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Biomedical subjects

G M Kostner

Publications and source records attributed to G M Kostner.

At least 217 records · Page 12Linked to original sources

In vitro modification of the chemical composition of human plasma low density lipoproteins: effects of morphology and thermal properties.

The effects of enzymatic action on human low density lipoproteins (LDL) occurring during in vitro incubation of plasma have been studied by chemical analysis, analytical ultracentrifugation, negative stain electron microscopy and X-ray small angle scattering. Chemically, the action of cholesteryl ester exchange and transfer proteins(s) (CEPT) leads to a relative increase in trigylcerides at the expense of cholesteryl esters. Morphologically, the particles maintain their characteristic features detectable by X-ray small angel scattering. Additional action of lecithin/cholesterol acyl transferase (LCAT) causes mainly a decrease in polar lipid contents and a reduction in particle size. The associated changes in the thermotropic transition were found to be strongly correlated to the triglyceride/cholesteryl ester ratio.

Carrier Proteins↗

Total plasma apo E and high density lipoprotein apo E in survivors of myocardial infarction.

Total apo E in plasma and the amount of apo E-HDL were measured in 40 normolipidemic male survivors of myocardial infarction and in 40 controls. LDL-C, Lp(a) and apo B were significantly higher and HDL-C and apo A-I were significantly lower in survivors than in controls. Total plasma apo E did not differ between patients and controls, but HDL-E and the ratio HDL-E/apo A-I were lower in survivors. The data support the view that atherosclerotic patients are often characterized by abnormalities in the concentration and distribution of lipoproteins as well as of apoproteins, even in the presence of normal total plasma lipids.

Adult↗

Structure of the cholesteryl ester core of human plasma low density lipoproteins: selective deuteration and neutron small-angle scattering.

The structural arrangement of cholesteryl esters in human plasma low density lipoproteins (LDL) has been studied by selective deuteration and neutron small-angle scattering. LDL were labeled by in vitro exchange with two different kinds of deuterated cholesteryl esters, one labeled in the fatty acyl chain (cholesteryl myristate-d27) and the other in the branched side chain of cholesterol (cholesteryl-25,26,27-d7 oleate). Neutron scattering data from deuterated and protonated LDL were compared to identify the locations of the fatty acyl and cholesterol side chain moieties. Below the thermotropic transition, radii of gyration of 60 A and 70 A were obtained for these two domains, respectively, indicating that the cholesteryl nuclei are situated more distantly from the center than the fatty acyl chains. At 37 degrees C, above the thermotropic transition of the cholesteryl esters in LDL, both parts have similar radii of gyration of approximately 56 A. This information is used in a discussion of possible structural models for the apolar lipid core of LDL.

Cholesterol Esters↗

The interaction of human apoB-containing lipoproteins with mouse peritoneal macrophages: a comparison of Lp(a) with LDL.

Cholesteryl ester accumulation in macrophages and foam cell formation is believed to play an important role in atherogenesis. The effect of Lp(a) on the incorporation of [14C]oleate into cholesteryl esters was studied in mouse peritoneal macrophages. In view of the physico-chemical similarities between Lp(a) and LDL, the results were compared with those obtained with LDL. Native Lp(a) and LDL did not stimulate cholesteryl ester formation. Incubation of macrophages with Lp(a)- or LDL-dextran sulfate complexes caused a significant increase in cholesteryl ester formation. A similar effect was observed when Lp(a) or LDL were incubated with macrophages in the presence of antibodies directed against the specific Lp(a) apoprotein or against LpB. Treatment of Lp(a) with acetic anhydride or malondialdehyde (MDA) was followed by precipitation of most of the lipoprotein. Therefore, these modifications were not suitable to study the uptake of modified Lp(a) by macrophages. Studies with acetyl-LDL or MDA-treated LDL caused the well-known stimulation of [14C]oleate incorporation into cholesteryl esters. Thus, the modification of Lp(a) by sulfated polysaccharides or by treatment with antibodies yields similar cholesteryl ester deposition in mouse peritoneal macrophages as observed with modified LDL. This might be one mechanism by which Lp(a) exerts its atherogenicity.

Animals↗

The determination of lipoprotein Lp(a) by rate and endpoint nephelometry.

Rate nephelometry is a fast and convenient method for Lp(a) quantification. The linearity in the range of the physiological serum concentrations is good and there is good correlation with other immunochemical methods. The antibody consumption is some 50% higher when compared to the Laurell procedure if light scatter is measured in the RU mode. Measurements in the SU mode on the other hand consume less antibody than the Laurell procedure. The endpoint method, however, is more time-consuming and considered to be less accurate. In addition, only clear samples should be used. Lipemic sera should be cleared by lipase treatment or by high speed centrifugation. The treatment of samples with detergents causes a pronounced reduction of the immunochemical response and thus should be avoided.

Electrophoresis, Agar Gel↗

A novel assay for the quantitative determination of lipoprotein-X (LP-X) in serum.

A new, fast and simple quantitative LP-X assay is described. The method selectively removes apoB-containing lipoproteins by precipitation with specific antibodies. To the supernatant, a precipitation system for LP-X, viz. SDS and MgCl2 in final concentrations of 0.5 g/l and 0.2 mol/l respectively, is added and the turbidity measured at 360 nm. The assay is linear from 0.2 g/l-5.0 g/l and shows a high precision (inter-assay CV of less than 3%). The test correlates favourably with other quantitative LP-X assays, but has the particular advantage that it can be automated and that the time required for one series is only 2 h.

Antibodies↗

Apolipoproteins (A-I, A-II, B), Lp(a) lipoprotein and lecithin: cholesterol acyltransferase activity in diabetes mellitus.

Concentrations of HDL cholesterol, apolipoprotein (apo) A-I and apo A-II were found to be significantly decreased in patients with insulin-dependent diabetes (IDD) and non-insulin-dependent diabetes (NIDD) compared with carefully selected controls matched for sex, age and body weight. LDL cholesterol and apo B levels did not differ significantly between diabetics and controls. Concentrations of lipoprotein Lp(a), an independent risk factor for coronary artery disease in non-diabetics, were above 20 mg/dl in only 14% of diabetics and in 5% of controls. LCAT activity was normal in diabetics, irrespective of type of diabetes, sex and age of patients. No correlation between HbA1 and either HDL cholesterol or A-I and A-II was found in IDD and NIDD. A positive correlation between HbA1 and either triglyceride or VLDL triglyceride was noted in IDD and NIDD. There was also a positive correlation between insulin dosage in IDD and HDL cholesterol, apolipoprotein A-I and A-II.

Adolescent↗

Studies on the role of specific cell surface receptors in the removal of lipoprotein (a) in man.

The binding of 125I-lipoprotein (a) [Lp(a)] to cell surface receptors was studied on cultured human fibroblasts. The results were compared with corresponding data obtained with 125I-low density lipoproteins (LDL). Equilibrium binding studies showed that Lp(a) is bound with high affinity by the cell surface receptors. The maximum binding capacity for Lp(a) was 37% lower than for LDL. For Lp(a) and LDL, the Scatchard plots displayed linearity, indicating a single category of binding sites. Half-maximal saturation occurred at a concentration of 9.52 +/- 1.04 nM for Lp(a) and 7.76 +/- 1.29 nM for LDL. Competition binding experiments revealed that Lp(a) and LDL are nearly equally potent in competing each other for the binding sites. Binding of Lp(a) and LDL were followed by suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity. Cyclohexanedione treatment of Lp(a) and LDL completely abolished receptor binding. Neither Lp(a) nor LDL were specifically bound by fibroblasts obtained from a patient with homozygous familial hypercholesterolemia (FH). The removal mechanisms for Lp(a) and LDL were further compared by in vivo studies. Radioiodinated Lp(a) and LDL were injected intravenously into 12 normolipemic individuals to measure kinetic parameters of these two lipoproteins simultaneously in each subject. Mean fractional catabolic rate (FCR) of Lp(a) was 0.260 +/- 0.060 and mean FCR of LDL was 0.377 +/- 0.077 (mean +/- SD). In each subject, FCR of Lp(a) was lower than the FCR of LDL; the mean difference was 31%. The absolute synthetic rate of Lp(a) was significantly lower than the corresponding value of LDL. In each individual, the percentage of total Lp(a) that was contained in the intravascular space was higher than the corresponding value of LDL; the mean difference was 19%. A highly significant positive correlation was found between FCR of LDL and FCR of Lp(a) (r = 0.853, P less than 0.01). No relationship was found between the serum concentration of LDL-apolipoprotein B and Lp(a). The serum level of Lp(a) was positively related to the absolute rate of Lp(a) synthesis (r = 0.979, P less than 0.01). The serum level of LDL-apolipoprotein B was inversely related to FCR of LDL (r = 0.613, P less than 0.05). In a patient with homozygous FH, FCR of LDL was 0.205 and FCR of Lp(a) was 0.210. The results of these studies show that Lp(a) is specifically bound with high affinity to the same receptors of human fibroblasts as LDL. The affinity and maximum binding capacity are slightly lower for Lp(a) than for LDL. The results of the turnover studies are consistent with the assumption that Lp(a) is removed from the plasma by similar mechanisms as LDL.

Adolescent↗

Apolipoproteins and lipoproteins of human plasma: significance in health and in disease.

When DeLalla and Gofman (1954) presented their work "Ultracentrifugal Analysis of Serum Lipoproteins" more than 25 years ago, we were thinking about lipoproteins in terms of density fractions. In the 1970s the electrophoresis concept was pushed by Fredrickson and his colleagues (Fredrickson et al., 1967). There is no doubt that both these lipoprotein research centers have fertilized entire investigations in this field and still have a tremendous impact on our current knowledge. It was, however, not until 1966, when Gustafson, Alaupovic, and Furmann first described the presence of a third lipoprotein family, LpC, that researchers in this area became aware of the dominant role of apolipoproteins in the transport and metabolism of plasma lipids. Lipoprotein density fractions and electrophoretic classes in the mean time have not lost their importance; they still exist and the application of methods yielding those fractions is still going on in lipoprotein laboratories. Yet we need to recognize that the whole lipid transport system is far more complex than was believed some 10 or 20 years ago. Lipoprotein density fractions consist of varying numbers of families; some of them comigrate upon electrophoresis, and the protein moiety of them is mostly composed of nonidentical polypeptides. There are a number of inborn errors of metabolism, for example, ABL, Tangier disease, and enzyme defects, which have taught us a lot about the functions and interplay of the complex apolipoprotein system. In dyslipoproteinemia, abnormal lipoproteins occur in the plasma and apolipoproteins, which are hardly recognized in normal fasting plasma, suddenly become prominent. There still exist, however, apolipoproteins and lipoproteins, one of which certainly is Lp(a), whose function and biological significance remains completely unknown. The structure and the molecular arrangement of lipids and apolipoproteins within a lipoprotein particle has been the subject of intensive investigations, and almost every physicochemical method available has been applied to reveal the morphology of individual lipoproteins in closest detail. Lipoproteins and apolipoproteins have often also served as model substances for cell membranes. After the purification of individual apolipoproteins succeeded in many laboratories and specific antibodies were available, clinical chemists and epidemiologists became interested in this area of research. Apolipoprotein quantification currently is most prominent for the prediction of atherosclerotic risk in preventive medicine.(ABSTRACT TRUNCATED AT 400 WORDS)

Abetalipoproteinemia↗

Immunoquantification of total apolipoprotein B in serum by nephelometry: influence of lipase treatment and detergents.

The immunoquantification of total apolipoprotein B in human serum has been evaluated by rate and equilibrium nephelometry. The presence of triglyceride-rich lipoproteins spoiled all immunochemical assays and yielded too-high values for apolipoprotein B. The use of detergents improved the results substantially, but results were inaccurate at high triglyceride concentrations. Of many detergents investigated, only Thesit, Kryo Ebo, and Apovax were useful, decreasing the light-scatter signals almost linearly with increasing detergent concentrations. The regression lines, however, were not parallel among the different apo B-containing lipoproteins. Incubating sera or apo B-containing lipoproteins with bovine milk lipoprotein lipase or bacterial triacylglycerol lipase, at concentrations of 100 kU/L, hydrolyzed all of the triglycerides and most of the phosphatidylcholine within 18 h at 37 degrees C Lipase-pretreatment of samples gave optimal correlation between apo B values as determined by nephelometry with those obtained gravimetrically. We also assessed the influence of sample storage, freezing, and thawing on the nephelometric apo B assays.

Apolipoproteins↗

Effect of bezafibrate on plasma lipids, lipoproteins, apolipoproteins AI, AII and B and LCAT activity in hyperlipidemic, non-insulin-dependent diabetics.

The effect of bezafibrate on plasma lipids, lipoproteins, apolipoproteins AI, AII and B, and LCAT activity was investigated in 16 hyperlipidemic, non-insulin-dependent diabetes, who were treated for 8 weeks with either placebo or bezafibrate in a double-blind cross-over design. Bezafibrate induced a significant decrease in plasma triglycerides (P less than 0.01), cholesterol (P less than 0.05), VLDL triglycerides (P less than 0.05) and VLDL cholesterol (P less than 0.01), and a significant increase in HDL cholesterol (P less than 0.01), whereas LDL cholesterol remained unchanged. The apolipoprotein AI/apolipoprotein B ratio increased significantly (P less than 0.05), although individual changes in these apolipoproteins were not significant. Apolipoprotein AII increased significantly (P less than 0.05), although individual changes in these apolipoproteins were not significant. Apolipoprotein AII increased significantly (P less than 0.0001) and the apolipoprotein AI/apolipoprotein AII ratio decreased (P less than 0.0001), indicating an increase in the HDL3 rather than the HDL2 fraction. No significant change in LCAT activity was observed.

Aged↗

Serum concentrations of Lp(a) and other lipoprotein parameters in heavy alcohol consumers.

The serum concentration of lipoprotein Lp(a) and of other lipoproteins of 64 individuals consuming more than 200 g of ethanol per day for several years was measured. 42 of the propositi with no clinical signs of alcoholic liver cirrhosis (ALC) had an apoAI/apoAII ratio twice as high as the control group. The apoAI values of the consumers of alcohol were higher and the HDL-C and Lp(a) levels were lower than those of the control group. 22 alcohol drinkers with ALC exhibited strikingly reduced values of all serum lipids and lipoproteins and the lowest Lp(a) concentrations of all. It is suggested that alcohol drinking lowers serum Lp(a) what might be an additional reason why this group of people seems to be a lower risk of atherosclerosis.

Adult↗

Neutron small angle scattering on selectively deuterated human plasma low density lipoproteins. The location of polar phospholipid headgroups.

Human plasma low density lipoproteins (LDL) were deuterated in the phospholipid headgroup region by exchange with phosphatidylcholine-N(CD3)3-apolipoprotein A complexes. The exchange was associated with a net transfer of phosphatidylcholine to LDL leading to an increase in total phospholipid content by 27%. Practically all of the endogenous phosphatidylcholine including lysophosphatidylcholine, and about one-third of the sphingomyelin pool was found to be exchangeable. Immunochemically, deuterated LDL was identical with native LDL. The hydrodynamic and ultrastructural properties were closely similar for the two particle species apart from a slight increase in overall particle size by about 2%. Both native and deuterated LDL were investigated by neutron small angle scattering at several representative contrasts in H2O/D2O buffers. Subtraction of the scattering amplitudes of native from deuterated LDL resulted in a radius of gyration of 103 +/- 5 A for the N(CD3)3 groups, and in a structure factor resembling that of a thin, spherical shell. Evaluation of the contrast variation experiments in combination with previous data from x-ray small angle scattering (Laggner, P., and Mueller, K. (1978) Q. Rev. Biophys. 11, 371-425) indicates that the phospholipids form a spherical monolayer shell in the radial range between 75 A and 103 A around the core of cholesteryl esters and triglycerides. For the protein moiety, a radius of gyration of 110 A was calculated, indicating that it is located, on average, 5 to 10 A from the polar phospholipid headgroups toward the aqueous environment.

Cholesterol↗

Studies on the composition of the protein part of triglyceride rich lipoproteins of human serum: isolation of polymorphic forms of beta 2-glycoprotein-I.

Isoelectric focusing (IEF) of the apoproteins of triglyceride rich human serum lipoproteins gives rise to the separation of some 15-20 protein bands. Three of these bands have been isolated in pure form and were characterized as isoelectric species of beta 2-glycoprotein-I (beta 2G-I). To compare the amino acid composition of these polymorphic forms with a representative specimen of beta 2G-I from total serum it was also necessary to apply a novel isolation procedure using Rivanol, perchloric acid and Heparin-Sepharose affinity chromatography. With the possible exception of the Pro content, the three isoforms were chemically and immunochemically identical. The isoelectric points of the polymorphic forms were 5.75, 6.0 and 6.2. Their molecular weight was identical by SDS polyacryl amide gel electrophoresis (54 000 D).

Amino Acids↗

[High density lipoproteins: the status quo (author's transl)].

The answer to the question which we are frequently asked as to whether there is a "good" cholesterol is most probably negative. There seems to be little doubt about the fact that a high serum level of cholesterol, irrespective of whether it is transported by HDL or LDL is undesirable. However, prospective studies have clearly shown that the risk of atherosclerosis is inversely proportional, at a given serum cholesterol concentration, to the HDL-transported fraction. On the other hand it must be borne in mind that HDL represent a very heterogeneous mixture of lipoprotein classes and families and generalizations should be avoided. Lp(a), for example, which is mostly found in the HDL density class may, even, represent an independent risk factor for atherosclerosis and vascular diseases. Thus, the present paper attempts to review current concepts of HDL structure, chemical and immunochemical composition and metabolism. Furthermore, factors are listed which are known to influence the serum HDL concentration. Finally, the most widely used methods for analysis and determination of HDL, alpha lipoprotein and LpA are listed and mean values obtained in our laboratory are given.

Arteriosclerosis↗

The determination of lecithin:cholesterol acyl transferase in the clinical laboratory: a modified enzymatic procedure.

A very simple and fast method for the determination of lecithin:cholesterol acyltransferase is described. The method is based on the enzymatic determination of free cholesterol in a serum or plasma sample before and after 40 min incubation at 37 degrees C. The proposed procedure is particularly useful for routine application in a clinical laboratory. It works with a coefficient of variation of less than 5%.

Cholesterol↗