whereas lysine is typically exposed on the protein surface. The hierarchy of amino acid reactivities examined here will aid in the prediction of residues in biological samples most likely to be modified by hypochlorite.
Asunto(s)
Aminoácidos/química , Ácido Hipocloroso , Lisina/química , Oligopéptidos/química , Acetilación , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Oxidación-Reducción , Espectrofotometría UltravioletaRESUMEN
BACKGROUND: Quantitative information on conversion of beta-carotene to vitamin A in humans is limited. AIM OF THE STUDY: Our laboratory has developed a stable isotope method for studying the conversion of beta-carotene (beta-C) to vitamin A. METHODS: Two dosage levels (a pharmacological dose, 126.0 mg beta-C-d8, and a physiological dose, 6.0 mg beta-C-d8) were used 2.5 y apart in an adult female volunteer to study dose effects on the conversion of beta-C to vitamin A. Blood samples were collected over 21 d. beta-C and retinol were extracted from serum and isolated by high performance liquid chromatography. The retinol fraction was derivatized to a trimethylsilyl ether which was analyzed by gas chromatograph/mass spectrometry with electron capture negative chemical ionization. RESULTS: The retinol-d4 response in the circulation peaked at 24 hours after the beta-C-d8 dose, with a higher percent enrichment after the pharmacological dose than after the physiological dose. By using retinyl acetate-d8 as the vitamin A reference, the retinol-d4 formed from 6 mg of beta-C-d8 (11.2 mumol) was calculated to be equivalent to 1.6 mg of retinol (i.e., 3.8 mg of beta-C was equivalent to 1 mg of retinol). However, the retinol-d4 formed from 126 mg of beta-C-d8 (235 mumol) was equivalent to 2.3 mg of retinol (i.e., 55 mg beta-C was equivalent to 1 mg retinol). CONCLUSION: These results provide evidence that it is feasible to use stable isotope reference method to study retinol equivalence of beta-C and that there may be a dose-dependence on bioconversion of beta-carotene to retinol.
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Deuterio/análisis , Vitamina A/metabolismo , beta Caroteno/farmacocinética , Adulto , Cromatografía Líquida de Alta Presión , Relación Dosis-Respuesta a Droga , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Estudios Longitudinales , Vitamina A/sangre , Vitamina A/química , beta Caroteno/administración & dosificación , beta Caroteno/químicaRESUMEN
Subjects with the apolipoprotein (apo) E4 allele have been shown to have higher low density lipoprotein (LDL) cholesterol and apoB levels than do subjects with the other alleles. To elucidate the metabolic mechanisms responsible for this finding, we examined the kinetics of apoB-48 within triglyceride-rich lipoproteins (TRLs) and of apoB-100 within very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL), and LDL by using a primed constant infusion of [5,5,5-(2)H(3)]leucine in the fed state (hourly feeding) during consumption of an average American diet in 18 normolipidemic subjects, 12 of whom had the apoE3/E3 genotype and 6, the apoE3/E4 genotype. Lipoproteins were isolated by ultracentrifugation and apolipoproteins, by sodium dodecyl sulfate gels; isotope enrichment was assessed by gas chromatography-mass spectrometry. Kinetic parameters were calculated by multicompartmental modeling of the data with SAAM II software. Compared with the apoE3/E3 subjects, the apoE3/E4 subjects had significantly higher levels of total apoB, 100. 1+/-17.8 versus 135.4+/-34.0 mg/dL (P=0.009), and significantly higher levels of LDL apoB-100, 88.1+/-19.2 versus 127.5+/-32.7 mg/dL (P=0.005), respectively. The pool size of TRL apoB-48 was 17.4% lower for apoE3/E4 subjects compared with apoE3/E3 subjects due to a 33.3% lower production rate (P=0.28). There was no significant difference in the TRL apoB-48 fractional catabolic rate (5.1+/-2.2 versus 5.0+/-2.1 pools per day). The pool size for VLDL apoB-100 was 36% lower for apoE3/E4 subjects compared with apoE3/E3 subjects due entirely to a 30% lower production rate (P=0.04). The LDL apoB-100 pool size was 57.8% higher (P=0.003) for apoE3/E4 subjects compared with apoE3/E3 subjects due to a 35.5% lower fractional catabolic rate of LDL apoB-100 (P=0.003), with no significant difference in production rate. In addition, 77% of VLDL apoB-100 was converted to LDL apoB-100 in apoE3/E4 subjects compared with 58% in apoE3/E3 subjects (P=0.05). In conclusion, the presence of 1 E4 allele was associated with higher LDL apoB-100 levels owing to lower fractional catabolism of LDL apoB-100 and a 33% increase in the conversion of VLDL apoB-100 to LDL apoB-100.
Asunto(s)
Apolipoproteínas B/metabolismo , Apolipoproteínas E/genética , Adulto , Anciano , Apolipoproteína B-100 , Apolipoproteína B-48 , Apolipoproteína E3 , Apolipoproteína E4 , LDL-Colesterol/sangre , Femenino , Genotipo , Humanos , Cinética , Leucina/farmacocinética , Masculino , Persona de Mediana Edad , Tritio/farmacocinéticaRESUMEN
F2-Isoprostanes in plasma and urine are generally determined by labor-intensive methods requiring sample purification by solid-phase extraction and thin-layer chromatography (TLC). A streamlined and more sensitive method for the measurement of esterified plasma F2-isoprostanes was developed by replacing these steps with high-performance liquid chromatography (HPLC) using an amino column with a hexane/2-propanol gradient. Pentafluorobenzyl esters of F2-isoprostanes were prepared and purified by HPLC, silylated, and then analyzed by gas chromatography (GC) with negative chemical ionization mass spectroscopy (NCI-MS). This method permits analysis with lower plasma volumes (100 microL) and greater sensitivity (to 10 pg; allowing detection to 50 pg/mL) than provided by other methods. Urinary F2-isoprostanes can also be efficiently quantified by this method, with 8-iso-PGF2alpha being identified as a major isomer. With this procedure, esterified plasma F2-isoprostanes were found to be 8.3-fold higher in an end-stage renal failure patient on hemodialysis and urinary 8-iso-PGF2alpha was 7.1-fold higher in a cigarette smoker than respective control subjects. This method, particularly the substitution of the TLC step common to other methods with HPLC, results in a more sensitive and reproducible assay.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dinoprost/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Dinoprost/sangre , Dinoprost/orina , Femenino , Humanos , Masculino , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The purpose of our study was to investigate high density lipoprotein (HDL) apolipoprotein (apo) A-I and apoA-II kinetics in a state of constant feeding after a primed-constant infusion of [5,5, 5-(2)H(3)]L-leucine in 32 normolipidemic older men and postmenopausal women (aged 41 to 79 years). ApoA-I and apoA-II were isolated from plasma HDL, and enrichment was determined by gas chromatography/mass spectrometry. The fractional secretion rate was obtained by using a monoexponential equation calculated with the SAAM II program (Department of Bioengineering, University of Washington, Seattle). Mean HDL cholesterol (HDL-C) and total triglyceride levels were 23% higher and 27% lower, respectively, in women than in men. Mean plasma apoA-I levels were 10% greater in women than in men, whereas mean apoA-II levels were similar. HDL size, estimated by gradient-sizing gels and by the HDL-C/apoA-I+apoA-II ratio, was significantly higher in women than in men. Mean apoA-I secretion rates (SRs) were similar in men and women (12.28+/-3.64 versus 11.96+/-2.92 mg/kg per day), whereas there was a trend toward a lower (-13%) apoA-I fractional catabolic rate (FCR) in women compared with men (0.199+/-0.037 versus 0. 225+/-0.062 pools per day, P=0.11). Mean apoA-II SRs (2.21+/-0.57 versus 2.27+/-0.91 mg/kg per day) and FCRs (0.179+/-0.034 versus 0. 181+/-0.068 pools per day) were similar in men and women. For the group as a whole, there was an inverse association between the HDL-C/apoA-I+apoA-II ratio and apoA-I FCR and between the ratio and triglyceride levels. Plasma levels of apoA-I and apoA-II were correlated with their respective SRs but not FCRs. These data suggest a major role for apoA-I and apoA-II SRs in regulating the plasma levels of these proteins, whereas apoA-I FCR might be an important factor contributing to the differences in apoA-I levels between men and postmenopausal women. Moreover, plasma triglyceride levels are important determinants of HDL size and apoA-I catabolism.
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Apolipoproteína A-II/farmacocinética , Apolipoproteína A-I/farmacocinética , Caracteres Sexuales , Adulto , Anciano , HDL-Colesterol/metabolismo , Femenino , Humanos , Leucina/farmacocinética , Masculino , Persona de Mediana Edad , Posmenopausia , TritioRESUMEN
The kinetics of apolipoprotein (apo) B-100 and apoB-48 within triglyceride-rich lipoproteins (TRLs) and of apoB-100 within IDL and LDL were examined with a primed-constant infusion of (5,5,5-(2)H(3)) leucine in the fed state (hourly feeding) in 19 subjects after consumption of an average American diet (36% fat). Lipoproteins were isolated by ultracentrifugation and apolipoproteins by SDS gels, and isotope enrichment was assessed by gas chromatography/mass spectrometry. Kinetic parameters were calculated by multicompartmental modeling of the data with SAAM II. The pool sizes (PS) of TRL apoB-48, VLDL apoB-100, and LDL apoB-100 were 17+/-10, 273+/-167, and 3325+/-1146 mg, respectively. There was a trend toward a faster fractional catabolic rate (FCR) for VLDL apoB-100 than for TRL apoB-48 (6.73+/-3.48 versus 5.02+/-2.07 pools/d, respectively, P=0.06). The mean FCRs for IDL and LDL apoB-100 were 10.07+/-7.28 and 0.27+/-0.08 pools/d, respectively. The mean production rate (PR) of TRL apoB-48 was 6.5% of VLDL apoB-100 (1. 3+/-0.90 versus 20.06+/-6.53 mg. kg(-1). d(-1), P<0.0001). TRL apoB-48 PS was correlated with apoB-48 PR (r=0.780, P<0.0001) but not FCR (r=-0.1810, P=0.458). VLDL apoB-100 PS was correlated with both PR (r=0.713, P=0.0006) and FCR (r=-0.692, P=0.001) of VLDL apoB-100 and by apoB-48 PR (r=0.728, P=0.0004). LDL apoB-100 PS was correlated with FCR (r=-0.549, P=0.015). These data indicate that (1) the FCRs of TRL apoB-48 and VLDL apoB-100 are similar in the fed state, (2) TRL apoB-48 PS is correlated with TRL apoB-48 PR, (3) VLDL apoB-100 PS is correlated with both PR and FCR of VLDL apoB-100 and PR of TRL apoB-48, and (4) LDL apoB-100 PS is correlated with LDL FCR.
Asunto(s)
Apolipoproteínas B/farmacocinética , Deuterio , Leucina/farmacocinética , Anciano , Apolipoproteína B-100 , Apolipoproteína B-48 , Colesterol/metabolismo , LDL-Colesterol/metabolismo , VLDL-Colesterol/metabolismo , Femenino , Humanos , Cinética , Lipoproteínas/metabolismo , Hígado/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: Vitamin A activity of plant provitamin A carotenoids is uncertain. OBJECTIVE: The objective was to determine whether plant carotenoids can sustain or improve vitamin A nutrition during the fall season in kindergarten children in the Shandong province of China. DESIGN: The serum vitamin A concentration of 39% of the children was <1.05 micromol/L and of 61% of the children was > or = 1.05 micromol/L. For 5 d/wk for 10 wk, 22 children were provided approximately 238 g green-yellow vegetables/d and 34 g light-colored vegetables/d. Nineteen children maintained their customary dietary intake, which included 56 g green-yellow vegetables/d and 224 g light-colored vegetables/d. Octadeuterated and tetradeuterated vitamin A were given before and after the interventions, respectively, and their enrichments in the plasma were determined by gas chromatography-mass spectrometry. Serum retinol and carotenoid concentrations were measured by HPLC. RESULTS: Carotenoid nutrition improved after consumption of green-yellow vegetables. Serum concentrations of retinol were sustained in the group fed green-yellow vegetables but decreased in the group fed light-colored vegetables (P < 0.01). The isotope-dilution tests confirmed that total-body vitamin A stores were sustained in the group fed green-yellow vegetables, but decreased 27 micromol (7700 microg retinol) per child, on average, in the group fed light-colored vegetables (P < 0.06). CONCLUSION: Green-yellow vegetables can provide adequate vitamin A nutrition in the diet of kindergarten children and protect them from becoming vitamin A deficient during seasons when the provitamin A food source is limited.
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Carotenoides/administración & dosificación , Dieta , Verduras , Deficiencia de Vitamina A/prevención & control , Vitamina A/sangre , Análisis de Varianza , Carotenoides/sangre , Niño , Preescolar , China , Cromatografía Líquida de Alta Presión , Femenino , Humanos , MasculinoRESUMEN
Stable isotope methodology was used to determine the kinetic behavior of apolipoprotein (apo) A-I within the triglyceride-rich lipoprotein (TRL) fraction and to compare TRL apoA-I kinetics with that of apoA-I in high density lipoprotein (HDL) and TRL apoB-48. Eight subjects (5 males and 3 females) over the age of 40 were placed on a baseline average American diet and after 6 weeks received a primed-constant infusion of [5,5,5-(2)H(3)]-l-leucine for 15 h while consuming small hourly meals of identical composition. HDL and TRL apoA-I and TRL apoB-48 tracer/tracee enrichment curves were obtained by gas chromatography;-mass spectrometry. Data were fitted to a compartmental model to determine the fractional secretion rates of apoA-I and apoB-48 within each lipoprotein fraction. Mean plasma apoA-I levels in TRL and HDL fractions were 0. 204 +/- 0.057 and 134 +/- 15 mg/dl, respectively. The mean fractional catabolic rate (FCR) of TRL apoA-I was 0.250 +/- 0.069 and HDL apoA-I was 0.239 +/- 0.054 pools/day, with mean estimated residence times (RT) of 4.27 and 4.37 days, respectively. The mean TRL apoB-48 FCR was 5.2 +/- 2.0 pools/day and the estimated mean RT was 5.1 +/- 1.8 h. Our results indicate that apoA-I is catabolized at a slower rate than apoB-48 within TRL, and that apoA-I within TRL and HDL fractions are catabolized at similar rates.
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Apolipoproteína A-I/sangre , Lipoproteínas HDL/sangre , Lipoproteínas/sangre , Triglicéridos/sangre , Adulto , Anciano , Apolipoproteína B-48 , Apolipoproteínas B/sangre , Femenino , Humanos , Cinética , Lipoproteínas/química , Lipoproteínas HDL/química , Masculino , Persona de Mediana EdadRESUMEN
We examined the mechanisms responsible for the decrease in HDL cholesterol (HDL-C) levels after the consumption of a diet low in total fat, saturated fat, and cholesterol. Twenty-one subjects with a mean age of 58+/-12 years were placed on a baseline isocaloric diet (15% protein, 49% carbohydrate, 36% fat, and 150 mg/1000 kcals of cholesterol) and then switched to an NCEP Step 2 diet (15% protein, 60% carbohydrate, 25% fat, and 45 mg/1000 kcals of cholesterol). After 6 or 24 weeks on each diet, subjects received a 15-hour primed-constant infusion of [5,5,5-2H3]-L-leucine. HDL apoA-I and apoA-II tracer curves were determined by gas chromatography-mass spectrometry and fitted to a monoexponential equation. Compared with the baseline diet, consumption of the Step 2 diet lowered HDL-C mean levels by 15% (1.03+/-0.23 to 0.88+/-0.16 mmol/L, P<0.001), apoA-I by 12% (1.25+/-0.15 to 1.10+/-0.13 g/L, P<0. 001) and the TC/HDL-C ratio by 5% (0.145+/-0.04 to 0.137+/-0.03). No significant changes were observed in apoA-II levels and HDL particle size with diet. HDL apoA-I fractional catabolic rate did not change (0.219+/-0.052 to 0.220+/-0.043 pools/day, P=0.91) but HDL apoA-I secretion rate decreased by 8% (12.26+/-3.07 to 10.84+/-2.11 mg. kg-1. day-1, P=0.03) during consumption of the Step 2 diet. There was no effect of diet on apoA-II fractional catabolic rate or secretion rate. Our results indicate that the decrease in HDL-C and apoA-I levels during the isocaloric consumption of a Step 2 diet paralleled the reductions in apoA-I secretion rate.
Asunto(s)
Apolipoproteína A-I/metabolismo , Colesterol en la Dieta/administración & dosificación , Dieta con Restricción de Grasas , Lipoproteínas HDL/sangre , Adulto , Anciano , Apolipoproteína A-I/sangre , Apolipoproteína A-II/metabolismo , HDL-Colesterol/sangre , LDL-Colesterol/sangre , VLDL-Colesterol/sangre , Femenino , Humanos , Lipoproteínas HDL/química , Masculino , Persona de Mediana Edad , Triglicéridos/sangreRESUMEN
Alterations in body composition and nutritional status are common in humans with heart failure and are related, in part, to increases in cytokine concentrations. Cytokines have not been studied previously in dogs with naturally occurring cardiac disease nor has fish oil administration been used in this population to decrease cytokine production. The purposes of this study were to characterize nutritional and cytokine alterations in dogs with heart failure and to test the ability of fish oil to reduce cytokines and improve clinical outcome. Body composition, insulinlike growth factor-1, fatty acids, and cytokines were measured in 28 dogs with heart failure and in 5 healthy controls. Dogs with heart failure then were randomized to receive either fish oil or placebo for 8 weeks. All parameters were measured again at the end of the study period. At baseline, 54% of dogs with heart failure were cachectic and the severity of cachexia correlated with circulating tumor necrosis factor-alpha concentrations (P = .05). Cytokine concentrations at baseline, however, were not significantly increased in dogs with heart failure compared to controls. Baseline plasma arachidonic acid (P = .02), eicosapentaenoic acid (P = .03), and docosahexaenoic acid (P = .004) concentrations were lower in dogs with heart failure than in controls. Fish oil supplementation decreased interleukin-1 beta (IL-1) concentrations (P = .02) and improved cachexia (P = .01) compared to the placebo group. The mean caloric intake of the heart failure dogs as a group was below the maintenance energy requirement (P < .001), but no difference was found in food intake between the fish oil and placebo groups. Insulinlike growth factor-1 concentrations (P = .01) and reductions in circulating IL-1 concentrations over the study period (P = .02) correlated with survival. These data demonstrate that canine heart failure is associated with cachexia, alterations in fatty acids, and reduced caloric intake. Fish oil supplementation decreased IL-1 concentrations and improved cachexia. In addition, reductions in IL-1 predicted survival, suggesting that anticytokine strategies may benefit patients with heart failure.
Asunto(s)
Enfermedades de los Perros/metabolismo , Aceites de Pescado/uso terapéutico , Cardiopatías/veterinaria , Animales , Composición Corporal/efectos de los fármacos , Agua Corporal/metabolismo , Caquexia/metabolismo , Caquexia/prevención & control , Caquexia/veterinaria , Cromatografía de Gases/veterinaria , Dinoprostona/sangre , Enfermedades de los Perros/dietoterapia , Perros , Ingestión de Alimentos/efectos de los fármacos , Ingestión de Alimentos/fisiología , Ácidos Grasos/análisis , Aceites de Pescado/farmacología , Cromatografía de Gases y Espectrometría de Masas/veterinaria , Cardiopatías/dietoterapia , Cardiopatías/metabolismo , Factor I del Crecimiento Similar a la Insulina/análisis , Interleucina-1/sangre , Estado Nutricional/fisiología , Valores de Referencia , Factor de Necrosis Tumoral alfa/químicaRESUMEN
Apolipoprotein (apo) E is a 299-amino acid glycoprotein that serves a number of functions in lipoprotein metabolism. Apo E binds to the triglyceride-rich lipoproteins (TRL), very-low-density lipoprotein (VLDL), and chylomicrons, as they are lipolyzed, mediating their removal from plasma via lipoprotein receptors. Apo E is also found associated with high-density lipoprotein (HDL) and has been suggested to play a role in reverse cholesterol transport. Studies on the kinetic behavior of apo E from the TRL and HDL fractions provide insights into the metabolic relationships between TRL and HDL in vivo. We sought to develop a compartmental model that can be used for analysis of kinetic data in studies on the metabolism of TRL and HDL apo E. Using radioactive tracers, it has been previously observed that, in some instances, a portion of VLDL apo E that is removed from plasma subsequently reappears in VLDL. Four multicompartmental models were considered that could account for this type of behavior: model A, in which there is transfer of apo E from HDL to VLDL; model B, in which there is a bidirectional extravascular exchange; model C, in which there is removal and subsequent reintroduction of TRL apo E into plasma; and model D, in which there is secretion of TRL apo E into plasma directly and via an extravascular pathway. Models C and D provided the best fit to the experimental data. While no physiologically plausible analog to model C could be found, an extravascular delay, analogous to newly secreted apo E that enters the lymphatic system before appearing in plasma, was postulated for model D. It was this model that was used to analyze kinetic data from metabolic studies of apo E. The model was able to provide a satisfactory fit to kinetic data in studies in which subjects were given a primed-constant infusion of 2H3-leucine. It was determined that TRL apo E from the six subjects studied had a mean residence time of 0.11 +/- 0.05 days and a mean production rate of 10.6 +/- 7.2 mg/kg/d, while HDL apo E had a mean residence time of 2.96 +/- 0.99 days and a mean production rate of 0.07 +/- 0.07 mg/kg/d. We conclude that this model describes a potential pathway for the metabolism of a portion of apo E in plasma and can be used to calculate the residence time and production rate of TRL and HDL apo E under a variety of conditions.
Asunto(s)
Apolipoproteínas E/metabolismo , HDL-Colesterol/metabolismo , Triglicéridos/metabolismo , Adulto , Anciano , Apolipoproteínas E/biosíntesis , Apolipoproteínas E/sangre , HDL-Colesterol/biosíntesis , HDL-Colesterol/sangre , Humanos , Cinética , Lípidos/sangre , Masculino , Persona de Mediana Edad , Fenotipo , Triglicéridos/biosíntesis , Triglicéridos/sangreRESUMEN
Bovine insulin was reacted at pH 4.0 with submolar amounts of hypochlorite. At least one molecule of insulin was modified per two molecules of hypochlorite added, as estimated by HPLC of native and modified insulin. About 5% of the hypochlorite-modified insulin reacted with dinitrophenylhydrazine (DNPH), a reagent which specifically labels carbonyl groups. The major DNPH-labeled product was isolated from the native insulin on reverse-phase HPLC, using trifluoroacetic acid/water/acetonitrile gradients. The UV spectrum of the major peak on the HPLC diode-array detector was representative of DNPH adducts, with lambda max = 365 nm. Several methods, including total amino acid analysis, tryptic digestion, and collision-induced dissociation-electrospray MS, indicate that the major carbonyl in the DNPH-labeled product was on the amino-terminal phenylalanine of the insulin B-chain. Amino acid analysis indicated that tyrosine was also degraded by hypochlorite, but we could not detect a carbonyl group formed at tyrosine. These findings suggest that the terminal amino groups of proteins are highly vulnerable to carbonyl formation during hypochlorite attack. The use of relatively low amounts of active oxygen species (such as hypochlorite), followed by chromatographic isolation of the protein labeled with a carbonyl-specific reagent, can be a useful approach to the study of reactive sites on proteins.
Asunto(s)
Ácido Hipocloroso/química , Insulina/química , Secuencia de Aminoácidos , Aminoácidos/análisis , Animales , Bovinos , Cromatografía Líquida de Alta Presión , Concentración de Iones de Hidrógeno , Hidrólisis , Indicadores y Reactivos , Espectrometría de Masas/métodos , Datos de Secuencia Molecular , Peso Molecular , Fenilhidrazinas , Espectrofotometría Ultravioleta , Tripsina/químicaRESUMEN
The effect of lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, on the kinetics of de novo cholesterol synthesis and apolipoprotein (apo) B in very-low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), and low-density lipoprotein (LDL) was investigated in five male patients with combined hyperlipidemia. Subjects were counseled to follow a Step 2 diet and were treated with lovastatin and placebo in randomly assigned order for 6-week periods. At the end of each experimental period, subjects were given deuterium oxide orally and de novo cholesterol synthesis was assessed from deuterium incorporation into cholesterol and expressed as fractional synthesis rate (C-FSR) and production rate (C-PR). Simultaneously, the kinetics of VLDL, IDL, and LDL apo B-100 were studied in the fed state using a primed-constant infusion of deuterated leucine to measure fractional catabolic rates (FCR) and production rates (PR). Drug treatment resulted in significant decreases in total cholesterol (-29%), VLDL cholesterol (-40%), LDL cholesterol (-27%), and apo B (-16%) levels and increases in HDL cholesterol (+13%) and apolipoprotein (apo) A-I (+11%) levels. Associated with these plasma lipoprotein responses was a significant reduction in both de novo C-FSR (-40%; P = .04) and C-PR (-42%; P = .03). Treatment with lovastain in these patients had no significant effect on the FCR of apoB-100 in VLDL, IDL, or LDL, but resulted in a significant decrease in the PR of apoB-100 in IDL and LDL. Comparing the kinetic data of these patients with those of 10 normolipidemic control subjects indicates that lovastatin treatment normalized apoB-100 IDL and LDL PR. The results of these studies suggest that the declines in plasma lipid levels observed after treatment of combined hyperlipidemic patients with lovastatin are attributable to reductions in the C-FSR and C-PR of de novo cholesterol synthesis and the PR of apoB-100 containing lipoproteins. The decline in de novo cholesterol synthesis, rather than an increase in direct uptake of VLDL and IDL, may have contributed to the decline in the PR observed.
Asunto(s)
Anticolesterolemiantes/uso terapéutico , Apolipoproteínas B/biosíntesis , Colesterol/biosíntesis , Hiperlipidemias/tratamiento farmacológico , Lovastatina/uso terapéutico , Adulto , Anciano , Apolipoproteína B-100 , Colesterol/sangre , LDL-Colesterol/sangre , VLDL-Colesterol/sangre , Humanos , Hiperlipidemias/metabolismo , Lipoproteínas/sangre , Lovastatina/farmacología , Masculino , Persona de Mediana EdadRESUMEN
In vivo kinetics of apoB were performed in three apoB-67/apoB-100 heterozygotes using a primed-constant infusion of (5,5,5-(2)H3)-leucine. Mean plasma VLDL and LDL apoB-67 concentrations were 0.05 +/- 0.01 mg/dl and 0.62 +/- 0.17 mg/dl, respectively, which were 0.2% and 2.8% of total plasma apoB concentrations. When cation exchange chromatography was used to separate plasma amino acids from fragments of polyacrylamide gels, the tracer/tracee ratio at plateau for VLDL apoB-67 was less than 50% of that observed when centrifugation was used. Mean fractional catabolic rate for LDL apoB-67 was 2-fold higher when the lower plateau was used in multicompartmental analysis compared to the higher plateau (0.70 +/- 0.21 versus 0.37 +/- 0.06 pools per day, P = 0.06). The lower plateau resulted from introduction of unlabeled leucine during cation exchange chromatography; therefore, all samples were processed with centrifugation. Mean fractional catabolic rates for VLDL and LDL apoB-67 were not significantly different from VLDL and LDL apoB-100 (VLDL: 9.7 +/- 3.4 versus 18.1 +/- 8.6 pools per day, respectively, P = 0.19; LDL: 0.37 +/- 0.06 versus 0.34 +/- 0.11 pools per day, respectively, P = 0.66). Mean secretion rate of VLDL apoB-67 was 5.6% of VLDL apoB-100 (0.20 +/- 0.04 versus 3.6 +/- 1.3 mg/kg/day, P = 0.01). Fifty-three % of apoB-67 was directly removed from VLDL compared to only 3.5% of apoB-100 (P = 0.003), thus accounting for the lower proportion of apoB-67 in LDL (3.3 +/- 1.8%) as compared to VLDL (11.2 +/- 2.5%). Mean LDL production rate for apoB-67 was 2.6% of LDL apoB-100 (0.09 +/- 0.02 versus 3.50 +/- 1.39 mg/kg/day, P = 0.05). Thus, decreased secretion of apoB-67 is responsible for low levels of apoB-67.
Asunto(s)
Apolipoproteínas B/biosíntesis , Apolipoproteínas B/genética , Heterocigoto , Hipobetalipoproteinemias/sangre , Hipobetalipoproteinemias/genética , Adulto , Apolipoproteína B-100 , Apolipoproteínas B/sangre , Cromatografía por Intercambio Iónico , Deuterio , Contaminación de Medicamentos , Femenino , Humanos , Cinética , Leucina , Lipoproteínas/sangre , Lipoproteínas IDL , Lipoproteínas LDL/sangre , Lipoproteínas VLDL/sangre , Masculino , Persona de Mediana EdadRESUMEN
Apolipoprotein (apo) B-67 is a truncated form of apoB-100 due to deletion of an adenine at cDNA 9327. Heterozygotes have one allele making apoB-100; therefore, plasma apoB levels would be predicted to be at least 50% of normal. However, apoB-67 heterozygotes have total plasma apoB levels that are 24% of normal. To determine the mechanisms responsible for the lower-than-expected levels of apoB, in vivo kinetics of apoB-100 were performed in three apoB-67/apoB-100 heterozygotes and compared with those of six control subjects by using a primed-constant infusion of [5,5,5-2H3]leucine in the fed state. Kinetic parameters were calculated by multicompartmental modeling of the data. The mean total apoB plasma concentration of the apoB-67 subjects was 21.8+/-6.1 mg/dL, or 24% of that of control subjects (89.6+/-24.1 mg/dL, P=.002). ApoB-67 subjects had lower mean VLDL apoB-100 production rates (3.6+/-1.2 versus 13.9+/-3.5 mg x kg(-1) x d(-1), P=.002) and lower mean transport rates of apoB-100 into LDL (3.5+/-1.4 versus 12.6+/-4.1 mg x kg(-1) x d(-1), P=.008) compared with control subjects. The transport rate into IDL was not significantly different (1.2+/-0.5 versus 6.2+/-4.0 mg x kg(-1) x d(-1), P=.07). The fractional catabolic rate of VLDL apoB-100 was significantly higher in apoB-67 subjects than in control subjects (18.1+/-8.6 versus 7.6+/-1.6 mg x kg(-1) x d(-1), P=.017). ApoB-100 IDL and LDL fractional catabolic rates were not significantly different. VLDL apoB-100 pool size in apoB-67 subjects was 11% of that of control subjects (15.8+/-7.7 versus 141.6+/-33.7 mg, P=.0004) due to a 74% lower production rate (26% of control values) and a 2.4-fold higher fractional catabolic rate. LDL apoB-100 pool size in apoB-67 subjects was 22% of that of control subjects (665.3+/-192.4 versus 2968.3+/-765.2 mg, P=.002) due primarily to a lower production rate (27% of control values). Thus, both decreased production of VLDL and LDL apoB-100 and increased catabolism of VLDL apoB-100 are responsible for the low levels of apoB-100 in apoB-67 subjects.