RESUMEN
Dietary fats are known to influence the fatty acid profile of plasma lipids, including phospholipids which are substrates of lecithin:cholesterol acyltransferase (LCAT; EC 2.3.1.43), an important enzyme in lipoprotein metabolism. We tested whether the dietary fatty acid profile has an effect on LCAT activity in an animal model. Rats were conditioned to eat two meals per day, which were enriched in either palmitic, oleic or linoleic acids, for 10 weeks. Serum was isolated from blood samples taken prior to the meal. The LCAT activity was determined in two ways: (1) by measuring serum cholesterol esterification rates, which are an estimate of LCAT action on endogenous lipoproteins, and (2) by measuring serum LCAT activity levels with excess exogenous substrates, an estimate of LCAT mass. Animals receiving the linoleic acid diet had lower serum concentrations of unesterified cholesterol and triglycerides, if compared with animals fed oleic acid or palmitic acid diets (p < 0.05). Serum LCAT activity levels (measured with excess exogenous substrates) were not different, but both the absolute and fractional rates of cholesterol esterification were highest on the linoleic acid rich diet (p < 0.01), showing that LCAT action on endogenous lipoproteins is improved. No differences were found in serum apolipoprotein B and A-IV concentrations between the dietary groups. Apolipoprotein A-I levels were lowest in the palmitic acid group (oleic and linoleic > palmitic; p < 0.05), and apolipoprotein E levels were highest in the palmitic acid group (palmitic > oleic and linoleic; p < 0.05). It is concluded that a linoleic acid rich diet may cause increased metabolism of serum cholesterol by LCAT in rats. This effect is not due to elevated serum concentrations of LCAT or of its apolipoprotein activators, but most likely to changes in the chemical composition of endogenous lipoprotein substrates. It remains to be established whether the serum cholesterol esterification rates measured in vitro are related to in vivo rates of reverse cholesterol transport.
Asunto(s)
Alimentación Animal , Ésteres del Colesterol/sangre , Grasas de la Dieta/administración & dosificación , Ácido Linoleico/administración & dosificación , Fosfatidilcolina-Esterol O-Aciltransferasa/sangre , Animales , Peso Corporal , Lípidos/sangre , Masculino , Ácido Oléico/administración & dosificación , Ácido Palmítico/administración & dosificación , Ratas , Ratas Wistar , Triglicéridos/sangreRESUMEN
Two-dimensional rotating-frame nuclear Overhauser enhancement spectroscopy has been applied to milk oligosaccharides containing the blood-group determinants Lea, Led, or Leb. The results indicate a large number of dipolar interactions that are consistent with the preferred conformations described for these blood-group determinants. Despite careful experimental conditions, coherent transfer processes are not suppressed completely, allowing only a qualitative evaluation of the nuclear Overhauser data. The 1H- and 13C-n.m.r. spectra are assigned by comparison with literature data and by application of two-dimensional 13C-1H shift-correlation spectroscopy.
Asunto(s)
Oligosacáridos/análisis , Análisis Espectral , Espectroscopía de Resonancia MagnéticaRESUMEN
A simple and efficient procedure is described for the preparation of cardiolipin sodium salt from beef heart. A crude phospholipid fraction is isolated by chloroform-methanol extraction of the homogenized tissue, followed by acetone precipitation and reprecipitation in 4% aqueous CaCl2-methanol. Cardiolipin is separated from the calcium salts of the acidic phospholipids by partition column chromatography on silica gel (Polygosil 60-63100) using 2-propanol-cyclohexane-water 50:43:7 (v/v/v) as eluent. Further purification of the cardiolipin is achieved by high performance liquid chromatography of the calcium salt on silica gel (Lichrosorb Si 60-5) with a neutral eluent (2-propanol-cyclohexane-water 45:50:5 (v/v/v], followed by quantitative conversion to the sodium salt. The yield of this procedure is 1.5-2.1 g of pure 99% sodium salt of cardiolipin per kg of moist ventricular tissue.