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.

Associations of lipoproteins and apolipoproteins with gradient gel electrophoresis estimates of high density lipoprotein subfractions in men and women.

We examined the relations of gender and lipoproteins to subclasses of high density lipoproteins (HDLs) in a cross-sectional sample of moderately overweight men (n = 116) and women (n = 78). The absorbance of protein-stained polyacrylamide gradient gels was used as an index of mass concentrations of HDL at intervals of 0.01 nm across the entire HDL particle size range (7.2-12 nm). At least five HDL subclasses have been identified by their particle sizes: HDL3c (7.2-7.8 nm), HDL3b (7.8-8.2 nm), HDL3a (8.2-8.8 nm), HDL2a (8.8-9.7 nm), and HDL2b (9.7-12 nm). Men had significantly higher HDL3b and significantly lower HDL2a and HDL2b than did women. Correlations of HDL subclasses with concentrations of other lipoprotein variables were generally as strong for gradient gel electrophoresis as for analytical ultracentrifugation measurements of HDL particle distributions. In both sexes, high levels of HDL3b were associated with coronary heart disease risk factors, including high concentrations of triglycerides, apolipoprotein B, small low density lipoproteins, intermediate density lipoproteins, and very low density lipoproteins and low concentrations of HDL2 cholesterol and HDL2 mass. Plasma concentrations of HDL3 cholesterol were unrelated to protein-stained HDL3b levels. HDL3 cholesterol concentrations also did not exhibit the sex difference or the relations with lipoprotein concentrations that characterized HDL3b. Thus, low HDL3b levels may contribute in part to the low heart disease risk in men and women who have high HDL cholesterol. Measurements of HDL3 cholesterol may not identify clinically important relations involving HDL3b.

Effect of a salmon diet on the distribution of plasma lipoproteins and apolipoproteins in normolipidemic adult men.

The effects of n-3 fatty acids on plasma lipids, lipoproteins and apoproteins have usually been studied in humans after feeding of purified fish oil. This study describes the effect of a natural diet, containing salmon as the source of n-3 fatty acids, on these parameters as compared to a diet very low in n-3 fatty acids. The subjects were nine normolipidemic, healthy males who were confined to a nutrition suite for 100 days. During the first 20 days of the study the participants were given a stabilization diet consisting of 55% carbohydrates, 15% protein, and 30% fat. The n-3 content of this diet was less than 1%, and it contained no 20- or 22-carbon n-3 fatty acids. After the stabilization period the men were split into two groups, one group continued on the stabilization diet while the other received the salmon diet that contained approximately 2.1 energy percent (En%) of calories from 20- and 22-carbon n-3 fatty acids. Both diets contained equal amounts of n-6 fatty acids. This regime continued for 40 days, then the two groups switched diets for the remainder of the study. Plasma triglycerides were lowered significantly (p less than 0.01) and high density lipoprotein cholesterol (HDL-C) was significantly elevated (p less than 0.01) after the men consumed the salmon diet for 40 days. The very low density lipoproteins (VLDL) were lowered, but the trend did not reach statistical significance during the intervention period.(ABSTRACT TRUNCATED AT 250 WORDS)

Colestipol-induced changes in LDL composition and metabolism. II. Studies in humans.

We investigated the effect of the bile acid sequestrant, colestipol hydrochloride, on the composition and metabolism of human low density lipoprotein (LDL). Colestipol treatment produced a disproportionate decrease in LDL cholesterol compared to LDL apoB, resulting in a significant decrease in the LDL cholesterol/apoB ratio. Electron microscopy revealed that LDL particles were smaller in size and analytical ultracentrifugation demonstrated that colestipol therapy selectively depleted larger, more buoyant LDL particles of Sf degrees 6-7. Thus, colestipol therapy produced LDL that were smaller in size, more dense, and characterized by a decreased cholesterol to protein ratio. To determine whether the altered LDL had different metabolic properties, autologous LDL was isolated from subjects before and during colestipol therapy and their fractional catabolic rates (FCR) were then simultaneously determined in the same patient while on therapy. Eight LDL turnover studies comparing the catabolism of LDL isolated during therapy (Rx-LDL) and LDL isolated off therapy (Con-LDL) were performed in six subjects. All subjects responded to colestipol treatment, with an average 29% fall in LDL cholesterol. In four of six subjects, and in six of eight studies, the FCR of Rx-LDL was substantially slower than that of Con-LDL. These studies demonstrate that a drug intervention may alter subpopulations of LDL particles in such a way that overall LDL composition is changed. This alteration may independently affect the intrinsic metabolic behavior of the LDL. We suggest that such drug- (or dietary-) induced changes in LDL composition need to be considered in kinetic studies designed to assess the overall impact of the perturbation being studied.

Human apolipoprotein E3 in aqueous solution. II. Properties of the amino- and carboxyl-terminal domains.

Hydrodynamic, chromatographic, and spectroscopic techniques were used to study the aqueous solution properties of the two structural domains of human apolipoprotein (apo) E3. An amino-terminal thrombolytic fragment of apoE (22 kDa, residues 1-191) and a carboxyl-terminal thrombolytic fragment of apoE (10 kDa, residues 216-299) were used as models for the two domains. Sedimentation equilibrium ultracentrifugation showed that apoE and the 10-kDa model domain self-associated predominantly as tetramers. The 22-kDa model domain was primarily monomeric. Molecular weights calculated from the weight average sedimentation and diffusion coefficients or from the sedimentation coefficients and Stokes radii were in agreement with the sedimentation equilibrium results. Derived frictional coefficients suggest larger axial ratios and/or more extensive hydration for the apoE and the 10-kDa domain tetramers as compared with the 22-kDa domain. Proteolysis of apoE followed by high performance liquid chromatography showed rapid production of free 22-kDa domain, whereas the free 10-kDa domain appeared as a tetramer late in the course of the hydrolysis. Assessment by circular dichroism demonstrated that both model domains and apoE had over 54% alpha-helical content, which changed little in a detergent (octyl-beta-D-glucopyranoside) or lipid (dimyristoylphosphatidylcholine) environment. In contrast to the circular dichroism results, apoE and the 10-kDa domain showed a marked blue shift in the fluorescence maximum in a lipid environment. The results suggest that the self-association of apoE in solution as a tetramer is mediated by the carboxyl-terminal domain and that the amino- and carboxyl-terminal domains do not associate with one another. The amino-terminal domain is most likely compact and globular, whereas the carboxyl-terminal domain is probably elongated. The isolated model domains appear to have structures that are similar to those of the domains in the intact protein.

Coordinate changes in levels of human serum low and high density lipoprotein subclasses in healthy men.

Measurement of serum lipoproteins by analytic ultracentrifugation revealed significant correlations involving subfractions of low density (LDL) and high density (HDL) lipoproteins in 81 men studied cross-sectionally at baseline and longitudinally during a 1-year exercise trial. One-year changes in lipoprotein levels in 38 exercising men and in 30 sedentary controls showed correlations that paralleled those observed at baseline. Positive correlations observed between plasma levels of larger, more buoyant LDL of flotation rate (Sof) 7 to 10 (LDL-I) and HDL2 may be due to processes that also coregulate changes in levels of these lipoprotein subclasses. Similarly, positive correlations among smaller, more dense LDL of Sof 2 to 6 (LDL-III), IDL, and HDL3 suggest that levels of these lipoprotein species are also coordinately regulated. An inverse correlation of change in LDL-I with change in LDL-III raises the possibility of precursor-product relationships between LDL in these categories. Thus, changes in lipoproteins which are related to coronary disease risk are not independent of one another, and the development of coronary disease may be influenced by processes linking the metabolism of individual IDL, LDL, and HDL components.

Lipoprotein metabolism during acute inhibition of lipoprotein lipase in the cynomolgus monkey.

To clarify the role of lipoprotein lipase (LPL) in the catabolism of nascent and circulating very low density lipoproteins (VLDL) and in the conversion of VLDL to low density lipoproteins (LDL), studies were performed in which LPL activity was inhibited in the cynomolgus monkey by intravenous infusion of inhibitory polyclonal or monoclonal antibodies. Inhibition of LPL activity resulted in a three- to fivefold increase in plasma triglyceride levels within 3 h. Analytical ultracentrifugation and gradient gel electrophoresis demonstrated an increase predominantly in more buoyant, larger VLDL (Sf 400-60). LDL and high density lipoprotein (HDL) cholesterol levels fell during this same time period, whereas triglyceride in LDL and HDL increased. Kinetic studies, utilizing radiolabeled human VLDL, demonstrated that LPL inhibition resulted in a marked decrease in the catabolism of large (Sf 400-100) VLDL apolipoprotein B (apoB). The catabolism of more dense VLDL (Sf 60-20) was also inhibited, although to a lesser extent. However, there was a complete block in the conversion of tracer in both Sf 400-100 and 60-20 VLDL apoB into LDL during LPL inhibition. Similarly, endogenous labeling of VLDL using [3H]leucine demonstrated that in the absence of LPL, no radiolabeled apoB appeared in LDL. We conclude that although catabolism of dense VLDL continues in the absence of LPL, this enzyme is required for the generation of LDL.

Effects of saturated and polyunsaturated dietary fat on the concentrations of HDL subpopulations in African green monkeys.

The effect of the type of dietary fat on the concentrations and compositions of high density lipoprotein (HDL) subpopulations was studied in groups of African green monkeys consuming 40% of calories as fat supplied as saturated fat (P/S = 0.3) or polyunsaturated fat (P/S = 2.2) in the presence of either 0.8 mg or 0.03 mg cholesterol/kcal. Plasma HDL cholesterol concentrations were lower in polyunsaturated fat-fed animals. The distribution of mass among HDL subfractions was assessed by analytic ultracentrifugation (AnUC), density gradient ultracentrifugation (DGUC), and polyacrylamide gradient gel electrophoresis (GGE). This made it possible to characterize and quantitate the HDL subpopulations HDL2b, HDL2a, HDL3a, HDL3b, and HDL3c (arranged in order of decreasing particle size and decreasing cholesterol content). Polyunsaturated fat-fed animals had lower concentrations of the large, cholesterol-rich HDL2b subpopulation, as well as higher concentrations of intermediate size HDL (HDL2a and HDL3a on the high cholesterol diet; HDL3a and HDL3b on the low cholesterol diet). Consistent with the observed fat-related redistribution of HDL mass, the saturated fat-fed monkeys had higher apo A-I/apo A-II ratios. The larger HDL often contained detectable apo E; however, the concentration of apo E in HDL was low in both saturated and polyunsaturated fat-fed animals. Thus, compared to saturated fat, dietary polyunsaturated fat induced the formation of smaller size HDL subpopulations and, therefore, an overall lower cholesterol content per particle for plasma HDL.

Intermediate-density lipoproteins and progression of coronary artery disease in hypercholesterolaemic men.

Lipoprotein mass concentrations were measured by analytical ultracentrifugation in a subset of 57 hypercholesterolaemic male participants in the National Heart, Lung, and Blood Institute Type II Coronary Intervention Study. 2-year changes in levels of intermediate-density lipoproteins (IDL) of flotation rate 12-20 were strongly predictive of progression of coronary artery disease at 5 years. Changes in serum mass concentrations of low-density lipoproteins (LDL; flotation rate 0-12), very-low-density lipoproteins (VLDL; flotation rate 20-400), high-density lipoproteins (HDL), and the HDL2 and HDL3 subfractions did not differ significantly between men with and without definite progression of coronary artery disease. The relation of IDL mass to disease progression remained significant (p less than 0.05) after adjustment for group assignment to cholestyramine treatment or placebo and was only slightly reduced (p less than or equal to 0.06) by adjustment for changes in LDL mass concentrations. Changes in IDL mass and ratios of HDL-cholesterol to total-cholesterol or LDL-cholesterol were inversely correlated and had a similar ability to predict progression. The findings are consistent with earlier evidence that IDL are directly involved in the development of coronary artery disease and suggest that ratios of HDL-cholesterol to total-cholesterol or LDL-cholesterol may be indicators of coronary disease risk partly owing to relations with IDL metabolism.

Evidence for heterogeneity of low-density lipoprotein metabolism in the cynomolgus monkey.

Preliminary studies were performed to establish whether there was kinetic heterogeneity in the metabolism of subclasses of low-density lipoproteins (LDL) in the cynomolgus monkey. Previous studies of the effects of inhibition of hepatic triglyceride lipase in this species had shown an increase in the mass of lighter LDL (Sf greater than 9) and a decrease in the mass of denser LDL. LDL (1.019 less than d less than 1.063) were subdivided into two subfractions LDL1 (1.019 less than d less than 1.035) and LDL2 (1.035 less than d less than 1.063) by ultracentrifugation. The lipoproteins in these two fractions could be shown to have different flotation by analytic and isopycnic ultracentrifugation. When tracer amounts of homologous 125I-labeled very-low-density lipoproteins (VLDL) were injected into chow-fed cynomolgus monkeys, apoB radioactivity appeared in LDL1 prior to its appearance in LDL2. [125I]LDL1 injected into the monkey was removed from the LDL1 density subclass with a half-life of 5.5-10.3 h. Much of the radioactivity injected as LDL1 was converted to denser LDL (LDL2). Labeled LDL2 injected into the monkey was not converted to LDL1. Thus, at least two kinetically distinct subpopulations of LDL circulate in the plasma of this species. The lighter LDL is to a large extent a metabolic precursor of the more dense LDL (LDL2).

Lack of relationship in humans of the parameters of body cholesterol metabolism with plasma levels of subfractions of HDL or LDL, or with apoE isoform phenotype.

The factors involved in regulating parameters of whole body cholesterol metabolism in humans have been explored in a series of investigations. Several physiological variables have been identified (weight, excess weight, plasma cholesterol, and age) that can predict 53-76% of the variation in production rate (PR) and in the sizes of the rapidly exchanging pool of body cholesterol (M1) and of the minimum estimates of the slowly exchanging pool of body cholesterol (M3min) and of total body cholesterol (Mtotmin). Surprisingly, measurements of the plasma levels of HDL cholesterol and of the major HDL apolipoproteins (apoA-I, A-II, and E) did not provide additional information useful in predicting parameters of whole body cholesterol metabolism. A study was therefore conducted to investigate possible relationships of the plasma levels of subfractions of lipoproteins, determined by analytic ultracentrifugation, and of apoprotein E phenotype, with the parameters of whole body cholesterol metabolism. Ultracentrifugal analysis of plasma lipoprotein subfractions was performed at the Donner Laboratory in 49 subjects; all of these subjects were currently undergoing whole body cholesterol turnover studies or had previously had such studies and were in a similar metabolic state as judged by plasma lipid and lipoprotein values. Apoprotein E phenotyping was carried out in 71 subjects. Differences in model parameters were sought among subjects with various apoprotein E phenotypes. Ultracentrifugal LDL subfractions Sof 0-2 (the region of LPa), Sof 0-7 (smaller LDL), Sof 7-12 (larger LDL), Sof 12-20 (IDL), and ultracentrifugal HDL subfractions Fo1.20 0-1.5 (smaller HDL3), Fo1.20 2-9 (larger HDL3 plus HDL2), and Fo1.20 5-9 (larger HDL2 or HDL2b) were examined for correlations with each other and with parameters of whole body cholesterol metabolism.

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.

Lipoprotein subfractions of runners and sedentary men.

Serum concentrations of lipoprotein mass by flotation rate were measured in 12 long-distance runners and 64 sedentary men by analytic ultracentrifugation. The runners had significantly lower serum mass concentrations of the smaller, denser low-density lipoprotein particles of flotation rates Sf 0-7 (including the LDL-II, LDL-III, and LDL-IV subspecies), very-low-density lipoprotein (VLDL) particles of Sf 20-400, and high-density lipoprotein (HDL) particles of flotation rates F1.20 0-1.5 (predominantly the HDL3 subspecies), and higher serum mass concentrations of HDL particles with flotation rates between F1.20 2.0-9.0 (including HDL2a and HDL2b and less dense particles belonging to HDL3) than did sedentary men. Lipoprotein lipase activity was higher, and hepatic lipase activity was lower in runners than in the sedentary men. Thus, the effects of endurance exercise training to lower LDL may be specific to the smaller, denser LDL particle region. Similarities in the lipoprotein mass profiles of the runners and the low-risk profiles of sedentary, middle-aged women suggest the effects of common metabolic factors possibly leading to reduced risk of coronary artery disease.

Lipoprotein abnormalities in primary biliary cirrhosis. Association with hepatic lipase inhibition as well as altered cholesterol esterification.

The nature and etiology of plasma lipoprotein abnormalities in patients with varying stages of primary biliary cirrhosis (PBC) were assessed. Two distinct lipoprotein patterns were observed. In patients with early and intermediate histologic stages of PBC, mild elevations of very low density lipoproteins and low density lipoproteins (LDLs), and marked increases in high density lipoproteins (HDLs) were often noted (group 1). In contrast, patients with advanced disease had marked elevations in LDLs with the presence of lipoprotein-X, and a significant decrease in HDLs (group 2). The lipoprotein pattern in group 1 was characterized by a normal ratio of free cholesterol to total cholesterol, whereas in group 2, this ratio was elevated. In plasma from group 1 patients only spherical micelles were observed on electron microscopy, whereas in plasma from group 2 patients bilayered disks could be seen in LDLs and HDLs. The increase in HDLs observed in group 1 subjects was associated with elevations in HDL2, whereas in group 2 subjects only very low amounts of HDL3 were observed on analytic ultracentrifugation. Mean plasma apolipoprotein B and C-II concentrations were increased in both groups; but apolipoprotein A-I and AII values were divergent, with group 1 subjects having elevated values,and group 2 subjects having decreases values. Mean postheparin hepatic lipase activity was decreased in both groups of patients with PBC due to the presence of an inhibitor in PBC plasma, while altered cholesterol esterification was only observed in group 2. These data indicate that hepatic lipase inhibition may play an important role in the pathogenesis of the lipoprotein abnormalities seen in early and intermediate PBC, and these abnormalities are then further modified by altered cholesterol esterification in advanced disease.

Familial apolipoprotein A-I and C-III deficiency, variant II.

The biochemical, clinical, and genetic features were examined in the proband (homozygote) and heterozygotes (n = 17) affected with familial apolipoprotein A-I and C-III deficiency, variant II (previously described as apolipoprotein A-I absence). The proband was a 45-year-old white female with mild corneal opacification and significant three-vessel coronary artery disease (CAD), who died shortly after bypass surgery. Autopsy findings included significant atherosclerosis in the coronary and pulmonary arteries and the abdominal aorta as well as extracellular stromal lipid deposition in the cornea. No reticuloendothelial lipid deposits in the liver, bone marrow, or spleen were noted (unlike Tangier disease). Laboratory features included marked high density lipoprotein (HDL) deficiency and undetectable plasma apolipoproteins (apo) A-I and C-III. The percentage of plasma cholesterol in the unesterified form was normal at 30%. The activity and mass of lecithin:cholesterol acyltransferase (LCAT) were 42% and 36% of normal, respectively, and the cholesterol esterification rate was 43% of normal. Deficiencies of plasma vitamin E and essential fatty acid (linoleic, C18:2) were also noted. Evaluation of plasma lipoproteins and apolipoproteins in 37 kindred members revealed 17 heterozygotes with HDL cholesterol values below the 10th percentile of normal. Of these, all had apoA-I levels more than one standard deviation below the normal mean, and 37.5% had a similar decrease in apoC-III values. Mean (+/- SD) plasma HDL cholesterol, apoA-I, and apoC-III values (mg/dl) in heterozygotes were 54.0%, 62.4%, and 79.2% of normal, respectively. No evidence of CAD was observed in 10 heterozygotes 40 years of age or less; however, CAD was detected in 3 of 7 heterozygotes over 40 years of age, one of whom died at age 56 years of complications of myocardial infarction and stroke. The inheritance pattern in this kindred was autosomal codominant. ApoA-I isolated from a heterozygote had an isoelectric focusing pattern and amino acid composition similar to normal. Utilizing DNA isolated from two obligate heterozygotes, no abnormalities in the apoA-I or apoC-III genes were detected by Southern blot analysis utilizing specific probes following restriction enzyme digestion. The data indicate that familial apolipoprotein A-I and C-III deficiency, variant II, is similar to variant I (described by Norum et al. 1982. N. Engl. J. Med. 306: 1513-1519), but differs at the clinical level (lack of xanthomas), the biochemical level (lack of detectable apoA-I, lower apoA-II level), and at the gene level.(ABSTRACT TRUNCATED AT 400 WORDS)

Plasma lipoprotein abnormalities associated with acquired hepatic triglyceride lipase deficiency.

Two enzymes, lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL), are released into human plasma after intravenous injection of heparin. LPL is the major enzyme responsible for initiating catabolism of chylomicrons and very-low-density lipoproteins (VLDL). The physiological role of HTGL is less certain. HTGL has been postulated to be an alternate enzyme to LPL in hydrolysis of triglyceride in VLDL and to be an important enzyme for removal of phospholipid from both low-density lipoproteins (LDL) and high-density lipoproteins (HDL). In this latter role, this enzyme would convert larger, lighter lipoprotein particles to smaller denser particles. HTGL deficiency has been found in severe liver disease and with a genetic deficiency of this enzyme. A unique patient is described with acquired hepatic triglyceride lipase deficiency and vitamin A intoxication. This patient developed hypercholesterolemia with an increase in both LDL and HDL. An increased proportion of lighter LDL (LDL1) and HDL (HDL2) was noted. In addition, after administration of heparin there was no shift in the distribution of apoE in plasma fractionated using a column containing 4% agarose. These findings are consistent with a postulated role of HTGL in metabolism of light LDL and HDL particles and some classes of apoE containing lipoproteins.

Characterization of plasma lipids and lipoproteins in patients with beta 2-glycoprotein I (apolipoprotein H) deficiency.

The fasting plasma lipids, lipoproteins, and apolipoproteins were evaluated in 5 subjects with undetectable levels of the plasma protein beta 2-glycoprotein I (apolipoprotein H). Family studies confirmed an autosomal co-dominant inheritance pattern for the concentrations of apo H. The total lack of this protein is rare and less than 0.3% of clinic patients demonstrated levels undetectable by radial immunodiffusion. Plasma lipoprotein evaluation in these subjects with beta 2-glycoprotein I absence by analytical ultracentrifugation and compositional analysis demonstrated low concentrations of HDL2b and HDL3. More striking, however, was the lack of a consistent marked effect on the plasma lipoproteins as is found in other apolipoprotein deficiency states. We conclude that the lack of apolipoprotein H does not result in a significant perturbation of normal lipoprotein metabolism as reflected by analysis of fasting plasma lipoproteins. Further study is required to evaluate the role of this glycoprotein in the metabolism of triglyceride-rich lipoproteins.

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.

Serum lipid and lipoprotein concentrations following exposure to ozone.

The effects of exposure to ozone (O3) on concentrations of serum lipids and lipoproteins were investigated. Male and female guinea pigs were exposed to O3 at 1 ppm for two weeks. Serum concentrations of cholesterol, triglycerides, low density (LDL) and very low density (VLDL) lipoproteins were elevated after O3 exposure, particularly in males. During O3 exposure the food intake per day decreased (for a constant body weight), suggesting that metabolic rate and possibly basal metabolic rate was lower. Lung wet weights increased during O3 exposure by 87% for males and 45% for females. When individual lung weight/body weight ratios were correlated with cholesterol and LDL values from the same animal, a high correlation is found for males (r = 0.81, P less than 0.05), suggesting that there may be a relationship between lipoprotein elevations and lung damage for males. Because elevated concentrations of lipids and lipoproteins in humans increase the risk of coronary heart disease (CHD), the lipoprotein results suggest that an epidemiological study of the incidence of CHD with metropolitan O3 levels may be warranted.