Subclinical atherosclerosis and its progression are modulated by PLIN2 through a feed-forward loop between LXR and autophagy.

BACKGROUND: Hyperlipidaemia is a major risk factor for cardiovascular disease, and atherosclerosis is the underlying cause of both myocardial infarction and stroke. We have previously shown that the Pro251 variant of perilipin-2 reduces plasma triglycerides and may therefore be beneficial to reduce atherosclerosis development. OBJECTIVE: We sought to delineate putative beneficial effects of the Pro251 variant of perlipin-2 on subclinical atherosclerosis and the mechanism by which it acts. METHODS: A pan-European cohort of high-risk individuals where carotid intima-media thickness has been assessed was adopted. Human primary monocyte-derived macrophages were prepared from whole blood from individuals recruited by perilipin-2 genotype or from buffy coats from the Karolinska University hospital blood central. RESULTS: The Pro251 variant of perilipin-2 is associated with decreased intima-media thickness at baseline and over 30 months of follow-up. Using human primary monocyte-derived macrophages from carriers of the beneficial Pro251 variant, we show that this variant increases autophagy activity, cholesterol efflux and a controlled inflammatory response. Through extensive mechanistic studies, we demonstrate that increase in autophagy activity is accompanied with an increase in liver-X-receptor (LXR) activity and that LXR and autophagy reciprocally activate each other in a feed-forward loop, regulated by CYP27A1 and 27OH-cholesterol. CONCLUSIONS: For the first time, we show that perilipin-2 affects susceptibility to human atherosclerosis through activation of autophagy and stimulation of cholesterol efflux. We demonstrate that perilipin-2 modulates levels of the LXR ligand 27OH-cholesterol and initiates a feed-forward loop where LXR and autophagy reciprocally activate each other; the mechanism by which perilipin-2 exerts its beneficial effects on subclinical atherosclerosis.

Integrative studies implicate matrix metalloproteinase-12 as a culprit gene for large-artery atherosclerotic stroke.

BACKGROUND: Ischaemic stroke and coronary heart disease are important contributors to the global disease burden and share atherosclerosis as the main underlying cause. Recent evidence from a genome-wide association study (GWAS) suggested that single nucleotide polymorphisms (SNP) near the MMP12 gene at chromosome 11q22.3 were associated with large-vessel ischaemic stroke. Here, we evaluated and extended these results by examining the relationship between MMP12 and atherosclerosis in clinical and experimental studies. METHODS AND RESULTS: Plasma concentrations of MMP12 were measured at baseline in 3394 subjects with high-risk for cardiovascular disease (CVD) using the Olink ProSeek CVD I array. The plasma MMP12 concentration showed association with incident cardiovascular and cerebrovascular events (130 and 67 events, respectively, over 36 months) and carotid intima-media thickness progression (P = 3.6 × 10-5 ). A GWAS of plasma MMP12 concentrations revealed that SNPs rs499459, rs613084 and rs1892971 at chr11q22.3 were independently associated with plasma MMP12 (P < 5 × 10-8 ). The lead SNPs showed associations with mRNA levels of MMP12 and adjacent MMPs in atherosclerotic plaques. MMP12 transcriptomic and proteomic levels were strongly significantly increased in carotid plaques compared with control arterial tissue and in plaques from symptomatic versus asymptomatic patients. By combining immunohistochemistry and proximity ligation assay, we demonstrated that MMP12 localizes to CD68 + macrophages and interacts with elastin in plaques. MMP12 silencing in human THP-1-derived macrophages resulted in reduced macrophage migration. CONCLUSIONS: Our study supports the notion that MMP12 is implicated in large-artery atherosclerotic stroke, functionally by enhancing elastin degradation and macrophage invasion in plaques.

Correlates of low physical activity levels in aging men and women: the DR's EXTRA Study (ISRCTN45977199).

Recognizing correlates of low physical activity (PA) can help in targeting PA interventions for individuals who would benefit most from increasing their PA. We studied the associations of demographic, social, health, and lifestyle factors with low PA by sex in a population sample of 1,303 Finnish individuals aged 57-78 years. We defined low PA as no moderate or vigorous leisure-time PA reported in an interview. Altogether, 39% of men and 48% of women reported low PA. Satisfactory or poor perceived health and high BMI were independently associated with low PA in both sexes. In men, factors such as age, being divorced or separated, still working, having a weak social network, poor diet, and a health professional's suggestion to increase PA were associated with low PA. In women, cardiovascular disease and depressive symptoms were associated with low PA. These results can be applied in targeting PA interventions.

Combined low-saturated fat intake and high fitness may counterbalance diabetogenic effects of obesity: the DR's EXTRA Study.

We report associations of saturated fat (SF) intake with impaired fasting glucose (IFG), impaired glucose tolerance (IGT), concurrent IFG+IGT and type 2 diabetes (T2DM) at different levels of cardiorespiratory fitness and body mass index (BMI). In a population-based sample (n=1261, age 58-78 years), oral glucose tolerance, 4-day food intake and maximal oxygen uptake were measured. High intake of SF (>11.4 E%) was associated with elevated risk for IFG (4.36; 1.93-9.88), concurrent IFG+IGT (6.03; 1.25-29.20) and T2DM (4.77; 1.93-11.82) in the category of high BMI (>26.5) and high fitness, whereas there was no significantly elevated risk in individuals reporting low intake of SF. Concurrent high BMI and low fitness were associated with elevated risks. In general, SF intake and fitness did not differentiate the risk of abnormal glucose metabolism among subjects with low BMI. Limited intake of SF may protect from diabetogenic effects of adiposity, but only in individuals with high level of fitness.

LDL oxidative modification and carotid atherosclerosis: results of a multicenter study.

OBJECTIVE: Serum LDL conjugated diene concentration is a marker of oxidative modification of LDL. We investigated the relationship between LDL conjugated dienes and cross-sectional subclinical atherosclerosis assessed by carotid IMT in high-risk subjects of a multicenter study. METHODS: Serum LDL conjugated dienes and ultrasonographically assessed carotid intima-media thickness (IMT(mean), IMT(max) and IMT(mean-max)) were available for 553 subjects from Finland, France, Italy, the Netherlands, and Sweden. RESULTS: In multivariate regression analysis, gender (p < 0.001), age (p < 0.001), systolic blood pressure (IMT(mean), p = 0.01; IMT(mean-max), p = 0.05) and serum LDL conjugated dienes (p = 0.02 for both IMT(mean) and IMT(mean-max)) were the strongest determinants of IMT variation, adjusted for study center, ultrasound videotape reader and serum LDL cholesterol. Pack-years of smoking, added into the regression model, did not destroy the significant association between increased serum LDL conjugated dienes and IMT. Ratio of LDL conjugated dienes to LDL particle cholesterol was higher in subjects of Northern recruiting centers than of Southern centers (r = 0.39, p < 0.0001). CONCLUSIONS: There was a cross-sectional association between in vivo increased LDL oxidative modification and subclinical atherosclerosis after adjustment for traditional risk factors. The subjects in Northern countries of Europe had more oxidatively modified lipids per cholesterol in LDL particle than subjects in Southern countries.

Dietary associations with prediabetic states--the DR's EXTRA Study (ISRCTN45977199).

BACKGROUND/OBJECTIVES: Impaired fasting plasma glucose (IFG) and impaired glucose tolerance (IGT) predict development of type 2 diabetes (T2D), but display different pathophysiology for T2D. We studied the association of selected food items and nutrients with IFG, IGT and combined IFG and IGT (IFG+IGT), independent of cardiorespiratory fitness (VO(2max)). SUBJECTS/METHODS: In a population-based sample of 1261 individuals, aged 58-78 years, we identified 126 subjects with IFG, 97 with IGT and 49 with simultaneous IFG and IGT by an oral glucose tolerance test. Dietary intake was assessed by 4-day food records. Cardiorespiratory fitness was assessed by defining maximal oxygen uptake (VO(2max)) from respiratory gas analysis during a maximal symptom-limited exercise stress test on a bicycle ergometer. RESULTS: Increased intake of saturated fat was associated with higher odds for IFG (OR 1.07; 1.01-1.14) after adjustment for age, gender, VO(2max) and energy misreporting variable. Consumption of additional whole-grain bread (50 g/1000 kcal) and intake of dietary fiber (g/1000 kcal) were inversely associated with IGT (OR 0.61; 0.41-0.92, OR 0.91; CI 0.85-0.97, respectively). CONCLUSION: Dietary fiber and sources of cereal fiber are negatively associated with IGT, and saturated fat intake is positively associated with IFG, but not with IGT. The present data give practical dietary means at the population level for the elimination of prediabetic conditions.

The current standard measure of cardiorespiratory fitness introduces confounding by body mass: the DR's EXTRA study.

OBJECTIVE: Cardiorespiratory fitness is currently estimated by dividing maximal oxygen consumption (VO(2max)) by body weight (per-weight standard). However, the statistically correct way to neutralize the effect of weight on VO(2max) in a given population is adjustment for body weight by regression techniques (adjusted standard). Our objective is to quantify the bias introduced by the per-weight standard in a population distributed across different categories of body mass. DESIGN: This is a cross-sectional study. SUBJECTS AND METHODS: Baseline measures from participants of the Dose-Responses to Exercise Training Study (DR's EXTRA), 635 men (body mass index (BMI): 19-47 kg m⁻²) and 638 women (BMI: 16-49 kg m⁻²) aged 57-78 years who performed oral glucose tolerance tests and maximal exercise stress tests with direct measurement of VO(2max). We compare the increase in VO(2max) implied by the per-weight standard with the real increase of VO(2max) per kg body weight. A linear logistic regression model estimates odds for abnormal glucose metabolism (either impaired fasting glycemia or impaired glucose tolerance or Type 2 diabetes) of the least-fit versus most-fit quartile according to both per-weight standard and adjusted standard. RESULTS: The per-weight standard implies an increase of VO(2max) with 20.9 ml min⁻¹ in women and 26.4 ml min⁻¹ in men per additional kg body weight. The true increase per kg is only 7.0 ml min⁻¹ (95% confidence interval: 5.3-8.8) and 8.0 ml min⁻¹ (95% confidence interval: 5.3-10.7), respectively. Risk for abnormal glucose metabolism in the least-fit quartile of the population is overestimated by 52% if the per-weight standard is used. CONCLUSIONS: In comparisons across different categories of body mass, the per-weight standard systematically underestimates cardiorespiratory fitness in obese subjects. Use of the per-weight standard markedly inflates associations between poor fitness and co-morbidities of obesity.

Diet, fitness and metabolic syndrome--the DR's EXTRA study.

BACKGROUND AND AIMS: To study the independent and combined associations of diet and cardiorespiratory fitness with the prevalence of the metabolic syndrome (MetS). METHODS AND RESULTS: We studied a population-based random sample of 663 men and 671 women 57-78 years of age at baseline of an ongoing randomised controlled trial. Based on a 4-day food record a diet score was created according to goals achieved (vegetables ≥400 g/day, fish ≥2 servings/week, fibre ≥14 g/1000 kcal, saturated fat <10 E%/day). Cardiorespiratory fitness was measured as maximal oxygen uptake (VO(2 max)) in a maximal symptom-limited bicycle ergometer test. MetS was defined by the National Cholesterol Education Program criteria. The lowest prevalence of MetS (5%) was observed among individuals in the highest VO(2 max) tertile and achieving 3-4 dietary goals. The highest prevalence (55%) was observed among those in the lowest VO(2 max) tertile and achieving none of the dietary goals. Among individuals in the highest VO(2 max) tertile, the odds ratio of having MetS was 0.04 (95% CI 0.02-0.10) for those achieving 3-4 dietary goals, 0.07 (0.04-0.14) for those achieving 1-2 dietary goals, and 0.16 (0.07-0.37) for those achieving none of the dietary goals compared with individuals in the lowest VO(2 max) tertile and achieving none of the goals after adjustment for confounding factors. CONCLUSION: Healthy diet and higher levels of cardiorespiratory fitness are associated with a reduced risk of having MetS. However, fitness seems to have a stronger association with MetS than diet.

Two-minute heart rate recovery after cycle ergometer exercise and all-cause mortality in middle-aged men.

BACKGROUND: A slow heart rate recovery (HRR) after an exercise test is associated with an increased risk of all-cause mortality in asymptomatic individuals, but the data regarding additional prognostic information provided by HRR beyond other exercise test variables are inconsistent. We investigated the prognostic significance of HRR for premature death, particularly in relation to other exercise test variables. METHODS: The study subjects were a representative population-based sample of 1102 men (42-61 years of age) without cardiovascular disease, cancer or diabetes. HRR was defined as the difference between maximal HR and HR 2 min after a maximal symptom-limited exercise test using a cycle ergometer. The association between HRR and premature mortality was examined with Cox regression models. RESULTS: During an average follow-up of 18 years, 238 deaths occurred. HRR was an independent predictor of death [for a decrease of 12 beats min(-1) , relative risk (RR) 1.16, 95% CI 1.02-1.33, P = 0.02] after adjustment for age and established risk factors. When added in a Cox model with chronotropic response (decrease of 12 beats min(-1) , RR 1.09, 95% CI 0.93-1.27, P = 0.26) or cardiorespiratory fitness (decrease of 12 beats min(-1) , RR 1.12, 95% CI 0.98-1.30, P = 0.08), the association between a slow HRR and an increased risk of death was clearly weaker. CONCLUSION: A slow 2-min HRR after a cycle ergometer exercise test was an independent predictor of death in healthy middle-aged men after accounting for demographic and clinical characteristics. However, it was no longer predictive after accounting for chronotropic response and exercise capacity.

Intensity of leisure-time physical activity and cancer mortality in men.

OBJECTIVE: there is a lack of evidence to show the role of exercise intensity in the prevention of cancer mortality because no previous studies have shown this relation. The relationship of leisure-time physical activity with cancer mortality was therefore assessed. METHODS: participants were from a population-based sample of 2560 men from eastern Finland with no history of cancer at baseline. Physical activity was assessed using the 12-month leisure-time physical activity questionnaire. During an average follow-up of 16.7 years, a total of 181 cancer related deaths occurred. RESULTS: an increase of 1.2 metabolic units (MET or metabolic equivalents of oxygen consumption; 1 SD in metabolic equivalents) in the mean intensity of leisure-time physical activity was related to a decrease (relative risk (RR) 0.85, 95% CI 0.72 to 0.99) in cancer mortality mainly due to lung and gastrointestinal cancers, after adjustment for age, examination year, alcohol consumption, smoking, body mass index and energy, fibre and fat intake. Men with leisure-time physical activity of more than 5.2 MET (highest quartile) had a lower (RR 0.63, 95% CI 0.40 to 0.99) cancer mortality compared with men whose mean intensity of physical activity was less than 3.7 MET (lowest quartile). The mean intensity of physical activity was related to the risk of cancer death among men who exercised at least 30 minutes per day on average. CONCLUSIONS: this prospective study indicates that the mean intensity of leisure-time physical activity is inversely associated with the risk of premature death from cancer in men.

Single nucleotide polymorphisms in the myostatin (MSTN) and muscle creatine kinase (CKM) genes are not associated with elite endurance performance.

Maximal oxygen uptake (VO2max) is one of the most important determinants of elite endurance performance. VO2max is determined by a whole range of genetic and environmental factors. Single nucleotide polymorphisms (SNPs) in muscle myostatin (MSTN) and creatine kinase (CKM) genes are candidates for VO2max and skeletal muscle performance phenotypes. Common MSTN (rs3791783, rs11681628 and rs7570532) and CKM (rs344816, rs10410448, rs432979, rs1133190, rs7260359, rs7260463 and rs4884) SNPs, selected from HapMap CEU data in order to tag the genetic variability of the proteins, were genotyped in 316 male Caucasian elite endurance athletes and 304 sedentary controls from the Genathlete study. Association with elite endurance performance was determined by logistic regression analysis. The P-value for statistical significance was set at <0.01. None of the SNPs or haplotypes showed a significant association with elite endurance status. We conclude that common variants of MSTN and CKM genes do not play a role in attaining high-level endurance performance in Caucasian populations.

Cardiorespiratory fitness in aging men and women: the DR's EXTRA study.

The aim of the study was to describe the levels and to create reference values of cardiorespiratory fitness, expressed as maximal oxygen consumption (VO(2max) ), maximal metabolic equivalents (METs) and maximal workload in aging men and women. We measured VO(2max) directly by a breath-by-breath method during a maximal exercise stress test on a bicycle ergometer with a linear workload increase of 20 W/min in a representative population sample of 672 men and 677 women aged 57-78 years. We presented the age and sex-specific categories of cardiorespiratory fitness (very low, low, medium, high and very high) based on variable distribution and non-linear regression models of VO(2max) , maximal METs and maximal workload. The linear age-related decrement of VO(2max) was -0.047 L/min/year (-2.3%) and -0.404 mL/kg/min/year (-1.6%) in men and -0.027 L/min/year (-1.9%) and -0.328 mL/kg/min/year (-1.6%) in women. After exclusion of diseased individuals, the rate of VO(2max) decrement remained similar. The number of chronic diseases (0, 1, 2 or ≥3) was inversely associated with VO(2max) in men (P<0.001) and women (P<0.001). The present study provides clinically useful reference values of cardiorespiratory fitness for primary and secondary prevention purposes in aging people.

Food consumption, nutrient intake and the risk of having metabolic syndrome: the DR's EXTRA Study.

BACKGROUND: The associations of different components of diet with metabolic syndrome (MetS) are largely unknown. We therefore studied the associations of intakes of selected food items and nutrients with the risk of having MetS. METHODS: The participants were a representative population sample of 1334 individuals (671 women, 663 men) 57-78 years of age. Dietary intake was assessed by a 4-day food record. MetS was defined by the National Cholesterol Education Program criteria. RESULTS: Consumption of vegetables, non-root vegetables, legumes and nuts berries and fish had an inverse and consumption of sausage had a direct association with the risk of having MetS in men after adjustment for age, smoking and alcohol consumption. Consumption of vegetables and non-root vegetables had an inverse and consumption of sausage had a direct association with the risk of having MetS in women after these adjustments. However, after further adjustment for maximal oxygen uptake (VO2(max)) most of these associations vanished. Men in the highest third of consumption of berries, fish, and legumes and nuts had 49, 37 and 44% lower risk of having MetS, respectively, than those in the lowest third after further adjustment for VO2(max). Women in the highest third of sausage consumption had a 72% higher risk of having MetS than non-consumers. CONCLUSIONS: Consumption of legumes and nuts, berries and fish was inversely associated with MetS in men. Consumption of sausage was directly associated with MetS in women. VO2(max) seems to be a strong confounding factor between food consumption and MetS.

Exercise workload, cardiovascular risk factor evaluation and the risk of stroke in middle-aged men.

OBJECTIVE: We investigated the prognostic significance of risk scores and exercise workload with respect to stroke. Background. There are no data on exercise workload combined with European Systematic Coronary Risk Evaluation (SCORE) in the prediction of stroke. METHODS: Exercise workload was measured by exercise test with an electrically braked cycle ergometer performed at baseline. The study is based on a random population-based sample of 1639 men (42-60 years) without history of type 2 diabetes or atherosclerotic cardiovascular disease including coronary heart disease, stroke or claudication. RESULTS: During an average follow-up of 16 years, a total of 97 strokes occurred, of which 71 were ischaemic strokes. Independent predictors for all strokes were European SCORE [for 1% increment, relative risk (RR): 1.12, 95% CI: 1.02 to 1.22, P=0.017), maximal workload (for 20 W increment, RR: 0.87, 95% CI: 0.80 to 0.95, P=0.003) and body mass index (for 5 kg m(-2) increment, RR: 1.08, 95% CI: 1.03 to 1.14, P=0.004), when adjusted for serum HDL, alcohol consumption, C-reactive protein, family history of coronary heart disease, exercise-induced ST changes and the use of medications for hypertension, dyslipidaemia or aspirin. The risk was 2.54-fold (95% CI: 1.27-5.09, P=0.008) for any strokes and 4.43-fold (95% CI 1.69-11.78, P=0.003) for ischaemic strokes amongst men with exercise capacity less than 162 W when compared with those with high exercise capacity over 230 W, after adjustment for risk factors. CONCLUSIONS: Low exercise workload predicts an especially high risk for stroke in the presence of high risk SCORE.

Downregulation of genes involved in NFkappaB activation in peripheral blood mononuclear cells after weight loss is associated with the improvement of insulin sensitivity in individuals with the metabolic syndrome: the GENOBIN study.

AIMS/HYPOTHESIS: The transcription factor nuclear factor-kappa-B (NFkappaB) is implicated in inflammatory responses, obesity and the metabolic syndrome, while immune cells appear to play a central role in mediating insulin resistance and can be used as a model to study inflammation and its relationship with insulin resistance. In peripheral blood mononuclear cells of overweight participants with the metabolic syndrome, we evaluated (1) the effect of diet-induced weight loss on the expression of genes involved in NFkappaB activation and (2) their association with insulin sensitivity. The genes studied were: TNF receptors TNFRSF1A and TNFRSF1B, and IL1R1, TLR4, TLR2, ICAM1, CCL5 and IKBKB. METHODS: We analysed data from 34 overweight participants with abnormal glucose metabolism and the metabolic syndrome, who were randomised to a weight-reduction (n = 24) or control group (n = 10) for 33 weeks. The mRNA expression was measured using real-time PCR. Measures of insulin and glucose homeostasis were assessed by IVGTT and OGTT. RESULTS: In general, the genes studied were downregulated after weight loss intervention. The changes in TLR4, TLR2, CCL5 and TNFRSF1A mRNA expression were associated with an increase in insulin sensitivity index independently of the change in waist circumference (p < 0.05). The change in IKBKB expression correlated with most of the changes in gene expression in the weight-reduction group. CONCLUSIONS/INTERPRETATION: These results suggest that proteins encoded by CCL5, TLR2 and TLR4, and TNFRSF1A might contribute to insulin-resistant states that characterise obesity and the metabolic syndrome. TRIAL REGISTRATION: ClinicalTrials.gov NCT 00621205.

Serum concentrations and expressions of serum amyloid A and leptin in adipose tissue are interrelated: the Genobin Study.

OBJECTIVE: Serum amyloid A (SAA) is a novel link between increased adipose tissue mass and low-grade inflammation in obesity. Little is known about the factors regulating its serum concentration and mRNA levels. We investigated the association between SAA and leptin in obese and normal weight subjects and analyzed the effect of weight reduction on serum SAA concentration and gene expression in adipose tissue of the obese subjects. METHODS: Seventy-five obese subjects (60+/-7 years, body mass index (BMI) 32.9+/-2.8 kg/m(2), mean+/-s.d.) with impaired fasting plasma glucose or impaired glucose tolerance and other features of metabolic syndrome, and 11 normal weight control subjects (48+/-9 years, BMI 23.7+/-1.9 kg/m(2)) were studied at the baseline. Twenty-eight obese subjects underwent a 12-week intensive weight reduction program followed by 5 months of weight maintenance. Blood samples and abdominal s.c. adipose tissue biopsies were taken at the baseline and after the follow-up. Gene expression was studied using real-time quantitative PCR. RESULTS: The gene expressions in women and serum concentrations of leptin and SAA were interrelated independently of body fat mass in the obese subjects (r=0.54, P=0.001; r=0.24, P=0.039 respectively). In multiple linear regression analyses, leptin mRNA explained 38% of the variance in SAA mRNA (P=0.002) in the obese women. Weight loss of at least 5% increased SAA mRNA expression by 48 and 36% in men and women, but serum SAA concentrations did not change. CONCLUSIONS: The association between SAA and leptin suggests an interaction between these two adipokines, which may have implications in inflammatory processes related to obesity and the metabolic syndrome.

Workload at the heart rate of 100 beats/min and mortality in middle-aged men with known or suspected coronary heart disease.

OBJECTIVE: To investigate whether a workload which an individual is able to perform at the heart rate (HR) of 100 beats/min (WL(100)) independently predicts mortality in middle-aged men with known or suspected coronary heart disease (CHD). DESIGN: Prospective population-based study based on 365 middle-aged men with known or suspected CHD at baseline. RESULTS: During an average follow-up of 11.1 years, there were 75 deaths (20.5%). In Cox multivariable models mortality increased by 72% (95% CI 32% to 122%, p<0.001) with 1 SD (34 Watts) decrement in WL(100) after adjustment for age, examination year, alcohol consumption, body mass index, cigarette smoking, cardiac insufficiency, history of myocardial infarction, diabetes, myocardial ischaemia during exercise test, serum low-density lipoprotein and high-density lipoprotein cholesterol, systolic and diastolic blood pressure at rest, testing protocol, and use of HR-lowering medication. The risk of death was 2.4 (95% CI 1.5 to 4.0, p<0.001) times higher in 130 men with WL(100) <55 W than in 235 men with WL(100) >or=55 W. In men using and not using HR-lowering medication the risk of death increased 72% (95% CI 14% to 163%, p = 0.01), and 54% (95% CI 14% to 108%, p = 0.005) with 1 SD decrement in WL(100), respectively. WL(100 )improved the predictive power of the adjusted Cox models including other HR and exercise test variables. CONCLUSIONS: WL(100) predicts mortality in men with known or suspected CHD. The association of WL(100) with mortality was not explained by other well-established HR and exercise test variables. WL(100) is derived from a submaximal test which avoids the cardiovascular risks associated with a high-intensity exertion.

Endothelial nitric oxide synthase gene polymorphism and elite endurance athlete status: the Genathlete study.

In the Genathlete study, we examined the contribution of three polymorphisms in the endothelial nitric oxide synthase (NOS3) gene to discriminate elite endurance athletes (EEA) from sedentary controls (SC). The EEA group included a total of 316 Caucasian males with a VO2max >75 mL/kg. The SC group comprised 299 unrelated sedentary Caucasian males who had VO2max values below 50 mL/kg. The polymerase chain reaction technique was used to amplify a microsatellite (CA)(n) repeat in intron 13, a 27 bp repeat in intron 4 and a third fragment in exon 7 containing the Glu298Asp SNP. No difference was found between the EEA and SC groups for the 27 bp repeat and the Glu298Asp polymorphism. Chi-square analysis of the overall allelic distribution of the (CA)(n) repeat revealed no significant difference between the two groups (P=0.135). However, comparing carriers and non-carriers for the most common (CA)(n) repeat alleles, we found significant differences between SC and EEA, with more EEA subjects carrying the 164 bp allele (P=0.007). In summary, we found suggestive evidence that the 164 bp allele of the (CA)(n) repeat in intron 13 is associated with EEA status and may account for some of the differences between EEA and SC.

Weight reduction modulates expression of genes involved in extracellular matrix and cell death: the GENOBIN study.

OBJECTIVE: Lifestyle and genetic factors interact in the development of obesity and the metabolic syndrome. The molecular mechanisms underlying the beneficial dietary modifications are, however, unclear. We aimed to examine the effect of the long-term moderate weight reduction on gene expression in adipose tissue (AT) and to identify genes and gene clusters responsive to treatment and thereby likely contributing to the development of the metabolic syndrome. DESIGN: Randomized controlled and individualized weight reduction intervention. SUBJECTS: Forty-six subjects with impaired fasting glycemia or impaired glucose tolerance and features of metabolic syndrome, aged 60+/-7 years were randomized either to a weight reduction (WR) (n=28) or a control (n=18) group lasting for 33 weeks. MEASUREMENTS: Oral and intravenous glucose tolerance tests and subcutaneous AT biopsies were performed before and after the intervention. Gene expression of AT was studied using microarray technology in subgroups of WR (with weight reduction > or =5%, n=9) and control group (n=10). The results were confirmed using quantitative PCR. RESULTS: In the WR group, glucose metabolism improved. Moreover, an inverse correlation between the change in S (I) and the change in body weight was found (r=-0.44, P=0.026). Downregulation of gene expression (P<0.01) involving gene ontology groups of extracellular matrix and cell death was seen. Such changes did not occur in the control group. The tenomodulin-gene was one of the most downregulated genes (-39+/-16%, P<0.0001). Moreover, its expression correlated with insulin sensitivity (r=-0.34, P=0.005) before the intervention and with body adiposity both before (r=0.42, P=0.007) and after (r=0.30, P=0.056) the intervention. CONCLUSION: Genes regulating the extracellular matrix and cell death showed a strong downregulation after long-term weight reduction. This likely reflects a new stable state at the molecular level in AT. Further studies are warranted to elucidate the mechanisms of these genetic factors.

The predictive value of cardiorespiratory fitness combined with coronary risk evaluation and the risk of cardiovascular and all-cause death.

BACKGROUND: There are no data on directly measured cardiorespiratory fitness combined coronary risk evaluation with respect to death from cardiovascular diseases and all-causes. We investigated the prognostic significance of risk scores and cardiorespiratory fitness with respect to cardiovascular disease and all-cause mortality. METHODS: Cardiorespiratory fitness (maximal oxygen uptake, VO2peak) was measured by exercise test with an electrically braked cycle ergometer. The study is based on a random population-based sample of 1639 men (42-60 years) without history of type 2 diabetes or atherosclerotic cardiovascular diseases. RESULTS: During an average follow-up of 16 years, a total of 304 deaths occurred. Independent predictors for all-cause death were European Score (for 1% increment, RR 1.15, 95% CI 1.10-1.20), VO2peak (for 1 MET increment, RR 0.84, 95% CI 0.78-0.89), when adjusted for C-reactive protein, alcohol consumption, serum high-density lipoprotein, waist-to-hip ratio, family history of coronary heart disease, exercise-induced ST changes and the use of medications for hypertension, dyslipidaemia or aspirin. Also, Framingham risk score was related to the risk of death (RR 1.05, 95% CI 1.03-1.07, P < 0.001). Subjects with high European or Framingham score and low VO2peak represent the highest risk group. CONCLUSION: An important finding is that the risk scores can be used to identify men for whom low cardiorespiratory fitness predicts an especially high risk for death from cardiovascular and any other cause.