scholarly journals Estimation of Low Density Lipoprotein Fraction Concentration in Blood Plasma of Laying Turkey Hens Using Density Gradient Centrifugation

1974 ◽  
Vol 53 (6) ◽  
pp. 2015-2019 ◽  
Author(s):  
Wayne L. Bacon ◽  
Margery A. Musser
Keyword(s):  
Blood Plasma ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Lipoprotein Fraction ◽  
Low Density ◽  
In Blood Plasma

scholarly journals STUDIES ON THE IMMUNOCHEMISTRY OF HUMAN LOW DENSITY LIPOPROTEINS UTILIZING AN HEMAGGLUTINATION TECHNIQUE

1959 ◽  
Vol 110 (1) ◽  
pp. 113-122 ◽  
Author(s):  
William W. Briner ◽  
Jackson W. Riddle ◽  
David G. Cornwell
Keyword(s):  
Serum Proteins ◽  
Gradient Centrifugation ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Normal Subjects ◽  
Low Density Lipoproteins ◽  
Lipoprotein Fraction ◽  
High Density Lipoproteins ◽  
Low Density ◽  
Chicken Erythrocytes

Hemagglutination is a specific and sensitive technique for investigating the purity of lipoproteins and the immunologic relationships between low density lipoprotein fractions. The Sf 10–400 and Sf 3–9 lipoprotein fractions, isolated from human serum by dextran sulfate-density gradient centrifugation procedure and repurified by centrifugation appeared to contain only lipoprotein antigens since these fractions did not stimulate the production of antibodies against other serum proteins. Cross-absorption experiments with lipoproteins carried on "tanned" cells demonstrated that the Sf 3–9 lipoprotein fraction contains all the antigenic components of the Sf 10–400 lipoprotein fraction together with additional antigenic components not found in the Sf 10–400 lipoprotein fraction. Thus Sf 3–9 and Sf 10–400 lipoprotein fractions are immunologically similar but not identical. Low density lipoproteins contain no antigens in common with the high density lipoproteins. An Sf 3–9 antiserum can be used to detect both Sf 3–9 and Sf 10–400 antigens. The Sf 3–9 lipoprotein fraction used as an antigen will detect antibodies against both Sf 3–9 and Sf 10–400 lipoprotein fractions. The Sf 3–9 and Sf 10–400 antisera did not contain immune antibodies against erythrocytes of the different blood groups or against sheep, guinea pig, dog, calf, pig, horse, and chicken erythrocytes. Normal subjects and subjects with recent myocardial infarctions had no circulating autoantibodies against the Sf 3–9 and Sf 10–400 lipoprotein fractions.

scholarly journals Effect of High-Density Lipoprotein from Healthy Subjects and Chronic Kidney Disease Patients on the CD14 Expression on Polymorphonuclear Leukocytes

2021 ◽  
Vol 22 (6) ◽  
pp. 2830
Author(s):  
Gerald Cohen
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
High Density Lipoprotein ◽  
Density Gradient ◽  
Healthy Subjects ◽  
Polymorphonuclear Leukocytes ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Density Lipoprotein ◽  
Surface Expression

In uremic patients, high-density lipoprotein (HDL) loses its anti-inflammatory features and can even become pro-inflammatory due to an altered protein composition. In chronic kidney disease (CKD), impaired functions of polymorphonuclear leukocytes (PMNLs) contribute to inflammation and an increased risk of cardiovascular disease. This study investigated the effect of HDL from CKD and hemodialysis (HD) patients on the CD14 expression on PMNLs. HDL was isolated using a one-step density gradient centrifugation. Isolation of PMNLs was carried out by discontinuous Ficoll-Hypaque density gradient centrifugation. CD14 surface expression was quantified by flow cytometry. The activity of the small GTPase Rac1 was determined by means of an activation pull-down assay. HDL increased the CD14 surface expression on PMNLs. This effect was more pronounced for HDL isolated from uremic patients. The acute phase protein serum amyloid A (SAA) caused higher CD14 expression, while SAA as part of an HDL particle did not. Lipid raft disruption with methyl-β-cyclodextrin led to a reduced CD14 expression in the absence and presence of HDL. HDL from healthy subjects but not from HD patients decreased the activity of Rac1. Considering the known anti-inflammatory effects of HDL, the finding that even HDL from healthy subjects increased the CD14 expression was unexpected. The pathophysiological relevance of this result needs further investigation.

Abstract 437: Optimizing a Prokaryotic Cell Free Expression System for the Generation of synthetic ApoB-Containing Lipoprotein Particles

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Humra Athar ◽  
Zhenghui G Jiang ◽  
Christopher J McKnight
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Expression System ◽  
Low Density Lipoproteins ◽  
Free Expression ◽  
Low Density ◽  
Prokaryotic Cell ◽  
Lipoprotein Particles ◽  
Cell Free Expression System

High serum levels of low density lipoproteins (LDL) is associated with increased risk of atherosclerosis. Apolipoprotein B (ApoB) is required for the assembly and secretion of chylomicrons and very low density lipoproteins (VLDL), the precursor of low density lipoproteins (LDL). Despite its clinical significance, the mechanism of the assembly of these ApoB containing lipoproteins is poorly understood. The assembly process is an interplay of several key components including but not limited to nascent ApoB, lipids, ER resident chaperones and importantly, microsomal triglyceride transfer protein (MTP). In the current study, we are trying to understand several unanswered questions in the mechanism of the lipoprotein assembly. We have used a novel prokaryotic cell-free expression system and lipids mimicking the ER membrane to produce particles that represent the early dense initiation particles formed in the ER. After optimizing several different conditions, we were able to make “synthetic” lipoproteins by cotranslational expression of constructs from the first 22% of ApoB tagged with a 6-histidine tag at the C-terminus (ApoB 22-His) with small unilamellar phosphatidylcholine (PC) vesicles and phosphatidylcholine:triolein (PC:TO) emulsions. After cotranslational interaction with lipids, these constructs migrate to a lower density in potassium bromide (KBr) density gradient centrifugation. Here we report a new ApoB 22 construct with a FLAG tag at the N-terminus in addition to the C-terminal His tag. The construct makes significant amount of soluble protein that is soluble in the cell free reaction. The two N- and C-terminal tags allow us to purify full length construct from any truncation products. In addition, the dual-tag approach will allow us to purify the synthetic lipoproteins directly from the cell free system, and thereby avoid the requirement for KBr density gradient centrifugation. This new strategy will provide far more efficient generation and purification of synthetic ApoB containing lipoprotein particles.

Single-spin density-gradient ultracentrifugation vs gradient gel electrophoresis: two methods for detecting low-density-lipoprotein heterogeneity compared

1991 ◽  
Vol 37 (6) ◽  
pp. 853-858 ◽  
Author(s):  
Tom P J Dormans ◽  
Dorine W Swinkels ◽  
Jacqueline de Graaf ◽  
Jan C M Hendriks ◽  
Anton F H Stalenhoef ◽  
...  
Keyword(s):  
Density Gradient ◽  
Spin Density ◽  
Gel Electrophoresis ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Density Gradient Ultracentrifugation ◽  
Low Density ◽  
Single Spin ◽  
Screening Programs ◽  
Gradient Gel Electrophoresis

Abstract Single-spin density-gradient ultracentrifugation (DUC) has proven to be a reproducible method for detection of low-density-lipoprotein (LDL) heterogeneity. Recently another method has been described for this: gradient gel electrophoresis (GGE) of serum, a method that might be more suitable for screening. To gain insight into the relationship of GGE to DUC and into their reproducibility, we determined LDL heterogeneity by DUC and GGE in 41 healthy individuals. In 90.2% (n = 37) of the subjects, the number of LDL subfractions found by both methods agreed. In addition, the density and the relative migration distance of the predominant LDL subfraction observed with the respective methods showed a strong correlation (Pearson correlation, r = 0.85, P less than 0.0001). Although it was not possible to compare for all aspects of LDL heterogeneity, these data suggest that GGE is a valid method of analysis for LDL heterogeneity. In screening programs, it may be necessary to store samples. Therefore, we studied in 24 sera the influence of storage at -80 degrees C for one, four, and 12 weeks on the LDL subfraction distribution detected by each method. LDL heterogeneity was maintained during storage under these conditions.

scholarly journals Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Barbara W Sódar ◽  
Ágnes Kittel ◽  
Krisztina Pálóczi ◽  
Krisztina V Vukman ◽  
Xabier Osteikoetxea ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density
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