Biomedical subjects

G Siest

Publications and source records attributed to G Siest.

At least 55 records · Page 3Linked to original sources

PubMed2000

Dietary intakes, eating style and overweight in the Stanislas Family Study.

OBJECTIVES: To describe the eating patterns of members of French families and to assess the relationships between dietary intakes, eating style and overweight. DESIGN: Cross-sectional analysis of nutritional and behavioural characteristics. SUBJECTS: 1,320 members of 387 families (age 11-65y) attending the Centre for Preventive Medicine for a routine medical check-up. MEASUREMENTS: Individual body weight and height were measured. Food intake was assessed using a three day dietary record. Eating style was measured using the French validated version of the Dutch Eating Behaviour Questionnaire. RESULTS: In each of the four groups (men, women, boys and girls), dietary restraint was positively correlated with overweight (P< or =0.001) and associated with lower energy intakes (P < or = 0.05-P < or = 0.001). A negative association between energy intake and overweight was found in girls only (P< or =0.001). In all cases, overweight and dietary restraint exaggerated any existing macronutrient imbalance in energy intake (ie higher protein and fat contributions, lower carbohydrate contribution). Emotional eating was positively correlated to body mass index in women only (P< or =0.01). External eating was mainly a characteristic of children (P< or =0.001). CONCLUSION: As in overweight subjects, clear relationships were found in this sample of general population between dietary intakes and eating style. The population will be followed up for 10y. In the long term, these results should have implications in the prevention of obesity.

Adolescent↗

PubMed2000

Lipoprotein lipase (C/G)447 polymorphism and blood pressure in the Stanislas Cohort.

OBJECTIVE: Association between blood pressure and triglyceride levels, and between lipoprotein lipase (LPL) (C/G)447 polymorphism and triglyceride levels has been described. We investigated whether the LPL (C/G)447 polymorphism was associated with blood pressure (BP) levels and longitudinal changes. DESIGN AND PARTICIPANTS: For cross-sectional analysis, 767 men and 816 women (29-55 years) were selected from the Stanislas Cohort, a cohort of volunteers for a free health check-up. Only subjects without anti-hypertensive or lipid-lowering medication were included in the study. A subset of this sample population, 359 men and 337 women, had been followed during the 11 years prior to recruitment in the Stanislas Cohort and was used for longitudinal analysis. RESULTS: The cross-sectional study showed that serum triglyceride levels differed significantly according to LPL genotypes in both genders, the G447 allele being associated with the lowest triglyceride levels (P < or = 0.01). Univariate and multivariate analysis found that LPL polymorphism was not related to BP levels in men. In contrast, women with the LPL-G447 allele had lower systolic (SBP) and pulse (PP) pressure levels than those with the LPL-CC genotype (P < or = 0.01 and P < or = 0.05, respectively); this association being independent of triglyceride level. The longitudinal study showed LPL genotype was an independent predictor of PP and SBP follow-up levels in women; changes over 11 years being lower for LPL-G447 allele carriers (P < or = 0.05). These associations were independent of triglyceride level. CONCLUSION: The LPL-G447 allele was found associated with lower PP and SBP independently of triglyceride level in women. This result suggests that the LPL gene may influence blood pressure.

Adult↗

PubMed2000

Effect of short- and long-term storage on human serum and recombinant apolipoprotein E concentration.

In order to assess the short- and long-term stability of apolipoprotein (apo) E concentration in serum, we compared the apo E concentrations measured in fresh human serum samples with those determined after storage at +4 degrees C, -20 degrees C or -80 degrees C. The serum apo E concentration was measured by immunoturbidimetry using an anti-human apo E polyclonal antibody from goats. One week storage at +4 degrees C did not significantly affect the serum apo E concentration. At -20 degrees C or -80 degrees C no significant change in apo E concentration occurred during up to three months of storage. Moreover, the concentration of apo E was not modified after long-term storage of serum samples kept at -196 degrees C in liquid nitrogen for up to four years. In addition, 15 freeze-thaw cycles, over a 3-week period, did not affect the apo E concentration in serum. A similar freeze-thaw procedure applied to purified human recombinant apo E showed that apo E2 isoform was the most stable in comparison with the apo E3 and apo E4 isoforms.

Apolipoproteins E↗

PubMed2000

Need for revisiting the concept of reference values.

The reference values concept has been adopted by health care professionals, including clinical chemists, laboratory scientists, and clinicians and simultaneously by all the official organizations in charge of the establishment of legislation. But the estimation of reference limits, and the evaluation of biological variability need to be improved at the level of the procedures, which are currently too long and too expensive and not feasible easily for all laboratories. The procedures for obtaining reference values, if we follow the original documents, are complex, and that is the main reason that clinical chemists or diagnostic kit manufacturers have not used them systematically. There is clearly a need that scientific societies and international organizations propose practical recommendations: 1) Recommendations to describe methods linked to systematic error. * How to transfer reference limits from one laboratory to another laboratory using different methods? * Should we determine reference limits for each method? * How can we differentiate bias due to the populations from these due to the method? Clear collaborations with manufacturers involved in kits and diagnostic systems are needed. 2) Practical recommendations linked to the reference population. * How to select a homogeneous population? (Careful recommendations on the choice between healthy individuals, blood donors and individuals hospitalised for other diseases should be given.) * How to estimate ethnic differences? * How to define the exclusion and inclusion criteria according to quantity? * How to deal with the question of reference limits for unstable periods, aging or old people particularly, when the difference between aging and disease is very difficult to define? 3) Practical recommendations on the statistical methods to be used. * How to make a good choice of the interquartile interval? Should we use and present only the centiles 2.5 or 97.5, or on the contrary should we give other centiles in addition, for example 5, 10, 75, 80, 85, 90? 4) Practical recommendations linked to the use of the concept of the reference values. * How to make this concept more concrete and to have official definitions which are better understandable and not only abstract? * How to demonstrate the value of using simultaneously reference limits and decision limits, and what does each of these limits bring to results interpretation? * How to improve the presentation of the results? How to give more information on biological variability in the laboratory data, taking into account the scientific validity of their determination? Should we use new information techniques and new communication systems for reaching these objectives? The responses to all these questions could only be provided if there is a concerted effort at the international level. Practical recommendations should be given, which would be very useful for a better understanding and use of reference values by laboratory scientists and clinicians.

Clinical Chemistry Tests↗

PubMed2000

Associations of apolipoprotein E concentration and polymorphism with lipids and apolipoprotein levels in Chinese from Beijing and Shanghai.

The association of apolipoprotein E concentration and common polymorphism (codons 112/158) with lipid and apolipoprotein concentrations was studied in two Chinese healthy population samples from Beijing (n=99) and Shanghai (n=67). Body mass index and apolipoproteins E and AI and triglyceride concentrations were significantly different between the two populations (0.001<p<0.05), but apolipoprotein E common allele frequencies were not significantly different. Apolipoprotein E concentration was independently associated with apolipoproteins B and AI, total cholesterol and triglyceride concentrations in the Beijing group, but only with total cholesterol and apolipoprotein AI concentrations in the Shanghai group. Apolipoprotein E polymorphism was significantly associated with apolipoproteins E and AI, and triglyceride concentrations in the Beijing subjects (0.001<p<0.01), whereas apolipoprotein E polymorphism was significantly related only to apolipoprotein E concentration in the Shanghai group (p<0.001). This preliminary study with a limited number of subjects shows that the relations between both apolipoprotein E genotypes and concentration with lipid traits are not the same in the two studied Chinese populations.

Adolescent↗

PubMed2000

Apolipoprotein E polymorphism and serum concentration in Alzheimer's disease in nine European centres: the ApoEurope study. ApoEurope group.

As part of the ApoEurope Project, apolipoprotein E (apo E) common polymorphism and serum concentration were determined in 489 Alzheimer's disease patients and 429 controls. Patients and controls were recruited through nine centres in eight European countries. Age, sex ratios and education levels of both case and control populations were similar, although discrete differences appeared between centres. The prevalence of the epsilon4 allele was higher in Alzheimer's disease than in controls (increased by 140%), while serum apo E concentration was lower by 11.2% (p<0.001). In addition, serum total cholesterol and triglyceride concentrations were lower in Alzheimer's disease (p<0.001), while that of apo Al was not affected. The decrease in serum apo E concentration was not accounted for by the epsilon4 allele, age or gender, suggesting that apo E concentration might represent an additional risk factor for Alzheimer's disease, complementary and independent of the epsilon4 allele. Further analysis will be aimed at determining whether the quantitative link between apo E concentration and Alzheimer's disease occurs through the effect of apo E genotype on lipid parameters or by other mechanisms.

Age Factors↗

PubMed2000

Familial studies on the genetics of cardiovascular diseases: the Stanislas cohort.

In a given individual, the level of cardiovascular risk results from the combination of and interactions between genetic and environmental components. We choose to investigate segregation analysis of intermediate phenotypes in healthy nuclear families, belonging to the Stanislas cohort, a large familial cohort composed of 1006 families, which will be followed for 10 years. We developed a panel of 35 genetic markers including genes involved in lipid metabolism, regulation of blood pressure, thrombosis, platelet function, and endothelial cell adhesion. The allele frequencies of the studied polymorphisms were in agreement with those reported in other Caucasian populations. As an example of segregation analysis, we investigated carotid intima-media thickness (CIMT) variability in a subset sample of the Stanislas cohort. We found that about 30% of CIMT variability was attributable to genetic factors. Associations between CIMT and polymorphisms in apo CIII, cholesteryl ester transfer protein, methylene tetrahydrofolate reductase, and fibrinogen genes were observed and explained about 20% of CIMT variability in men. Furthermore, as another example of association studies, we investigated the relations between E-selectin polymorphisms and blood pressure interindividual variability and longitudinal changes in unrelated adults of this familial population. The E-selectin Phe554 allele was found associated with lower systolic blood pressure and diastolic blood pressure.

Adult↗

PubMed2000

Apolipoprotein E polymorphisms and concentration in chronic diseases and drug responses.

Apolipoprotein (apo) E is an important circulating and tissue protein involved in cholesterol homeostasis and many other functions. The common polymorphism in the coding region of the gene, four polymorphisms in the promoter region, other additional single nucleotide polymorphisms, as well as several apo E variants have been identified. The common coding polymorphism strongly influences the lipid metabolism and the circulating concentration of apo E itself. This polymorphism is at the origin of the implication of apo E in cardiovascular and neurodegenerative diseases, but also of the relation of apo E with longevity. Probably due to its many metabolic and functional consequences, apo E polymorphism has been shown to influence the responses of patients to several drugs (fibrates, statins, hormone replacement therapy, anti-Alzheimer drugs) or environmental interventions (black tea, alcohol, diet). Apo E genotyping may be clinically helpful in defining the risk of patients and their responses to therapeutics. Finally, circulating apo E concentration appears to be altered in diseases and can be modulated by some of the drugs cited above. This parameter can thus also give interesting clinical information and could be a therapeutic target, providing it is validated. At the present time, we cannot exclude that apo E concentration may be the most prominent apo E parameter to be considered in health and disease, while apo E polymorphisms would represent only secondary parameters influencing apo E concentration.

Alleles↗

PubMed2000

High sensitivity C-reactive protein: biological variations and reference limits.

Serum C-reactive protein (CRP) concentration was determined for 3605 subjects using an immunonephelometric assay improved to provide greater sensitivity. Subjects were from 5 to 75 years old and belonging to 1003 nuclear families recruited from the Stanislas Cohort Study between January 1994 and August 1995. Sample values for CRP ranged from 0.17 mg/l to 100 mg/l. Geometric means (mean - SD; mean +/- SD) were in the 5-14 years old group 0.37 (0.17-1.07) mg/l, in the 15-28 years old group 0.47 (0.17-1.38) mg/l and in the 29-75 years old group 0.98 (0.34-2.85) mg/l. For women, the geometric means were 0.38 (0.17-1.10) mg/l, 0.62 (0.20-1.90) mg/l and 0.98 mg/l (0.31-3.13) mg/l respectively. The interindividual variability ranged from 138% to 759% among different age classes. Biological factors associated with CRP concentration variations were examined and accounted for 25% of the CRP variability in men and 40% in women. The main biological factors statistically associated with CRP concentration variations in men were: drugs, leukocyte count, body mass index, tobacco consumption, age, and in women: drugs, leukocyte count, age, body mass index and hemoglobin concentration. These factors were used to define the exclusion and partition criteria when obtaining the reference samples. Medians for reference values ranged from 0.20 to 0.68 mg/l in males and from 0.20 to 0.78 mg/l in women.

Adolescent↗

PubMed1999

Kinetics of apolipoprotein E isoforms-binding to the major glycosaminoglycans of the extracellular matrix.

Apolipoprotein E (apoE), a key lipid transport protein, displays a heparin-binding property that is critical in several apoE functions. The kinetics of the interaction between apoE isoforms and glycosaminoglycans (GAGs) were studied using surface plasmon resonance. The dissociation constant of equilibrium K(D) for apoE3-heparin interaction was estimated to be 12 nM for apoE3 and three common apoE isoforms revealed similar affinities for heparin. ApoE binds to GAGs in the following order: heparin>heparan sulfate>dermatan sulfate>chondroitin sulfate. The affinity parameter of the binding of low molecular weight heparins to apoE is correlated with the chain length. The effective number Z of electrostatic interactions between plasma apoE3 and heparin was assessed to be three. Metal chelators were able to diminish apoE-binding to heparin, suggesting some stabilizing effect of metal ions while reconstitution with lipids did not affect binding affinities for heparin, suggesting that the N-terminal heparin-binding site is responsible for apoE-containing lipoprotein interactions with heparin.

Apolipoproteins E↗

PubMed1999

Wine, beer, and mortality in middle-aged men from eastern France.

OBJECTIVE: To evaluate prospectively the health risk of wine and beer drinking in middle-aged men in the area of Nancy, France. DESIGN: Prospective cohort study. SUBJECTS: A total of 36 250 healthy men who underwent comprehensive health appraisals in a center of preventive medicine between January 1, 1978, and December 31, 1983. MAIN OUTCOME MEASURES: Education, professional and leisure activities, and smoking and drinking habits were evaluated using a questionnaire. Blood pressure and mean corpuscular volume and gamma-glutamyltransferase, glucose, and serum cholesterol levels were routinely measured, and electrocardiography was routinely performed. We recorded mortality from all causes and specific causes during a 12- to 18-year follow-up across categories of baseline alcohol consumption. RESULTS: Of the subjects, 28% drank beer, 61% drank wine but no beer, and 11% were abstainers; there was not much difference between social classes. During the follow-up, 3617 subjects died. The relative risk of death was estimated by the Cox proportional hazards model using nondrinkers as the reference and adjusting for 4 or 5 covariables. Moderate intake of both wine and beer was associated with lower relative risk for cardiovascular diseases; the risk was more significant with the intake of wine. For all-cause mortality, only daily wine intake (22-32 g of alcohol) was associated with a lower risk (0.67; 95% confidence interval, 0.58-0.77; P<.001) due to a lower incidence of cardiovascular diseases, cancers, violent deaths, and other causes. CONCLUSION: In eastern France, moderately drinking only wine was associated with a lower all-cause mortality, although drinking both wine and beer reduced the risk of cardiovascular death.

Beer↗

PubMed1999

Glycation of apolipoprotein E impairs its binding to heparin: identification of the major glycation site.

The increased glycation of plasma apolipoproteins represents a possible major factor for lipid disturbances and accelerated atherogenesis in diabetic patients. The glycation of apolipoprotein E (apoE), a key lipid-transport protein in plasma, was studied both in vivo and in vitro. ApoE was shown to be glycated in plasma very low density lipoproteins of both normal subjects and hyperglycemic, diabetic patients. However, diabetic patients with hyperglycemia showed a 2-3-fold increased level of apoE glycation. ApoE from diabetic plasma showed decreased binding to heparin compared to normal plasma apoE. The rate of Amadori product formation in apoE in vitro was similar to that for albumin and apolipoproteins A-I and A-II. The glycation of apoE in vitro significantly decreased its ability to bind to heparin, a critical process in the sequestration and uptake of apoE-containing lipoproteins by cells. Diethylenetriaminepentaacetic acid, a transition metal chelator, had no effect on the loss of apoE heparin-binding activity, suggesting that glycation rather than glycoxidation is responsible for this effect. In contrast, glycation had no effect on the interaction of apoE with amyloid beta-peptide. ApoE glycation was demonstrated to be isoform-specific. ApoE(2) showed a higher glycation rate and the following order was observed: apoE(2)>apoE(4)>apoE(3). The major glycated site of apoE was found to be Lys-75. These findings suggest that apoE is glycated in an isoform-specific manner and that the glycation, in turn, significantly decreases apoE heparin-binding activity. We propose that apoE glycation impairs lipoprotein-cell interactions, which are mediated via heparan sulfate proteoglycans and may result in the enhancement of lipid abnormalities in hyperglycemic, diabetic patients.

Aged↗

PubMed1999

Capillary electrophoretic analysis of recombinant human apolipoprotein E. Calibration mode of a protein reference material.

Actual lack of standardization of serum apolipoprotein (apo) E measurements prevents intensive studies on the significance of apoE concentration in clinical chemistry. For this purpose the use of standards calibrated against a common reference material is essential in order to obtain reliable results. The assignment of a certified value to this reference material involves the use of an accurate and non immunological technique. We demonstrate here that capillary electrophoresis in sodium dodecyl sulfate containing gel (SDS-CGE) can be a method of choice. ApoE quantification was performed according to the peak area and a standard solution of purified apoAI. We obtained very close results to that obtained by HPLC of phenylalanine apoE content measurement i.e., 81.6 mg/l versus 84.6 mg/l and concluded that the SDS-CGE analysis is an accurate and reliable method for the certification of an apoE reference material to be used in serum apoE concentration measurements. The SDS-CGE combines advantages of SDS-polyacrylamide gel electrophoresis (high resolving power, rapidity and tolerance of complex sample) with that of quantitative HPLC analysis (accuracy, precision, linearity and speed).

Apolipoprotein E3↗

PubMed1999

Reversal of hyperlipidaemia in apolipoprotein C1 transgenic mice by adenovirus-mediated gene delivery of the low-density-lipoprotein receptor, but not by the very-low-density-lipoprotein receptor.

We have shown previously that human apolipoprotein (apo)C1 transgenic mice exhibit hyperlipidaemia, due primarily to an impaired clearance of very-low-density lipoprotein (VLDL) particles from the circulation. In the absence of at least the low-density-lipoprotein receptor (LDLR), it was shown that APOC1 overexpression in transgenic mice inhibited the hepatic uptake of VLDL via the LDLR-related protein. In the present study, we have now examined the effect of apoC1 on the binding of lipoproteins to both the VLDL receptor (VLDLR) and the LDLR. The binding specificity of the VLDLR and LDLR for apoC1-enriched lipoprotein particles was examined in vivo through adenovirus-mediated gene transfer of the VLDLR and the LDLR [giving rise to adenovirus-containing (Ad)-VLDLR and Ad-LDLR respectively] in APOC1 transgenic mice, LDLR-deficient (LDLR-/-) mice and wild-type mice. Remarkably, Ad-VLDLR treatment did not reduce hyperlipidaemia in transgenic mice overexpressing human APOC1, irrespective of both the level of transgenic expression and the presence of the LDLR, whereas Ad-VLDLR treatment did reverse hyperlipidaemia in LDLR-/- and wild-type mice. On the other hand, Ad-LDLR treatment strongly decreased plasma lipid levels in these APOC1 transgenic mice. These results suggest that apoC1 inhibits the clearance of lipoprotein particles via the VLDLR, but not via the LDLR. This hypothesis is corroborated by in vitro binding studies. Chinese hamster ovary (CHO) cells expressing the VLDLR (CHO-VLDLR) or LDLR (CHO-LDLR) bound less APOC1 transgenic VLDL than wild-type VLDL. Intriguingly, however, enrichment with apoE enhanced dose-dependently the binding of wild-type VLDL to CHO-VLDLR cells (up to 5-fold), whereas apoE did not enhance the binding of APOC1 transgenic VLDL to these cells. In contrast, for binding to CHO-LDLR cells, both wild-type and APOC1 transgenic VLDL were stimulated upon enrichment with apoE. From these studies, we conclude that apoC1 specifically inhibits the apoE-mediated binding of triacylglycerol-rich lipoprotein particles to the VLDLR, whereas apoC1-enriched lipoproteins can still bind to the LDLR. The variability in specificity of these lipoprotein receptors for apoC1-containing lipoprotein particles provides further evidence for a regulatory role of apoC1 in the delivery of lipoprotein constituents to different tissues on which these receptors are located.

Adenoviridae↗

PubMed1999

Bivariate familial correlation analysis of quantitative traits by use of estimating equations: application to a familial analysis of the insulin resistance syndrome.

Familial correlation analysis involving two traits may give a better insight into the etiology of multifactorial syndromes than familial analysis focused on single traits. Significant cross-trait correlations between biological relatives but not between spouses suggest that the two traits share common transmissible factors whereas correlations between spouses additionally suggest the influence of shared lifestyle factors. We apply the Estimating Equations (EE) technique to the estimation of intra-trait and cross-trait familial correlations on two quantitative traits. Unlike maximum likelihood methods, the EE method does not require one to specify the joint distribution of the traits. Estimation of correlations and of their variance involves an iterative three-stage algorithm which converges rapidly. The generalized Wald test can be used to test any specific hypothesis of familial resemblance. This method has great flexibility for handling covariates and incomplete family data. A simulation study indicated that the EE technique performed well in large samples (100 families), both in terms of type I error and coverage probability . However, in small samples (50 families), an increase of the type I error and a decrease of the coverage probability was observed. As an illustration, we applied this technique to a family study of metabolic factors involved in the Insulin Resistance Syndrome (body mass index, insulin, triglycerides, HDL-cholesterol, and diastolic blood pressure). The study was carried out in a sample of 216 healthy nuclear families with > or =2 offspring. The results suggested the existence of a common transmissible (genetic or cultural) factor influencing both body mass index and insulin, whereas the weak clustering of triglycerides and HDL-cholesterol would be more compatible with the influence of shared lifestyle factors.

Algorithms↗

PubMed1999

Simultaneous measurement of reactive oxygen species and reduced glutathione using capillary electrophoresis and laser-induced fluorescence detection in cultured cell lines.

A capillary zone electrophoretic (CZE) method coupled with laser-induced fluorescence (LIF) was developed for the simultaneous determination of two important intracellular parameters related to oxidative stress (i.e. reactive oxygen species, ROS, and reduced glutathione, GSH). This rapid and sensitive method was applied to the study of oxidative stress in cultured V79 fibroblast cells. The fluorogenic reagents selected were: (i) dihydrorhodamine-123 (DHR-123) which is converted intracellularly by ROS to the fluorescent rhodamine-123 dye (Rh-123), and (ii) naphthalene-2,3-dicarboxaldehyde (NDA), which reacts quickly with GSH in cell extracts to produce a fluorescent adduct. Separation of Rh-123, GSH-NDA and gamma-glutamylcysteine-NDA adducts was performed using an uncoated fused-silica capillary and a 100 mM borate buffer, pH 9.2, at 20 degrees C and at an applied voltage of 25 kV; LIF detection was operated using an argon laser. The cell line was also tested for its ability to alleviate oxidative stress induced by tert-butylhydroperoxide (t-BuOOH). Exposure to t-BuOOH (up to 3 mm for 2 h) did not affect the intracellular ROS and GSH concentrations. At higher (4-10 mM) t-BuOOH concentrations, an inverse relationship between the concentrations of ROS and GSH was obtained, showing that the present method can readily evaluate the gradual consumption of the primary cellular scavenger of ROS which occurs simultaneously with the increase of oxidative insult.

Animals↗

PubMed1999

Measurement of thiols in human plasma using liquid chromatography with precolumn derivatization and fluorescence detection.

A liquid chromatography (LC) method for the simultaneous measurement of the main low molecular mass thiols (i.e., cysteine, cysteinylglycine, homocysteine, and glutathione) in human plasma is described. The sample treatment consists of the reduction of disulfide bounds with tri-n-butylphosphine and protein precipitation with trichloroacetic acid followed by precolumn derivatization with a thiol-selective fluorogenic reagent (7-fluoro-2,1,3-benzoxadiazole-4-sulfonamide). The structure of thiol derivatives is assessed using electrospray ionization-mass spectrometry (MS). The stability of resulting adducts in acidic medium (24 h at 10 degrees C) allows the automation of the technique and a high throughput of samples (approximately 50 per day). Separation is complete within 12 min using isocratic reversed-phase mode, and detection is operated by spectrofluorimetry (lambda ex = 385 nm and lambda em = 515 nm). Quantitation is performed by an internal standardization mode using thioglycolic acid. The LC method is fully validated, and homocysteine concentrations obtained in plasma samples are compared with values measured using either fluorescence polarization immunoassay or capillary gas chromatography-MS; a good correlation is observed between LC and both methods. The method has been applied in daily use to a large-scale study in a human healthy population, and some resulting data are discussed.

Adolescent↗

PubMed1999

A multilocus genotyping assay for candidate markers of cardiovascular disease risk.

A number of chronic diseases, including cardiovascular disease, appear to have a multifactorial genetic risk component. Consequently, techniques are needed to facilitate evaluation of complex genetic risk factors in large cohorts. We have designed a prototype assay for genotyping a panel of 35 biallelic sites that represent variation within 15 genes from biochemical pathways implicated in the development and progression of cardiovascular disease. Each DNA sample is amplified using two multiplex polymerase chain reactions, and the alleles are genotyped simultaneously using an array of immobilized, sequence-specific oligonucleotide probes. This multilocus assay was applied to two types of cohorts. Population frequencies for the markers were estimated using 496 unrelated individuals from a family-based cohort, and the observed values were consistent with previous reports. Linkage disequilibrium between consecutive pairs of markers within the apoCIII, LPL, and ELAM genes was also estimated. A preliminary analysis of single and pairwise locus associations with severity of atherosclerosis was performed using a composite cohort of 142 individuals for whom quantitative angiography data were available; evaluation of the potentially interesting associations observed will require analysis of an independent and larger cohort. This assay format provides a research tool for studies of multilocus genetic risk factors in large cardiovascular disease cohorts, and for the subsequent development of diagnostic tests.

Adult↗