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Objective Hormone replacement therapy (HRT) has generally been documented to reduce plasminogen activator inhibitor-1 (PAI-1) and fibrinogen levels in plasma of postmenopausal women. We used a wash out protocol to study whether stopping long-term HRT with estrogen alone or a combination of estrogen–progestin have different effects on these markers of hemostasis. Study design Thirty healthy postmenopausal women on HRT participated. Fifteen had estradiol valerate, and 15 had estradiol valerate and levonorgestrel. Each was studied after long-term HRT (period 1), four weeks after cessation of the treatment (period 2, wash out), and three weeks after reintroducing the therapy (period 3). Results In the estrogen group, PAI-1 increased by 18% during the wash out period ( P =0.013) and decreased by 22% after reintroduction of therapy ( P =0.001). In the combined therapy group, there was a trend of PAI-1 to increase by 18% when therapy was discontinued ( P =0.17), and it decreased by 25% after reintroduction of hormone replacement therapy ( P =0.036). Fibrinogen was initially lower in the estrogen group compared with the combined therapy group ( p =0.014), and did not change during wash out. Conclusion This wash out study shows that cessation of long-term HRT unfavorably increases PAI-1, but appears to have no adverse effect on fibrinogen.

OBJECTIVES: To establish whether the relationship between interleukin-6 (IL-6) and plasma lipid and C-reactive protein (CRP) concentrations is different in Finnish nonagenarians than in middle-aged subjects with lower inflammatory status. DESIGN: Cross-sectional. SETTING: Observational cohort study concentrating on the oldest old. PARTICIPANTS: Nonagenarians (n = 291, mean age ± standard deviation 90 ± 1; 68 men, 223 women) who lived in the Tampere municipality in southern Finland and a middle-aged control population from the same area (n = 227, aged 44 ± 8). MEASUREMENTS: Plasma high sensitive CRP and lipid concentrations were analyzed using an automatic analyzer and IL-6 levels using enzyme-linked immunosorbent assay. RESULTS: Plasma concentrations of IL-6 (4.39 ± 5.25 vs 1.88 ± 1.98 pg/mL) and CRP (3.54 ± 4.98 vs 1.53 ± 1.91 mg/L) were significantly higher in nonagenarians than in middle-aged subjects (P

Insulin resistance, LDL particle size, and LDL susceptibility to oxidation in pediatric kidney and liver recipients.BackgroundDyslipidemia is common after solid organ transplantation. We have described hypertriglyceridemia in about 50% of our pediatric kidney, and in about 30% of our liver recipients. The aim of the present study was to find out whether this post-transplantation hypertriglyceridemia after pediatric solid organ transplantation is associated with insulin resistance and the occurrence of small, dense low-density lipoprotein (LDL).MethodsFifty kidney and 25 liver recipients (aged 4 to 18 years) on triple immunosuppression, and 181 control children participated in the study for an average of 5.3 and 6.4 years after kidney and liver transplantation (range 1 to 11 years), respectively. Homeostasis model assessments for insulin resistance (HOMA) were calculated and fasting lipoprotein lipid profile, apolipoprotein A-I and B concentrations, LDL particle diameter, and indices of LDL susceptibility to copper-induced oxidation determined.ResultsKidney patients had significantly higher serum total, high-density, and low-density lipoprotein cholesterol, triglyceride, apolipoprotein A-I and B concentrations than liver patients or control subjects (P < 0.003 for all). HOMA indices higher than the 95th percentile of Canadian normal children were seen in 50.0% of kidney (of liver 41.2%) recipients younger than 11 years, and in 27.3% of older recipients (of liver 37.5%). Smaller sized LDL or LDL of increased oxidizability was not more frequent in patients than in control children.ConclusionPediatric kidney recipients had significantly higher lipid and insulin concentrations than healthy control children. Combined hyperlipidemia and features of the dysmetabolic syndrome were common in children after kidney and liver transplantation. However, no small, dense LDL, or LDL prone to oxidation was seen in either group.

Trans-fatty acids (TFA) have been classified as atherogenic dietary constituents but the effect of hormone replacement therapy (HRT) on their concentrations is not known. We used a washout protocol to study the effect of long-term estrogen and combined estrogen-progestin HRT on plasma elaidate (18:1t), which is the trans isomer of oleate and the major TFA in the diet. The study group comprised 15 women receiving estradiol valerate HRT and 15 women receiving combined HRT with estradiol valerate and levonorgestrel. The concentrations of elaidate in plasma phospholipids, cholesteryl esters and triglycerides were determined by gas chromatography. At baseline, the total plasma elaidate concentration was lower in the combined HRT group than in the estradiol valerate HRT group (p < 0.01). In the combined HRT group, the concentration of elaidate increased significantly after withdrawal of HRT (p < 0.001) and decreased again to the baseline level after restart of therapy (p < 0.001). These changes were due to decreases in the concentrations of phospholipids and triglycerides; in phospholipids there was also a proportional decrease of elaidate. There were no changes in elaidate in women receiving estradiol valerate alone. Our results suggest that long-term combined HRT treatment decreases plasma TFA, which is not achieved by estrogen alone.

Increased concentrations of serum triglyceride and low-density lipoprotein (LDL) cholesterol are common after heart transplantation (HTx). These abnormalities may promote transplant vascular disease and atherosclerosis, especially if LDL is small, dense, and oxidized. There have been no previous studies of LDL particle size and LDL susceptibility to oxidation in children after HTx.Twenty-three HTx recipients (aged 3-19 years) who received triple-drug immunosuppression therapy after HTx and 181 controls within the same age range participated in the study. Total, high-density lipoprotein, and LDL-cholesterol concentrations; triglyceride concentration, and glucose and insulin concentrations during oral glucose tolerance tests were determined an average of 3 years after HTx (range, 1-7 years). Moreover, we determined serum lipoprotein (a) concentration, apolipoprotein E phenotype, LDL particle size, and indices of LDL susceptibility to copper-induced oxidation in 12 HTx recipients.We found hypertriglyceridemia in 56.5% and hyperinsulinemia in 30.4% of patients. Triglyceride concentration and body mass index were associated significantly with insulin concentration (p0.008 for both). Low-density lipoprotein particle size, LDL susceptibility to in vitro oxidation, and lipoprotein (a) concentrations did not differ significantly between HTx patients and controls. Low-density lipoprotein particle size was associated inversely with cyclosporine through level (Neoral, r = -0.59, p = 0.045), whereas weight-adjusted dosage of cyclosporine correlated positively with longer lag time of LDL oxidation (r = 0.69, p = 0.013).Hypertriglyceridemia and hyperinsulinemia were common in children receiving triple-drug immunosuppression therapy after HTx. Increased cyclosporine through concentration was associated with small LDL particle size but did not increase LDL susceptibility to oxidation.

Although dyslipidaemia is common after solid organ transplantation (Tx), there are few long-term studies in children. We investigated the prevalence of dyslipidaemia up to 5 years after Tx in 125 children on triple immunosuppression with one of three different well-functioning grafts, kidney, liver, and heart, and 181 controls. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) concentrations were measured annually. Low-density lipoprotein cholesterol concentrations were also calculated. The risk factors for dyslipidaemia were determined at 3 years. There was a high prevalence of hypertriglyceridaemia in all three groups, 50% in the kidney transplantation (KTx) and heart transplantation (HTx) groups and 30% in the liver transplantation (LTx) group. In addition, 50% of KTx patients had high TC. In the Tx groups taken together, the following independent associations were observed: KTx and high pre-Tx TC were associated with high TC, high trough concentration of blood cyclosporine with low HDL-C, and older age at Tx accounted for higher TG. Dyslipidaemia, especially hypertriglyceridaemia, was common 3–5 years after Tx. The aetiology is multifactorial and depends on the transplanted organ.

Estrogens modulate lipid metabolism and the increased risk of atherosclerosis in postmenopausal women is at least partly due to the reduction of estrogen production after menopause. We studied the effect of menopause on the contents of long-chain fatty acids, free cholesterol (FC) and cholesterol ester (CE) in uterine artery wall.The uterine artery intima samples were obtained in connection with surgery of 21 postmenopausal and 51 premenopausal women. The amount of FC, CE and phospholipid fatty acids were measured by gas chromatography after extraction and fractionation and these lipid values were related to menopausal status, age and serum total and low-density lipoprotein (LDL) cholesterol levels.Premenopausal females had significantly less intimal FC (161 +/- 50 vs. 407 +/- 276 microg/100 mg wet weight, P = 0.003) and CE (19 +/- 34 vs. 305 +/- 348 microg/100 mg wet weight, P = 0.050) and smaller proportion of linoleic acid out of all phospholipid fatty acids (4.2 vs. 7.2%, P = 0.002) than postmenopausal women after adjustment with age. The content of CE (r = 0.34, P = 0.025) and the FC-to-CE ratio (r = -0.45, P = 0.002) correlated with age in premenopausal but not in postmenopausal women. Moreover, the intimal content of CE correlated with the percentage of intimal phospholipid linoleic acid in postmenopausal women (r = 0.79, P = 0.020). The same was true for FC (r = 0.73, P0.001).These results indicate that CE and FC accumulation into the wall of uterine artery depends on menopausal status, independently of age, and that the phospholipid long-chain fatty acid composition differs significantly between premenopausal and postmenopausal women. This suggests that estrogens may be involved in the regulation of artery wall lipid composition.

The atheroprotective action of estrogen is mediated by estrogen receptors (ESR) 1 and 2, expressed in atherosclerotic lesions. The effects of hormone replacement therapy (HRT) and ESR1 PvuII genotypes on atherosclerosis have not previously been studied prospectively in postmenopausal women.We investigated the effect of HRT on the progression of atherosclerosis in a 5-year follow-up study of 88 postmenopausal women aged 45-71 years at baseline allocated into three groups based on the use of HRT. The HRT-EVP group (n=26) used sequential estradiol valerate (EV) plus progestin (P), the HRT-EV group EV alone (n=32), and a control group (n=30) was without HRT. The atherosclerosis severity score (AS) of the abdominal aorta and carotid arteries were determined by sonography and the ESR1 PvuII genotypes (P/P, P/p and p/p) by PCR.HRT, time and ESR1 PvuII genotype had a statistically significant or borderline significant main effect on AS during 5-year follow-up (P=0.004, P0.001 and P=0.090, respectively), when analyzed by repeated measures analysis of variance. There was a significant genotype-by-treatment (HRT-EVP and control groups) interaction for AS (P=0.034). In response to HRT-EVP, subjects with P/P, compared with those with P/p and p/p genotypes, had a less increase in AS (1.61+/-1.14 vs. 1.71+/-1.27 vs. 2.43+/-1.27). Baseline AS as covariate in similar model does not change the significant interaction effect between HRT-EVP and control groups (P=0.036). But this effect was not found between HRT-EV and control groups.Our results suggest that the effect of HRT-EVP in postmenopausal women on progression of AS may be determined in part by the genotype of ESR1 PvuII.

Background Impairment of coronary blood flow reserve has been shown to be an early manifestation of atherosclerosis and coronary artery disease (CAD). We studied more closely the contribution of various risk factors on early deterioration of coronary function. Materials and methods Fifty-one young, apparently healthy adults, with normal or mildly elevated serum cholesterol levels but without other major risk factors for CAD, such as diabetes or hypertension, underwent positron emission tomography (PET) studies. Coronary flow reserve (CFR) was measured using O15-water. In addition to the classical risk factors, the role of several new risk indicators, such as low-density lipoprotein (LDL) oxidation, infection (Chlamydia pneumoniae antibodies), and inflammation parameters (adhesion molecules, ICAM, VCAM, selectin, and C-reactive protein), homocysteine and body iron stores were investigated. Results Elevated lipid and lipoprotein levels were not associated with reduced coronary reactivity. However, high autoantibody titers against oxidized LDL (oxLDL) were associated with 21% lower CFR than low oxLDL (P

Background: In epidemiologic studies, the incidence of atherosclerosis rises soon after menopause in women, and hormone replacement therapy (HRT) has proved to be useful in preventing onset of clinical manifestations of the disease. However, it is not known how HRT affects sonographically determined atherosclerotic severity (AS) and number of atherosclerotic plaques (NAP) in large arteries. Furthermore, it is not clear how HRT affects oxidation of low density lipoproteins (LDL), which obviously has an important role in the pathogenesis of atherosclerosis. Objectives: The purpose of the study was to determine whether HRT has a beneficial effect on sonographically determined AS and NAP in large arteries of 101 postmenopausal women compared to 40 controls without HRT. We also studied the interaction of HRT and antibodies against oxidized LDL on AS and NAP progression. Results: Estradiol valerate alone, combined estradiol valerate–levonorgestrel and combined estradiol valerate–medroxyprogesterone acetate therapy are each associated with lower NAP and AS as compared to controls without HRT. In a multiple regression model explaining NAP in the whole study population, the strongest predictors were HRT ( P =0.0006) and copper-oxidized LDL cholesterol autoantibodies ( P =0.0491). Discussion: Our findings indicate that postmenopausal HRT is associated with a lower total number of atherosclerotic plaques and less severe atherosclerotic lesions, as compared to controls without HRT, and that this outcome may be associated with the effect of HRT on LDL cholesterol oxidation.

Background Elevated serum homocysteine concentrations have been related to coronary heart disease. However, the association has not indisputably been proven, and the mechanisms by which homocysteine may be atherogenic have only partially been elucidated. The objective of the present study was to investigate whether serum homocysteine is associated with angina pectoris and myocardial infarction. Methods We compared serum homocysteine concentrations in subjects with clinical evidence of angina pectoris or history of myocardial infarction to age-matched controls. The study included 248 males, who participated in a large cross-sectional risk factor survey carried out in five geographic areas in Finland. Results Serum homocysteine concentration was significantly higher in subjects with a history of myocardial infarction compared to controls (15·3 μmol L−1 and 13·9 μmol L−1 respectively, P = 0·037). In a logistic regression model including several cardiovascular risk factors, serum homocysteine was significantly associated with myocardial infarction (95% CI 1·0157–1·2990, P = 0·027). Serum homocysteine concentrations did not differ between subjects with angina pectoris and age-matched controls (13·9 μmol L−1 and 14·2 μmol L−1 respectively). Conclusions Our results suggest that elevated serum homocysteine is associated with myocardial infarction but not with uncomplicated coronary heart disease.

The aim of the study was to compare the recently introduced laboratory markers trypsinogen-2 and trypsin-2-alpha1 antitrypsin complex (trypsin-2-AAT) in serum with lipase and amylase in the diagnostic and prognostic evaluation of patients with acute pancreatitis (AP).The analytes were measured on admission in 64 consecutive patients with AP and in 30 controls with acute abdominal disease of extrapancreatic origin. Twenty-one patients had severe and 43 mild AP. As reference methods we used serum amylase and C-reactive protein.In subjects with AP, elevated trypsinogen-2 values (or = 90 microg/L) were observed in 63 patients (98%), trypsin-2-AAT values (or = 12 microg/L) in 64 patients (100%), lipase values (or = 200 U/L) in 64 patients (100%), and amylase values (or = 300 IU/L) in 62 patients (97%). The diagnostic accuracy of the markers was evaluated by receiver operating characteristic (ROC) analysis. On admission, trypsinogen-2, trypsin-2-AAT, lipase, and amylase differentiated patients with AP from controls with high accuracy and ROC analyses showed similar areas under the ROC curves (AUC) for trypsinogen-2 (AUC 0.960), trypsin-2-AAT (0.948), lipase (AUC 0.947), and amylase (AUC 0.930). For differentiation between severe and mild AP, trypsin-2-AAT (AUC 0.805) was slightly better than trypsinogen-2 (AUC 0.792), and they were both clearly better than lipase (AUC 0.583), C-reactive protein (AUC 0.519), or amylase (AUC 0.632) (p0.05).All the markers studied showed high accuracy for differentiating between AP and extrapancreatic diseases. However, trypsinogen-2 and trypsin-2-AAT displayed the best accuracy for predicting a severe AP already at admission, which makes these markers superior for clinical purposes.

Human serum paraoxonase (PON) is an antioxidative enzyme, which circulates on high-density lipoproteins and appears to use oxidized phospholipids as physiological substrates. PON M/L55 substitution changes the ability of PON to prevent lipid oxidation. Urinary 8-iso-PGF(2alpha) (one of F2 -isoprostanes) may represent a non-invasive in vivo index of free radical generation and we propose that PON might influence the biosynthesis of 8-iso-PGF(2alpha) in the vasculature. We studied the urinary excretion of 8-iso-PGF(2alpha) and related it to PON M/L55 genotypes in patients with type 2 diabetes mellitus (n = 55) and non-diabetic control subjects (n = 55). Urinary 8-iso-PGF(2alpha) was determined by competitive ELISA and the PON genotype by a PCR based restriction enzyme digestion method. LL homozygotes were compared to M-allele carriers (ML heterozygotes and MM homozygotes). The urinary excretion of 8-iso-PGF(2alpha) among non-diabetic non-smoking LL homozygotes was 3995.5 +/- 3352.8 ng/24-hour and among M-allele carriers 1689.8 +/- 1051.3 ng/24-hour (p = 0.017, ANCOVA; gender, hypertension, total cholesterol, triglycerides and LDL cholesterol as covariates). The excretion of 8-iso-PGF(2alpha), was increased in type 2 diabetes mellitus compared to non-diabetic control subjects. PON may thus protect against oxidative stress by destroying some biologically active lipids. Excretion of 8-iso-PGF(2alpha) is increased in type 2 diabetes, which may reflect oxidant injury.

To study the association of alcohol consumption and lipid-based cardiovascular risk factors among middle-age women, cross-sectional analysis among 274 middle-aged healthy women with different drinking habits and a follow-up analysis of alcoholic women during abstinence was performed. Serum total cholesterol, low and high-density lipoprotein cholesterol (LDL and HDL cholesterol), triglycerides (TG), apolipoproteins A1 (Apo A1) and B (Apo B), and HDL-cholesterol subfractions 2 (HDL(2)) and 3 (HDL(3)) were measured. All lipid values except LDL cholesterol positively correlated with self-reported alcohol consumption. When alcoholics were excluded the correlation was significant only for HDL cholesterol, HDL(3), and Apo A1. The increasing trend of HDL cholesterol, HDL(3) and Apo A1 were clearly seen first in women consuming >20-40 g/day of absolute alcohol. Alcohol consumption >40 g/day increased all lipid values except LDL cholesterol. Abstinence for 2 weeks caused a significant decrease in HDL(3) cholesterol, and an increase in LDL cholesterol and Apo B. The results indicate that among middle-aged women the Apo A1 and HDL cholesterol via its HDL(3) but not HDL(2) subfraction might play a role in the beneficial coronary consequences associated with moderate alcohol consumption. However, the increasing beneficial trend first appears when daily drinking exceeds 20 g/day.

The effect of 1 wk of supervised fasting on plasma lipid concentrations in subjects with different apolipoprotein E (apo E) phenotypes was studied in 58 healthy free-living volunteers. The participants consumed an 870-kJ(208 kcal)/d liquid diet containing fruit and berry juices, tea, and water. The decline in plasma total cholesterol during 1 wk of fasting was 0.46 mmol/L in women and 0.35 mmol/L in men. The decreases were significant in both women and men. The response patterns of plasma total cholesterol were not significantly different between the sexes. In men, the changes in plasma low-density-lipoprotein cholesterol during the fast differed significantly (P = 0.0181) between the apo E phenotypes, whereas in women there were no differences due to phenotype (P = 0.695). The magnitude of the change in plasma triacylglycerol during the fast was different between the sexes (P = 0.0099). The changes in plasma triacylglycerols differed significantly between apo E phenotype groups in men (P = 0.0295) but not in women (P = 0.0661). Statistical comparison between different apo E phenotypes was performed with and without the small apo E3,2+E2,2 group, with essentially similar results. During fasting, plasma high-density-lipoprotein cholesterol concentrations decreased slightly but not significantly. The study shows significant differences in the associations of apo E alleles and sex on plasma lipid responses during fasting and illustrates the importance of gene-diet interactions in the regulation of lipid metabolism in humans.

aDepartment of Obstetrics and Gynecology, University Hospital of Tampere, Tampere, Finland bDepartment of Clinical Chemistry, University Hospital of Tampere, Tampere, Finland CDepartment of Diagnostic Radiology, University Hospital of Tampere, Tampere, Finland dDepartment of Pathology, University Hospital of Tampere, Tampere, Finland eDepartment of Public Health/Biometry Unit, University of Tampere, Tampere, Finland

Some epidemiological studies have shown that serum total cholesterol increases with age. especially in women. On the other hand, the risk of coronary artery disease is smaller in women than in men. Earlier studies have shown that a small dense low density lipoprotein (LDL) is more atherogenic than a large LDL. We studied LDL size and apolipoprotein E (apo E) phenotypes in premenopausal and postmenopausal women and in men at the same age. In this study 342 subjects participating in a health screening study were examined. There were four subgroups: 40-year-old men (n = 85), 40-year-old women (n = 80), 70-year old men (n = 88) and 70-year-old women (n = 89). In the present study LDL size was larger (P0.01) in women (26.39 +/- 0.07 nm) than in men (25.95 +/- 0.07 nm). We found that LDL size correlated highly positively (r = 0.606; P0.001) with serum high density lipoprotein (HDL) concentration and inversely with serum triglyceride concentration (r = -0.627; P0.001). Measuring serum HDL cholesterol and triglycerides in health screening studies gives information indirectly about LDL size and its atherogenicity. Apo E phenotype was not significantly associated with serum triglycerides, but was associated with LDL size, LDL cholesterol, total cholesterol and HDL cholesterol. In our sample LDL size decreased and LDL cholesterol and total cholesterol increased according to the most prevalent apo E phenotypes in the order E2/3, E3/3, E3/4 and E4/4. Subjects with phenotype apo E4/4 had the smallest LDL size (25.70 +/- 0.19 nm), the highest total cholesterol (6.53 +/- 0.35 mmol/l) and the lowest HDL cholesterol values (1.28 +/- 0.04 mmol/l). We conclude that there was a significant interaction between sex and age in serum total cholesterol which was highest in older women. However, their LDL size was larger and their LDL is less atherogenic. Apo E phenotype had a significant influence on LDL size.

In several populations, the apolipoprotein E (apo E) allele epsilon 4 is associated with high concentration of plasma total and low density lipoprotein (LDL)-cholesterol and coronary artery disease (CAD). We determined the apo E phenotypes of 309 patients with angiographically verified CAD and 38 patients without CAD by isoelectric focusing and Western blotting. In men with CAD, the plasma total and LDL-cholesterol increased according to apo E phenotype in the following order: E3/2E3/3E4/3E4/4 (P = 0.03 for total cholesterol, P = 0.007 for LDL-cholesterol). In women, there was a similar trend (P = 0.22 for total cholesterol, P = 0.15 for LDL-cholesterol). The relative frequency of men with three vessel CAD increased (P = 0.43) together with LDL-cholesterol levels (P = 0.05) according to apo E phenotype E3/2, E3/3, E4/3, E4/4. Total and LDL-cholesterol levels were higher in patients with three vessel CAD than in patients with less serious types of CAD (P = 0.02 for total cholesterol, P = 0.007 for LDL-cholesterol). The relative frequency of patients with myocardial infarction increased according to apo E phenotype (P = 0.51). Both in men and women, there were no differences between apo E phenotypes in age at occurrence of the first myocardial infarction. The apo E allele frequencies of patients with CAD vs. without CAD were 2.3% vs. 1.3% for epsilon 2, 79.0% vs. 76.3% for epsilon 3 and 18.7% vs. 22.4% for epsilon 4. There were no statistically significant differences in apo E allele or phenotype frequencies between patients with CAD and without CAD or between patients with CAD and the general Finnish population. Our results support previous studies in suggesting that the apo E allele epsilon 4 is a risk factor for atherosclerosis, which affects plasma total and LDL-cholesterol. In addition, our results suggest that the apo E allele determines the severity of CAD.

High plasma homocysteine (Hcy) has been linked to impaired endothelial function. We investigated whether treatment with pravastatin affects the Hcy levels. Moreover, we studied whether the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism affects coronary vasomotion at baseline and during the treatment with pravastatin. Fifty-one healthy, mildly hypercholesterolemic men (mean age 35 ± 4 years) attended this randomised, double-blind, placebo-controlled study. The volunteers were randomised into groups with 6-month treatment with pravastatin (40 mg/day, n = 25) or placebo ( n = 26). Coronary blood flow measurements with positron emission tomography at rest and during adenosine infusion as well as biochemical analyses were done at baseline and at the end of the treatment period. The Hcy concentration decreased significantly during the pravastatin therapy (− 0.81 ± 1.46 μmol/l, p = 0.01), but not during placebo (0.02 ± 2.39 μmol/l, p = 0.97). The MTHFR polymorphism did not affect the Hcy concentration or coronary flow indices. Neither did the MTHFR polymorphism modulate the effects of pravastatin on coronary vasomotion. In conclusion, a 6-month therapy with pravastatin decreases Hcy concentration in Finnish healthy young men. The MTHFR genotype is neither a determinant of baseline coronary flow indices nor does it modulate the effect of pravastatin on coronary reactivity.

The myeloperoxidase enzyme (MPO) is a potent precursor of low-density lipoprotein (LDL) oxidation in atherosclerotic lesions. The MPO gene has a promoter polymorphism, 463G/A, which leads to high (GG) and low-expression (AG, AA) genotypes. Hormone replacement therapy (HRT) is known to affect MPO activity and LDL oxidation. The purpose of this study was to test whether the effect of HRT on the levels of oxLDL-ab varies according to MPO genotype.Eighty-seven postmenopausal women aged 45-71 years were divided into three groups based on the use of HRT. The HRT-EVP group (n = 25) used sequential estradiol valerate (EV) plus progestin, the HRT-EV group (n = 32) used EV alone, and the control group (n = 30) no HRT. MPO genotypes were determined by polymerase chain reaction (PCR) and oxLDL-ab by ELISA.We found a significant HRT group by MPO genotype interaction (p = 0.021) in plasma oxLDL-ab levels. In subjects with the GG genotype, the oxLDL-ab titer increased in the order of 2.13 in controls, 2.53 in the EV group and 3.21 in the EVP group (ANOVA for trend p = 0.006).The effects of HRT on LDL oxidation can vary according to MPO genotype and the concurrent progestin therapy with EV may counteract the more neutral effect of EV on LDL oxidation in subjects with the MPO high-expression genotype.

Background Oxidative modification of low-density lipoprotein (LDL) is a key event in the oxidation hypothesis of atherogenesis. Some in vitro experiments have previously suggested that high-density lipoprotein (HDL) co-incubated with LDL prevents Cu2+-induced oxidation of LDL, while some other studies have observed an opposite effect. To comprehensively clarify the role of HDL in this context, we isolated LDL, HDL2 and HDL3 from sera of 61 free-living individuals (33 women and 28 men). Results When the isolated LDL was subjected to Cu2+-induced oxidation, both HDL2 and HDL3 particles increased the rate of appearance and the final concentration of conjugated dienes similarly in both genders. Oxidation rate was positively associated with polyunsaturated fatty acid content of the lipoproteins in that it was positively related to the content of linoleate and negatively related to oleate. More saturated fats thus protected the lipoproteins from damage. Conclusion We conclude that in vitro HDL does not protect LDL from oxidation, but is in fact oxidized fastest of all lipoproteins due to its fatty acid composition, which is oxidation promoting.

To establish whether the relationship between interleukin-6 (IL-6) and plasma lipid and C-reactive protein (CRP) concentrations is different in Finnish nonagenarians than in middle-aged subjects with lower inflammatory status.Cross-sectional.Observational cohort study concentrating on the oldest old.Nonagenarians (n=291, mean age+/-standard deviation 90+/-1; 68 men, 223 women) who lived in the Tampere municipality in southern Finland and a middle-aged control population from the same area (n=227, aged 44+/-8).Plasma high sensitive CRP and lipid concentrations were analyzed using an automatic analyzer and IL-6 levels using enzyme-linked immunosorbent assay.Plasma concentrations of IL-6 (4.39+/-5.25 vs 1.88+/-1.98 pg/mL) and CRP (3.54+/-4.98 vs 1.53+/-1.91 mg/L) were significantly higher in nonagenarians than in middle-aged subjects (P.001). In nonagenarians, plasma CRP levels increased (P.001) and plasma total cholesterol (P=.006), low-density lipoprotein cholesterol (P=.02), and high-density lipoprotein cholesterol (P=.002) levels decreased according to IL-6 quartiles. In middle-aged subjects, similar associations were not found.The relationship between IL-6 and plasma CRP and cholesterol levels in nonagenarians with enhanced systemic inflammation differs from that of middle-aged subjects.

Hepatic lipase (HL) is a lipolytic enzyme that hydrolyzes triglycerides and phospholipids in almost all major classes of lipoproteins. The HL gene has a functional promoter polymorphism at position 480, which affects transcription and leads to CC, CT, and TT genotypes. We investigated the effect of long-term hormone replacement therapy (HRT) on the progression of atherosclerosis in a 5-yr follow-up observational study of 88 postmenopausal women with different HL genotypes (CC, n 49; CT, n 34; TT, n 5). These women, aged 45–71 yr, were divided into three groups based on the use of HRT. The HRT-EVP group (n 26) used sequential estradiol valerate (EV) plus progestin (levonorgestrel), the HRT-EV group used EV alone (n32), and the control group (n 30) used no HRT. The HRT-EV and HRT-EVP groups started estrogen at menopause for estrogendeficiency symptoms, whereas the control group took no estrogen due to either the absence of such symptoms or a dislike of estrogen therapy. In addition to serum lipid concentration and HL genotype, the atherosclerosis severity score (ASC) for the abdominal aorta and carotid arteries was determined by ultrasonography. There was a significant interaction between HRT therapy and HL genotypes on the increaseinASC(P0.046)afteradjustmentforage,bodymassindex, changes in high-density lipoprotein cholesterol and baseline ASC. In subjects with the T allele, the progression of ASC was significantly faster in the control group than the HRT group (P 0.0006), whereas in the CC genotype, there were no significant differences in ASC progression between the control and HRT groups. Our results suggest that the beneficial effect of HRT on atherosclerosis progression was restricted to women with the T allele, in whom the progression of ASC was slower by half. These results may help us understand in greater detail the benefits and possible risks associated with HRT in atherosclerotic diseases. (J Clin Endocrinol Metab 90: 3786–3792, 2005)

Background Polyunsaturated fatty acids (PUFA) are thought to play important roles in inflammation. The n-3 series is considered as anti-inflammatory, and some studies have reported increased plasma n-3 polyunsaturated fatty acid pattern in chronic inflammatory conditions. In this study we sought to clarify relationships of the levels of arachidonic acid and the polyunsaturated n-3 fatty acid compositions of isolated LDL, HDL2 and HDL3 particles with matrix metalloproteinase-9 (MMP-9), a marker of inflammation. Results The subjects were divided into two groups: those with lower and those with higher than the median serum MMP-9 concentration. In all lipoprotein fractions, the mean percentage of docosapentaenoic acid (C22:5n-3) was higher in the group of subjects with higher MMP-9 level than in those with lower serum MMP-9 concentration (P < 0.01 for all). Likewise, the ratio of docosapentaenoic acid to arachidonic acid (C20:4n-6) was higher in the subjects with higher MMP-9 compared with the lower MMP-9 group (P < 0.001 for all). Conclusion So far, the evidence for an anti-inflammatory role of the n-3 PUFA has come from dietary interventions. Our results were obtained from a free-living population and indicate that there is a positive correlation between n-3 docosapentaenoic acid and MMP-9. What had triggered the rise in MMP-9 is not known, since serum level of MMP-9 is raised in many inflammatory conditions. These findings may indicate an increased biosynthesis of n-3 polyunsaturated fatty acids in subclinical inflammation.

Antibody titer against malondialdehyde (MDA)-modified low-density lipoprotein (LDL) has been found to be associated with atherosclerosis, but it has not been established whether it would detect subjects with coronary artery disease (CAD). In the present study, receiver-operating characteristic (ROC) analysis was used to compare the diagnostic accuracy of the antibody titer against MDA-modified LDL and high-density lipoprotein (HDL) and LDL cholesterol levels in discrimination between subjects with (n=51) and without (n=35) angiographically verified 3-vessel CAD. As a result, the antibody titer against MDA-modified LDL was lower in subjects with CAD compared with subjects without CAD (p

Myeloperoxidase (MPO) is an oxidative enzyme present in phagocytes and atherosclerotic lesions. The MPO gene has a promoter polymorphism -463G/A, which leads to high (GG) and low expression (AG, AA) genotypes. We investigated the effect of long-term hormone replacement therapy (HRT) on the progression of atherosclerosis in a 5-yr follow-up study of postmenopausal women with different MPO genotypes. Eighty-seven nonsmoking postmenopausal women, aged 45-71 yr, were divided into three groups based on the use of HRT. The HRT-EVP group (n = 25) used sequential estradiol valerate plus progestin, the HRT-EV group used estradiol valerate alone (n = 32), and the control group (n = 30) used no HRT. The atherosclerosis severity score (ASC) for abdominal aorta and carotid arteries was determined by ultrasonography, and the MPO genotype was analyzed. In subjects with the GG genotype, the progression of ASC was significantly faster in the control group than in the HRT group (genotype by time interaction, P = 0.042), whereas in A allele carriers there were no significant differences in ASC progression between control and HRT. The effects of HRT on atherosclerosis progression in subjects with the GG genotype seem to be especially beneficial compared with controls with the same genotype but without HRT. These results may help us understand in greater detail the benefit and possible risk of HRT in atherosclerotic diseases.

Apolipoprotein (apo)E gene epsilon4 allele carrier status modulates the responses of lipoprotein metabolism to hormone-replacement therapy (HRT). We investigated the effect of long-term HRT on the progression of atherosclerosis in postmenopausal women with or without apoE epsilon4 allele. One hundred forty-one nonsmoking postmenopausal women, 45-71 yr old, were divided into 3 groups based on the use of HRT. The HRT-EVP group (n = 61) used sequential estradiol valerate (EV) plus progestin (P), the HRT-EV group used EV alone (n = 40), and a control group had no HRT. Of these 141 women, 93 participated in a 5-yr follow-up study in 1998. In addition to serum lipid concentration and apoE genotype, the atherosclerosis severity score of the abdominal aorta and carotid arteries was determined by sonography. In apoE4-negative subjects, the progression of atherosclerosis severity score was significantly faster in control than in the HRT groups (genotype-by-time interaction P = 0.0026); whereas in apoE4-positive subjects, there were no significant differences in atherosclerosis severity score progression between the control and HRT groups. The effects of HRT on atherosclerosis progression in subjects with no apo epsilon4-allele seems to be especially beneficial, compared with controls with same phenotype status but without HRT. These results may help us to understand, in more detail, the benefit and possible risk of HRT on atherosclerotic diseases.

This study examined the relationships between paraoxonase genotypes, coronary artery reactivity, and indices of low-density lipoprotein oxidation in healthy men. Impairment in coronary flow reserve, as assessed by positron emission tomography, is associated with lipoprotein oxidation, which is affected by high-density lipoprotein bound enzyme, paraoxonase. Paraoxonase has two common polymorphisms (M/L55 and R/Q192) that change the activity of the enzyme. Forty-nine healthy men (mean age 35 +/- 4 years) were divided by paraoxonase genotype into low (Q192/Q192, or M55/M55, M55/L55) and high-active (R192/Q192, R192/R192, or L55/L55) groups and related to the myocardial blood flow, to the susceptibility of low-density lipoprotein to oxidation, and the autoantibody titer against oxidized low-density lipoprotein. The blood flow was measured by positron emission tomography at rest and during adenosine infusion. The low-active Q192/Q192 genotype was associated with higher resting blood flow corrected for rate-pressure product compared to the high-active R192/R192 and R192/Q192 genotypes (P=0.011). The blood flow stimulated by adenosine was not significantly different in the low- and high-active genotype groups. Paraoxonase genotypes had no effect on low-density lipoprotein susceptibility to oxidation or autoantibody formation against oxidized low-density lipoprotein. Genotypes of paraoxonase may not clearly contribute to the early changes in coronary reactivity. Coronary vasomotor tone at rest appears to be modulated by paraoxonase R/Q192 polymorphism through mechanism(s) unrelated to low-density lipoprotein oxidation.

We describe an HPLC method for quantifying creatinine, separating the analyte from other compounds in serum and urine by cation-exchange chromatography and measuring its absorbance at 234 nm. The precision of the method (CV) varied from 2.9% (mean creatinine concentration, 31 mumol/L) to 1.7% (361 mumol/L) within a series of assays and from 3.9% (34 mumol/L) to 2.4% (391 mumol/L) between series. A comparison with the Jaffé method, as performed with a Technicon SMA analyzer, gave the regression line yHPLC = 1.00xJaffé - 12.0 (n = 141, r = 0.998, and Syx = 19). Results also are comparable with those of an enzymatic method, if the enzymatic method is standardized with a serum-based standard when serum samples are measured. An aqueous standard has to be used for enzymatic determination of creatinine in urine.

Apolipoprotein A-IV (apoA-IV) is a glycoprotein constituent of triglyceride-rich and high-density lipoproteins (HDL) and may thus play an important role in lipid metabolism. In Finland two common isoforms (A-IV-1 and A-IV-2) of apoA-IV have been found. The isoforms are the result of the G to T substitution in the third base of the codon 360 in the apoA-IV-2 allele of the apoA-IV gene. The purpose of the study was to determine the apoA-IV allele frequencies in the Saami and the Finns, and to relate the apoA-IV phenotypes to serum lipids. The sample was drawn in connection with a Reindeer Herders' Health Survey performed in northern Finland in 1989. The study group included 248 men with known ethnic origin, Saami and Finns, who lived in the area of the nine northernmost municipalities of Finland. ApoA-IV phenotypes from 71 Saami (both parents Saami) and 177 Finns (both parents Finns) were determined by isoelectric focusing and Western blotting. Serum lipids were determined enzymatically. ApoA-IV allele frequencies in the Saami and the Finns were for A-IV-1 0.894 vs 0.944 and for A-IV-2 0.106 vs 0.056, respectively (chi2-test, P0.05). The effect of the apoA-IV phenotype on serum HDL-cholesterol levels differed significantly between the Saami and the Finns (two-way ANCOVA, interaction between ethnicity and apoA-IV phenotype, P0.02). In the Saami, HDL-cholesterol levels were significantly higher in the apoA-IV-2/1 than in the apoA-IV-1/1 phenotypes (ANCOVA, P0.05). Mean total cholesterol, low-density lipoprotein (LDL)-cholesterol, apolipoprotein B, HDL-cholesterol and triglyceride levels did not differ statistically significantly between the Saami and the Finns. Yet, there was a trend in the Saami of having higher mean total cholesterol, LDL-cholesterol and apolipoprotein B levels than the Finns among the apoA-IV-2/1 phenotypes, while there was only a small difference in these parameters between the Saami and the Finns among the apoA-IV-1/1 phenotypes. In conclusion, the Saami have a higher frequency of the apoA-IV-2 allele than the Finns and most of the other studied populations.

Objectives : The aim of the study was to obtain information on the possible relationship between impaired ovarian function and risk factors for cardiovascular disease. Material and methods : Serum lipid levels, plasma fibrinolytic potential and histological and biochemical changes in the intima of the uterine artery were investigated in premenopausal women with irregular menstrual cycles, and the results were compared with those from regularly menstruating women. In addition, the same parameters were studied in postmenopausal women on hormone replacement therapy (HRT) and in postmenopausal women who had never used HRT. In total 64 patients undergoing hysterectomy for benign reasons were included the study. Results : Plasma fibrinogen concentration was significantly higher in irregularly menstruating women as compared with women with regular cycles. In women with irregular cycles thickened or sclerotic arterial intima was a significantly more common finding as compared with regularly menstruating women. A significant positive correlation was observed between plasma fibrinogen concentration and intimal esterified cholesterol content in women with thickened or sclerotic uterine artery. Conclusions : These data suggest an important role for normal ovarian function in the prevention of atherosclerosis.

137 Russians living in Estonia was screened by isoelectric focusing and immunoblotting procedures to determine the distribution of genetic variations in apolipoprotein E (apoE) and apolipoprotein A-IV (apoA-IV) genes. The apoA-IV-2 allele and epsilon4 allele frequency of the Russians tended to be lower than in most other European populations.

The incidence of atherosclerotic plaques in the aorta and in the carotid and iliac arteries was investigated using high resolution B-mode ultrasonography in 40 postmenopausal women using sequentially combined oestradiol valerate 2 mg--levonorgestrel 0.25 mg replacement therapy. In addition, serum lipid, lipoprotein and apolipoprotein levels were measured. Similar studies were performed for 40 postmenopausal women using oestradiol valerate alone and for 40 women who had never used hormone replacement therapy (HRT). Ultrasonographically observed atherosclerotic plaques were less common (P = 0.011) in oestradiol valerate--levonorgestrel users than in women without HRT. More than three atherosclerotic plaques were observed in 62% of the women without HRT but only in 30% of the women using combined therapy (P = 0.003). The total number of plaques did not differ between the two hormone using groups. The mean serum triglyceride, total cholesterol and apolipoprotein B levels were lowest in oestradiol valerate--levonorgestrel users when compared with those of women with oestradiol valerate monotherapy and those of women without HRT. Combined oestradiol valerate--levonorgestrel replacement therapy, like oestradiol valerate monotherapy, prevents the progression of atherosclerosis in postmenopausal women. The lipid patterns were not adversely affected by the addition of the androgenic levonorgestrel.

Open studies in hypercholesterolaemia with guar gum, a dietary fibre obtained from Cyamopsis tetragonolobus, have shown 10% to 15% reductions in serum concentration of total cholesterol and 10% to 20% in serum concentration of low density lipoprotein cholesterol after short term treatment. The results from long term studies have been conflicting. To assess long term efficacy of guar gum we treated 40 patients with moderate hypercholesterolaemia for 24 months. This randomised double blind, placebo controlled trial with two parallel groups was approved by the university's ethics committee, and subjects' consent was obtained. Between August 1987 and November 1989, 40 consecutive patients without diabetes were operated on for a haemodynamically measurable carotid stenosis. Both groups comprised 16 men and four women, whose mean age was 63 years (range 49 to 81) and mean body weight 75.8 kg (55.0 to 97.0). There were no group differences in age, sex, or body weight. …

Epidemiological studies have consistently shown an apparent protective effect of moderate alcohol consumption on coronary artery disease (CAD). This has been considered to be due to the rise in the high-density cholesterol lipoprotein (HDL-cholesterol). Since the response of the HDL-subfractions to moderate or heavy dose of alcohol is less clear, we now compared the high-density lipoprotein cholesterol status between groups consuming different amounts of alcohol. In this population-based survey serum total high-density lipoprotein cholesterol and its HDL2 and HDL3 subfractions were blindly compared between 264 consecutive middle-aged men (37 teetotallers, 137 moderate drinkers, 90 heavy drinkers) participating in a voluntary health screening and 104 male alcoholics. Alcohol consumption correlated significantly (P

The traditional method for DNA separation is slab gel electrophoresis. Because slab gel electrophoresis, which involves the casting, loading, running, and staining of the gel is labor-intensive and time-consuming, high-resolution capillary electrophoresis (CE) has become an attractive alternative. CE, by contrast to conventional electrophoretic techniques, is capable of rapid, automated, reproducible, and high-resolution separation of minute amounts of DNA samples. To separate DNA fragments by CE, polyacrylamide gels or liquid buffers containing soluble polymers are used. Soluble polymers, such as hydroxyethyl cellulose, hydroxypropylmethyl cellulose, and methyl cellulose act as effective molecular sieves and allow for separation of DNA according to size (1). CE has recently shown promising results for the analysis of double-stranded DNA such as restriction fragment length polymorphisms and polymerase chain reaction (PCR) products (2)(3). The insertion (I)/deletion (D) of a 287-bp sequence polymorphism within intron 16 of the gene for angiotensin-converting enzyme (ACE) is strongly associated with serum ACE concentrations (4). The D allele has been identified as a risk factor for the development of coronary heart disease and myocardial infarction (5). In addition, the deletion polymorphism has been associated with either microalbuminuria or overt nephropathy in diabetic patients (6)(7). Here we report a sensitive, simple, rapid, and nonisotopic procedure for identification of …

The paraoxonase enzyme (PON) gene polymorphism causes a change of methionine (M-allele) to leucine (L-allele). PON may reduce low density lipoprotein oxidation and prevent atherosclerosis. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive index of oxidative DNA damage. We have studied the association between the PON genotypes and the urinary excretion of 8-OHdG. The study population consisted of 93 Finnish type 2 diabetes patients and 106 non-diabetic control subjects. The 24-h excretion of 8-OHdG was significantly higher in diabetic patients than in control subjects (P < 0.001). In control subjects, the ratio of the 8-OHdG/glomerular filtration rate increased in order of genotype from MM to ML to LL (P < 0.0412). These results suggest that lipid peroxidation may have an effect on DNA oxidation.

Serum lipids were measured in 30 Finnish and Japanese postmenopausal women. Total cholesterol, HDL cholesterol, LDL cholesterol, apo B and the HDL cholesterol/apo A1 ratio were higher in Finnish than in Japanese women. The LDL cholesterol/apo B and apo A1/apo B ratios were lower in Finns than in Japanese. In serum phospholipids the percentage of arachidonic acid was higher and the percentage of the n - 3 fatty acids, eicosapentaenoic acid and docosahexaenoic acid, and the eicosapentaenoic acid/arachidonic acid ratio were lower in Finnish than in Japanese women. No significant correlations were found between serum oestrone levels and lipid parameters.

Serum albumin was assayed with bromcresol purple using routine clinical chemical photometric analysers. Bromcresol purple and bromcresol green dyes were compared, and albumin assays using these dyes were compared to those made by radial immunodiffusion and rocket immunoelectrophoresis. Bromcresol purple seems to be insensitive to all serum proteins except albumin, and this dye is correlated linearly with radial immunodiffusion and with rocket electrophoresis.

The material consisted of 32 patients, 12 of whom were selected for control. All were cases approaching the menopause and had been subject to oophorectomia bilateralis and hysterectomia for myomas. All 20 investigation patients received estradiol valerianate orally 4 mg/day for 6 mth. During the course the serum estradiol level increased 5–6-fold from the postmenopausal levels. No significant changes occurred in the serum triglycerides and total cholesterol, whereas in the control group the triglycerides decreased. The proportional concentration of β-lipoproteins decreased. There was also a slight decrease in the pre-β-lipoproteins. The proportional concentration of β-lipoproteins increased, as also serum high density lipoprotein (HDL) cholesterol. In the liver enzymes no changes occurred. Blood pressure increased in one patient. There were no other complications.

The quality control of insulin radioreceptor assay for human erythrocytes is based on the storage of erythrocyte preparations in Hepes buffer of pH 8.0, containing 10 g/l of albumin and 20 mmol/l of glucose. The change of erythrocytes into spherocytes and crenated cells reduces the apparent number of insulin receptors in a relatively constant way by less than 8% a week after 10 days of storage. At the same time the dissociation constants of the insulin-receptor complex increase rapidly. Thus the use of a preparation must be limited to controlling the determination of the insulin binding sites of erythrocytes, and not to the measurement of the affinities of the receptors. When mono-125I-Tyr-A14-insulin gets old, a slow decrease in the insulin binding sites can be measured, but the dissociation constants of the insulin receptor complex are not affected.

Citrate is a normal constituent of urine which combines with calcium to form a soluble salt. Urinary citrate excretion was examined in patients with urolithiasis and normal subjects by a specific enzymatic technique. There was a considerable overlap in the urinary citrate excretion between normal subjects and stone-formers, but the citrate-creatinine ratio, the citrate-calcium ratio and the citrate-magnesium-calcium ratio, which were all highly significantly lower (p less than 0.001) in stone-formers than in controls, proved most reliable in discriminating between these groups.

The series comprises 70 women with climacteric symptoms; 14 of them were oophorectomized. The first group of patients received 4 mg of oestriol succinate per day or a placebo in a double-blind experiment. In the second part of the study oophorectomized women were given 8 mg of oestriol succinate per day or 16 mg/day divided into two doses. Serum total cholesterol, HDL-cholesterol and serum triglycerides were estimated before drug treatment and after drug administration for three and six months. Administration of 4–16 mg/day of oestriol succinate was without effect on serum total cholesterol. A significant increase of 25–30% in HDL-cholesterol was observed following administration of oestriol succinate (8 mg or 16 mg divided into two doses) with a rising tendency in total triglyceride value.