J Lipid Res
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Free fatty acids (FFA) released during the lipolysis of triglyceride (TG)-rich lipoproteins in vivo are generally believed to be bound to serum albumin. When hypertriglyceridemic (HTG) sera were lipolyzed in vitro by purified bovine milk lipoprotein lipase (LpL), there was an 11- to 18-fold increase in serum FFA levels, and a major portion (> 80%) of the FFA in serum was partitioned to lipoprotein fractions. The greatest portion (33%) of FFA in lipolyzed HTG serum was associated with newly formed flocculent remnants that banded just below low density lipoproteins (LDL) in the density gradient tube. ⋯ The cytotoxic potencies of FFA bound to lipoproteins and albumin were further compared after in vitro incorporation of FFA (oleic acids) into LDL and to albumin. FFA bound to LDL but not to albumin were cytotoxic to cultured MPM; the cytotoxicity of FFA bound to LDL was more closely related to the FFA to LDL-cholesterol molar ratio than to the total FFA concentration in the culture dish. The ability of FFA bound to LDL and albumin to induce foam cell formation was studied in THP-1 monocyte-derived macrophages, which were less susceptible to cytotoxicity produced by FFA bound to LDL than MPM.(ABSTRACT TRUNCATED AT 400 WORDS)
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We have developed a sandwich-enzyme immunoassay (EIA) for the quantification of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) in human postheparin plasma (PHP) using monoclonal antibodies (MAbs) directed against the corresponding enzymes purified from human PHP. The sandwich-EIA for LPL was performed by using the combination of two distinct types of anti-LPL MAbs that recognize different epitopes on the LPL molecule. The immunoreactive mass of LPL was specifically measured using a beta-galactosidase-labeled anti-LPL MAb as an enzyme-linked MAb, an anti-LPL MAb linked with the bacterial cell wall as an insolubilized MAb, and purified human PHP-LPL as a standard. ⋯ Both assay methods yielded a highly significant correlation in either normolipidemic (r = 0.945 for LPL; r = 0.932 for HTGL) or hypertriglyceridemic subjects (r = 0.989 for LPL; r = 0.954 for HTGL). The normal mean (+/- SD) level of lipoprotein lipase mass and activity in postheparin plasma was 223 +/- 66 ng/ml and 10.1 +/- 2.9 mumol/h per ml, and that of hepatic triglyceride lipase mass and activity was 1456 +/- 469 ng/ml and 26.4 +/- 8.7 mumol/h per ml, respectively. The present sandwich-enzyme immunoassay methods make it possible to study the molecular nature of LPL and HTGL in PHP from patients with either primary or secondary hyperlipoproteinemia.
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Comparative Study
Comparison of apolipoprotein B to cholesterol in low density lipoproteins of patients with coronary heart disease.
This study was carried out to determine whether patients with coronary heart disease (CHD) have an unusually high level of apolipoprotein B (apoB) relative to cholesterol (C) in low density lipoproteins (LDL). Seven groups of men were studied. Seventy-two with normolipidemia (NLP) had CHD documented on clinical grounds; another 34 NLP patients had proven coronary artery disease (CAD) by angiography (greater than 50% occlusion of two or three coronary arteries). ⋯ HTG patients with CHD and CAD however tended to have higher LDL-apoB levels, and their LDL-apoB/C ratios were higher on average than normal. Nevertheless, among all coronary groups, there were no sizable subgroups with elevated LDL-apoB; only about 11% of all coronary patients had LDL-apoB levels over 120 mg/dl (compared to 8% for normo- and hypertensive controls of middle age). The data of this study therefore suggest that LDL-apoB may not prove to be a better indicator of coronary risk in normolipidemic people, but LDL-apoB could be a superior predictor of risk in HTG patients.
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The aim of this study was to define the role of the alpha-adrenergic receptor in the regulation of lipolysis by human adipocytes. Glycerol production by isolated human adipocytes was stimulated by the pure beta-adrenergic agonist isoproterenol in a dose-dependent fashion. ⋯ Whereas the attenuation of beta-adrenergic agonist-stimulated lipolysis by alpha-adrenergic agonists was reversed completely by the alpha 2-adrenergic antagonist yohimbine, the alpha 1-antagonist prazosin did not reverse such attenuation. It is concluded that alpha-adrenergic agonists act as antilipolytic agents in human adipocytes and that this action may result from the interaction of these compounds with a population of alpha 2-adrenergic receptors.
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A diet supplemented with cholesterol and coconut oil is atherogenic in dogs. The purpose of the present study was to examine the effects of this diet on red cells in pure-bred beagles and greyhounds. Within 3 days after the initiation of this diet red cell cholesterol/phospholipid increased and membrane fluidity decreased, with maximum changes attained by 12 weeks. ⋯ Red cell morphology resembled acanthocytes or spur cells. All red cell parameters returned to normal within 4 weeks after stopping the diet. These studies demonstrate that a cholestrol-enriched, atherogenic diet causes profound and reversible changes in the lipid composition, membrane fluidity, and morphology of red cells in dogs.