Heterogeneity of molecular weight and apolipoproteins in low density lipoproteins of healthy human males.

The molecular weights of five low density lipoprotein (LDL) subfractions from four normal healthy males were determined by analytic ultracentrifuge sedimentation equilibria. Protein content of each subfraction was determined by elemental CHN analysis, and weights of apoprotein peptides were calculated. Molecular weights in subfractions of increasing density were 2.92 +/- 0.26, 2.94 +/- 0.12, 2.68 +/- 0.09, 2.68 +/- 0.28 and 2.23 +/- 0.22 million Da, and protein weight percentages were 21.05, 21.04, 22.05, 23.10 and 29.10, in subfractions 1, 2, 3, 4 and 5, respectively. Total mean apoprotein weights for respective subfractions were 614 +/- 53, 621 +/- 45, 588 +/- 9, 637 +/- 83 and 645 +/- 62 KDa. In addition to a single apoprotein B-100 (apo B-100) peptide with a mean carbohydrate content of 7.1% and a molecular weight of 550 KDa per LDL particle, there may be one or more apoprotein E peptides of 34 KDa and/or apoprotein C-III of 9 KDa. In addition, subfractions 4 and 5 may contain 3-7% apolipoprotein (a). There is considerable heterogeneity among LDL subfractions as well as within the same fraction from different individuals. This heterogeneity may relate to differences in origin, metabolism and/or atherogenicity as a result of their content of apoproteins other than apo B-100.

Cholesterol-lowering in hamsters fed rice bran at various levels, defatted rice bran and rice bran oil.

This study was conducted to determine the relative cholesterol-lowering effects of several levels of full-fat rice bran in hamsters. In addition, the separate effects of defatted rice bran and/or crude rice bran oil were investigated at levels equivalent to those present in 43.7% full-fat rice bran. Diets containing 10.9, 21.8, 32.8 or 43.7% full-fat rice bran, 35% defatted rice bran and/or 9% rice bran oil were fed to 4-wk-old male hamsters. All diets contained 10% total dietary fiber, 9% fat and 3.2% nitrogen; hypercholesterolemic diets contained 0.3% cholesterol; two diets were cholesterol-free, i.e., 10% cellulose and 43.7% full-fat rice bran. After 21 d, plasma and liver cholesterol, plasma triglycerides and liver weights were significantly greater in hamsters fed the 10% cellulose diet with 0.3% cholesterol compared with those fed the cholesterol-free cellulose diet. In animals fed cholesterol-free diets, plasma cholesterol values were significantly lower in those fed the 43.7% full-fat rice bran diet than in those fed the cellulose diet. In animals fed cholesterol-containing diets, plasma and liver cholesterol were significantly lower in animals fed the 43.7% full-fat rice bran diet than in those fed the cellulose diet. Plasma cholesterol reductions were significantly correlated to the level of rice bran in the diet. In cholesterol-fed hamsters, total liver cholesterol content was significantly lower in those fed the defatted rice bran diet with rice bran oil compared with those fed the cellulose diet. Full-fat rice bran was the only treatment that significantly lowered both plasma and liver cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)

Partial specific volume and preferential hydration of low density lipoprotein subfractions.

We have determined the partial specific volume (v-) for five low density lipoprotein (LDL) subfractions (n = 5-7) and evaluated preferential hydration (n = 2) for LDL subfraction 3 in normolipoproteinemic subjects in order to characterize these highly atherogenic components of the human plasma lipoprotein spectra. Values for v- at 1 g were determined by sixth place density measurements of the solvent and lipoprotein solutions and carbon, hydrogen and nitrogen (CHN) absolute mass of the lipoprotein concentrations. Mean values for v- were 0.9757 +/- 0.0019, 0.9701 +/- 0.0007, 0.9674 +/- 0.0016, 0.9616 +/- 0.0016 and 0.9550 +/- 0.0025 ml/g for subfractions 1, 2, 3, 4 and 5, respectively. However, molecular densities (sigma) obtained from rho (rho) = 1/v- for respective LDL subfractions were 1.0249, 1.0308, 1.0337, 1.0399 and 1.0471 g/ml, respectively. The preferential hydration of lipoprotein subfraction 3 (n = 2) in NaCl/H2O solutions was 2.9-4.8 wt percent, whereas values were much lower (0.3-0.6 wt percent) in NaCl/NaBr/H2O solvent system. Unhydrated densities for LDL subfraction 3 (n = 2) at 1 g (sixth-place density meter) were 1.0287 and 1.0269 g/ml, whereas at 200,000 X g (used in D2O flotation eta F degrees vs rho determinations) both values were 1.0308 g/ml, indicating that these similar LDL fractions have 23 and 53% higher compressibility than the solvent at 200,000 X g force. It was observed that the linearity of eta F degrees vs rho may not be valid for solvents NaCl/NaBr/H2O of density as high as 1.4744 g/ml. Thus, flotation velocity data using extreme salt concentrations (1.4744 g/ml and higher) may be viewed with caution.

Analytic ultracentrifuge calibration and determination of lipoprotein-specific refractive increments.

Accurate quantification of the major classes and subfractions of human serum lipoproteins is an important analytical need in the characterization and evaluation of therapy of lipid and lipoprotein abnormalities. For calibrating the analytic ultracentrifuge (AnUC), we routinely use a Beckman calibration wedge cell with parallel scribed lines 1 cm apart. Such a cell gives a rectangular pattern in the schlieren diagram, which determines magnification and also provides an area corresponding to an invariant refractive increment. We have independently validated this wedge calibration cell using a special boundary-forming cell in which 1.174% sucrose is overlayered with distilled water. Comparing wedge cell area with extrapolated zero time boundary area refractive increment gives agreement to within less than 1%, corresponding to a refractive increment error of +/- 0.00002 delta n. Complete calibration for AnUC analysis of lipoproteins also requires accurate determination of the specific refractive increments (SRI) of the major lipoprotein classes, namely low density lipoprotein (LDL) and high density lipoprotein (HDL). These are measured in the density in which they are analyzed, i.e., 1.061 g/ml for LDL and 1.200 g/ml for HDL. Five fresh serum samples were fractionated for total LDL and total HDL and their SRI determined. Total lipoprotein mass was determined using precise CHN elemental analysis and compositional analyses. The results yielded corrected SRI of 0.00142 and 0.00135 delta n/g/100 ml for LDL and HDL. Thus, our current values using 0.00154 and 0.00149 delta n/g/100 ml underestimate LDL and HDL by 9% and 11%. Corrections of all previous LDL and HDL AnUC data can be made using appropriate factors of 1.087 and 1.106.

Sedimentation equilibrium of human low density lipoprotein subfractions.

The molecular weights of low density lipoprotein (LDL) subfractions were determined precisely by meniscus depletion sedimentation equilibrium. Equilibrium speeds ranged from 9743 to 5896 rpm. The average molecular weights of various LDL subfractions of Sf values 9.49, 7.94, 6.42, 5.17, and 3.71 determined by sedimentation equilibrium were 2.97 X 10(6) ; 3.13 X 10(6); 2.89 X 10(6); 2.45 X 10(6); and 2.61 X 10(6) daltons, respectively; and their respective densities were 1.0267, 1.0306, 1.0358, 1.0422, and 1.0492 g/ml. Minimal hydrated molecular weights for this fractions determined by flotation velocity at 37,020 rpm were 2.57 X 10(6); 2.37 X 10(6); 2.09 X 10(6); 1.94 X 10(6); and 1.81 X 10(6) daltons; whereas similar molecular weights determined at 52,640 rpm were 2.53 X 10(6); 2.27 X 10(6); 1.99 X 10(6); 1.86 X 10(6); and 1.74 X 10(6) daltons for the respective LDL subfractions. Higher molecular weights of fractions 2 and 5 compared to their adjacent fractions 1 and 4 by sedimentation equilibrium are of great interest. The calculated fractional ratio f/f O from sedimentation equilibrium and flotation velocity data ranges from 1.10 to 1.31, suggesting complexity and asymmetry of LDL subfraction molecules. There is also evidence that compressibility of LDL molecules may be different than that for the salt solution under high g-force. Assuming that redistributed LDL molecules at equilibrium under low g-force are spherical, it is possible that the shape of LDL molecules undergoing flotation velocity determinations may be distorted in high g-force conditions. Such distortion may be consistent with the high f/f O values obtained and may also be a basis for structural rearrangement and/or lipoprotein degradation with prolonged preparative ultracentrifugation at high g-force and pressure.

Free choice consumption of minerals by lambs fed calcium-adequate or calcium-deficient diets.

Four growth trials were conducted to determine whether lambs have the ability to recognize a dietary Ca deficiency and to correct that deficiency by consuming minerals offered free choice. In trial 1, lambs were fed a control or Ca-deficient diet with 0, 1, 5 or 10 mineral choices, only one of which provided Ca. Daily gains and feed efficiencies of lambs fed control diets tended to be superior to those of lambs fed Ca-deficient diets. Consumption of free choice calcium carbonate was greater (P less than .05) for lambs fed Ca-deficient diets than for those fed control diets. However, total Ca intake was greatest (P less than .05) for lambs fed control diets. In trial 2, lambs were fed diets containing .35, .20 or .06% Ca with zero or four mineral choices in either a constant or varied location. Daily gains were highest (P less than .05) for lambs fed the control diet. Performance of lambs fed diets deficient in Ca was not improved by providing free choice minerals. Although free choice Ca intakes were higher (P less than .05) for lambs fed Ca-deficient diets, total intake of Ca decreased with severity of dietary Ca deficiency. In trial 3, lambs were fed a (1) Ca-adequate diet, (2) Ca-deficient diet, (3) Ca-deficient diet with four mineral choices (one of which contained Ca) or (4) Ca-deficient diet with seven times the daily Ca requirement offered once weekly. Lambs fed control diets gained faster (P less than .05) than those fed deficient diets with free choice minerals. Ca intakes were greater (P less than .05) for control lambs than for those fed Ca-deficient diets with Ca available free choice. In trial 4, lambs were fed either a control or a Ca-deficient diet for 42 d. All lambs were then offered both control and Ca-deficient diets in separate compartments of the feed bunks. Although performance, bone and serum data followed trends similar to those observed in trials 1, 2 and 3, differences between treatment groups were not significant. Data from these trials support the recommendation that, when possible, required minerals should be provided in the diet rather than on a free choice basis.