Use of a dual-precipitation procedure for measuring high-density lipoprotein 3 (HDL3) in normolipidemic serum.

1989 ◽  
Vol 35 (7) ◽  
pp. 1390-1393 ◽  
Author(s):  
T A Cloey ◽  
P S Bachorik
Keyword(s):  
High Density Lipoprotein ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Cholesterol Concentration ◽  
Precipitation Method ◽  
Serum Samples ◽  
Precipitation Procedure ◽  
The Difference ◽  
Hdl2 Cholesterol

Abstract We compared results by a dual-precipitation method (J Lipid Res 1982;23:1206-23) for measuring high-density lipoprotein 3 (HDL3) cholesterol with those by ultracentrifugation at d 1.125, using 56 fresh and 105 frozen-stored serum samples. For both methods, HDL2-cholesterol was calculated as the difference between total HDL-cholesterol and HDL3-cholesterol. In general, for pooled serum samples, agreement was closest with ultracentrifugation when we used a dextran sulfate concentration of 5.0 mg/L to precipitate the HDL2-rich fraction, although the optimal concentration varied from 3.0 to 6.8 mg/L for different pools. For individual samples, the values for HDL3 by dual precipitation averaged 12.8% lower than by ultracentrifugation. The coefficients of correlation between the two methods were HDL3, r = 0.70; and HDL2, r = 0.92. The dual-precipitation method reflected the expected sex-related differences in HDL2-cholesterol concentration and inverse relationship with triglyceride concentration.

Simplified methods for measuring cholesterol concentrations of high-density lipoprotein subclasses in serum compared.

1986 ◽  
Vol 32 (7) ◽  
pp. 1274-1278 ◽  
Author(s):  
C F Whitaker ◽  
S R Srinivasan ◽  
G S Berenson
Keyword(s):  
High Density Lipoprotein ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Precipitation Method ◽  
Affinity Column ◽  
Affinity Precipitation ◽  
Lipoprotein Subclasses ◽  
Affinity Column Chromatography ◽  
Hdl2 Cholesterol

Abstract We used a combination of heparin micro-affinity column chromatography and heparin-Mn2+/dextran sulfate (DS) precipitation procedures to measure directly the total high-density lipoprotein (HDL)-cholesterol and HDL3-cholesterol in serum. The value for HDL2-cholesterol was obtained by subtracting the value for HDL3-cholesterol from that for total HDL-cholesterol. Results of this methodology and of the original heparin-Mn2+/DS double-precipitation method were compared with those of preparative ultracentrifugation. Concentrations of HDL- and HDL2-cholesterol by the original double-precipitation method were respectively 26 and 33 mg/L lower than by the ultracentrifugation method (p less than 0.001). Corresponding values by the micro-affinity/precipitation method were 36 and 29 mg/L higher than by the ultracentrifugation method (p less than 0.001). The values for HDL3-cholesterol by ultracentrifugation and by precipitation differed only by 7 mg/L (p greater than 0.05). Micro-affinity/precipitation and double-precipitation results both correlated well with those by ultracentrifugation (r = 0.84 to 0.94); the y-intercept of the comparison with the micro-affinity/precipitation method was close to zero. Use of the micro-affinity/precipitation method with samples from black or white adolescents showed that the blacks had higher concentrations of HDL2-(p less than 0.01) and HDL3-cholesterol (p less than 0.02) than did the whites.

A dual-precipitation method evaluated for measurement of cholesterol in high-density lipoprotein subfractions HDL2 and HDL3 in human plasma.

1989 ◽  
Vol 35 (2) ◽  
pp. 265-270 ◽  
Author(s):  
W Patsch ◽  
S A Brown ◽  
J D Morrisett ◽  
A M Gotto ◽  
J R Patsch
Keyword(s):  
High Density Lipoprotein ◽  
Density Lipoprotein ◽  
Cholesterol Content ◽  
Epidemiological Studies ◽  
High Density ◽  
Cholesterol Concentration ◽  
Precipitation Method ◽  
Lipoprotein Subfractions ◽  
High Density Lipoprotein Subfractions ◽  
Hdl2 Cholesterol

Abstract The dual-precipitation method for measurement of cholesterol in high-density lipoprotein subfractions HDL2 and HDL3 (Warnick et al., Clin Chem 1982;28:1574) was compared with quantification of cholesterol in HDL2 and HDL3 by zonal ultracentrifugation (Patsch et al., J Lipid Res 1974;15:356-66). For 39 plasma specimens differing widely in their HDL subfraction cholesterol concentration, the coefficient of correlation between the two methods was 0.94 for HDL2-cholesterol, 0.82 for HDL3-cholesterol. Storage of plasma specimens at -70 degrees C decreased the apparent content of HDL3-cholesterol by 5%; no significant changes in HDL2-cholesterol were observed. In frozen plasma, interference by apoB-containing lipoproteins and by lipoprotein(a) was negligible. Mean weight ratios of apoA-I to cholesterol were twice as high for HDL3 as for HDL2, reflecting the increased cholesterol content of HDL2. The study suggests that quantification of HDL2 and HDL3 cholesterol by precipitation is appropriate for use in epidemiological studies.

Modification of the dextran-Mg2+ high-density lipoprotein cholesterol precipitation method for use with previously frozen plasma

1994 ◽  
Vol 40 (2) ◽  
pp. 233-239 ◽  
Author(s):  
J R McNamara ◽  
C Huang ◽  
T Massov ◽  
E T Leary ◽  
G R Warnick ◽  
...  
Keyword(s):  
High Density Lipoprotein ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
Sample Volume ◽  
High Density ◽  
Precipitation Method ◽  
Modified Method ◽  
Fresh Plasma ◽  
Frozen Plasma ◽  
Clear Supernatant

Abstract Although dextran-Mg2+ precipitation produces accurate and precise results for high-density lipoprotein (HDL) cholesterol in fresh plasma and serum, precipitation of frozen specimens with triglycerides > 2.26 mmol/L (> 200 mg/dL) is difficult. We developed a modification that dilutes thawed samples by 35% and increases dextran-Mg2+ reagent to 15% of sample volume. Standard precipitations were performed on 62 fresh EDTA-treated plasma specimens; supernatant solutions were analyzed fresh and after freezing. Standard and modified methods were also performed on thawed, paired plasmas. In specimens with triglycerides < or = 2.26 mmol/L, HDL cholesterol results for all methods were similar. For triglycerides > 2.26 mmol/L, however, bias and precision were significantly affected by freezing, and 38.5% of samples with standard precipitation required additional procedures to produce clear supernatant solutions. HDL cholesterol concentrations for thawed samples with standard precipitation were significantly greater than for fresh samples (P < 0.02), but those for the modified method were not different from fresh samples, and only one specimen required additional steps to produce a clear supernate.

scholarly journals Usefulness of apolipoprotein B-depleted serum in cholesterol efflux capacity assays using immobilized liposome-bound gel beads

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Yuna Horiuchi ◽  
Ryunosuke Ohkawa ◽  
Shao-Jui Lai ◽  
Shitsuko Shimano ◽  
Michio Hagihara ◽  
...  
Keyword(s):  
Serum Protein ◽  
Cholesterol Efflux ◽  
Serum Proteins ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
Cholesterol Concentration ◽  
Precipitation Method ◽  
Serum Samples ◽  
Apo B ◽  
Cholesterol Efflux Capacity

Abstract Cholesterol efflux capacity (CEC) in atherosclerotic lesions is the main anti-atherosclerotic function of high-density lipoprotein (HDL). In recent studies, apolipoprotein (apo) B-depleted serum (BDS) obtained with the polyethylene glycol (PEG) precipitation method is used as a cholesterol acceptor (CA) substitution for HDL isolated by ultracentrifugation. However, the suitability of BDS as a CA is controversial. In the present study, CEC obtained from BDS (BDS-CEC) was evaluated based on a parameter, defined as whole-CEC, which was calculated by multiplying CEC obtained using fixed amounts of HDL by cholesterol concentration to HDL-cholesterol (HDL-C) levels in the serum. Significant correlation (r = 0.633) was observed between both CECs. To eliminate systematic errors from possible contamination with serum proteins and low-density lipoprotein (LDL) or very-LDL (VLDL) in BDS-CEC, the deviation of each CEC-BDS from the regression equation was compared with serum protein, LDL, and triglyceride (TG) levels. No correlation was observed between the deviation and the levels of each of these serum components, indicating that the deviations do not derive from systematic error. Further, to evaluate the effects of serum protein on the results, we measured BDS-CEC of reconstituted serum samples prepared using combinations of five levels of serum proteins with five levels of HDL-C. No significant change in BDS-CEC was observed in any combination. These results indicate that BDS-CEC reflects not only the function of HDL but also its concentration in serum.

Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2.

1983 ◽  
Vol 29 (12) ◽  
pp. 2026-2030 ◽  
Author(s):  
G Assmann ◽  
H Schriewer ◽  
G Schmitz ◽  
E O Hägele
Keyword(s):  
High Density Lipoprotein ◽  
High Performance ◽  
Phosphotungstic Acid ◽  
Density Lipoprotein ◽  
High Density ◽  
Precipitation Method ◽  
Lipoprotein Subfractions ◽  
Precipitation Procedure ◽  
High Density Lipoprotein Subfractions ◽  
Layer Chromatography

Abstract We evaluated the use of a modified phosphotungstic acid/MgCl2 precipitation procedure for the precipitation of apolipoprotein B-containing lipoproteins. Precipitation of these lipoproteins [very-low- and low-density lipoproteins, and lipoprotein (a)] is complete, with negligible coprecipitation of high-density lipoprotein subfractions (HDL1, HDL2, HDL3), even in hypertriglyceridemic sera. In comparison with ultracentrifugation, the precipitation method yields, on the average, values that are 0.17 mmol/L lower for cholesterol values but almost identical for apolipoprotein A-I and phosphatidylcholine. Looking for delta 3,5-cholestadiene formed from cholesterol in the precipitation residue, we used "high-performance" liquid chromatography and "high-performance" thin-layer chromatography and found none.

Falsely high high-density lipoprotein triglyceride values by the heparin-manganese precipitation method.

1988 ◽  
Vol 34 (10) ◽  
pp. 2127-2129 ◽  
Author(s):  
I E Simo ◽  
Z Kiss ◽  
T C Ooi
Keyword(s):  
High Density Lipoprotein ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
High Density ◽  
Manganese Chloride ◽  
Precipitation Method ◽  
Low Density ◽  
High High Density Lipoprotein ◽  
The Difference ◽  
Artery Disease

Abstract Recent evidence indicates that high-density lipoprotein triglyceride (HDL-Tg) may be a predictor of coronary artery disease. We examined three methods for HDL-Tg measurement, comparing results obtained by measurement of Tg in the supernate after heparin-manganese chloride (heparin-Mn) precipitation of EDTA-treated plasma (I) with results obtained after preparative ultracentrifugation (II and III). In II, we used heparin-Mn precipitation of low-density lipoprotein (LDL) from the infranate after ultracentrifugation at d 1.006 to remove very-low-density lipoprotein (VLDL). In III, we performed sequential flotation ultracentrifugation at d 1.006 and 1.063, then measured Tg in the d greater than 1.063 fraction. Method I gave significantly higher HDL-Tg results than II and III, which gave essentially identical results. The difference in results between I and II was not caused by the presence of heparin or manganese chloride, because these were used in both methods. Prior removal of VLDL in II and III resulted in lower HDL-Tg values, and subsequent removal of LDL by precipitation or ultracentrifugation did not alter final HDL-Tg values. The higher values obtained in I were the result of the presence of VLDL-rich unsedimented precipitate in the supernate.

Cholesterol distribution between high-density-lipoprotein subfractions HDL2 and HDL3 determined in serum by discontinuous gradient gel electrophoresis

1991 ◽  
Vol 37 (7) ◽  
pp. 1149-1152 ◽  
Author(s):  
Véronique Atger ◽  
Denise Malon ◽  
Marie Claude Bertiere ◽  
Françoise N'Diaye ◽  
Anik Girard-Globa
Keyword(s):  
High Density Lipoprotein ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Lipoprotein Subfractions ◽  
Cylindrical Tubes ◽  
High Density Lipoprotein Subfractions ◽  
Gradient Gel Electrophoresis ◽  
Hdl2 Cholesterol

Abstract We used discontinuous gradients of polyacrylamide gel to determine the high-density-lipoprotein (HDL) subfractions HDL2 and HDL3 of serum lipoproteins. Serum (40 microL) prestained with Sudan Black was electrophoresed in cylindrical tubes over successive layers of 3.5%, 6%, 13%, and 17.5% acrylamide gels in a Tris.glycine buffer (3-4 h, 300 V). Very-low- (VLDL) and low-density lipoprotein (LDL) were retained by the 3.5% and 6% gels. HDL2 was concentrated at the interface between the 13% and 17.5% gels, and HDL3 migrated into the 17.5% gel. The distribution between HDL2 and HDL3 was obtained by densitometric scanning. Application of the respective percentages to HDL cholesterol assayed after phosphotung-state-Mg2+ precipitation of VLDL and LDL gave calculated concentrations of HDL2 and HDL3 cholesterol. The calculated values for HDL2 cholesterol were in excellent agreement with those for HDL2 isolated by ultracentrifugation (r = 0.920 for n = 120 sera; differences nonsignificant by Student's paired t-test). Besides being highly discriminating, the method is rapid, easily performed, and economical.

High-density lipoprotein composition and paraoxonase activity in Type I diabetes

2001 ◽  
Vol 101 (6) ◽  
pp. 659-670 ◽  
Author(s):  
Jonathan VALABHJI ◽  
Avril J. McCOLL ◽  
Michael SCHACHTER ◽  
Surinder DHANJIL ◽  
William RICHMOND ◽  
...  
Keyword(s):  
High Density Lipoprotein ◽  
Particle Density ◽  
Type I Diabetes ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Cholesterol Concentration ◽  
Pon1 Activity ◽  
Type I ◽  
Carotid Imt

Type I diabetes is associated with a high incidence of coronary heart disease (CHD), despite a normal or even increased concentration of high-density lipoprotein (HDL) cholesterol. This paradox may be explained by changes in the antioxidant capacity of HDL, related to paraoxonase (PON1) activity. HDL compositional changes in subjects with Type I diabetes may result in changes in PON1 activity that are associated with a higher incidence of CHD. Single-vertical-spin density-gradient ultracentrifugation was used to isolate seven HDL fractions from serum according to density. PON1 activity was measured in serum and in the HDL fractions using phenyl acetate as substrate. The mean recovery of PON1 activity in the HDL fractions was 87% (S.D. 12%). CHD risk was assessed using B-mode ultrasound to measure carotid artery intima-media thickness (IMT). Groups of 35 subjects with Type I diabetes {duration of diabetes 18 years (12-32 years) [median (interquartile range)]; glycated haemoglobin 7.67% (1.17%)} and 24 non-diabetic control subjects were studied. Carotid IMT was greater in the diabetic subjects [0.60 (0.55-0.70) compared with 0.55 (0.45-0.64) mm; P = 0.042] and HDL cholesterol concentration was higher [1.53 (0.36) compared with 1.32(0.34)mmol/l; P = 0.031]. There were qualitative differences in HDL in subjects with Type I diabetes: HDL particles were triacylglycerol-deplete, and there were greater numbers of the larger, more buoyant HDL particles. These properties were not those found to determine PON1 activity. PON1 activity increased as HDL particle density increased and particle size decreased; the increase in PON1 activity was associated with an increase in the ratio of the two HDL surface lipid components, phospholipid and unesterified cholesterol, as particle density increased. PON1 activity was similar in diabetic and non-diabetic subjects [121 (28) and 120 (36)μmolċmin-1ċml-1 respectively; P = 0.887]. PON1 activity was not associated with carotid IMT in either group. Our results suggest that the PON1 activities of HDL particles relate to the density, size and composition of the particles. However, PON1 activity does not appear to contribute to the greater risk of CHD in subjects with Type I diabetes.

scholarly journals Pengaruh suplementasi serat Psyllium husk dan diet rendah kalori seimbang terhadap berat badan, kadar kolesterol high-density lipoprotein, dan trigliserida serum pada obes I

2014 ◽  
Vol 11 (1) ◽  
pp. 1
Author(s):  
Irnawaty Rasyid ◽  
Rachmad Soegih ◽  
Dante Saksono Harbuwono
Keyword(s):  
Body Weight ◽  
High Density Lipoprotein ◽  
Dietary Fiber ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Cholesterol Concentration ◽  
Soluble Fiber ◽  
Psyllium Husk ◽  
Insignificant Difference

Background: The increased prevalence of obesity will bring a great impact in the health sector, due to the effect of the influence of organ in the body such as type 2 diabetes mellitus (T2DM) and cardiovascular disease (CVD). Reduced energy diet and exercise are effective for management weight loss. During the restriction diet, an obese person should increase the amount of dietary fiber up to 20−35 g/day, specifically of soluble fiber, to more effective fat loss and improve serum high-density lipoprotein (HDL) and triglyceride (TG) cholesterol concentration. Psyllium husk (PH) is a source of natural soluble fiber obtained from Plantago ovate Forssk seed.Objective: The aim of the study have investigated the change of body weight, serum HDL cholesterol, and TG concentration in obese I after supplemented PH 8.4 g/day and balanced deficit calories diet (BDCD) for 4 weeks.Method: The survey used double-blind randomized clinical trial with parallel design. Subjects were randomly divided into two groups; treatment (T) group and placebo (P) group. The T subjects received psyllium husk (PH) 8.4 g/day and BDCD 1200 kcal/day and the P subjects received placebo and BDCD 1200 kcal/day. The analyzed used independent t-test and Mann-Whitney.Results: A total 28 subjects (14 subjects in each group) had completed the intervention. There were no serious adverse effects reported during the intervention. Intake of dietary fiber in T group was 17.2 ± 2.8 g/day had significantly higher than P group 8.6 (5.2−15.2) g/day, although supplemented with PH didn’t meet the recommendation of fiber intake (20-35 g/day). Decrease of body weight was -1,8 ± 0,8 kg and triglyceride level was -1,5 (-416−77) in T group that statistically insignificant difference (p=0,39 and p=0,84) with P group -1,6 ± 0,9 kg and -10,0 ± 31,3. Soluble supplementation (P group) increased serum HDL cholesterol concentration was 0,0 ± 4,3 mg/dL that statistically insignificant difference (p=0,86) with T group -0,4 ± 5,9.Conclusion: PH supplementation 8.4 g/day and BDCD 1200 kcal/day in obese I can not reduce body weight, serum high-density lipoprotein cholesterol, and triglyceride concentration level in 4 weeks.

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