Immunoturbidimetry of serum apolipoproteins A-I and B on the Cobas Bio centrifugal analyzer: Method validation and reference values

Eugui, J; Logroño, María Jesús; Ruíz, R.; Zugaza, Carmen; Mirabel, J; Martínez, Consuelo Sánchez

doi:10.1016/0009-9120(94)90035-3

Cited by 16 publications

(10 citation statements)

References 27 publications

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“…The process was monitored with the inbuilt spectrophotometer at an optical density of 280 nm. Fractions were analyzed using an AU480 Chemistry Analyzer (Beckman Coulter) to determine the concentrations of cholesterol, triglyceride, protein, apoA-I, apoA-II, and apoB as previously described ( Stahler et al, 1977 ; Smith et al, 1985 ; Eugui et al, 1994 ). Phosphatidylcholine (the most abundant phospholipid in HDLs and LDLs) was assayed as previously described ( Takayama et al, 1977 ).…”

Section: Methodsmentioning

confidence: 99%

Altered High Density Lipoprotein Composition in Behavioral Variant Frontotemporal Dementia

et al. 2018

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Frontotemporal dementia (FTD) is a common cause of early onset dementia with behavioral variant FTD (bvFTD) being the most common form. bvFTD is characterized clinically by behavioral and personality changes, eating abnormalities, and pathologically, by systemic lipid dysregulation that impacts on survival. As lipoprotein metabolism is at the core of lipid dysregulation, here, we analyzed the composition, both proteins and lipids, of the two major lipoprotein classes in blood – high density lipoproteins (HDLs) and low density lipoproteins (LDLs). Fasted plasmas from bvFTD and Alzheimer’s disease (AD) patients and controls were fractionated using fast protein liquid chromatography (FPLC) and samples analyzed by lipid assays, ELISA and western blotting. We found that apolipoprotein A-I (apoA-I) and apolipoprotein A-II (apoA-II) levels in HDLs were decreased in bvFTD compared to controls, whereas apolipoprotein B (apoB) levels in LDLs were unaltered. We also found that cholesterol and triglyceride levels in FPLC fractions were altered in bvFTD compared to controls. The apoB:apoA-I ratio and the standard lipid ratios were significantly increased in bvFTD compared to AD and controls. Furthermore, we found that plasma apolipoprotein C-I and paraoxonase 1 levels were significantly altered in bvFTD and AD, respectively, compared controls. This study represents the first apolipoprotein analysis of bvFTD, and our results suggest altered HDL function and elevated cardiovascular disease risk in bvFTD.

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Section: Methodsmentioning

confidence: 99%

Altered High Density Lipoprotein Composition in Behavioral Variant Frontotemporal Dementia

et al. 2018

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“…An enzymatic kit was used to measure total cholesterol concentrations (Roche Diagnostics GmbH). ApoA-I, apoA-II, phospholipid, and UC concentrations were determined as described previously (26)(27)(28).…”

Section: Chemical Analysesmentioning

confidence: 99%

Evidence that endothelial lipase remodels high density lipoproteins without mediating the dissociation of apolipoprotein A-I

Jahangiri

Marchadier

et al. 2005

Journal of Lipid Research

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“…CE concentrations were calculated as the difference between the total and UC concentrations. The concentrations of apoA-I and apoA-II were determined by an immunoturbidometric assay (26). The size of the rHDL and native HDL 2 was determined by electrophoresis on 3-35% nondenaturing polyacrylamide gradient gels (Gradipore, Sydney, Australia) (27).…”

Section: Determination Of Hl-mediated Hydrolysis In Unlabeled (A-i/cementioning

confidence: 99%

The Influence of Apolipoproteins on the Hepatic Lipase-mediated Hydrolysis of High Density Lipoprotein Phospholipid and Triacylglycerol

Hime¹,

Barter²,

Rye³

1998

Journal of Biological Chemistry

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This study describes the influence of apolipoproteins on the hepatic lipase (HL)-mediated hydrolysis of phospholipids and triacylglycerol in high density lipoproteins (HDL). HL-mediated hydrolysis was assessed in well characterized, homogeneous preparations of spherical reconstituted high density lipoproteins (rHDL). Hepatic lipase (HL)1 is a 476-amino acid glycoprotein of molecular weight 64,000 -69,000 (1) that is bound to liver sinusoidal endothelial cells (2). HL hydrolyzes acyl ester bonds of triacylglycerol and the sn-1 acyl ester bond of phospholipids. The main plasma substrates for HL are very low density lipoproteins and high density lipoproteins (HDL). The role of HL in HDL metabolism is of considerable importance, as shown by strong negative associations between HL activity and plasma HDL 2 levels (3-5) and the dramatic reduction in the HDL levels of rabbits that have been made transgenic for human HL (6). Unlike lipoprotein lipase (LPL), which requires apolipoprotein C-II (apoC-II) for maximal activity, there is no known protein cofactor for HL. However, there is some conflicting evidence to suggest that the apoA-II in HDL may influence the HL-mediated hydrolysis of triacylglycerol in HDL (7-10). Some investigators have reported that apoA-II enhances (7, 8), while others have concluded that it inhibits, the HL-mediated hydrolysis of triacylglycerol in HDL (9, 10).The present study was carried out in order to determine whether there are significant differences in the HL-mediated hydrolysis of phospholipids and triacylglycerol in HDL that differ in their apolipoprotein composition. This has been achieved by using well defined, homogeneous preparations of spherical reconstituted HDL (rHDL) as substrates for HL. The rHDL were comparable in size and lipid composition and contained either apoA-I or apoA-II as their sole apolipoprotein constituent. The results show that apolipoproteins not only have a major influence on the HL-mediated hydrolysis of the triacylglycerol and phospholipids in rHDL but also regulate the affinity of HL for the rHDL surface. EXPERIMENTAL PROCEDURESPurification of ApoA-I and ApoA-II-ApoA-I and apoA-II were prepared from pooled human plasma donated by the Transfusion Service, Royal Adelaide Hospital. HDL were isolated from the plasma by sequential ultracentrifugation in the 1.07 Ͻ d Ͻ 1.21 g/ml density range (11). The isolated HDL were delipidated (12), and the resulting apoHDL was subjected to anion exchange chromatography on Q Sepharose Fast Flow (Amersham Pharmacia Biotech, Uppsala, Sweden) (13). The purified apoA-I and apoA-II appeared as single bands following electrophoresis on a homogeneous 20% SDS-polyacrylamide PhastGel (Amersham Pharmacia Biotech) and Coomassie staining.

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Immunoturbidimetry of serum apolipoproteins A-I and B on the Cobas Bio centrifugal analyzer: Method validation and reference values

Cited by 16 publications

References 27 publications

Altered High Density Lipoprotein Composition in Behavioral Variant Frontotemporal Dementia

Altered High Density Lipoprotein Composition in Behavioral Variant Frontotemporal Dementia

Evidence that endothelial lipase remodels high density lipoproteins without mediating the dissociation of apolipoprotein A-I

The Influence of Apolipoproteins on the Hepatic Lipase-mediated Hydrolysis of High Density Lipoprotein Phospholipid and Triacylglycerol

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