Asunto(s)

RESUMEN

BACKGROUND: LDL-cholesterol (LDL-C) concentrations currently are determined in most clinical laboratories using the Friedewald calculation. This approach has several limitations and may not always meet the current total error recommendation in LDL-C measurement of

Asunto(s)

RESUMEN

We evaluated a new liquid homogeneous assay for the direct measurement of high density lipoprotein cholesterol (HDL-C Plus) in seven laboratories. The assay includes two reagents which can be readily used in most available clinical chemistry analyzers. The total CVs of the new method were below 4.6% and the bias in relation to the designated comparison method was below 3.9%. The total error ranged between 4 to 7%. HDL-C values determined by this method were in good agreement with those obtained by the old homogeneous assay using lyophilized reagents, and other homogeneous and precipitation assays (0.944 < r < 0.996). The assay was linear up to at least 3.89 mmol/l HDL-C. Hemoglobin did not interfere, whereas in icteric samples slight deviations were observed. Lipemia up to 11.3 to 22.6 mmol/l triglycerides did not interfere with this homogeneous HDL-C assay. In samples of patients with paraproteinemia, discrepant results were seen. This liquid homogeneous HDL-C assay was easy to handle and produced similar results in all laboratories participating in this study. This method will enable clinical laboratories to reliably measure HDL-C for risk assessment of coronary heart disease.

Asunto(s)

RESUMEN

The values of apolipoproteins (apo) A-I and B were determined in a population sample of hospital outpatients with a standardized method to verify if the cutpoints calculated in a cross-sectional study in the US are usable with other populations. We also tested the apolipoproteins' ability to discriminate between healthy people and survivors of myocardial infarction. In the studied population the apo A-I value corresponding to the HDL-cholesterol decisional centile is 1.12 g/L for males and 1.17 g/L for females; the apo B value corresponding to the LDL-cholesterol decisional centile is 1.23 g/L for males and 1.14 g/L for females. These values are quite close to the cutpoints proposed for the American population (1.20 g/L for both apolipoproteins). In comparison with the LDL- and HDL-cholesterol decisional concentrations, the cutpoints for apolipoproteins allow a correct classification of a greater percentage of postmyocardial infarction patients (16% higher for apo B and 5% for apo A-I). Standardized assays coupled with a reference database allow a better clinical use of apolipoprotein measurements.

Asunto(s)

RESUMEN

Serum protein electrophoresis and typing of monoclonal components (MCs) are routine but time-consuming and technically demanding assays. We evaluated capillary electrophoresis (Paragon CZE 2000) for automation of the two assays. CZE and cellulose acetate electrophoresis gave similar data on 794 samples. Within-run and between-run CVs were < 2% for albumin and gamma-globulins and 4-7% for alpha 1-, alpha 2-, and beta-globulins. Bilirubin, hemoglobin, triglycerides, and fibrinogen were found not to interfere. No carryover by capillaries was detected. The detection limit for MC was < 0.5 g/L. MC assessment by immunosubtraction on 403 samples identified the monoclonal type in all samples with peak concentrations > 10 g/L; only 50% of MCs that could not be quantified by densitometric scan were typed.

Asunto(s)

RESUMEN

We studied the penetration of dapsone into the epithelial lining fluid (ELF) of sixteen human immunodeficiency virus type 1-infected patients who had received the drug at a dose of 100 mg twice weekly as primary prophylaxis for Pneumocystis carinii pneumonia. Bronchoscopy, bronchoalveolar lavage (BAL), and venipuncture were performed for each patient at a specific time after administration of the last dose of dapsone. Dapsone concentrations in plasma and BAL were determined by high-performance liquid chromatography. The apparent volume of ELF recovered by BAL was determined by using urea as an endogenous marker. The mean concentrations of dapsone in ELF at 2 h (five patients), 4 h (three patients), 12 h (two patients), 24 h (three patients), and 48 h (three patients) were 0.95, 0.70, 1.55, 0.23, and 0.45 mg/liter, respectively, while concentrations in plasma were 1.23, 0.79, 1.31, 0.83, and 0.18 mg/liter, respectively. Dapsone concentrations in ELF were 76, 79, 115, 65, and 291% of those observed in plasma at the same times, respectively. These data show that dapsone is well distributed into ELF and that a twice-weekly 100-mg prophylactic regimen results in sustained concentrations in this compartment.

Asunto(s)

RESUMEN

We report the results of an external quality assessment scheme for serum total cholesterol measurement involving about 100 Italian laboratories participating in an epidemiological study of post myocardial infarction. Two frozen human serum pools with Abell-Kendall assigned values are distributed quarterly at the laboratories (up to now seven events occurred); the obtained results are evaluated and discussed. In one exercise (# 5) duplicated measurements were repeated on three different days. Eighty-five to 98% of the laboratories obtained results within the total error limits (+/- 8.9%). But, while precision (calculated on the six replicates of exercise # 5) is good (90% of the laboratories obtained CV < 3%), inaccuracy problems are evident in every event. Indeed the mean bias from the reference method value ranged from 1.54 and 3.49% in the various events.

Asunto(s)

RESUMEN

Some studies have reported an inverse correlation between serum cholesterol level and risk of cancer. This correlation might be due to a decrease in serum retinol, a lipid-soluble vitamin that controls cell proliferation and differentiation. We evaluated the influence of cholesterol-lowering therapy on serum retinol in 102 subjects (mean +/- SE: aged 47.1 +/- 4.1 years; body mass index, 23.8 +/- 0.6 kg/m2) with primary hypercholesterolemia treated for 2 years with different therapeutic protocols. Twenty-two subjects had been treated with diet alone, 35 with diet and fibrates, 37 with diet and hepatic hydroxymethyl glutaryl coenzyme A (HMG CoA) reductase inhibitors (statins), and eight with diet and cholestyramine. Postabsorptive serum retinol, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglyceride levels were determined at baseline and every 3 months. Baseline TC and LDL-C were significantly lower in the diet-treated group than in other groups. No intergroup differences were found in pretreatment levels of triglycerides and serum retinol. After 2 years of treatment, TC and LDL-C serum levels were not significantly decreased in the diet-alone group, whereas they were decreased by 20% and 24%, respectively, in the gemfibrozil group, 28% and 34% in the statins group; and 21% and 27% in the cholestyramine group. In the entire population (N = 102), serum retinol was 3.46 +/- 0.08 mumol/L before therapy and 3.76 +/- 0.07 after 2 years of therapy (P < .001). Serum retinol increased in diet- and statin-treated groups, but not in fibrate- and resin-treated groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Asunto(s)

RESUMEN

We describe a method for measuring apolipoprotein (apo) C distribution between apo B-containing lipoprotein (apo B-LP) and non-apo B-LP. The procedure requires the precipitation of apo B-LP, the redissolution of the pellet, and the quantification of C peptides in the redissolved pellet. The ratio of apo C in non-apo B-LP to apo C in apo B-LP has been calculated for both CII and CII (R-CII and R-CIII, respectively). R-CII (0.49 +/- 0.25) and R-CIII (0.84 +/- 0.54) in patients on maintenance dialysis are significantly lower than in the control group (1.14 +/- 0.57 and 1.45 +/- 0.92, respectively), indicating that hypertriglyceridemia in these patients results from a reduced catabolism of triglyceride-rich LP (TGRLP). Patients with coronary artery disease (CAD) show a distribution of C peptides no different from the control group. Analysis of covariance reveals that the patterns of R-CII and R-CIII are not entirely predictable from the serum concentration of triglycerides. This result seems to support the hypothesis that the underlying metabolic defects involving TGRLP in dialysis patients are not the same as those in patients with CAD.

Asunto(s)

RESUMEN

It is generally accepted that Lp(a) is an independent risk factor of cardiovascular diseases. Since the apolipoprotein component of Lp(a) shows some homologies to plasminogen, it is, however, unclear as to whether the pathological effect is due to the role played by the lipoprotein in lipid metabolism or in the fibrinolytic system. We compared two groups of patients with myocardial infarction, with and without angiographically documented coronary artery disease. In the latter group, imbalances in the clotting system are very likely, while members of the former group may also display disturbances of lipid metabolism. The results show that the two groups display differences in lipid metabolism, whereas they have similar patterns of thrombogenicity indices and Lp(a) values. This study seems to support the hypothesis that Lp(a) does play a role in the fibrinolytic system, since even those myocardial infarctions without obstructive coronary artery disease have a high frequency of Lp(a) concentrations above 300 mg/l, i.e. similar to the situation found in the myocardial infarctions with angiographically documented coronary artery disease. Whether the high Lp(a) concentrations in the two groups are related to an impaired fibrinolysis will be the subject of further investigation.

Asunto(s)

RESUMEN

The Reflotron HDL Cholesterol test (Boehringer Mannheim GmbH) directly separates and analyzes high-density lipoprotein (HDL) cholesterol in plasma collected with EDTA in an integrated dry-reagent system suitable for alternative site testing of lipoproteins. We describe a multicenter evaluation of this test by two US and six European laboratories experienced in lipid analysis. Each laboratory compared the Reflotron with the same conventional wet-chemistry method, Boehringer phosphotungstate-Mg2+ precipitation with enzymatic cholesterol assay. Imprecision was within accepted guidelines, with CVs of < or = 8% for fresh and frozen plasmas (median CV 1.7-3.9%) and for lyophilized sera (median CV 3.8-4.7%), similar to those of the conventional method. Results of linear-regression analysis were as follows: Reflotron HDL Cholesterol = 1.03 conventional - 3.9 mg/L, r = 0.987. The Reflotron results were somewhat low in the two US laboratories, demonstrating the need for general standardization of methods for measuring HDL cholesterol. Results from capillary fingerstick plasma agreed well with those from venous-derived plasma; capillary = 1.04 venous + 4.5 mg/L, r = 0.967. The system is relatively insensitive to interference from hemoglobin (< or = 0.75 g/L), ascorbic acid (< or = 0.3 g/L), bilirubin (< or = 50 mg/L), cholesterol (< or = 3.5 g/L), and triglycerides (< or = 4 g/L). The relative ease of operation and the rapid availability of results (within 90 s for plasma collected in EDTA) make the method appropriate for use by well-trained, but not necessarily technical, operators in the physician's office or other alternative sites.

Asunto(s)

RESUMEN

The concentrations of Fe, Mn, Pb and Cr have been determined in broncho-alveolar lavage (BAL) fluid of 25 subjects without occupational or abnormal environmental exposure to metals, using the AAS method. The numerous factors which can interfere with the results in pre-analytical and in analytical phases are stressed. Metals concentrations in BAL are expressed in micrograms/l. They were not correlated with the volume of fluid recovered, the total cells, alveolar macrophages and erythrocytes. The results were not modified by stratification considering age and sex. Iron concentrations were higher than others, probably due to higher environmental exposure and partly to its essential role in humans. The diagnostic significance of element determination in BAL fluid and the relationship with exposure and lung load is discussed.

Asunto(s)

RESUMEN

Reference methods for serum low density lipoprotein (LDL) separation are time-consuming and the salts used in density-gradient ultracentrifugation may cause chemical and/or immunological changes in the lipoprotein structure. A method has been developed to provide native LDL suitable for chemical and immunochemical studies. The three step procedure involves firstly the separation of very low density lipoproteins by non-density adjusted ultracentrifugation, secondly the separation of low from high density lipoproteins by LDL precipitation with PEG-6000, and finally the re-dissolution of the pellet in NaCl 150 mmol/l. The effectiveness of LDL separation, as well as the preservation of the electrophoretic mobility of the LDL molecules, was verified by electrophoresis in agarose gel, and the maintenance of the immunochemical reactivity of apolipoprotein B was verified by an immunochemical assay.

Asunto(s)

RESUMEN

Lowering blood pressure is not totally effective in preventing the atherosclerotic complications of systemic hypertension. In hypertensive patients both platelet hyperaggregation and dyslipidemia have been suggested as important risk factors. The effect of 8 weeks' treatment with ketanserin on blood pressure, serum lipid parameters (cholesterol, triglycerides, LDL, HDL-C, apolipoprotein A1 and B) and platelet aggregation, induced by collagen, ADP, arachidonic acid, was evaluated in 10 patients with essential hypertension. Ketanserin was effective in lowering blood pressure in all patients, 6 of whom became normotensive. Both CHOL and TG levels and APO B were significantly reduced, whereas HDL-C and APO A1 were significantly increased after treatment. These results might be attributed to the antagonistic activity of ketanserin on alpha-1 adrenoceptors with a consequent inhibition of phosphodiesterase. Platelet aggregation, after stimulation with collagen and arachidonic acid, was significantly reduced secondary to the inhibition of intraplatelet serotonin synthesis and release. These results suggest that keranserin is effective in reducing blood pressure and in achieving normal serum lipid pattern and platelet aggregation. Therefore, this drug might be helpful in controlling the main risk factors for cardiovascular damage.

Asunto(s)

Asunto(s)

Asunto(s)

RESUMEN

Ten patients with essential hypertension and normal renal function were treated with ketanserin (20-40 mg twice a day), administered for 8 weeks. In all patients, the changes in systemic and renal hemodynamics, and in urine albumin excretion, were assessed. Ketanserin monotherapy effectively lowered blood pressure in all patients. No change in cardiac output, pulse rate and stroke volume was observed; peripheral vascular resistance was significantly decreased. Plasma volume was unaltered. Renal plasma flow, glomerular filtration rate and filtration fraction were stable, with a slight but not significant reduction in renal vascular resistance. Urine albumin excretion remained unchanged. No relevant side effects were observed during the treatment period. In conclusion, our results confirm that ketanserin alone is an effective antihypertensive agent in patients with uncomplicated essential hypertension. The blood pressure lowering effect is mainly due to the systemic vasodilatation; renal hemodynamics and function are well preserved.

Asunto(s)

Asunto(s)

RESUMEN

The aim of this study was to evaluate the ability of plasma levels of apo A-I and apo B to discriminate between male patients with and without angiographically determined coronary artery disease (CAD) in comparison to the levels of cholesterol, HDL-cholesterol and triglycerides. The plasma apo A-I and B levels were measured by a radial immunodiffusion assay that made use of well characterized monoclonal antibodies. Univariate statistical analysis showed that the mean values for HDL-cholesterol and apo A-I were significantly lower in patients with single-, double- and triple-vessel disease than in the control subjects. The mean values for plasma cholesterol and apo B were significantly higher in the group with double- and triple-, but not single-vessel disease, than in the group without CAD. No statistically significant difference among groups was found for plasma triglyceride values. Since the considered variables failed to assess the severity of the disease, as determined by the number of vessels involved, the 3 groups of patients with CAD were combined into one group. A multiple logistic analysis, performed in order to assess the independent role of the variables and to estimate the corresponding odds ratio, indicated an independent association of HDL-cholesterol, apo A-I and apo B with CAD. When the multiple logistic analysis was repeated with a stepwise procedure, only apo A-I and B entered into the model. The results of our study indicate that plasma levels of apo A-I and B are more useful than plasma lipid in detecting the presence or absence of coronary artery disease in a group of male patients undergoing coronary angiography.

Asunto(s)

RESUMEN

A procedure is presented and evaluated for separation of plasma high density lipoprotein from either capillary or venous whole blood. The lipoprotein is separated by adding 50 microliter of sample to 250 microliter of 0.15 M NaCl solution containing 99.9 g/l polyethyleneglycol 6000, 0.0374 g/l dextran sulfate (Mr 15,000) and 2.6 mM Mg2+. After gentle mixing for a few minutes and standing 10 min at room temperature, mixtures are centrifuged (1,500 g) for 10 min and cholesterol is measured on 200 microliter of supernatant by an enzymatic-colorimetric method. Comparison studies demonstrate a good correlation between high density lipoprotein cholesterol in plasma and capillary or venous whole blood. The procedure is simple, has the advantage of using either K3-EDTA-anticoagulated whole blood, without the need of centrifugation, or capillary whole blood which can also be collected away from the laboratory.

Asunto(s)