STANISLAS Cohort Research Articles

Association between EGF and lipid concentrations: A benefit role in the atherosclerotic process?

Context and objective Previous in vitro studies have shown a relationship between epidermal growth factor (EGF) and lipid metabolism. Indeed, EGF is able to modulate lipoprotein fractions in human fetal intestine and hepatic-derived cell lines. The aim of this study was to search for potential associations between EGF concentrations and lipid parameters in both plasma and peripheral blood mononuclear cells (PBMCs) among healthy individuals. Design EGF concentrations were quantified in plasma of 273 men and 279 women and in PBMCs of 57 men and 62 women from the STANISLAS cohort. In addition, basic blood constituents including lipid parameters (apoliporoteins A1, and B, total and high-density lipid (HDL)-cholesterols and triglycerides) were measured. Results Plasma EGF concentrations were significantly and positively correlated with apolipoprotein A1 and HDL-cholesterol concentrations after adjustment for age, sex, and body mass index ( r = 0.155, P = 0.0003; and r = 0.112, P = 0.0086, respectively). They were also negatively correlated with the apolipoprotein B/A1 ratio ( r = − 0.107, P = 0.012). In addition, EGF concentrations in PBMCs were negatively correlated with apolipoprotein B, total and low-density lipid-cholesterols, and triglyceride concentrations, and apolipoprotein B/A1 ratio ( P = 0.0018, P = 0.0137, P = 0.0142, P = 0.0162, and P = 0.0077, respectively, with r ranging between − 0.287 and − 0.222). No interactions were found with gender. Conclusion New associations between EGF and lipid concentrations have been reported, providing new perspecctives with regard to the role of EGF in atherosclerosis.

Visfatin, low‐grade inflammation and body mass index (BMI)

Visfatin is an adipokine with revealing roles in inflammatory mechanisms but its implication in inflammation related to excessive adiposity/obesity is not studied yet. Our aim was to investigate the relations of visfatin with inflammation markers and body mass index (BMI) in the peripheral blood mononuclear cells (PBMCs), a type of cells closely related to inflammatory mechanisms. Cross-sectional study, quantification of visfatin, TNF-alpha, IL-6 mRNA in PBMCs. Eighty-three supposed healthy individuals from the STANISLAS cohort, belonging in three BMI categories: BMI < 25 kg/m(2) (lean), 25 kg/m(2) <or= BMI < 30 kg/m(2) (overweight) or BMI >or= 30 kg/m(2) (obese). We measured visfatin gene expression (by real-time quantitative PCR), in relation to gene expression of the pro-inflammatory cytokines TNF-alpha, IL-6 in PBMCs and to anthropometric parameters (weight, BMI, waist : hip ratio), blood pressure, lipid profile, glucose and inflammatory markers (C-reactive protein, lymphocyte count). Visfatin expression in PBMCs was significantly associated with BMI in a negative way (r = -0.21, P = 0.05). Global anova analysis test for lean and over-weight/obese individuals showed a negative significant association between visfatin expression in PBMCs and BMI both for men and women (P = 0.05 and P = 0.01, respectively) and these associations remained significant after separating subjects in three groups (lean, overweight, obese) for men and women (P = 0.02 and P = 0.05, respectively). Correlation analysis between levels of expression of visfatin and TNF-alpha showed a significant positive linear association (r(2) = 0.27, P < 0.0001). These findings reveal a probable new role of visfatin in inflammation reflected in PBMCs, in the context of obesity.

P-selectin polymorphisms' influences on P-selectin serum concentrations and on their familial correlation: the STANISLAS family study.

P-selectin is an adhesion molecule known to be involved in the pathogenesis of several diseases through its major role in the initial phase of leukocytes recruitment during inflammation. However, genetic characterization of soluble P-selectin remains unclear. In the STANISLAS cohort, we study the familial correlations of P-selectin levels and investigate the association of six P-selectin polymorphisms (C-2123G, A-1969G, S290N, N562D, V599L and T715P) and cardiovascular risk factors with P-selectin concentrations. Full phenotypic and genotypic information was available for 136 healthy families composed of both natural parents and at least one child (boys, n = 125; and girls, n = 139) aged more than 4 years. While no correlation was observed between spouses, family correlations of P-selectin concentrations were highly significant for sibling (0.50 +/- 0.12, P < 10(-3)) and child-parent pairs (0.42 +/- 0.04, P < 10(-3)). P-selectin haplotypes explained about 25% of the variability of P-selectin concentrations, this effect being mainly due to the additive effects of two polymorphisms, V599L and T715P. After adjusting for the effect of the P-selectin polymorphisms, the sibling and child-parent correlations decreased to (0.39 +/- 0.08, P < 10(-4)) and (0.32 +/- 0.06, P < 10(-4)), respectively. In the present study, we showed that two P-selectin polymorphisms, V599L and T715P, explained about 25% of the variability of P-selectin concentrations and accounted for about 40% of their family resemblance. These results would suggest a genetic influence on P-selectin concentrations beyond the contribution of the P-selectin gene.

Genetic Polymorphism of CYP2C19 Gene in the Stanislas Cohort. A link with Inflammation

CYP2C19, a member of the cytochrome P450 family, metabolises arachidonic acid to produce epoxyeicosanoid acids, which are involved in vascular tone and inflammation. Thus, this study describes the possible relationship between a CYP2C19 polymorphism (681G>A) and three inflammatory markers: interleukin (IL)-6, tumor necrosis factor-alpha (TNF-alpha) and high sensitivity C-reactive protein (hs-CRP) in healthy individuals. In a sub-sample of 178 men and 181 women from the Stanislas study, we quantified plasma IL-6 and TNF-alpha concentrations by using an enzyme-linked immunosorbent assay, and serum hs-CRP concentration by immunonephelometry. The CYP2C19 681G>A polymorphism was genotyped using the kinetic thermocycling allele specific PCR method. In the Stanislas cohort, the frequency of the allele CYP2C19*2 (681A) was 17.8%. Circulating levels of inflammatory factors were increased in individuals homozygous for the defective allele CYP2C19*2 (A) notably IL-6 in the whole sample (P= 0.0008) and hs-CRP only in women (P= 0.008), with a significant interaction with sex (P= 0.005), in comparison to carriers of one copy or more of the wild type allele CYP2C19*1 (G). Only a trend of association (P= 0.089) was found between this polymorphism and TNF-alpha concentration in the whole sample. The association between CYP2C19*2 polymorphism and inflammatory markers' concentrations could suggest that CYP2C19 may be considered as a new candidate gene for cardiovascular risks via inflammation.

The STANISLAS Cohort: a 10-year follow-up of supposed healthy families. Gene-environment interactions, reference values and evaluation of biomarkers in prevention of cardiovascular diseases

The description of this familial longitudinal cohort was published in this journal 10 years ago, in 1998. To date, 117 publications on the STANISLAS Cohort (SC) have appeared, corresponding to five main categories of results: familial resemblance and heritability; genetics and gene-environment interactions; mRNA and proteins as gene products; reference values and biological variations of proteins; and finally preventive medicine and prepathological epidemiological data. More than 600 data values on demographic and laboratory data have been collected on each individual taking part out of the 1006 families at the beginning and for all three recruitments. Serum and plasma are stored in liquid nitrogen for all participants for all three recruitments. DNA has been extracted from all participants and mRNA from 357 families. They are stored at -80 degrees C. Owing to the SC study, heritability and many gene-environment interactions have been described. The expression of 166 genes related to cardiovascular diseases was measured in peripheral blood mononuclear cells RNA. Reference values for proteins and vitamins have been established in addition to reference values for the carotid and femoral intima media thickness in adults and children. The data obtained contribute to a better understanding of the relation between the studied polymorphisms (161 polymorphic sites) and health, and predisposition to obesity, high blood pressure and metabolic syndrome. To the best of our knowledge, the SC study is internationally the only longitudinal family cohort of subjects who are presumed to be healthy, which enables the study of the chain DNA-RNA-proteins.

Genetic profiling in healthy subjects from the Stanislas cohort based on 24 polymorphisms: effects on biological variables

The association of genetic profiles with biological or clinical assessments is not clearly established especially among apparently healthy subjects. A multivariate statistical analysis was performed on 24 polymorphisms related to the main metabolic pathways involved in cardiovascular diseases (CVDs). They were collected among 1551 healthy subjects of the Stanislas cohort to obtain genetic profiles. Association with biological variables was then studied at baseline (t0) and 5 years later (t5). Six genetic clusters were identified with relevant profiles and five polymorphisms from the selectin, apolipoprotein C3 and lipoprotein lipase genes (SELE-98G/T, APOC3-3175C/G, APOC3-482C/T, APOC3-1100C/T, LPL-93T/G) were sufficiently characteristic to associate 99.6% of the subjects with their corresponding cluster. A 5-year follow-up showed that clinical and biological measurements in relation to CVD risk factors already differ with triglyceride (p=0.009 for t0 and p=0.005 for t5) and high-density lipoprotein cholesterol (p=0.014 for t0 and p=0.003 for t5) for these previous genetic clusters. This study presents the hypothesis that SELE could be protective, whereas APOC3 could be associated with risk. It remains to be seen whether these polymorphisms will be predictive of CVD events among the selected clusters of different metabolic subtypes after a 10-year follow-up.

Transcription Factor and Drug-Metabolizing Enzyme Gene Expression in Lymphocytes from Healthy Human Subjects

We aimed to measure simultaneously the expression of drug-metabolizing enzymes (DME) and transcription factors (TF) with high importance in cardiovascular physiopathology in lymphocytes from healthy subjects. RNA was isolated from peripheral blood mononuclear cells (PBMC) of 20 subjects from the Stanislas Cohort. We used a microarray approach to measure 16 DME and 13 TF. Cytochromes P450 (P450s), including CYP2C19, CYP2C9, CYP2J2, CYP2D6, CYP1A1, CYP4F2, CYP4A11, CYP2E1, CYP11B2, CYP2C18, and CYP2A6, were expressed in all the subjects. CYP3A4 and CYP3A5 were not expressed. Glutathione S-transferases (GST) were expressed, but GSTM1 was seen only in some subjects. Pregnane X receptor (PXR), myocyte enhancer factor 2, vitamin D receptor, liver X receptor (LXR)-alpha, aryl hydrocarbon receptor (AHR), T-cell factor 7, constitutive androstane receptor, and aryl hydrocarbon receptor nuclear translocator (ARNT) were expressed in the majority of the subjects. Glucocorticoid receptor, peroxisome proliferator-activated receptor (PPAR)-gamma, and LXRbeta were expressed only in some individuals. PPARalpha mRNA was found in one subject only, and farnesoid X-activated receptor was not expressed. In addition, we found significant correlations between the expression of AHR, ARNT, and CYP1A1 and between PXR and P450 involved in leukotriene metabolism (CYP2C, CYP4F2, CYP4A11, CYP2J2, and CYP11B2). We describe here for the first time the presence of the majority of TF and DME in PBMC of healthy subjects without previous induction. The expression of these genes in lymphocytes could be a useful tool for further studying the physiological and pathological variations of DME and TF related to environment, to drug intake, and to cardiovascular metabolic cycles.

Une étude prospective de la prévalence du syndrome métabolique dans des familles françaises supposées saines: Importance du polymorphisme du TNF-alpha et de sa concentration plasmatique

Metabolic syndrome (MS) is a cluster of synergistically interacting cardiovascular risk factors which may have serious consequences for the development of cardiovascular disease and diabetes. In this study, we aimed to estimate the prevalence of MS within presumably healthy French families of the STANISLAS cohort, and to observe biological parameters involved in cardiovascular diseases among the offspring of MS subjects. 371 apparently healthy families (1366 individuals) were examined at two visits with a five-year interval (t0 and t+5). MS prevalence was assessed among parents following the ATP-III definition. Our results show that MS is present in presumably healthy adults of the STANISLAS cohort and increases with age. Moreover, low HDL-C and TNF-alpha may play an important role in the development of MS in childhood, at least in our population. Therefore, a systematic tracking of MS appears to be all the more important as it will permit early management of MS in parents and the installation of efficient preventive measures in children including specific advice for diet and physical activity.

Ambulatory Blood Pressure for Cardiovascular Risk Stratification

Ambulatory blood pressure (ABP) monitoring is increasingly recognized as a valuable tool to refine prediction of cardiovascular risk related to blood pressure (BP).1 After the first landmark study published by Perloff and colleagues 24 years ago,2 several longitudinal event-based studies provided unequivocal evidence of an independent association between ABP and risk of cardiovascular disease. Although experimental procedures and statistical analyses varied from study to study, ABP generally improved cardiovascular risk stratification over and beyond traditional risk factors, including clinic BP.3 The Table, obtained through an electronic search of literature using the terms “ambulatory blood pressure” and “prognosis,” shows a list of longitudinal event-based studies performed by independent groups. It is worth noting that the list of available studies is longer because each group generally published other analyses of their database. Only the first-appearing or main contribution from each group has been included in the Table. View this table: Longitudinal Event-Based Studies From Independent Groups That Addressed the Prognostic Value of ABP Article p 2145 Three aspects of available investigations deserve special mention. First, the prognostic value of ABP has been examined not only in subjects with clinical diagnosis of hypertension but also in the general population and in a variety of settings, including diabetes mellitus, renal failure, and cerebrovascular disease. Second, subjects could be untreated or treated at the time of ABP monitoring. This point may raise concerns, because drug treatment could exert unpredictable effects on 24-hour ABP profile and, consequently, interpretation and applicability of results. Third, although a continuous relation emerged in most studies between ABP and cardiovascular risk, several investigators tried to define clinical categories based on arbitrary thresholds of ABP. Although such categories are potentially useful to make diagnostic and therapeutic decisions in clinical practice, their prognostic role requires confirmation from large and independent cohort …

Association of classical and related inflammatory markers with high-sensitivity C-reactive protein in healthy individuals: results from the Stanislas cohort

Although high-sensitivity C-reactive protein (hs-CRP) has emerged as a cardiovascular marker, questions arise regarding the relative information provided by other inflammatory molecules. Therefore, as a first step, we examined interrelationships between serum hs-CRP concentrations and inflammatory, adhesion and growth factors in healthy adults. Circulating concentrations of hs-CRP, haptoglobin, orosomucoid, interleukin-6 (IL-6), IL-8, IL-18, tumor necrosis factor-alpha (TNF-alpha), TNF-receptor II (TNF-RII), E-, P-, and L-selectins, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1, endothelial growth factor (EGF), vascular EGF (VEGF), insulin-like growth factor-1 (IGF-1) and IGF-binding protein (IGFBP-3) were measured in 154 men and 161 women of the Stanislas cohort. Leukocyte and platelet counts were also determined. Correlations were significant between hs-CRP concentrations and leukocyte and platelet counts, as well as haptoglobin, orosomucoid, IL-6, and ICAM-1 concentrations (p< or =0.001). Correlation coefficients for ICAM-1 were higher in men than in women (p< or =0.05). When stratifying subjects according to hs-CRP levels, the group with high hs-CRP levels had significantly higher haptoglobin and orosomucoid concentrations than the others, in addition to higher leukocyte counts and IL-6 concentrations in women, and platelet counts and ICAM-1 concentrations in men. Further studies are warranted to explain the association pattern for hs-CRP. Partition of these factors according to their association with hs-CRP concentration opens a new perspective for choice of the best factors in terms of cardiovascular risk in relation to hs-CRP, while non-associated markers could be used to give additional information.

Interaction between CYP1A1 T3801C and AHR G1661A polymorphisms according to smoking status on blood pressure in the Stanislas cohort

CYP1A1, one of the key enzymes in detoxifying toxic components produced during cigarette smoking, is regulated by aromatic hydrocarbon receptor (AHR). A CYP1A1 T3801C polymorphism, associated with a higher CYP1A1 inducibility and enhanced catalytic activity, has been linked to stroke, triple vessel disease and may, therefore, be associated with blood pressure (BP). The relation of the widely studied G1661A polymorphism of the human AHR gene with BP is unknown. To investigate the genetic influence of CYP1A1 T3801C and AHR G1661A polymorphisms on BP in relation to tobacco consumption. Study participants were selected from a French longitudinal cohort of volunteers for a free health check-up. These individuals (302 men and 311 women) were not taking medication that can affect blood pressure. Information about active smoking status was obtained by a self-administered questionnaire. After multiple regression analysis, systolic blood pressure (SBP) and diastolic blood pressure (DBP) did not differ significantly according to their tobacco status excepted for DBP in men. In addition, neither CYP1A1 T3801C nor AHR G1661A polymorphism was linked to blood pressure. However, systolic and diastolic blood pressures differed significantly according to CYP1A1 T3801C genotype between ex-smokers and smokers. Finally, the interaction between CYP1A1 T3801C and AHR G1661A polymorphisms explained a significant difference of SBP and DBP between carriers of both CYP1A1-C3801 and AHR-A1661 alleles. This study is the first to show an interaction between the CYP1A1 T3801C and AHR G1661A polymorphisms. This interaction could explain the difference in blood pressure level between smokers and non-smokers/ex-smokers but needs to be confirmed in a large sample.

Analysis of the effect of multiple genetic variants of cardiovascular disease risk on insulin concentration variability in healthy adults of the STANISLAS cohort: The role of FGB −455 G/A polymorphism

Introduction Given the hypothesis of a common soil for atherosclerosis, type 2 diabetes and metabolic syndrome, we tested the contribution of gene polymorphisms involved in cardiovascular diseases on fasting insulin concentration (FIC). Methods The polymorphisms were investigated by a multiplex assay in 308 apparently healthy French middle-aged men and women, taken from the STANISLAS cohort. FIC was measured by a microparticular enzymatic immunoassay. Results After a series of regression analyses involving 34 polymorphisms, FGB − 455G/A was the only polymorphism that remained significantly associated with FIC when adjusting the analyses for multiple testing. Stepwise models showed that FGB polymorphism accounted for 4.39% of FIC variability in men. Additionally, interactions between FGB and with environmental factors (alcohol and smoking in men, and BMI in women) were found. Discussion To our knowledge, this is the first study reporting an influence of FGB polymorphism on FIC in a healthy population. Our results concord with the already shown link between fibrinogen concentration and FIC, and support the hypothesis of a relationship between fibrinogen and endothelium in FIC homeostasis whose alteration may induce several metabolic disorders. The contribution of this gene, although modest, is consistent with the polygenic nature of insulin levels.

Association of CYP2A6*1B genetic variant with the amount of smoking in French adults from the Stanislas cohort

This study was designed in order to investigate the influence of the genetic polymorphism of CYP2A6 on the amount of smoking. In all, 463 French adults included in the Stanislas cohort were studied and underwent two examinations at 5 years distance (t0 and t(+5) years). Information on their smoking habits was collected. They were genotyped by RFLP for the CYP2A6*1A, CYP2A6*1B and CYP2A6*4 alleles. CYP2A6*1B and CYP2A6*4 allele frequencies were 32 and 4%, respectively. The subjects carrying the CYP2A6*1B allele oxidize nicotine to cotinine faster than subjects with the CYP2A6*1A allele. The number of cigarettes smoked per day was significantly higher in the CYP2A6*1B/*1B group as compared to the CYP2A6*1A/*1A group (P = 0.01 at t0; P = 0.001 t(+5) years), with a larger increase in their daily cigarettes consumption over the 5-year period (P = 0.006). No significant difference in the smoking status was observed according to the CYP2A6 genotype. These data suggest that the CYP2A6*1B is associated with the number of cigarettes smoked per day.

A Prospective Study on the Prevalence of Metabolic Syndrome Among Healthy French Families

The purpose of this study was to estimate the longitudinal variation of prevalence of metabolic syndrome within French families and to observe biological parameters involved in cardiovascular disease among their offspring. Three hundred seventy-one apparently healthy families (1,366 individuals) taken from the STANISLAS cohort were studied. The subjects were examined at two time points with a 5-year interval (t(0) and t(+5)). The crude prevalence of metabolic syndrome was assessed among parents according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP) definition. The prevalence of metabolic syndrome was 5.9% in men and 2.1% in women at t(0), rising to 7.2 and 5.4% in men and women, respectively, at t(+5). Children of parents having metabolic syndrome showed higher levels of tumor necrosis factor-alpha (TNF-alpha), whereas their HDL cholesterol and apolipoprotein (apo) E concentrations were lower compared with those of age- and sex-matched control subjects (P </= 0.05). When applying NCEP ATP definitions that included either antihypertensive drugs only or all the drugs involved in metabolic syndrome, we found that the three parameters shared by the three different versions of the definition were TNF-alpha, HDL cholesterol, and an interaction between alcohol consumption and parental metabolic syndrome on HDL cholesterol concentration. Metabolic syndrome increases with age in supposedly healthy families from the STANISLAS cohort. In offspring of affected people, it seems to be predictive of higher values of TNF-alpha and low HDL cholesterol levels, which are two major cardiovascular factors. Therefore, in terms of prevention, it is important to identify and follow subjects with metabolic syndrome as well as their offspring, even in apparently healthy populations, to enable early disease management.

Biological and genetic factors influencing plasma factor VIII levels in a healthy family population: results from the Stanislas cohort

The mechanisms underlying the variability of factor VIII (FVIII) levels are still poorly understood. The only receptor of FVIII identified so far is the lipoprotein receptor-related protein (LRP), which is thought to be involved in FVIII degradation. We aimed to characterize biological and genetic factors related to FVIII variability, focusing on coding polymorphisms of the LRP gene and polymorphisms potentially detected by molecular screening of the LRP-binding domains of the FVIII gene. Plasma FVIII coagulant activity (FVIII:C) and von Willebrand factor (VWF:Ag) antigen levels were measured in a sample of 100 healthy nuclear families (200 parents and 224 offspring). The ABO blood group and the three coding polymorphisms of the LRP gene (A217V, D2080N and C766T) were genotyped. Lipids and anthropometric factors poorly contributed to the variability of FVIII:C (<5%). A strong effect of ABO blood groups on FVIII:C levels was observed that remained significant after adjustment for VWF:Ag levels (P = 0.02). These two factors explained more than 50% of FVIII:C variability. After adjustment for VWF:Ag and ABO blood groups, a residual resemblance for FVIII:C persisted between biological relatives (rho = 0.13 +/- 0.06 between parents and offspring, rho = 0.24 +/- 0.09 between siblings) compatible with an additional genetic influence. The N allele of the LRP/D2080N polymorphism was associated with decreased levels of FVIII:C (90.4 +/- 8.7 vs. 102.2 +/- 3.5 IU/dl, P = 0.03) and VWF:Ag levels (109.1 +/- 11.2 vs. 125.4 +/- 4.4 IU/dl, P = 0.02). No polymorphism was detected in the LRP-binding domains of the FVIII gene. This study reinforces the hypothesis of a genetic influence of FVIII levels beyond the influence of VWF:Ag and ABO blood groups. The D2080N polymorphism of the LRP gene weakly contributed to the variability of FVIII:C levels in this healthy population.

Biological variations, genetic polymorphisms and familial resemblance of TNF-α and IL-6 concentrations: STANISLAS cohort

Cytokines are involved in the development of several inflammatory diseases and atherosclerosis. Their variations in healthy individuals are not well defined. The aims of this study were: firstly, to identify factors affecting biological variation of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha); secondly, to study their family resemblance; and thirdly, to evaluate the effect of two TNF-alpha (-308G/A and -238G/A) and two IL-6 polymorphisms (174G/C and -572G/C) on their corresponding circulating levels. A total of 171 healthy families selected from the STANISLAS cohort were studied. Age was negatively related to TNF-alpha concentrations in offspring only (both sons and daughters). Additionally, IL-6 and TNF-alpha levels were differently influenced by gender, white blood cells, tobacco consumption, and HDL-cholesterol level. A weak significant familial resemblance for TNF-alpha concentration was observed in siblings only. There was no significant familial resemblance for IL-6 levels. The TNF-alpha -308A allele was associated with decreased TNF-alpha concentrations in both offspring aged less than 18 and males without overweight (BMI<25 kg/m(2)). Fathers carrying the IL-6 -174CC genotype had higher IL-6 levels than those with the IL-6 -174G allele. Parents with the IL-6 -572GG genotype had higher IL-6 concentrations than the C allele carriers. In this sample of healthy families, plasma levels of IL-6 and TNF-alpha were differently affected by biological parameters including age, gender and smoking, and the impact of their respective polymorphisms was influenced by gender, age and BMI.

Association between factor VII polymorphisms and blood pressure: the Stanislas Cohort.

The purpose this study was to determine whether Arg353Gln and -323Del/Ins polymorphisms of factor VII (FVII) are related to blood pressure levels and hypertension. Subjects were drawn from the Stanislas Cohort, a longitudinal, familial French cohort examined twice since 1994. The "blood pressure study" included 1342 subjects free of medication use that could affect blood pressure. The "hypertension study" included 645 normotensive and 77 hypertensive adult subjects. Association with hypertension was also studied in 547 hypertensives enrolled in a clinical trial and in 624 normotensives drawn from the Stanislas Cohort. In the "blood pressure study," parents with the 353Gln or -323Ins allele had lower blood pressures than did noncarriers at each examination, independent of covariates (0.01< or =P< or =0.05, except for diastolic blood pressure [DBP] at baseline, where P=0.103). Similarly significant relations were observed in their offspring (P< or =0.05, except for systolic blood pressure [SBP] at 5 years, where P=0.186). In a representative subgroup of 267 individuals, the -323Del/Ins polymorphism was significantly associated with plasma FVII levels in both parents and offspring (P<0.001). FVII levels in plasma were significantly correlated with SBP and DBP in parents but not in offspring. After inclusion of both FVII levels and the -223Del/Ins in the same model in parents, only FVII levels remained significantly associated with SBP and DBP. The "hypertension study" revealed that the 353Gln and -323Ins alleles were related to decreased risk (odds ratio [OR]=0.554, 95% confidence interval [CI], 0.362 to 0.848, and OR=0.475, 95% CI, 0.299 to 0.755, respectively). These results suggest that the FVII gene may be a susceptibility locus for hypertension.

The Leu554Phe polymorphism in the E-selectin gene is associated with blood pressure in overweight people.

Associations between circulating concentrations of E-selectin, blood pressure and obesity, and between the Leu554Phe (L/F554) polymorphism and blood pressure have been documented. To investigate how the E-selectin L/F554 polymorphism is involved in longitudinal blood pressure changes, and how this polymorphism interacts with body mass index (BMI) on blood pressure. For this study, 478 men and 546 women were selected from the Stanislas cohort, a French longitudinal study of volunteers for a free health check-up. These individuals underwent two examinations (t(0) and t(+5)) and were not taking medication that can affect blood pressure. At t(0), no relationship was observed between L/F554 polymorphism and blood pressure. However at t(+5), systolic blood pressure (SBP) was greater in individuals carrying the F allele, and the L/F554 polymorphism was associated with SBP in interaction with BMI (P < 0.001 in men and P < 0.05 for women). There was a steeper increase in SBP with BMI greater than 25 kg/m2 in carriers of the F allele than in LL homozygotes. Similar results were observed for diastolic blood pressure in men (P = 0.0103). These results suggest a BMI-specific effect of L/F554 polymorphism of the E-selectin gene on blood pressure, and strengthen the hypothesis that E-selectin is implicated in hypertension.

Genetic influences on blood pressure within the Stanislas Cohort.

To investigate the association of 21 polymorphisms within 13 genes, APOE, APOB, APOC3, CETP, LPL, PON1, MTHFR, FGB, F5, GPIIIa, SELE, ACE and AGT, with inter-individual blood pressure (BP) variation. Seven hundred and seventy-six men and 836 women, free of antihypertensive and lipid-lowering medications, were selected from the Stanislas Cohort. ANOVA on blood pressure values after adjustment for covariates [age, body mass index (BMI), contraceptive pill, tobacco and alcohol] showed that lipoprotein lipase (LPL) Ser447Ter and glycoprotein IIIA (GpIIIa) Pl polymorphisms were significantly associated with BP in women (0.01 < or = P < or = 0.05), whereas BP levels in men were significantly different according to apolipoprotein CIII (APOC3) 3206T/G and -482C/T polymorphisms (P < or = 0.05). In women, compared to the most common allele, the GpIIIa Pl allele was associated with increased mean arterial pressure (MAP) (P < 0.05) and pulse pressure (PP) (P < 0.001), and the LPL 447Ter allele was associated with decreased systolic blood pressure (SBP) and PP levels (0.001 < or = P < or = 0.05). These two polymorphisms appeared to act independently. In men, the APOC3 3206GG genotype was related to decreased diastolic blood pressure (DBP) and MAP levels (P < or = 0.01), and the APOC3 -482T allele with decreased PP levels (P < or = 0.05). The presence of both the -482C allele and the 3206GG genotype was related to decreased DBP, suggesting that specific haplotypes might be involved. The APOC3, LPL and GpIIIa genes were found to be associated with BP levels. The contributions of these genes, although modest, are consistent with the polygenic nature of BP levels.

Study of reference values and biological variation: a necessity and a model for Preventive Medicine Centers

Laboratory tests were at the origin of the modern approach to the evaluation of the health status in many Preventive Medicine Centers. Guided by the theory of reference values and its applications, years of efforts have been devoted to defining the health status of people, and understanding the biological variation of the population. At the Nancy Center for Preventive Medicine, 40,000 people are invited every year for health examination; many coming with their families. French legislation authorized every citizen to have full health examination every five years. Annually, 600,000 people are seen at 70 centers. The visit includes answering questionnaires, physiological and biological testing modified for age, risks, and social criteria. The process concludes with consultation with a general practitioner. Data accumulated over 30 years have led to the identification of primary factors influencing biological variation of common laboratory tests. For example, body mass index (BMI) has to be considered, in addition to age and gender, for measurement of gamma-glutamyltransferase, alanine aminotransferase, and other analytes. More recently, the familial Stanislas cohort has shown that apolipoprotein E (apoE) genetic polymorphism has to be considered as a factor for variation in some individuals. For example, there was greater than 100% difference in the mean concentration of apoE between epsilon2/epsilon2 and epsilon4/epsilon4 individuals. This example nicely illustrates the relationship between genotype and phenotype. Demonstrating the effects of multiple gene polymorphisms on phenotypes will be a valid approach to predictive laboratory medicine.