Selection of TNF-alpha binding affibody molecules using a beta-lactamase protein fragment complementation assay.

Protein fragment complementation assays (PCAs) based on different reporter proteins have been described as powerful tools for monitoring dynamic protein-protein interactions in living cells. The present study describes the construction of a PCA system based on genetic splitting of TEM-1 beta-lactamase for the selection of proteins specifically interacting in the periplasm of Escherichia coli bacterial cells, and its application for the selection of affibody molecules binding human tumour necrosis factor-alpha (TNF-alpha) from a combinatorial library. Vectors encoding individual members of a naïve 10(9) affibody protein library fused to a C-terminal fragment of the beta-lactamase reporter were distributed via phage infection to a culture of cells harbouring a common construct encoding a fusion protein between a non-membrane anchored version of a human TNF-alpha target and the N-terminal segment of the reporter. An initial binding analysis of 29 library variants derived from surviving colonies using selection plates containing ampicillin and in some cases also the beta-lactamase inhibitor tazobactam, indicated a stringent selection for target binding variants. Subsequent analyses showed that the binding affinities (K(D)) for three selected variants studied in more detail were in the range 14-27 nm. The selectivity in binding to TNF-alpha for these variants was further demonstrated in both a cross-target PCA-based challenge and the specific detection of a low nm concentration of TNF-alpha spiked into a complex cell lysate sample. Further, in a biosensor-based competition assay, the binding to TNF-alpha of three investigated affibody variants could be completely blocked by premixing the target with the therapeutic monoclonal antibody adalimumab (Humira), indicating overlapping epitopes between the two classes of reagents. The data indicate that beta-lactamase PCA is a promising methodology for stringent selection of binders from complex naïve libraries to yield high affinity reagents with selective target binding characteristics.

Quantitative analysis of DNA methylation after whole bisulfitome amplification of a minute amount of DNA from body fluids.

Cell-free circulating DNA isolated from the plasma of individuals with cancer has been shown to harbor cancer-associated changes in DNA methylation, and thus it represents an attractive target for biomarker discovery. However, the reliable detection of DNA methylation changes in body fluids has proven to be technically challenging. Here we describe a novel combination of methods that allows quantitative and sensitive detection of DNA methylation in minute amounts of DNA present in body fluids (quantitative Methylation Analysis of Minute DNA amounts after whole Bisulfitome Amplification, qMAMBA). This method involves genome-wide amplification of bisulphite-modified DNA template followed by quantitative methylation detection using pyrosequencing and allows analysis of multiple genes from a small amount of starting DNA. To validate our method we used qMAMBA assays for four genes and LINE1 repetitive sequences combined with plasma DNA samples as a model system. qMAMBA offered high efficacy in the analysis of methylation levels and patterns in plasma samples with extremely small amounts of DNA and low concentrations of methylated alleles. Therefore, qMAMBA will facilitate methylation studies aiming to discover epigenetic biomarkers, and should prove particularly valuable in profiling a large sample series of body fluids from molecular epidemiology studies as well as in tracking disease in early diagnostics.

Multi-factor dimensionality reduction applied to a large prospective investigation on gene-gene and gene-environment interactions.

It is becoming increasingly evident that single-locus effects cannot explain complex multifactorial human diseases like cancer. We applied the multi-factor dimensionality reduction (MDR) method to a large cohort study on gene-environment and gene-gene interactions. The study (case-control nested in the EPIC cohort) was established to investigate molecular changes and genetic susceptibility in relation to air pollution and environmental tobacco smoke (ETS) in non-smokers. We have analyzed 757 controls and 409 cases with bladder cancer (n=124), lung cancer (n=116) and myeloid leukemia (n=169). Thirty-six gene variants (DNA repair and metabolic genes) and three environmental exposure variables (measures of air pollution and ETS at home and at work) were analyzed. Interactions were assessed by prediction error percentage and cross-validation consistency (CVC) frequency. For lung cancer, the best model was given by a significant gene-environment association between the base excision repair (BER) XRCC1-Arg399Gln polymorphism, the double-strand break repair (DSBR) BRCA2-Asn372His polymorphism and the exposure variable 'distance from heavy traffic road', an indirect and robust indicator of air pollution (mean prediction error of 26%, P<0.001, mean CVC of 6.60, P=0.02). For bladder cancer, we found a significant 4-loci association between the BER APE1-Asp148Glu polymorphism, the DSBR RAD52-3'-untranslated region (3'-UTR) polymorphism and the metabolic gene polymorphisms COMT-Val158Met and MTHFR-677C>T (mean prediction error of 22%, P<0.001, mean CVC consistency of 7.40, P<0.037). For leukemia, a 3-loci model including RAD52-2259C>T, MnSOD-Ala9Val and CYP1A1-Ile462Val had a minimum prediction error of 31% (P<0.001) and a maximum CVC of 4.40 (P=0.086). The MDR method seems promising, because it provides a limited number of statistically stable interactions; however, the biological interpretation remains to be understood.

High incidence of vascular reconstructions in socioeconomically deprived areas of an urban Swedish population.

BACKGROUND: In Malmö approximately 250,000 citizens live in 17 administrative areas with substantial socioeconomic differences. At the single centre for arterial reconstruction, Malmö University Hospital, all procedures are registered prospectively. METHODS: Between 1987 and 2002, 1832 Malmö citizens underwent reconstruction for peripheral arterial disease, either intermittent claudication (IC) or critical leg ischaemia (CLI). A socioeconomic score based on migration rate, percentage of residents with foreign citizenship/residents with foreign background, social welfare support dependency and unemployment rate has previously been developed and validated for each area. The relationship between socioeconomic score and area-specific standardized morbidity ratios (SMRs) after vascular reconstruction was analysed by population-weighted linear regression. RESULTS: The mean incidence of vascular reconstruction was 76 (range 27-106) per 100,000 person years. Age- and sex-adjusted SMRs ranged from 0.57 to 1.39. A strong correlation between SMR and socioeconomic score was found overall (R=0.63; P=0.007), in men (R=0.63; P=0.007) and in women (R=0.58; P=0.039), and for IC (R=0.58; P=0.015) and CLI (R=0.58; P=0.015). CONCLUSION: In an urban population with similar access to medical care, vascular reconstruction rates varied substantially. High-rate areas were characterized by inferior socioeconomic circumstances and a higher prevalence of smoking, hypertension and obesity.

Reduced lung function predicts increased fatality in future cardiac events. A population-based study.

OBJECTIVE: Moderately reduced lung function in apparently healthy subjects has been associated with incidence of coronary events. However, whether lung function is related to the fatality of the future events is unknown. This study explored whether reduced forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV(1)) in initially healthy men is related to the fatality of the future coronary events. DESIGN: Prospective cohort study. SETTING: Population-based study from Malmö, Sweden. SUBJECTS: A total of 5452 healthy men, 28-61 years of age. MAIN OUTCOME MEASURES: Incidence of first coronary events was monitored over a mean follow-up of 19 years. The fatality of the future events was studied in relation to FEV and FVC. RESULTS: A total of 589 men suffered a coronary event during follow-up, 165 of them were fatal during the first day. After risk factors adjustment, low FEV or FVC were associated with incidence of coronary events (fatal or nonfatal) and this relationship was most pronounced for the fatal events. Amongst men who subsequently had a coronary event, the case-fatality rates were higher in men with low FEV or FVC. Adjusted for risk factors, the odds ratio for death during the first day was 1.00 (reference), 1.63 (95% CI: 0.9-3.1), 1.86 (1.0-3.5) and 2.06 (1.1-3.9), respectively, for men with FVC in the 4th, 3rd, 2nd, and lowest quartiles (trend: P < 0.05). FEV showed similar relationships with the fatality rates. CONCLUSION: Apparently healthy men with moderately reduced lung function have higher fatality in future coronary events, with a higher proportion of coronary heart disease deaths and less nonfatal myocardial infarction.

Weight gain in relation to plasma levels of complement factor 3: results from a population-based cohort study.

AIMS/HYPOTHESIS: Mice that are deficient for complement factor 3 (C3) have shown resistance to weight gain, despite increased food intake. Cross-sectional studies of humans have reported correlations between C3 and obesity. This longitudinal study explored whether C3 predicts a large weight gain in middle-aged men. METHODS: Plasma concentrations of C3 and complement factor 4 (C4) were measured in 2,706 non-diabetic healthy men aged between 38 and 50 years, who were re-examined after a mean period of 6.1 years. RESULTS: After adjustments for initial weight, age, height and follow-up time, the odds of incurring large weight gain (75th percentile, > or =3.8 kg) were 1.00 (reference), 0.96 (95% CI:0.7-1.2), 1.1 (CI:0.9-1.5) and 1.4 (CI:1.1-1.8), respectively, among men with C3 levels in the first, second, third and fourth quartiles (p for trend=0.01) respectively. This relationship remained significant after further adjustments for lifestyle factors (physical inactivity, alcohol, smoking), metabolic factors (glucose or homeostasis model assessment values, cholesterol, triglycerides), inflammatory markers (fibrinogen, haptoglobin, ceruloplasmin, orosomucoid, alpha1-antitrypsin) and for C4. C4 was associated with weight gain after adjustments for initial weight, height, follow-up time and lifestyle factors, but not after adjustments for C3. CONCLUSIONS/INTERPRETATION: C3 is a risk factor for incurring large weight gain in middle-aged men.

Dietary habits after myocardial infarction - results from a cross-sectional study.

OBJECTIVE: Comparing habitual nutrient intakes in persons with a history of acute myocardial infarction (AMI), and age-matched controls. Design. Cross-sectional study. Subjects. Men and women (525 cases and 1890 matched controls), aged 47-73 years, of the population-based Malmö Diet and Cancer cohort. METHODS: Nutrient intakes were assessed by a validated modified diet history method. Body fatness was assessed by bioimpedance analysis. Case ascertainment was provided by national and regional registries. Men and women were analysed separately. Median time since AMI was 5.5 years in men and 3.8 years in women. Cases reported lower energy intakes (EIs) than controls, despite having similar basal metabolic rates. After adjustment for total EI, both male and female cases had lower fat intake and higher intake of several micronutrients, such as ascorbic acid, folate, and vitamin E, than controls, the difference being largest in men. Most of the cases reporting dietary change quoted 'disease' as their main reason for change. They had lower EI and lower energy-adjusted intake of fat than other cases. CONCLUSIONS: Survivors of AMI reported dietary habits more in line with current recommendations, particularly those who afterwards reported having changed their dietary habits. The possible bias introduced by social desirability is discussed.

Prevalence and prognostic significance of asymptomatic peripheral arterial disease in 68-year-old men with diabetes. Results from the population study 'Men born in 1914' from Malmö, Sweden.

OBJECTIVE: To assess the prevalence of asymptomatic peripheral arterial disease (PAD) in older men with diabetes and to compare the incidence of cardiac events and deaths in diabetic and non-diabetic men with abnormal and normal systolic ankle-brachial pressure index, respectively. RESEARCH DESIGN AND METHODS: Population-based cohort of 68-year-old men (n = 474). Diabetes was defined as history of diabetes or a fasting blood glucose > or = 6.1 mmol/l. PAD was defined as an ankle-brachial pressure index (ABI) < 0.9 in either leg. Fourteen-year mortality and cardiac event rates were based on record linkage with regional and national registers. RESULTS: The prevalence of PAD in men with and without diabetes was 29 and 12%, respectively (p = 0.003). The incidence of cardiac events was 22.9/1000 person years in men free from both diabetes and PAD. In the absence of an abnormal pressure index, diabetes was associated with an event rate of 28.4 (p = 0.469). In the presence of an abnormal index the incidence was 102 (p < 0.001). This pattern remained in the multivariate analysis when other atherosclerotic risk factors were taken into account. Cardiovascular mortality rates similarly differed substantially between diabetic men with and without PAD. CONCLUSIONS: A fasting blood glucose value above 6.1 mmol/l even in the absence of symptoms indicating diabetes was associated by an increased prevalence of asymptomatic PAD. The cardiovascular risk in diabetes varied widely between men with and without abnormal ankle-brachial pressure index.

Risk of myocardial infarction and stroke in smokers is related to plasma levels of inflammation-sensitive proteins.

BACKGROUND: The extent to which differences in cardiovascular risk between smokers with similar daily tobacco consumption may be related to plasma levels of inflammation-sensitive proteins (ISP) and whether these proteins are associated with levels of carboxyhemoglobin (COHb%) have not been clarified. METHODS AND RESULTS: In a population-based cohort of 1489 never smokers, 1685 former smokers, and 2901 current smokers, aged 28 to 61 years, plasma levels of orosomucoid (alpha(1)-acid glycoprotein), alpha(1)-antitrypsin, haptoglobin, fibrinogen, and ceruloplasmin were measured. COHb% levels were available for 2098 of them. Incidence of myocardial infarction, stroke, and death were monitored over 18.7+/-4.7 years. The proportion with high ISP levels (ie, > or =2 ISP in the top quartile) increased progressively with daily tobacco consumption (P<0.01) and COHb% (P<0.01). In all smoking categories, the incidence of stroke, cardiac events, and death was related to ISP. In heavy smokers, high ISP levels were associated with adjusted relative risks of 1.57 (1.05 to 2.35) and 1.50 (1.11 to 2.03) for cardiac events and death, respectively. Corresponding figures for moderate and light smokers were 1.59 (1.13 to 2.24) and 1.14 (0.87 to 1.49), respectively, and 1.32 (0.95 to 1.85) and 1.48 (1.10 to 1.98), respectively. CONCLUSIONS: ISP levels are related to COHb% in smokers. High levels are associated with an increased cardiovascular risk.

Leg blood flow and long-term cardiovascular prognosis in men with typical and atypical intermittent claudication.

OBJECTIVES: To compare leg blood flow and the long-term cardiovascular prognosis in men with typical and atypical intermittent claudication (IC) according to Rose's questionnaire. METHODS: Leg blood flow during reactive hyperaemia was assessed at 55 years of age by calf plethysmography. Measurement of the systolic ankle-arm pressure index was used to assess the prevalence of peripheral arterial disease (PAD) at 68 years of age. Mortality and incidence of cardiac events is based on record linkage with regional and national registers. RESULTS: Twenty-one (3%) out of 700 men had typical IC at 55 years of age. Sixteen (76%) of these had normal plethysmography, with a mean peak flow (95% CI) of 24.6 (19.3-30.0) ml/min/100 ml. Peak flows were similar and normal in men with atypical IC (mean 22.5; 95% CI 21.3-23.6) and men having no pain (mean 23.8; 95% CI 23.1-24.4). Mortality rates were increased in men with typical IC but who had normal leg blood flow (49.5 deaths/1000 person years; p = 0.008), and men having atypical IC (35.3 deaths/1000 person years; p = 0.007) in comparison with men having no leg pain (27.4 deaths/1000 person years). This could not be accounted for by an increased cardiac event rate. The prevalence of PAD at 68 years of age was not increased in either of these two groups. CONCLUSIONS: Only 12% of subjects with exertional pain fulfilled all criteria for typical IC. Typical and atypical claudication in absence of objective evidence of PAD was associated with a reduced life expectancy, but not with an increased cardiovascular risk. To properly assess the prognosis, non-invasive assessment of PAD should be used in addition to questionnaires.

Influence of obesity on cardiovascular risk. Twenty-three-year follow-up of 22,025 men from an urban Swedish population.

OBJECTIVE: To assess to what extent the incidence of coronary events and death related to smoking, hypertension, hyperlipidemia and diabetes is modified by obesity. DESIGN: Prospective cohort study. SUBJECTS: A total of 22 025 men aged 27 to 61-y-old at entry. MEASUREMENTS: Incidence of coronary events (CE, ie acute myocardial infarctions and deaths due to chronic ischaemic heart disease) and death during 23 y of follow-up was studied in relation to body mass index (BMI), heart rate, blood pressure, blood lipids, glucose and insulin, lifestyle factors, history of angina pectoris, history of cancer, self-reported health and socio-economic conditions. RESULTS: At the end of follow-up 20% of the obese men were no longer alive, and 13% had had a coronary event. Incidence of CE was 16% lower (RR (relative risk) 0.84; 95% confidence interval (CI) 0.65-1.10) among underweight (n=1171), 24% higher (RR 1.24; CI 1.12-1.37) among overweight (n=7773), and 76% higher (RR 1.76; 95% CI 1.49-2.08) among obese men (n=1343) than it was among men with normal BMI (n=11 738). The risk associated with overweight and obesity remained statistically significant after adjustment for potential confounders (RR 1.18; CI 1.07-1.31; and 1.39; 1.17-1.65, respectively). The association between BMI and mortality was J-shaped. In all, 1.7% of the obese men were smokers with hypertension, hyperlipidaemia and diabetes, 16.3% were not exposed to any of these risk factors. The cardiovascular risk associated with obesity was small in the absence of other risk factors. Between smoking and obesity there was a statistically significant synergistic effect. CONCLUSIONS: Obesity is associated with an increased incidence of coronary events and death. The risk associated with obesity is substantially increased by exposure to other atherosclerotic risk factors, of which smoking seems to be the most important. The preventive potential of these associations should be evaluated in controlled trials.

Insulin resistance in non-diabetic subjects is associated with increased incidence of myocardial infarction and death.

AIMS: To compare the incidence of myocardial infarction and death in non-diabetic subjects with and without insulin resistance. METHODS: Population-based prospective cohort study, in Malmö, Sweden, of 4748 non-diabetic subjects (60% women), aged 46-68 years, with no history of myocardial infarction or stroke. The prevalence of insulin resistance was established by the homeostasis model assessment (HOMA) and defined as values above the sex-specific 75th percentile (1.80 for women and 2.12 for men). Incidence of myocardial infarction and death is based on record linkage with local and national registers. Cox's proportional hazards model was used to assess the influence of insulin resistance after adjustment for age, sex, hyperglycaemia, raised arterial blood pressure, dyslipidaemia, central obesity, smoking and leisure-time physical activity. RESULTS: Sixty-two subjects suffered a coronary event, and 93 subjects died during the 6-year follow-up period. Insulin resistance was after adjustment for other factors included in the insulin resistance syndrome and other potential confounders, associated with an increased incidence of coronary events (relative risk (RR) 2.18; 95% confidence interval (CI) 1.22-3.87; P = 0.008) and deaths (RR 1.62; 1.03-2.55; P = 0.038). CONCLUSIONS: Insulin resistance, as assessed by the HOMA method, was in this cohort of middle-aged non-diabetic subjects associated with an increased incidence of myocardial infarction and death. This risk remained when smoking, low physical activity and factors included in the insulin resistance syndrome were taken into account in a stepwise regression model.

Risk of developing diabetes is inversely related to lung function: a population-based cohort study.

AIM: To investigate whether reduced lung function is a risk factor for developing diabetes. METHODS: Non-diabetic men (n = 382) from the population-based cohort 'Men Born in 1914' were examined with spirometry at age 55 years. The cohort was re-examined at 68 years. Diabetes and fasting plasma glucose at follow-up were studied in relation to vital capacity (VC) and forced expiratory volume (FEV1.0) at baseline. RESULTS: Fifteen men developed diabetes during the follow-up. The percentage with diabetes in the 1st, 2nd, 3rd and top quartile of vital capacity were 7%, 5%, 2%, and 1%, respectively (P for trend = 0.01). Fasting glucose (log transformed, mmol/l) at follow-up was 1.63 +/- 0.16, 1.62 +/- 0.18, 1.61 +/- 0.11 and 1.60 +/- 0.11, respectively (P for trend = 0.11). The longitudinal associations between VC and diabetes (P = 0.001) and log glucose (P = 0.036) were significant after adjustments for several potential confounders. FEV(1.0) at baseline showed similar associations with diabetes at follow-up. CONCLUSIONS: The risk of developing diabetes is inversely associated with pulmonary function among middle-aged men.

Meat consumption in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts: results from 24-hour dietary recalls.

OBJECTIVE: To evaluate meat intake patterns in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts. DESIGN AND SETTING: 24-Hour dietary recalls were assessed within the framework of a prospective cohort study in 27 centres across 10 European countries by means of standardised computer-assisted interviews. SUBJECTS: In total, 22 924 women and 13 031 men aged 35-74 years. RESULTS: Mean total meat intake was lowest in the 'health-conscious' cohort in the UK (15 and 21 g day-1 in women and men, respectively) and highest in the north of Spain, especially in San Sebastian (124 and 234 g day-1, respectively). In the southern Spanish centres and in Naples (Italy), meat consumption was distinctly lower than in the north of these countries. Central and northern European centres/countries showed rather similar meat consumption patterns, except for the British and French cohorts. Differences in the intake of meat sub-groups (e.g. red meat, processed meat) across EPIC were even higher than found for total meat intake. With a few exceptions, the Mediterranean EPIC centres revealed a higher proportion of beef/veal and poultry and less pork or processed meat than observed in central or northern European centres. The highest sausage consumption was observed for the German EPIC participants, followed by the Norwegians, Swedish, Danish and Dutch. CONCLUSIONS: The results demonstrate distinct differences in meat consumption patterns between EPIC centres across Europe. This is an important prerequisite for obtaining further insight into the relationship between meat intake and the development of chronic diseases.

Variability of fish consumption within the 10 European countries participating in the European Investigation into Cancer and Nutrition (EPIC) study.

OBJECTIVE: To describe and compare the consumption of total fish (marine foods) and the fish sub-groups - white fish, fatty fish, very fatty fish, fish products and crustacea, in participants from the European Investigation into Cancer and Nutrition (EPIC) study. DESIGN: Cross-sectional analysis of dietary intake using a computerised standardised 24-hour recall interview. Crude means, means and standard errors adjusted by age, season and day of the week were calculated, stratified by centre and gender. SETTING: Twenty-seven redefined centres in the 10 European countries participating in the EPIC study. SUBJECTS: In total, 35 955 subjects (13 031 men and 22 924 women), aged 35-74 years, selected from the main EPIC cohort. RESULTS: A six- to sevenfold variation in total fish consumption exists in women and men, between the lowest consumption in Germany and the highest in Spain. Overall, white fish represented 49% and 45% of the intake of total fish in women and men, respectively, with the greatest consumption in centres in Spain and Greece and the least in the German and Dutch centres. Consumption of fatty fish reflected that of total fish. However, the greatest intake of very fatty fish was in the coastal areas of northern Europe (Denmark, Sweden and Norway) and in Germany. Consumption of fish products was greater in northern than in southern Europe, with white fish products predominating in centres in France, Italy, Spain, The Netherlands and Norway. Intake of roe and roe products was low. The highest consumption of crustacea was found in the French, Spanish and Italian centres. The number of fish types consumed was greater in southern than in northern Europe. The greatest variability in consumption by day of the week was found in the countries with the lowest fish intake. CONCLUSIONS: Throughout Europe, substantial geographic variation exists in total fish intake, fish sub-groups and the number of types consumed. Day-to-day variability in consumption is also high.

Food patterns and components of the metabolic syndrome in men and women: a cross-sectional study within the Malmö Diet and Cancer cohort.

This study examined the relations between food patterns and five components of the metabolic syndrome in a sample of Swedish men (n = 2,040) and women (n = 2,959) aged 45-68 years who joined the Malmö Diet and Cancer study from November 1991 to February 1994. Baseline examinations included an interview-administered diet history, a self-administered questionnaire, blood pressure and anthropologic measurements, and blood samples donated after an overnight fast. Cluster analysis identified six food patterns for which 43 food group variables were used. Logistic regression analysis was used to examine the risk of each component (hyperinsulinemia, hyperglycemia, hypertension, dyslipidemia, and central obesity) and food patterns, controlling for potential confounders. The study demonstrated relations, independent of specific nutrients, between food patterns and hyperglycemia and central obesity in men and hyperinsulinemia in women. Food patterns dominated by fiber bread provided favorable effects, while food patterns high in refined bread or in cheese, cake, and alcoholic beverages contributed adverse effects. In women, food patterns dominated by milk-fat-based spread showed protective relations with hyperinsulinemia. Relations between risk factors and food patterns may partly depend on gender differences in metabolism or food consumption and on variations in confounders across food patterns.

Geographic distribution of stroke incidence within an urban population: relations to socioeconomic circumstances and prevalence of cardiovascular risk factors.

BACKGROUND AND PURPOSE: Geographic differences in stroke incidence indicate a potential for prevention. The present study from the city of Malmö, Sweden, sought to investigate whether incidence of stroke in residential areas is related to prevalence of cardiovascular risk factors and socioeconomic circumstances. METHODS: The Stroke Register in Malmö, Sweden, was used for retrieval of the 3540 patients who suffered a first stroke between 1989 and 1998. The Malmö Diet and Cancer cohort (n=28 466) was used to assess area specific prevalence of hypertension, diabetes, smoking, and being overweight and for computation of a cardiovascular risk score. Socioeconomic circumstances for the 17 administrative areas were expressed in terms of a composite score. RESULTS: Standardized stroke incidence ranged among areas from 437 to 743 per 100 000 for men and from 223 to 518 per 100 000 for women. Socioeconomic score correlated significantly with area-specific stroke rates among men (r=-0.62, P=0.008) and women (r=-0.67, P=0.004). Incidence of stroke was significantly associated with cardiovascular risk score for each area (men, r=0.53, P<0.05; women, r=0.76, P<0.001). The cardiovascular score and the socioeconomic score together accounted for 44% of the geographic variance among men and 63% among women. CONCLUSIONS: Marked differences occurred in stroke incidence among residential areas within this urban population. High-rate areas were characterized by a higher prevalence of smoking, hypertension, diabetes, and being overweight and by inferior socioeconomic circumstances. These risk factors accounted for a substantial proportion of the geographic variance in incidence of stroke.

Breast cancer incidence in ex-smokers in relation to body mass index, weight gain and blood lipid levels.

According to several studies breast cancer is more common among former smokers. This study explores whether this association has any relationship with anthropometric measurements or blood lipid levels. The 2082 ex-smokers (mean age 49.9 years) in the Malmö Preventive Cohort were followed for an average of 13.3 years using official cancer registries. This yielded 93 incident breast cancer cases. Oestrogen receptor (ER) status was assessed by an immunological method. Incidence of breast cancer covaried with height, body mass index, weight gain and cholesterol levels. None of these associations reached statistical significance. Incidence of breast cancer increased over quartiles of serum triglycerides, Ptrend: 0.02, relative risk (RR) for triglycerides as a continuous variable: 1.46 (1.21-1.77). Nineteen tumours were ER negative; this subgroup was similarly related to high triglycerides, 1.76 (1.40-2.21). All results were similar when BMI and cholesterol levels were entered into the model. It is concluded that breast cancer incidence covaries with triglyceride levels in ex-smokers.