Very-low-density lipoprotein subfraction composition and metabolism by adipose tissue.

Lipoprotein lipase (LPL) plays a pivotal role in very-low-density lipoprotein (VLDL) metabolism. Within the circulation, the VLDL population is heterogeneous with respect to both size and composition. Several studies have investigated the action of LPL in vitro on different VLDL subfractions, but little is known of the action of LPL in vivo. To investigate this, arterial and adipose tissue venous plasma samples were obtained from 16 normal male healthy volunteers (aged 24.4 +/- 1.8 years; body mass index, 23.5 +/- 0.7 kg.m-2) following an overnight fast. VLDL subfractions were isolated (VLDL1 of Sf 60 to 400 and VLDL2 of Sf 20 to 60) and characterized in terms of triacylglycarol (TAG) and apolipoprotein (apo) B, E, CI, CII, and CIII content. The apolipoprotein content of VLDL1 differed from that of VLDL2: the VLDL2 fraction contained significantly more apo B (0.018 +/- 0.004 v 0.011 +/- 0.003 mumol.L-1, p = .001) but the ratios of TAG:apo B and apo CI:B, and CII:B, and CIII:B were significantly higher in VLDL1 (48,200 +/- 7,980 v 13,860 +/- 2,420, 22.7 +/- 5.5 v 12.5 +/- 2.2, 45.0 +/- 6.3 v 14.9 +/- 2.0, and 0.434 +/- 0.077 v 0.357 +/- 0.054, respectively, molar ratios, all P < .05). The venous blood draining an adipose tissue depot contained less VLDL1-TAG than arterial blood (328 +/- 68 v 381 +/- 83 mumol.L-1, respectively, P < .01), whereas VLDL2-TAG exhibited an opposite tendency (199 +/- 46 v 172 +/- 31 mumol.L-1, NS). Concentrations of VLDL1-apo B, -apo CII, and -apo CIII were significantly less in adipose tissue venous blood compared with arterial blood (0.011 +/- 0.004 v 0.013 +/- 0.004, 0.38 +/- 0.08 v 0.43 +/- 0.10, and 1.33 +/- 0.35 v 1.58 +/- 0.38 mumol.L-1, respectively, all P < .05). These studies demonstrated novel differences in VLDL1 and VLDL2 in terms of composition and metabolism by human adipose tissue LPL in vivo.

The effect of the treatment of hypothyroidism and hyperthyroidism on plasma lipids and apolipoproteins AI, AII and E.

OBJECTIVE: Although lipid abnormalities are well described in hypothyroidism, effects on apolipoproteins are less well understood. The aim of this study was to examine the effects of thyroid dysfunction on plasma lipids and apolipoproteins. DESIGN: A prospective study of lipids and apolipoproteins before and after treatment of hypothyroidism and hyperthyroidism. PATIENTS: Eighteen patients with hypothyroidism and 5 patients with hyperthyroidism were included. MEASUREMENTS: Plasma cholesterol, triglycerides, HDL cholesterol, apo AI, apo AII, and apo E were measured before and after treatment of the thyroid abnormality. RESULTS: Total and HDL cholesterol, apo AI and apo E decreased with treatment of hypothyroidism, while triglycerides and apo AII levels were unchanged. The total/HDL cholesterol and LDL/HDL cholesterol ratios also decreased with treatment of hypothyroidism. In contrast, treatment of hyperthyroidism was associated with an increase in total and HDL cholesterol, and apo AI. Triglycerides, apo AII and Apo E were unchanged by treatment of hyperthyroidism. The total/HDL cholesterol and the LDL/HDL cholesterol ratios increased with treatment of hyperthyroidism. CONCLUSIONS: Hypothyroidism and hyperthyroidism have opposite effects on plasma lipids and apolipoproteins. In hypothyroidism, total and HDL cholesterol, total/HDL cholesterol ratio, apo AI and apo E are elevated. The increase in apo AI without a concomitant increase in apo AII suggests selective elevation of HDL2. In contrast, hyperthyroidism is associated with decreased total and HDL cholesterol, total/HDL cholesterol ratio, and apo AI levels. These effects are reversible with treatment of the underlying thyroid disorder.

HDL subparticles and coronary artery disease in NIDDM.

Decreased HDL cholesterol levels are associated with an increased risk of coronary artery disease (CAD) in non insulin dependent diabetes mellitus (NIDDM). The aim of this study was to compare HDL subparticles with apo A-I (LpAI) and those with apo A-I and apo A-II (LpAI/AII) in subjects with and without NIDDM and to study the relationship between HDL subparticles and CAD in NIDDM. Lipids, apo A-I and HDL subparticles were measured in 240 subjects with NIDDM and in 248 age and gender matched controls. Subjects with NIDDM had higher triglyceride levels (2.5 +/- 1.8 vs. 1.4 +/- 0.8 mmol/1, P < 0.001), lower HDL cholesterol (0.9 +/- 0.3 vs. 1.2 +/- 0.3 mmol/l, P < 0.001), apo A-I (124.7 +/- 22.4 vs. 139.8 +/- 24.1 mg/dl, P < 0.001) and LpAI/AII (82.4 +/- 18.2 vs. 94.9 +/- 16.7 mg/dl, P < 0.001) in comparison to controls. LpAI levels were similar in both groups. Diabetic subjects with CAD (n = 109) had higher triglycerides (2.7 +/- 1.9 vs. 2.3 +/- 1.8 mmol/l, P = 0.02) and lower HDL cholesterol (0.8 +/- 0.2 vs. 1.0 +/- 0.3 mmol/l, P <0.001), apo A-I (115.5 +/- 20.1 vs. 132.3 +/- 21.4 mg/dl, P < 0.001), LpAI (40.2 +/- 9.1 vs. 44.4 +/- 12.4 mg/dl, P = 0.06), and LpAI/AII levels (75.4 +/- 18.0 vs. 88.3 +/- 16.2 mg/dl, P < 0.001) in comparison to diabetic subjects without CAD (n = 131). In a multivariate analysis, apo A-I was found to be the best predictor of CAD in subjects with NIDDM. In conclusion, reduced HDL cholesterol levels found in NIDDM are, principally, due to reduced concentrations of apo A-I and apo A-II-containing particles (LpAI/AII). While LpAI and LpAI/AII levels were lower in NIDDM subjects with CAD, plasma apo A-I is the best predictor of CAD in NIDDM.

Overview of lipoprotein metabolism: mechanisms involved and conveying this information to patients.

The major classes of lipoprotein particles are chylomicrons, very-low-density lipoproteins, low-density lipoproteins, and high-density lipoproteins. Each of these particles has distinct characteristics, including a distinct apolipoprotein composition. The basic functions of these major apolipoproteins are briefly summarized for clinicians, and disorders of lipoprotein metabolism are discussed. For the effective management of lipid disorders, the therapy should be tailored to the specific defect. Compliance may be improved if the patient understands the nature of the particular lipid problem. To that end, we have developed an analogy in which the various classes of lipoprotein particles are equated to different trucking systems, and the liver is considered a warehouse for storage and distribution of fat. With use of this explanation, patients may understand the rationale for selection of various therapeutic strategies, and treatment of the hyper-lipoproteinemia is likely to be more successful.

The role of lipoprotein A-I and lipoprotein A-I/A-II in predicting coronary artery disease.

The aim of this study was to examine the role of HDL subparticles with apolipoprotein (apo) A-I alone (LpA-I) and with apoA-I and apoA-II (LpA-I/A-II) in predicting coronary artery disease. Concentrations of these HDL subparticles were compared in 184 subjects with angiographically confirmed significant coronary artery disease (> 50% stenosis of at least one vessel) and 191 age- and sex-matched control subjects without clinical coronary artery disease. LpA-I and LpA-I/A-II were measured with magnetic beads coated with anti-apoA-II antibodies to separate particles containing apoA-II from plasma. Total plasma cholesterol and triglyceride levels were similar in both groups. Although subjects with coronary artery disease had lower HDL cholesterol, plasma apoA-I, LpA-I, and LpA-I/A-II than age- and sex-matched control subjects without coronary artery disease, plasma apoA-I was the best predictor of coronary artery disease. In conclusion, LpA-I and LpA-I/A-II are lower in subjects with coronary artery disease but do not add to plasma apoA-I in predicting the presence of coronary artery disease.

Increase in total plasma homocysteine concentration after cardiac transplantation.

OBJECTIVE: To determine whether plasma homocysteine concentrations are increased in patients after cardiac transplantation. DESIGN: Total plasma homocysteine concentration was measured in 44 consecutive patients before and at 3, 6, and 12 months after orthotopic heart transplantation between June 1, 1988, and Oct. 15, 1992, and the data were analyzed statistically. RESULTS: Mean homocysteine concentrations (normal range, 4 to 17 mumol/L) increased 70% from 12.5 mumol/L before cardiac transplantation to 21.2 mumol/L (P < 0.002) 3 months after transplantation, at which time the concentrations were above normal in 14 of 26 patients (54%). Homocysteine concentrations remained elevated 6 and 12 months after transplantation (20.4 and 22.6 mumol/L, respectively) but did not increase further. Mean concentrations of plasma folic acid and vitamin B12, cofactors in homocysteine metabolism, decreased 20% and 49%, respectively, within 3 months after transplantation (11.6 to 9.3 micrograms/L [P = 0.04] and 584 to 295 ng/L [P = 0.01]). The mean glomerular filtration rate decreased 25% during this same interval (81 to 61 mL/min; P = 0.0001). Linear regression analysis revealed an association between the increase in homocysteine concentration and the folic acid concentration that approached statistical significance (P = 0.07); we found no statistically significant correlates of the increase in homocysteine concentration. CONCLUSION: The homocysteine concentration increases in most patients within 3 months after cardiac transplantation to levels previously associated with premature atherosclerotic coronary artery disease, and it remains increased for at least 1 year. Further investigation into the mechanism for the increase in homocysteine concentration and the relationship between homocysteine and coronary artery disease after transplantation is warranted.

Serum lipid responses to a eucaloric high-complex carbohydrate diet in different obesity phenotypes.

OBJECTIVE: To determine whether the eucaloric substitution of complex carbohydrates for dietary fat (15% of daily energy intake) affects plasma lipid concentrations differently in upper-body obese, lower-body obese, and nonobese women. DESIGN: We studied 23 premenopausal women before and after dietary intervention. MATERIAL AND METHODS: After the 23 subjects achieved weight maintenance on their usual high-fat diet (43% fat, 37% carbohydrates, and 20% protein), the 7 upper-body obese, 8 lower-body obese, and 8 nonobese women consumed a eucaloric, high-complex carbohydrate, low-fat diet (27% fat, 53% carbohydrates, and 20% protein) for 4 weeks in the Clinical Research Center. Before and after the high-carbohydrate diet, body composition and plasma lipids and apoproteins were measured. RESULTS: After the high-carbohydrate diet, fasting plasma triglyceride concentrations increased (from 1.50 +/- 0.14 mmol/L to 2.00 +/- 0.25 mmol/L; P = 0.04) in upper-body obese women but were not significantly changed in lower-body obese (1.37 +/- 0.28 mmol/L and 0.96 +/- 0.12 mmol/L) or nonobese (0.70 +/- 0.08 mmol/L and 0.73 +/- 0.08 mmol/L) women. The hypertriglyceridemia was present before the evening meal and throughout the night in upper-body obese women. Plasma cholesterol and high-density lipoprotein cholesterol were not significantly affected by the change in diet. No changes in plasma apoprotein concentrations or body composition occurred that could account for the dietary-induced hypertriglyceridemia in the women with upper-body obesity. CONCLUSION: The hypertriglyceridemic response to a high-complex carbohydrate, low-fat diet may be obesity phenotype specific. These findings suggest that further studies of this phenomenon should be focused on this obesity phenotype and further emphasize the importance of assessing body fat distribution when treatment outcomes are determined.

A fluorescence-conjugated immunobinding assay for the detection of P-selectin on platelets.

P-selectin (GMP-140 or PADGEM) is translocated to the plasma membrane of platelets after platelet activation. P-selectin, therefore, may be a potential marker for evaluating platelet activation. A fluorescence-conjugated immunobinding assay (FCIBA) has been developed to detect specifically P-selectin on platelets. Platelets were isolated from fresh blood by centrifugation and stimulated with various doses of ADP before being fixed with 1% of paraformaldehyde. Fixed platelets were incubated with fluorescence-conjugated anti-P-selectin monoclonal antibody in the wells of fluoricon microtiter plates, and the fluorescence intensity was read on a fluorescence concentration analyzer. Once platelets were fixed, the procedures were completed in < 2 hours. The intra-assay coefficient of variation (CV) was 6.97% (n = 40), the time-based interassay CV was 8.11% (n = 16), and the sample-based inter-assay CV was 6.17% (n = 16). The FCIBA had an excellent correlation (r = 0.936, p < 0.001) with flow cytometry in the measurement of expressed P-selectin in platelets of 20 normal donors. Translocation of P-selectin in plasma-suspended platelets in response to increasing doses of adenosine diphosphate (ADP) occurred in a dose-dependent manner and correlated positively with ADP-induced platelet aggregation in terms of both stimulating doses of ADP (r = 0.99, p < 0.01) and time intervals (r = 0.92, p < 0.05). The findings show that FCIBA is a fast and convenient assay with good precision for the determination of P-selectin expression of human platelets.

Effect of heparin-binding growth factor-1 on bronchial healing in canine lung allograft transplantation.

Failure of airway healing complicates lung transplantation. Local application of heparin-binding growth factor-1 induces neovascularization in vivo. To determine the effect of direct application of heparin-binding growth factor-1 on bronchial healing, we performed single left lung allotransplantations in 12 dogs, and wrapped the bronchial anastomosis with Gelfoam impregnated with either recombinant heparin-binding growth factor-1 (100 micrograms, n = 6) or saline solution (n = 6) in a blinded fashion. After 21 days, the bronchial anastomosis was studied by gross examination, light and electron microscopy, bromodeoxyuridine uptake, measurement of bronchial breaking strength, and high-speed computerized tomography to assess bronchial cross-sectional area and distensibility. The transplanted lung was examined histologically for concomitant lung rejection. No significant difference was found between the two groups in histologic bronchial healing scores, proliferation index, bronchial breaking strength, cross-sectional area, anastomotic distensibility, or local blood flow as estimated by high-speed computerized tomography scan. In the growth factor group, both increased neovascularization and lung rejection were found at histologic evaluation. We conclude that, although the use of heparin-binding growth factor-1 increased perianastomotic neovascularization, it did not contribute to improved bronchial healing, possibly because of increased lung rejection.

Lipids and Lp(a) lipoprotein levels and coronary artery disease in subjects with non-insulin-dependent diabetes mellitus.

OBJECTIVE: To determine whether increased Lp(a) lipoprotein levels are associated with either non-insulin-dependent diabetes mellitus (NIDDM) or coronary artery disease (CAD) in patients with NIDDM and to examine the relationship between Lp(a) levels and glycemic control. DESIGN: We conducted a cross-sectional study of subjects with NIDDM who were participants in the Rochester Diabetic Neuropathy Study and healthy control subjects from the population of Rochester, Minnesota. MATERIAL AND METHODS: Lipids and Lp(a) lipoprotein levels were compared in 227 subjects with NIDDM and 163 control subjects and, among the subjects with NIDDM, in those with (N = 96) and without (N = 131) CAD. The correlation between Lp(a) levels and glycosylated hemoglobin was investigated. RESULTS: Subjects with NIDDM had higher triglyceride and lower high-density lipoprotein cholesterol levels than did control subjects. Subjects with NIDDM and CAD had higher total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels and lower high-density lipoprotein cholesterol levels than did subjects with NIDDM without CAD. Subjects with NIDDM had significantly higher Lp(a) levels than did control subjects, but subjects with NIDDM and CAD did not have significantly higher Lp(a) levels than did those without CAD. Among subjects with NIDDM, the level of Lp(a) was not significantly correlated with glycosylated hemoglobin. CONCLUSION: Although subjects with NIDDM have higher Lp(a) levels than do control subjects, Lp(a) does not seem to be associated with CAD in subjects with NIDDM. In this study, no association was found between Lp(a) level and glycemic control.

Immunomagnetic separation of subpopulations of apolipoprotein A-I.

OBJECTIVE: This study was undertaken to measure the subfractions of high-density lipoprotein (HDL) in patients with diabetes or coronary artery disease and in normal control subjects. DESIGN: A new immunomagnetic separation technique was used to characterize the lipid profile in four groups: (1) control subjects, (2) patients with diabetes but no coronary artery disease (CAD), (3) those with CAD only, and (4) those with both diabetes and CAD. MATERIAL AND METHODS: To study the individual roles of the two discrete HDL subpopulations of particles--LpAI/AII (apolipoprotein [apo] A-I associated with A-II) and LpAI (apo A-I without A-II)--in lipoprotein metabolism, we developed an immunomagnetic separation technique using magnetic beads coated with antibodies to human apo A-II. The beads bind particles that contain both apo A-II and apo A-I and are precipitated by a magnetic field. LpAI levels were measured in the supernatant by performing an apo A-I radioimmunoassay. LpAI/AII levels were determined by subtracting the LpAI levels from total plasma apo A-I. RESULTS: In comparison with control subjects, patients with diabetes, CAD, or both had significantly decreased levels of LpAI/AII. LpAI levels were normal in patients with diabetes without CAD but significantly lower than control values in those with diabetes and CAD. CONCLUSION: Our findings suggest that both subpopulations of HDL particles have implications in the development of atherosclerosis in patients with and without diabetes.

Lipoproteins and apolipoproteins. Making use of their metabolic properties to individualize therapy.

Plasma contains four major lipoprotein particles: chylomicrons, VLDLs, LDLs, and HDLs. How cholesterol is metabolized depends on the particle with which it is associated. Thus, treatment for hypercholesterolemia is most effective when it is designed to fit the lipoprotein features of a given patient. Dr Kottke describes the five common forms of hyperlipoproteinemia and provides specific therapeutic approaches for each.

A particle concentration fluorescence immunoassay for Lp(a).

The quantitation of Lp(a) by immunoassay presents a major technical problem, because the molecular mass of the (a) protein of Lp(a) can vary between 419,000 and 838,000 Da (Gaubatz et al. (1990) J. Lipid Res. 31, 603-612), and this variability is determined by at least 24 alleles of the (a) gene. In an attempt to overcome this problem, we have developed an assay that is independent of variation of the size of (a). The assay utilizes a mixture of monoclonal antibodies to (a) which do not react to plasminogen or to apolipoprotein (apo) B. These antibodies are bound to inert microscopic beads to capture the Lp(a) particles. Subsequently, a fluorescein-labeled monoclonal antibody to apo B is used for detection and quantitation. The assay is done with special microtiter plates containing filters so that the particles can be thoroughly washed after capture on the microbeads. Because Lp(a) particles contain only one apo B particle and the molecular weight of apo B is constant, the assay is not affected by variation in the size of apo(a). By binding the mixture of monoclonal antibodies to inert beads, it is possible to greatly increase the amount of antibody bound to an exposed surface and thus increase the sensitivity of the assay. A mixture of monoclonal antibodies can be used to increase the affinity of the capture step of the assay. The assay can be completed in 4 h and has a wide working range.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification and quantification of apolipoproteins in addition to apo[a] and apo B-100 in human lipoprotein[a].

The protein moiety of Lp[a] is widely believed to consist of one molecule of apo B-100 and one molecule of apo[a] per particle, linked by at least one disulfide bond. In this study we have re-examined the composition of Lp[a] to determine if other less abundant apolipoproteins might be present. Analysis of Lp[a] by sodium dodecyl sulfate-polyacrylamide electrophoresis under reducing conditions showed bands corresponding to < 200 kD but > 50 kD, 40 kD, 26 kD, 23 kD and 9 kD when stained with silver. Western immunoblot analysis of three preparations of Lp[a] revealed the presence of apoE and apoD. Enzyme-linked immunoassays were used to quantify apoA-I, apoA-II, apoC-I, apoC-II, apoC-III, apoE and apo B-100 in Lp[a] and autologous LDL isolated from three healthy males. There is a significant amount of apoA-I in the Lp[a], although the levels varied widely among the different samples. ApoE concentrations were consistent in the three Lp[a] samples and were between 22 and 26% of relative apo B-100 concentrations. Relatively minor amounts of apoA-II and no apoCs were detectable in the three Lp[a] preparations. In contrast, the autologous LDL preparations contained relatively higher amounts of apoA-I, apoA-II, apoE, apoC-I, apoC-II and apoC-III. The identity of the multiple bands corresponding to < 200 kD and > 54 kD and 9 kD is not established.

Lipoprotein compositional changes in the fasting and postprandial state on a high-carbohydrate low-fat and a high-fat diet in subjects with noninsulin-dependent diabetes mellitus.

Our aims were 1) to examine the effects of a high-carbohydrate low-fat diet on fasting and postprandial plasma lipids, apolipoproteins (apo), and lipoprotein composition in noninsulin-dependent diabetes mellitus, and 2) to determine whether postprandial shift of apo between triglyceride-rich lipoproteins (TRLP) and high density lipoproteins (HDL) is affected by diet. A cross-over study, of 4 weeks duration, of a high-carbohydrate (60% carbohydrate, 20% fat) and a high-fat (40% carbohydrate, 40% fat) diet was performed in seven subjects with noninsulin-dependent diabetes mellitus. TRLP, low density lipoproteins (LDL), and HDL were separated by fast protein liquid chromatography. The high-carbohydrate diet resulted in a decrease of fasting total, HDL, and LDL cholesterol and a trend toward an increase in plasma triglycerides. The apo composition of fasting TRLP and HDL was similar on both diets. TRLP apo CII, CIII, and E increased whereas HDL apo CII, CIII, and E decreased postprandially on both diets. In contrast, TRLP apo CI increased, and HDL apo CI decreased only after the high-carbohydrate diet. We conclude that 1) a high carbohydrate diet results in a decrease in total, LDL, and HDL cholesterol and a trend toward an increase in plasma triglycerides; 2) fasting TRLP and HDL apo composition was similar on a high-carbohydrate or a high-fat diet; and 3) on both diets, apo CII, CIII, and E transfer from HDL to TRLP postprandially. However, only the high-carbohydrate diet induced postprandial transfer of apo CI from HDL to TRLP. This may explain in part the changes in lipoproteins observed with this diet.

Current understanding of the mechanisms of atherogenesis.

Atherogenesis is a complex process involving several cell types, including endothelial cells, platelets, and smooth muscle cells. The development of atherogenesis depends on the modification of the function of these cells due to the interaction of cellular receptors with a variety of peptide hormones as well as with lipoprotein particles. Appropriate treatment of risk factors for atherogenesis depends on this mechanism and must be individualized to fit the major mechanisms present in each patient. New tools are emerging to improve the ability to tailor risk management to fit the needs of particular patient subgroups.