Cholesterol determination in serum after fractionation of lipoproteins by immunoprecipitation.

1985 ◽  
Vol 31 (2) ◽  
pp. 252-256 ◽  
Author(s):  
C C Heuck ◽  
I Erbe ◽  
D Mathias
Keyword(s):  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Phosphotungstic Acid ◽  
Cholesterol Concentration ◽  
Apo B ◽  
Cholesterol Determination ◽  
Apo Ai

Abstract We investigated the immunoprecipitation of apolipoprotein B-binding lipoproteins, or of apo A-I- and apo C-binding lipoproteins, by delipidated antiserum for measuring cholesterol in the nonprecipitated lipoprotein fractions. After immunoprecipitation of serum with delipidated anti-apo B, we determined by immunoelectrophoresis that no beta- or pre-beta lipoproteins were present, whereas alpha-lipoproteins remained in the supernate. Conversely, after immunoprecipitation with an antiserum against apo A-I + apo C, only lipoproteins with beta-mobility were detected and no apo B from beta-lipoproteins was in the precipitate. The concentration of cholesterol in the supernate after immunoprecipitation with anti-apo B correlated highly (r = 0.93, n = 118) with cholesterol measured after precipitation with phosphotungstic acid/MgCl2. The cholesterol concentration after immunoprecipitation with anti-apo AI and anti-apo C correlated similarly well (r = 0.94, n = 145) with the LDL-cholesterol calculated by Friedewald's formula in serum specimens reflecting moderate hyperlipoproteinemia.

scholarly journals Compound Heterozygous Familial Hypercholesterolemia and Familial Defective Apolipoprotein B-100 Produce Exaggerated Hypercholesterolemia

2001 ◽  
Vol 47 (3) ◽  
pp. 438-443 ◽  
Author(s):  
E Shyong Tai ◽  
Evelyn S C Koay ◽  
Edmund Chan ◽  
Tzer Jing Seng ◽  
Lih Ming Loh ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Genetic Analysis ◽  
Familial Hypercholesterolemia ◽  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Index Case ◽  
Ldl Receptor ◽  
Cholesterol Concentration ◽  
Apo B ◽  
First Degree Relatives

Abstract Background: Familial hypercholesterolemia (FH) and familial defective apolipoprotein B-100 (FDB) represent ligand-receptor disorders that are complementary. Individuals with both FH and FDB are unusual. We report a family with both disorders and the impact of the mutations on the phenotypes of the family members. Methods: We used single strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) for genetic analysis of all 18 exons and the promoter region of the LDL receptor and DGGE for genetic analysis of the apolipoprotein B-100 (apo B-100) gene. The functional significance of the apo B-100 mutation was studied using a U937 cell proliferation assay. Fasting serum lipid profiles were determined for the index case and seven first-degree relatives. Results: One of the patient’s sisters had a missense mutation (Asp407→Lys) in exon 9 of the LDL receptor and a serum LDL-cholesterol concentration of 4.07 mmol/L. Four other first-degree relatives had hyperlipidemia but no LDL-receptor mutation. However, these subjects had a mutation of the apo B-100 gene (Arg3500→Trp). The cell proliferation rate of U937 cells fed with LDL from other subjects with the same mutation was fourfold less than that of controls. The index case had both FH- and FDB-related mutations. Her serum LDL-cholesterol (9.47 mmol/L) was higher than all other relatives tested. Conclusions: Existence of both FH and FDB should be considered in families with LDL-receptor mutations in some but not all individuals with hypercholesterolemia or when some individuals in families with FH exhibit exaggerated hypercholesterolemia.

Screening for familial defective apolipoprotein B-100 with improved U937 monocyte proliferation assay

1994 ◽  
Vol 40 (3) ◽  
pp. 395-399 ◽  
Author(s):  
A J Van den Broek ◽  
L Hollaar ◽  
H I Schaefer ◽  
A Van der Laarse ◽  
H Schuster ◽  
...  
Keyword(s):  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Human Monocyte ◽  
Cell Number ◽  
Cholesterol Concentration ◽  
U937 Cells ◽  
Proliferation Assay ◽  
Apo B

Abstract Frostegård et al. (J Lipid Res 1990;31:37-44) demonstrated that the proliferation of the human monocyte cell line U937 is critically dependent on the uptake of low-density lipoprotein (LDL) via the apo B, E (LDL) receptor, a characteristic that was used to detect patients with familial defective apolipoprotein B-100 (FDB). Here we applied this principle to develop a simple and reproducible assay for the detection of patients with functionally defective LDL. We added serum to U937 cells in cholesterol-free incubation medium and determined the increase in cell number after a 72-h incubation at 37 degrees C by using an electronic cell counter. Sera from 10 normolipidemic individuals and from 34 patients with type IIa hyperlipoproteinemia stimulated the growth of U937 cells in proportion to the exogenous cholesterol concentration (r = 0.83, P < 0.001) and the LDL-cholesterol concentration (r = 0.81, P < 0.001). However, sera from 16 patients with FDB stimulated less cell proliferation than did sera from patients with type IIa hyperlipoproteinemia with equal LDL-cholesterol concentrations. With a 15% reduction in growth as the cutoff value, this test had a sensitivity and specificity for diagnosis of FDB of 87.5% and 100%, respectively. The improved U937 monocyte proliferation assay can be used for screening hypercholesterolemic patients to detect individuals with functionally defective LDL.

scholarly journals Lipoprotein Signatures of Cholesteryl Ester Transfer Protein and HMG-CoA Reductase Inhibition

2018 ◽  
Author(s):  
Johannes Kettunen ◽  
Michael V. Holmes ◽  
Elias Allara ◽  
Olga Anufrieva ◽  
Pauli Ohukainen ◽  
...  
Keyword(s):  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Triglyceride Composition ◽  
Cholesterol Concentration ◽  
Resonance Spectroscopy ◽  
Low Density ◽  
Triglyceride Content ◽  
Cetp Inhibition

AbstractBackgroundCETP inhibition reduces vascular event rates but confusion surrounds its low-density lipoprotein (LDL)-cholesterol effects. We sought to clarify associations of genetic inhibition of CETP on detailed lipoproteins.Methods and ResultsWe used variants associated withCETP(rs247617) andHMGCR(rs12916) expression in 62,400 Europeans with detailed lipoprotein profiling from nuclear magnetic resonance spectroscopy. Genetic associations were scaled to 10% lower risk of coronary heart disease (CHD). Associations of lipoprotein measures with risk of incident CHD in three population-based cohorts (770 cases) were examined.CETPandHMGCRhad near-identical associations with LDL-cholesterol concentration estimated by Friedewald-equation.HMGCRhad a relatively consistent effect on cholesterol concentrations across all apolipoprotein B-containing lipoproteins.CETPhad stronger effects on remnant and very-low-density lipoprotein cholesterol but no effect on cholesterol concentrations in LDL defined by particle size (diameter 18–26 nm) (-0.02SD 95%CI: -0.10, 0.05 forCETPversus -0.24SD, 95%CI -0.30, -0.18 forHMGCR).CETPhad profound effects on lipid compositions of lipoproteins, with strong reductions in the triglyceride content of all highdensity lipoprotein (HDL) particles. These alterations in triglyceride composition within HDL subclasses were observationally associated with risk of CHD, independently of total cholesterol and triglycerides (strongest HR per 1-SD higher triglyceride composition in very-large HDL 1.35; 95%CI: 1.18, 1.54).ConclusionCETP inhibition does not affect size-specific LDL cholesterol but may lower CHD risk by lowering cholesterol in other apolipoprotein-B containing lipoproteins and lowering triglyceride content of HDL particles. Conventional composite lipid assays may mask heterogeneous effects of lipid-altering therapies.

Comparison of methods for measurement of apolipoprotein B and cholesterol in low-density lipoproteins

1997 ◽  
Vol 43 (2) ◽  
pp. 390-393 ◽  
Author(s):  
Ljubica Vrga ◽  
Christine Contacos ◽  
Stephen C H Li ◽  
David R Sullivan
Keyword(s):  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Low Density Lipoproteins ◽  
Plasma Triglyceride ◽  
Low Density ◽  
Comparison Of Methods ◽  
Apo B ◽  
Simple Alternative ◽  
Cholesterol Measurement ◽  
Commercial Kit

Abstract We describe a new method for the direct measurement of LDL-apolipoprotein (apo) B by using a commercial kit that isolates LDL by immunoseparation. We evaluated immunoseparation of LDL for apo B and cholesterol measurement in 46 dyslipidemic patients with LDL-cholesterol (LDL-C) between 1.5 and 8.2 mmol/L, 11 of whom had plasma triglyceride (TG) concentrations >4.0 mmol/L. There was a reasonable correlation (r = 0.94, n = 40) between LDL-apo B obtained after immunoseparation and d >1.006 kg/L apo B obtained after ultracentrifugation. LDL-C by the immunoseparation method also correlated well (r = 0.98, n = 46) with the d >1.006 kg/L cholesterol after ultracentrifugation. These results show that immunoseparation can be used to determine LDL-apo B, even in hypertriglyceridemic samples. This method may provide a quick and simple alternative for the identification of hyperapobetalipoproteinemia, even when TG concentrations are high.

scholarly journals Calculation of LDL-cholesterol by using apolipoprotein B for classification of nonchylomicronemic dyslipemia

1997 ◽  
Vol 43 (5) ◽  
pp. 808-815 ◽  
Author(s):  
Teresa Planella ◽  
Mariano Cortés ◽  
Cecília Martínez-Brú ◽  
Francesc González-Sastre ◽  
Jordi Ordóñez-Llanos
Keyword(s):  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Good Alternative ◽  
Curve Analysis ◽  
Total Serum ◽  
Roc Curve Analysis ◽  
Apo B ◽  
Cutoff Value ◽  
Automated Methods

Abstract In this paper we propose a calculation of LDL-cholesterol (LDL-C) not affected by hypertriglyceridemia by using lipid quantities directly measured in total serum. We also propose an algorithm for the classification of nonchylomicronemic dyslipemias. Plasma apolipoproteins (apo) A-I, B, total cholesterol (TC), triglycerides (TG), and cholesterol of lipoproteins were measured in a group of 38 normolipemic and 120 dyslipemic patients (42 phenotype IIa, 38 IIb, and 40 IV) classified according to TG and LDL-C values. Discriminant analysis was applied to obtain the best classification with the lowest number of quantities directly measured from total serum (TC, TG, and apo B), and multiple regression analysis was performed to find an equation to calculate LDL-C from these quantities. Apo B seems to be a useful discriminator between normolipemic and phenotype IIa patients, by using a cutoff value of 1.35 g/L obtained by ROC curve analysis. The proposed algorithm, based on lipid quantities measured by easily automated methods, is shown to be a good alternative for the classification of nonhyperchylomicronemic dyslipemia. LDL-C calculated from TC, TG, and apo B proved a better estimate of true LDL-C than the estimate obtained with Friedewald’s formula.

scholarly journals Correlation of Small Dense LDL Cholesterol and Apolipoprotein B with LDL Cholesterol and its Clinical Significance in Overweight, Type 2 Diabetes Mellitus and Coronary Artery Disease

2016 ◽  
Vol 14 (2) ◽  
pp. 36-40 ◽  
Author(s):  
Arabinda Mohan Bhattarai ◽  
HS Batra ◽  
Suchit Bandyopadhyay ◽  
Pratibha Misra ◽  
Manushri Sharma ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Apolipoprotein B ◽  
Ldl Cholesterol ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Low Density Lipoprotein Cholesterol ◽  
Apo B ◽  
Artery Disease

Introduction: Atherosclerotic Coronary Artery Disease (CAD) is fundamentally related to disorders of lipid metabolism. Health problems like obesity, glucose intolerance and metabolic syndrome increase atherosclerotic CAD risk.  A fraction of Low density lipoprotein cholesterol (LDL) is called small dense low density lipoprotein cholesterol (sdLDL). These particles are more atherogenic because they are taken up more easily by arterial wall, readily oxidized and not easily cleared from plasma. Every LDL particle contain an Apo B molecule.Methods: In this cross sectional study we recruited 100 known cases each of CAD, type 2 diabetes, overweight and 100 age and sex matched healthy controls. We took a detailed case summary along with anthropometric measurements. We measured sdLDL by heparin magnesium precipitation method followed by direct estimation of the LDL in the supernatant.Result: Linear regressive analysis showed positive correlation between sdLDL and Apolipoprotein B (Apo B) with LDL cholesterol (r=0.61, p=0.004), (r=0.754, p=0.0034) respectively. Multiple Comparisons after Kruskalwallis test of sdLDL and Apo B levels of  type 2 diabetes, CAD and overweight with controls were significant (p<0.001).Conclusion: Our findings suggest that the estimation of sdLDL and Apo B provide a complimentary benefit in assessment of cases with CAD, type 2 diabetes and overweight.

scholarly journals Dyslipidemia in subclinical hypothyroidism

2003 ◽  
Vol 56 (5-6) ◽  
pp. 276-280 ◽  
Author(s):  
Zorica Caparevic ◽  
Gradimir Bojkovic ◽  
Dragos Stojanovic ◽  
Vesna Ilic
Keyword(s):  
Thyroid Hormone ◽  
Serum Cholesterol ◽  
Subclinical Hypothyroidism ◽  
Ldl Cholesterol ◽  
Hormone Replacement ◽  
Cholesterol Concentration ◽  
Overt Hypothyroidism ◽  
Apo B ◽  
Thyroid Hormone Replacement ◽  
Number Of Patients

Introduction Subclinical hypothyroidism is defined as an increased serum TSH and normal serum FT4 concentration. In subclinical hypothyroidism, thyroid peroxidase and thyroglobulin antibodies are frequently present. Subclinical hypothyroidism may have endogenous or exogenous causes. The prevalence of subclinical hypothyroidism is rather high. The number of patients progressing to overt hypothyroidism may be higher. These patients may be asymptomatic, or have only mild symptoms or a single symptom. Material and methods We investigated 35 patients with subclinical hypothyroidism in order to establish the type and degree of dyslipidemia and effects of therapy with L-thyroxine (50 micrograms/d) during three months. Results Serum cholesterol LDL-cholesterol and apo B were increased. A significant reduction of serum cholesterol, LDL-cholesterol and apo B concentrations was established during thyroid hormone replacement. Discussion and conclusion Only a few studies reported higher LDL and lower HDL-cholesterol values in subclinical hypothyroidism. Much interest was thus aroused to evaluate whether or not subclinical hypothyroidism is associated with hypercholesterolaemia. Only patients with serum thyrotropin (TSH) concentration above 10 mU/L had a significant reduction of serum cholesterol concentration during thyroid hormone replacement. Most patients with subclinical hypothyroidism should be treated with thyroxine to prevent progression to overt hypothyroidism Thyroid hormone replacement therapy may slow the progression of coronary heart disease, because of its beneficial effects on lipids. These findings and especially high rate of progression towards overt hypothyroidism suggest early thyroxine treatment.

scholarly journals Serum Apolipoprotein B/apolipoprotein A1 Ratio in Relation to Intervertebral Disc Herniation : A Cross-sectional Frequency-Matched Case-Control Study

2020 ◽  
Author(s):  
Fei Chen ◽  
Tongde Wu ◽  
Song Guo ◽  
Junwen Huang ◽  
Qiang Fu ◽  
...  
Keyword(s):  
Apolipoprotein B ◽  
Serum Lipid ◽  
Density Lipoprotein ◽  
Apolipoprotein A1 ◽  
Lipoprotein Cholesterol ◽  
Control Group ◽  
Lipid Levels ◽  
Apo B ◽  
Serum Lipid Levels ◽  
Apo Ai

Abstract BackgroundIn recent decades, the serum lipid profile of apolipoprotein(a) (Lp(a)) level and apolipoprotein B/apolipoprotein A1 ratio (Apo B/ApoA1) ratio were found more representative for serum lipid level and were recognized as the independent risk factors for various diseases. Although the serum lipid levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) were found associated with symptomatic IDH, no studies have been conducted to date for the evaluation of the association of Apo AI, Apo B, Lp(a) and Apo B/Apo AI levels with symptomatic IDH.MaterialsA total of 1,839 Chinese patients were recruited in the present study. 918 patients were diagnosed as IDH cases and were enrolled in the experimental group. A control group of 921 patients underwent a physical examination during the same period. The serum lipid levels of TC, TG, LDL-C, HDL-C, Lp(a), Apo B and Apo B/Apo AI were examined and analyzed.ResultsThe patients in the control group were collected randomly from patients who were matched with the baseline levels of the aforementioned lipid molecular. The patients with IDH exhibited significantly higher TC, TG, LDL, Apo B and Lp(a) levels compared with the control subjects. The percentage of high-TC, high-TG, high-LDL, high-Apo B and high-Lp(a) were significantly higher in the IDH group. However, hyperlipidaemia was not associated with the degenerated segment of the IDH (P=0.201). The odds ratios (OR) for the incidence of IDH with an elevated LDL-C, TC, TG, Lp(a), Apo B and Apo B/Apo AI were 1.583, 1.74, 1.62, 1.58, 1.49 and 1.39, respectively. The correlation analysis revealed the correlation between elevated LDL-C, TC, TG, Apo B, Lp(a) and incidence of IDH was significant (R2LDL=0.017; R2TC=0.004; R2TG=0.015; R2Apo B=0.004; R2LP(a)=0.021) (P<0.05). ConclusionsThe present study suggests that elevated levels of serum TC, TG, LDL, Apo B, Lp(a) and Apo B/Apo AI are associated with a higher risk for IDH.

scholarly journals Association of Apolipoprotein B with Incident Type 2 Diabetes in an Aboriginal Canadian Population1

2010 ◽  
Vol 56 (4) ◽  
pp. 666-670 ◽  
Author(s):  
Sylvia H Ley ◽  
Stewart B Harris ◽  
Philip W Connelly ◽  
Mary Mamakeesick ◽  
Joel Gittelsohn ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Apolipoprotein B ◽  
Odds Ratio ◽  
Ldl Cholesterol ◽  
Hdl Cholesterol ◽  
Apo B ◽  
Incident Type ◽  
Incident Type 2 Diabetes

Abstract Background: Expanding evidence indicates that apolipoprotein B (apo B) is superior to LDL cholesterol as a marker of vascular disease. Although traditional lipid measures are known to predict type 2 diabetes, limited data are available regarding apo B. We assessed the association of apo B with incident type 2 diabetes and compared it with traditional lipid variables as a risk predictor in aboriginal Canadians. Methods: Of an initial cohort of 606 individuals without diabetes in 1993–1995, 540 were contacted for the 10-year follow-up evaluation in 2003–2005. Fasting and 2-h postload glucose concentrations were obtained at baseline and follow-up to determine incident type 2 diabetes. Baseline fasting serum lipids were measured with standard laboratory procedures. Results: The cumulative 10-year incidence of type 2 diabetes was 17.5%. High concentrations of apo B, triglycerides, and LDL cholesterol, and low concentrations of HDL cholesterol were individually associated with incident type 2 diabetes in univariate analyses. Comparing C statistics of univariate models showed apo B to be a superior determinant of incident diabetes compared with LDL (P = 0.026) or HDL (P = 0.004) cholesterol. With multivariate adjustment including waist circumference, apo B (odds ratio, 1.50; 95% CI, 1.11–2.02) and triglycerides (odds ratio, 1.49; 95% CI, 1.12–1.98) remained associated with incident diabetes, whereas LDL and HDL cholesterol became nonsignificant. Conclusions: The association of plasma apo B with incident type 2 diabetes and its better prediction of risk compared with LDL or HDL cholesterol suggest the potential for the use of apo B in type 2 diabetes risk communication and prevention.

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