Adhesion and migration of leukocytes into the surrounding tissue are crucial steps in inflammation, immunity and atherogenesis. Expression of cell adhesion molecules by endothelial cells plays a role in these processes. Propionate is a naturally occurring short chain fatty acid produced by bacterial fermentation of dietary fibre. High intake of dietary fibre has been associated with an improved bowel function and with a reduced risk of cardiovascular disease. However, the molecular mechanisms responsible for these effects remain unknown. In this study, the effects of propionate on the expression of endothelial leukocyte adhesion molecules by cytokine-stimulated human umbilical vein endothelial cells (HUVEC) were investigated. Pretreatment of HUVEC with propionate significantly inhibited the tumor necrosis factor alpha (TNF-alpha)-induced expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) in a time- and dose-dependent manner. At 10 mM, propionate also inhibited the interleukin-1 (IL- 1)-mediated VCAM-1 and ICAM-1 expression, with the latter effect being more pronounced, as well as decreased the TNF-alpha-induced VCAM-1 and ICAM-1 mRNA expression in a similar manner. The decrease in VCAM-1 and ICAM-1 expression was associated with a reduction of adherence of monocytes and lymphocytes to the cytokine-stimulated HUVEC. In addition, propionate significantly inhibited the TNF-alpha-induced activation of nuclear factor-kappa B (NF-kappaB) and significantly increased the expression of peroxisome proliferator-activated receptor alpha (PPARalpha) in HUVEC. These results demonstrate that propionate may have antiinflammatory and possibly antiatherogenic properties. Our findings warrant further investigation into the therapeutic effects of propionate on a number of pathological events nvolving leukocyte recruitment.

Superoxide anion is produced in human platelets predominantly by Nox2-dependent NADPH oxidases. In vitro experiments have shown that it might play a role in modulating platelet functions. The relationship between platelet superoxide production and aggregation remains poorly defined. Accordingly, we aimed to study superoxide production and aggregation in platelets from subjects with significant cardiovascular risk factors (hypertension, hypercholesterolemia, smoking and diabetes mellitus) and from control individuals. Moreover, we studied the effects of novel polyphenol-rich extracts of Aronia melanocarpa (chokeberry) berries on platelet function in vitro. Superoxide production was significantly increased in patients with cardiovascular risk profile when compared to controls, while platelet aggregation in response to either collagen or thrombin were borderline higher, and did not reach statistical significance. Interestingly, no relationship was observed between platelet aggregation ex vivo and platelet superoxide production in either of studied groups. No correlation was found between endothelial function (measured by FMD) and platelet aggregation ex vivo either. Polyphenol-rich extracts of A. melanocarpa berries caused a significant concentration dependent decrease in superoxide production only in patients with cardiovascular risk factors, while no effect was observed in the control group. A. melanocarpa extracts abolished the difference in superoxide production between risk factor patients and controls. A. melanocarpa extracts exerted significant concentration dependent anti-aggregatory effects in both studied groups, which indicated that these effects may be independent of it's ability to modulate superoxide production. The anti-aggregatory effects of chokeberry extracts were similar irrespective of aggregation inducing agent (collagen or thrombin). Moreover, they appear to be independent of platelet NO release as NOS inhibition by L-NAME did not lead to their abrogation.

The reverse transcription polymerase chain reaction (RT-PCR) is one of the most useful molecular biology methods in opening the way to understanding of the mechanisms of atherosclerosis on the gene structure and/or expression level. We optimized this technique for assaying expression of the monocyte chemotactic protein type 1 (MCP-1) gene in rabbit aorta with respect to the temperature profile, yield to cycle number, interference of genomic DNA with the RNA matrix, and repeatability. Variability of expression of the constitutive GAPDH gene was also examined. The study was done in 18 New Zealand rabbits allocated to two groups and fed a standard chow for 2 (S1) or 3 (S2) months. The experiment ended with removal of part of the ascending rabbit aorta, from which RNA was isolated. The optimal temperature for binding of specific primers to the MCP-1 and GAPDH genes was 63 degrees C, and the optimal number of cycles for PCR amplification was 22 for MCP-1 and 26 for GAPDH. The GAPDH amplicon size was 465 base pairs in the presence or absence of reverse transcriptase showing contamination of the RNA matrix with genomic DNA. Repeatability of the RT-PCR method was 8.7%, and variability of expression of the GAPDH gene was 7.7%. Thus, RT-PCR adjusted for contaminating genomic DNA provides a reliable way of assaying expression of the MCP-1 gene in rabbit aorta.

The present study was designed to investigate the hypothesis that trans fatty acids can induce apoptosis of human umbilical vein endothelial cells (HUVEC). To test this hypothesis apoptosis was measured in HUVEC treated with 0.1, 1.0 or 5.0 mM trans elaidic acid (t-18:1) or linoelaidic acid (t,t-18:2) for 24 hours. For the detection of apoptosis, TdT-mediated dUTP nick end labelling assay (TUNEL), cell binding of annexin V and propidium iodide uptake were measured. Active Caspase-3 and cleaved PARP (poly-ADP-ribose polymerase) were also measured in the cell lysate. Moreover, cellular ability to produce ROS (reactive oxygen species) was measured by DCF fluorescence Both acids studied induce both early (annexin-positive cells) and late stages of apoptosis (cells stained by propidium iodide) in a dose-dependent manner. Also the appearance of TUNEL-positive cells was induced by both trans fatty acids tested, in a dose dependent manner. Both trans acids induce apoptosis through their effect on Caspase-3 activity and on intracellular ROS production. It is worth emphasising that linoelaidic acid proved to be a more potent inducer of apoptosis and ROS production in endothelial cells than elaidic acid. The present studies suggest that trans fatty acids may play a role in damaging and death of vascular endothelial cells in atherosclerosis.

The basic parapharmaceuticals in the Polish diet include natural anti-oxidants - bioflavonoids found in berry fruit. They were proven to have the ability to regulate genetic transcription and increase the synthesis of nitric oxide which counteracts dysfunction of the vascular endothelium. They also display anti-oxidant action through the inhibiting effect on cyclooxygenase - COX-2, and increase the level of adiponectin. We have also more and more proof of the important biological role of short-chained fatty acids formed as a result of fermentation of fibre by probiotic bacteria. Through their effects on peroxisome proliferators activated receptors (PPAR), butyric and propionic acids may reduce the expression of adhesion molecules and exert anti-inflammatory action both in the gastrointestinal tract as well as systemically.

BACKGROUND AND AIM: An imbalance in the hemostatic system is a frequent finding in untreated essential hypertension (HT), and it has been shown that treatment with angiotensin converting entyme (ACE) inhibitors improves hemostatic function. In order to elucidate the role of genetic factors, we studied hemostasis in patients with untreated and treated HT and correlated the results with ACE I/D and plasminogen activator enhibitor-1 (PAI-1) 4G/5G gene polymorphisms. METHODS AND RESULTS: Forty-three males with HT (mean age 31.7 +/- 6.8 years) were compared with 34 age and gender-matched controls. All of the patients were treated with perindopril (4 mg/day) and, after one and six months of therapy, their levels of plasma fibrinogen (Fb), t-PA antigen, PAI-1 antigen, von Willebrand factor (vWF), ACE activity and blood pressure were measured. ACE and PAI-1 genotypes were identified by means of the polymerase chain reaction on DNA isolated from peripheral blood lymphocytes. Untreated patients had significantly higher levels of Fb, PAI-1 (p < 0.01) and t-PA (p < 0.05) regardless of their ACE or PAI-1 genotypes. Perindopril reduced blood pressure regardless of ACE or PAI-1 genotype (p < 0.001). ACE II homozygotes showed the greatest decrease in ACE activity (p < 0.01) and a significant reduction in Fb levels (p < 0.05) after just one month of treatment. Analysis of the group as a whole revealed an increase in t-PA antigen levels after six months of treatment, regardless of ACE or PAI-1 genotype (p < 0.01). CONCLUSIONS: Our results show that essential hypertension predisposes to the procoagulant state characterized by hyperfibrinogenemia and hypofibrinolysis. Perindopril reduced fibrinogen levels in ACE II homozygotes due to its more potent inhibitory action on the renin-angiotensin system in such patients. It improved fibrinolysis by increasing t-PA levels regardless of ACE and PAI-1 genotype.

Adhesion and transendothelial migration of leukocytes into the vascular wall is a crucial step in atherogenesis. Expression of cell adhesion molecules by endothelial cells plays a leading role in this process. We investigated the effect of simvastatin, an inhibitor of HMG-CoA reductase administered to reduce plasma levels of LDL-cholesterol, on the expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1) by human umbilical vein endothelial cells (HUVEC) stimulated with tumor necrosis factor alpha (TNFalpha). We found the expression to be significantly inhibited by the drug in a time and concentration-dependent manner and to a greater extent in the case of VCAM-1 as compared with ICAM-1. In TNFalpha-stimulated HUVEC, simvastatin decreased VCAM-1 and ICAM-1 mRNA levels, inhibited TNFalpha-induced activation of nuclear factor kappaB (NF-kappaB) and enhanced expression of peroxisome proliferator-activated receptor alpha (PPARalpha). These effects were associated with reduction of adherence of monocytes and lymphocytes to HUVEC. The present findings suggest that the benefits of statins in vascular disease may include the inhibition of expression of VCAM-1 and ICAM-1 through effects on NF-kappaB.

BACKGROUND AND AIM: Von Willebrand factor (vWF) and fibrinogen (Fb) have recently emerged as plausible familial determinants of atherothrombosis. We investigated whether the vWF and Fb levels in patients with ischemic cerebrovascular stroke (ICS) correlate with those in their children. METHODS AND RESULTS: The study group consisted of 28 families (56 parents and 34 children) with one parent who had suffered an ICS at least three months before the study. All of the ICS patients had hyperlipoproteinemia and most arterial hypertension. The control group consisted of 15 families (30 parents and 20 children). The age of the parents and children did not exceed 55 and 16 years. The ICS parents had significantly higher vWF, Fb and protein C (PC) levels than the controls (vWF--fathers: 121.0 +/- 42.5% vs 79.2 +/- 23.4%; vWF--mothers: 110.7 +/- 40.1% vs 82.4 +/- 20.9%; Fb--fathers: 4.12 +/- 0.74 g/L vs 3.01 +/- 0.54 g/L; Fb--mothers: 3.64 +/- 0.84 gL vs 2.98 +/- 0.35 g/L; PC--fathers: 116.0 +/- 12.3% vs 105.6 +/- 13.7%; PC--mothers: 114.4 +/- 15.8% vs 105.0 +/- 12.2%). The children of the ICS parents had significantly higher PC and body mass index (BMI) values than the controls (PC: 102.6 +/- 13.7% vs 92.7 +/- 10.7%; BMI: 20.6 +/- 3.8 vs 17.8 +/- 3.5 Kg/m2), as well as an atherogenic lipid profile, higher blood pressure (BP) and a tendency toward higher vWF levels. Correlations between the ICS parents and their children were found for vWF, factor VIIc, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and BP, which were closer in the case of fathers. CONCLUSION: Regardless of gender, the parents with a history of ICS had a procoagulant state, with high levels of vWF, Fb and PC. In terms of inheritance, the most adverse risk factor profile was found in the children of ICS fathers.

BACKGROUND AND AIM: High plasma lipoprotein(a)[Lp(a)] and homocysteine (HCY) levels are now considered to be independent risk factors for cerebro- and cardiovascular atherosclerotic occlusive disease, but little is known about the influence of Lp(a) and HCY on the early events of ischemic disease or their significance in subjects with a positive family history of ischemia. The aim of this study was to evaluate the relationship between HCY levels and the severity of ischemic cerebral stroke, and investigate whether there was a correlation between Lp(a) and HCY levels in the stroke patients and their children. METHODS: The study involved 35 patients with early ischemic cerebral stroke aged 46.1 +/- 6.6 years and their 50 children aged 17.2 +/- 5.5 years. The patients were grouped on the basis of the form of the stroke (transient, progressive or complete stroke), and their levels of Lp(a), HCY, uric acid (UA), fibrinogen (Fb) and factor VII (FVII) activity were measured. RESULTS: HCY and Lp(a) concentrations increased with the severity of the ischemia, being highest in the patients with complete stroke (15.1 +/- 2.9 mumol/L and 32.9 +/- 37.6 mg/dL respectively). A similar trend was found in the offspring, with the highest HCY and Lp(a) values in the children of complete stroke patients (12.6 +/- 4.4 mumol/L and 23.0 +/- 24.6 mg/dL). The control values were respectively 8.7 +/- 1.6 mumol/L and 5.35 +/- 7.05 mg/dL. The following correlations between the parents and children were noted: Lp(a)(r = 0.87 p < 0.0001), UA (r = 0.71 p < 0.001), HCY (r = 0.45 p < 0.05), FVII (r = 0.45 p < 0.05), and Fb (r = 0.42 p = 0.06). Correlations between Lp(a) and HCY (r = 0.47 p < 0.05) and Fb and FVII (r = 0.60 p < 0.01) were found in the children. Multiple regression analysis revealed that only Lp(a) and Fb significantly influenced HCY levels in the offspring with a positive family history. CONCLUSIONS: HCY levels correlate with the severity of ischemic cerebral stroke and, in families with a history of ischemic cerebral stroke, the levels of the risk factors in children are determined by the levels in their parents.

Increasing evidence suggests the role of hemostatic risk factors in the development of ischemic heart disease (IHD). A raised plasma fibrinogen has been related to increased risk of IHD. The aim of the study was to determine the relationship between plasma fibrinogen and the coronary vessels state based on the coronary angiogram. 119 patients undergoing coronary angiography were classified into 5 groups according the severity of IHD: Group 0 without significant atherosclerotic lesions (control group), Group 1 with single vessel disease, Groups 2, 3 with multivessel disease (two and three affected arteries, respectively) and Group 4 with positive history of myocardial infarction. A statistically nonsignificant rise in fibrinogen levels in Groups 1, 2, 3 (3.9 +/- 0.8 g/l, 4.0 +/- 0.9 g/l, 4.1 +/- 0.9 g/l, respectively) as compared to control Group 0 (3.7 +/- 0.7 g/l) was found. In Group 4 plasma fibrinogen was significantly lower (2.8 +/- 0.6 g/l) comparing to Group 0 (p < 0.05). In addition plasma fibrinogen was positively correlated with blood pressure. These results supports the role of raised plasma fibrinogen in the pathogenesis and development of IHD.

"Functional foods" supplemented with plant sterols are advertised and added to regular meals to reduce serum cholesterol concentrations. The effects of increased phytosterol levels on cardiovascular diseases, however, are not known. Findings in patients with sitosterolemia, data from epidemiological studies, and experimental data from animal studies suggest that plant sterols may potentially exert negative cardiovascular effects. Additional studies investigating relevant clinical endpoints are needed before a diet supplemented with plant sterols can be recommended in the prevention of cardiovascular diseases.

Increased serum total and LDL-cholesterol concentrations are major risk factors for cardiovascular diseases. Many clinical trials have proven that plant sterol and stanol esters can effectively decrease high serum total and LDL cholesterol. They reduce the intestinal absorption of cholesterol by decreasing the incorporation of dietary and biliary cholesterol into micelles displacing cholesterol from these micelles. They also increase LDL receptor activity on liver cells causing a higher uptake of LDL cholesterol and thus decreasing the serum LDL cholesterol concentration. Animal studies have indicated that plant sterols and stanols may also lower atherosclerotic lesions development. However, the evidence from human studies to confirm this is still lacking. Anyhow, plant sterol and stanol esters can be considered as an effective and safe cholesterol-lowering functional food ingredient. To achieve additional effects they can be combined with statin therapy, and this combination is also well tolerated and safe.

The review deals with results of recent studies on biological activity of C29- and C28-sterols of plant origin (phytosterols) in mammals and in cultured mammalian cells. The review considers the following problems: phytosterols and nutrition; phytosterols and cholesterol level; phytosterols and intestinal absorption of lipids; the role of phytosterols in lipid metabolism regulation; phytosterols and mammalian cells in culture; products of phytosterols oxydation; phytoecdysteroids and induced gene expression.