Preparation of a stable fresh frozen primary lipoprotein[a] (Lp[a]) standard.

LP[a] is one of the most atherogenic lipoproteins consisting of an LDL-like core particle and a covalently linked glycoprotein of variable size. Due to its structural features, its heterogeneity and instability, there are great difficulties in standardizing quantitative immunochemical Lp[a] assays. One particular problem is the preparation of a pure primary standard, which is sufficiently stable to be used for value assignment of secondary reference material.

Cholesterol metabolism in cells with different peroxisomal defects.

We showed previously that cholesterol biosynthesis in dermal fibroblasts from patients with metabolic disorders of peroxisomal origin is increased in steps prior to mevalonate, whereas low-density-lipoprotein(LDL)-receptor activities were not different from control fibroblasts. Here, the suppression of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity by lovastatin was studied both in dermal fibroblasts from patients with different peroxisomal defects and in a Chinese hamster ovary (CHO) cell line lacking morphologically intact peroxisomes. In addition, the formation of intracellular cholesteryl esters (a measure of acyl-CoA:cholesterol acyltransferase(ACAT)-activity) stimulated by exogenous LDL was investigated.

Immunoturbidimetric determination of lipoprotein(a): improvement in the measurement of turbid and triglyceride-rich samples.

Immunoturbidimetry (IT), a widely used method in clinical chemical laboratories, was checked for its suitability for lipoprotein(a) (Lp(a)) quantification. When the conventional sample diluents were used, turbidimetry gave false results particularly with frozen or lipemic sera which correlated poorly with electroimmunodiffusion (EID). L-Proline which is known to dissociate Lp(a) from other apo B-containing lipoproteins improved the results considerably.

Transfer of phospholipase A-resistant pyrene-dialkyl-glycerophosphocholine to plasma lipoproteins: differences between Lp[a] and LDL.

1-O-Hexadecyl-2-O-pyrenedecanyl-sn-glycero-3-phosphocholine, a non-hydrolyzable fluorescent diether analog of phosphatidylcholine (PC), was synthesized as a probe for studying phospholipid transfer to different lipoprotein classes with potential phospholipase activities. After incubation of total human plasma with the new probe at 37 degrees C for 4.5 h, a characteristic partition between the main lipoprotein fractions was observed.

The interaction of Lp(a) with normal and LDL-receptor-deficient human skin fibroblasts.

The role of LDL receptors in the in vivo catabolism of Lp(a) is still a matter of controversy. Since Lp(a) binds LDL with high affinity, it was essential for this study to separate Lp(a) quantitatively from all other apo-B and apo-E-containing lipoproteins. This was achieved by the addition of proline as a dissociating agent to all buffers during Lp(a) preparation.