Atherosclerosis is being nowadays defined as chronic subclinical inflammatory disease. Recently published clinical and laboratory studies have shown that subclinical inflammation represents main role in initiation of creation, in progress and destabilization of atherosclerotic plaque. Screening including traditional cardiovascular risk factors fails in identification in more than 50% of individuals with later development of acute coronary syndrome. According to above mentioned reason indicators are being searched for, which would be usable to monitor the activity of atherosclerotic process. According to role of subclinical inflammatory process in pathogenesis of atherosclerosis, the determination of C-reactive protein using ultrasensitive method is being showed as perspective marker. Ultrasensitive C-reactive protein represents a strong, independent predictor of future cardiovascular events in apparently heal-thy individuals and has also prognostic utility in patients with acute coronary syndromes. Predictive capacity of C-reactive protein determination is independent of traditional risk factors and offers prognostic advantage as opposed to determination of lipids alone. The paper provides a review of currently available knowledge of possibilities for utilization of C-reactive protein laboratory assessment, as the main representative of acute phase proteins, in monitoring of creation and severity of coronary atherosclerosis, in possibilities of the disease prognosis determination and prediction of its acute complications, and also in prediction of prognosis in patient with already existing acute complication.

O219 A discussion is ongoing about an optimal method of glomerular filtration rate (GFR) estimation after kidney transplantation. Creatinine clearance (CrCl) as a measure of GFR is compromised not only outside the normal GFR range, but also by inaccurate urinary collection and by inaccuracy in creatinine measurement. Accepted biochemical difference ± 10% connotes that patients with creatinine clearance 60 ml/min fall in fact in range 53-67 ml/min, additionally, some laboratories are working with a difference that is much higher. Aim: to compare measured CrCl and predicted GFR (Cocroft- Gault) with Nankivell formula as a reference modality. Patients and methods: 120 single measurement in 20 patients divided in two groups: 3-6 months and more than 1 year after Tx. Bland-Altman difference plot was used for statistical analysis. Results: Variability of differences between methods was not constant across the range of measurement. During the period 3-6 months after Tx: measured CrCl was lower than values predicted by Nankivell form in 78% of cases (mean difference -0,18 ml/s, SD 0,23). Cocroft form overestimated the Nankivell GFR by 0,07 ml/s mean, SD 0,13, however a positive bias 0,17 ml/s, SD 0,07 was present in 50%, negative bias -0,06 ml/s, SD 0,09 in 33% of patiens. Different results were seen after 1 year. With GFR up to 1 ml/s, difference between measured CrCl and Nankivell GFR was negligible, however with GFR more than 1 ml/s the measured CrCl values were significantly higher than the Nankivell estimated GFR values (0,17 ml/s mean, SD 0,24). Cocroft form overestimated Nankivell GFR in 0,11 ml/s mean, SD 0,14 in full range of GFR. Conclusion: Measured CrCl as well as predicted GFR Cocroft-Gault and Nankivell formula can not be used interchangeably for longitudinal follow up of kidney transplant recipients. Another parameters, less charged by biochemical determination, sampling or calculation errors would be an advantage.

Because genetic predisposition to atherothrombosis in systemic lupus erythematosus (SLE) remains to be determined, the most common genetic prothrombotic factors, prothrombin G20210A and factor V Leiden mutations, were studied. Seventy-four SLE patients with vascular ischemia (SLE cases) were studied and stratified into myocardial infarction and/or cerebrovascular accident subgroup (MI/CVA), and coronary heart disease subgroup without overt arterial thrombotic events (CHD). Seventy-one SLE patients without atherothrombosis were investigated as SLE controls. Factor V Leiden was detected in six cases (five in MI/CVA, one in CHD group) and three controls (OR 2.00, 95%CI 0.48-8.32). Two cases (both CHD patients) had prothrombin G20210A mutation vs. three controls (OR 0.63, 95%CI 0.1-3.88). Anticardiolipin antibodies (aCL) were increased in cases vs. controls (39/74 vs. 27/71); however, this was not statistically significant (OR 1.82, 95%CI 0.94-3.52). Neither univariate nor multivariate analysis indicated that investigated mutations are risk factors for atherothrombosis in SLE cases, MI/CVA, or CHD subgroups. Overall, disease activity was the strongest risk factor for atherothrombosis (p=0.0014) in SLE cases. Combination of disease activity+gender was the best predictor of atherothrombotic process (p=0.00045) in this cohort. In MI/CVA subgroup, disease activity was the only predictor (p=0.0058). In CHD patients, the best predictive value was conferred by combination of hypertension+gender+disease activity (p=0.00077). No other investigated risk factor (including aCL) conferred an increased risk individually or potentiated the other risk factors. The results deny the role of investigated mutations in atherothrombosis in SLE, but they underscore the importance of disease activity (i.e., ongoing inflammation) in pathogenesis of atherosclerosis and arterial thrombosis.

The resistance to activated protein C (APC-resistance) based on the presence of factor V Leiden (F V Leiden) is the most frequent thrombophilic condition in the white race population. It contributes to the origin of thrombosis especially in the venous part of blood vessels. Significant geographic differences have been detected within Europe. The aim of this retrospective study was to determine the frequency in the occurrence of F V Leiden: 1. in healthy (asymptomatic) Slovak population, 2. in their consanguineously unrelated members with thrombosis and 3. in patients with myocardial infarction (IM) without or with other known risk factors of this disease (nicotinism, obesity, hypertension, dyslipoproteinemia, diabetes mellitus), respectively. The detection of FV Leiden was made by molecular biology methods. The occurrence in a group of 152 healthy individuals was four % (6 persons) and this frequency corresponds to the geographic localization of the Slovak Republic in Europe. In a group of 349 patients with thrombosis in anamnesis, FV Leiden was detected in 103 persons (29.5%). The occurrence was higher than the usually reported incidence in these patients (20%). Likewise, in a group of 35 patients with IM without risk factors in anamnesis, the occurrence of FV Leiden (8.6%) was significantly higher in comparison with healthy population and the incidence further increased significantly in a group of 41 patients with IM and the presence of at least one risk factor (14.6%). The authors therefore suppose an active role of the Leiden mutation of FV gene in the pathogenesis of this disease.

Bukovsky I, Pullmann R, Lang M, Orszagh M, Hybenova J, Koskova E: Endothelial Dysfunction and Plasma Lipids: the EDO Study Analyses Bratisl Lek Listy 1999; 100 (3): 149ñ155 On the basis of experimental as well as clinical observa- tions, endothelial dysfunction is (defined as impaired or ab- sent endothelium-dependent relaxation) considered to be an important factor in the atherogenesis. Serum lipids ab- normalities have been accepted as an epidemiological risk factor of atherosclerosis. In vitro, experimental as well as epidemiological studies revealed the fact, that lipoprotein oxidation plays an important role in atherogenesis. Recen- tly invented non-invasive methods to test and measure the endothelial function in vivo opened the opportunity to stu- dy the influence of different serum lipids on the endothelial function directly. Therefore, we decided to employ this non- invasive method for studying the endothelial function and observe the influence of various levels of plasma lipids and lipoprotein oxidation on the endothelial function of arteries in middle-aged men, since they are the most endangered part of population. In our study we used a†method of measuring the diameter of a. radialis by high-resolution ultrasound (Sonoline 450, Sie- mens, Japan) and further mathematical and statistical ana- lysis of functional as well as relative vasodilation reserve fol- lowed these measurements. Blood samples were taken within 24 hours of ultrasonography to study serum lipids (total cho- lesterol, HDL, triglycerides) and parameters of oxidation/an- tioxidation (superoxid dismutase ó SOD, malondialdehyde ó MDA). Sixty men, 25ó45 years old, from an area of basically the same level of pollution were examined. We found a†negative correlation between FVDR and TCH (p=0.01), FVDR and Tg (p=0.002) and FVDR a†TCH/HDL (p=0.015). Positive correlation exists (p<0.001) between TCH, Tg levels and TCH/HDL ratio and MDA level in all cases. Analysing fur- ther data from the EDO Study, we can conclude, that increa-

OBJECTIVE:ACE takes part in the renin-angiotensin and kallikrein-kininogen systems by creating angiotensin-II and inactivating bradykinin. ACE gene insertion/deletion polymorphism is associated with the level of circulating enzymes--subjects with the DD genotype have higher levels of circulating ACE than subjects with the II genotype and show an increased tendency towards impaired vascular function and structure. Patients with systemic lupus erythematosus (SLE) suffer from differentially expressed vascular pathology. We attempted to determine whether the type of ACE polymorphism could contribute to this pathology.METHODS:101 SLE patients fulfilling the ACR criteria were investigated. The I/D polymorphism was ascertained by PCR, followed by electrophoresis of the amplified fragments and UV visualization.RESULTS:The frequency of the D allele was higher in the SLE group (0.623) than in the controls (0.520) (chi 2 test, p < 0.025). The distribution of the ACE genotype in SLE group was different from that in the control group (p < 0.05). An association between the DD genotype and visceral damage (p < 0.006) was observed.CONCLUSION:Our results suggest that in the multifactorially determined vascular pathology of SLE, changes associated with I/D polymorphism could influence vessel wall inflammation (monocyte adhesion and activation with cytokine release, T-lymphocyte metabolism), a tendency towards vascular impairment (neointimal proliferation, vasospasm, platelet activation, myocyte proliferation) and lead to the subsequent ischemia. The ACE gene could serve as the visceral damage indicator in SLE.

The laboratory diagnosis of resistance to activated C protein (APC-resistance) involves examination of the phenotype and genotype of this thrombophilia. For examination of the phenotype coagulation and chromogenic tests are used. Their essence is examination in the presence and absence of exogenous APC. While the result of the original coagulation examination of APC-resistance which uses the APTT principle is influenced by a number of factors, the sensitivity and specificity of the modification of this examination (dilution of the examined plasma sample by FV deficient plasma before making the test) in relation to detection of FV Leiden is almost 100% and eliminates the majority of limitations of the original examination. The chromogenic assessment of APC-resistance has similar advantages, however, it cannot differentiate between the heterozygous and homozygous form of FV Leiden. During examination of the genotype of subjects with APC-resistance the mutation of FV Leiden is detected in as many as 90%. The group of subjects with the phenotype of APC-resistance comprises in particular subjects with acquired APC-resistance caused by conditions which lead to a disbalance between procoagulation and anticoagulation proteins of haemostasis which influence the reactions of the applied laboratory examinations. The acquired phenotype of APC-resistance can be also associated with an increased risk of thrombosis and the clinical manifestations of this thrombophilia resemble the classical, FV Leiden conditioned APC resistance. Rarely also congenital causes of the phenotype of APC-resistance are encountered caused by another mutation than the Leiden mutation of gene FV. The concurrent examination of the patient's plasma with the original and modified coagulation test makes it possible to assess the inborn cause of APC-resistance (positive finding also in modified examination). The presence of FV Leiden is then confirmed by examination of the genotype by the polymerase chain reaction.

OBJECTIVE Genetic susceptibility to systemic lupus erythematosus (SLE) is conferred not only by various genes within the major histocompatibility complex (MHC) region, but also by several other non-MHC linked genes. The negatively signalling molecule CTLA-4 is involved in establishing and maintaining of peripheral T cell tolerance, which controls T cell activation and reactivity. Its attenuating action helps to prevent an inappropriate initiation of T cell responses to self antigens and to terminate ongoing T cell responses. We tested if there was an association between CTLA-4 and SLE, a disease with B and T cell hyperreactivity and impaired peripheral T cell tolerance. METHODS Using the polymerase chain reaction--restriction fragment length polymorphism method with Bbv I digestion, we assessed an exon 1 transition dimorphism (49 A/G) of the CTLA-4 gene in 102 SLE patients and in 76 healthy controls. RESULTS The distribution of CTLA-4 exon 1 genotypes in the SLE group was significantly different from that in the controls (chi 2 = 6.178, p < 0.05). 17.6% of the SLE patients were G/G homozygotes compared to 5.3% of the controls; 36.3% were A/G heterozygotes vs 40.8% of controls; and 46.1% were A/A homozygotes vs 53.9% of the controls. The frequency of the G allele was significantly higher in SLE patients (35.8%) than in controls (25.7%; chi 2 = 4.142, p = 0.042). CONCLUSION Our results indicate that the non-MHC linked CTLA-4 gene could confer susceptibility in SLE, as it does in various other autoimmune diseases (Hashimoto thyroiditis, Graves' disease, IDDM).

BACKGROUND:The PlA1/A2 polymorphism of the human platelet membrane glycoprotein IIIa gene cause T-->C transition in the exon ii (position 1565) resulting in the leucine-->proline substitution in amino-acid sequence. This polymorphism was shown to be associated with increased risk of myocardial infarction (MI).AIM:To test genetic parameters of the PlA1/A2 polymorphism in our population and to assess the relation between mutant PlA2 allele and MI.METHODS:DNA was isolated from peripheral blood, collected from 40 patients with MI and with present risk factors (hypertension, hypercholesterolaemia, diabetes, obesity, smoking...), 33 patients with MI without risk factors, 34 controls with equivalent average age to both groups of the MI-patients, 58 control probands without MI in their family history and 33 healthy controls randomly recruited. After PCR amplification the resulting 267 bp fragment was digested with the restriction endonuclease NciI and subfragments were separated electrophoretically in 12% polyacrylamide gel.RESULTS:The frequency of the PlA2 allele was 0.121 in patients with MI without risk factors, 0.205 in patients with present risk factors, 0.162 in controls of the equivalent average age to the MI-patients, 0.172 in controls without MI in their family history and 0.20 in healthy controls randomly recruited. Genotype frequencies were in all groups in genetic equilibrium. Although the groups differed significantly (p < 0.01) in serum concentrations of total cholesterol, HDL-cholesterol, LDL-cholesterol, apolipoprotein A-1, apolipoprotein B and malondialdehyde, no significant differences in the serum concentrations of these metabolites between A1/A1, A1/A2 and A2/A2 genotypes were observed.CONCLUSIONS:PLA1/A2 polymorphism is associated with MI, however not as a dominant risk factor, but as a part of environmentally influencable multigene system. There is no relation between genotypes of the PLA1/A2 polymorphism and the lipoproteins plasma concentrations. (Tab. 4, Ref. 17.)

On the basis of experimental as well as clinical observations, endothelial dysfunction is (defined as impaired or absent endothelium-dependent relaxation) considered to be an important factor in the atherogenesis. Serum lipids abnormalities have been accepted as an epidemiological risk factor of atherosclerosis. In vitro, experimental as well as epidemiological studies revealed the fact, that lipoprotein oxidation plays an important role in atherogenesis. Recently invented non-invasive methods to test and measure the endothelial function in vivo opened the opportunity to study the influence of different serum lipids on the endothelial function directly. Therefore, we decided to employ this non-invasive method for studying the endothelial function and observe the influence of various levels of plasma lipids and lipoprotein oxidation on the endothelial function of arteries in middle-aged men, since they are the most endangered part of population. In our study we used a method of measuring the diameter of a. radialis by high-resolution ultrasound (Sonoline 450, Siemens, Japan) and further mathematical and statistical analysis of functional as well as relative vasodilation reserve followed these measurements. Blood samples were taken within 24 hours of ultrasonography to study serum lipids (total cholesterol, HDL, triglycerides) and parameters of oxidation/antioxidation (superoxid dismutase--SOD, malondialdehyde--MDA). Sixty men, 25-45 years old, from an area of basically the same level of pollution were examined. We found a negative correlation between FVDR and TCH (p = 0.01), FVDR and Tg (p = 0.002) and FVDR a TCH/HDL (p = 0.015). Positive correlation exists (p < 0.001) between TCH, Tg levels and TCH/HDL ratio and MDA level in all cases. Analysing further data from the EDO Study, we can conclude, that increased plasma lipids are more likely to be oxidized, which, in turn, is the probable reason of endothelium-dependent vasodilation impairment.

There are at least two groups of issues connected with an impact of a realization of the "Human Genome Project": a) philosophical; b) ethical, which can be divided into four groups: 1) the influence of DNA technologies on everyday applications of bioethical principles; 2) ethical aspects of genetic diversity; 3) ethical aspects of genetic screening; 4) somatic and germ-cell gene therapy; Unlike essential philosophical issues practical realization of issues in question can be largely expressed as only revitalization of old ones and concerns: the principle of justice-equal access and priorities; protection of reproductive choices; disclosure to patients and to relatives at genetic risk (disclosure and exclusion tests); prenatal diagnosis for "mild to moderate" diseases with and without genetic indication-commercialization; insurance policy; non-directive and directive genetic counseling. Maybe there are regional and other differences, but in practice, the main ethical issues are likely to involve screening for genetic risk of common diseases of adult life e.g. hypertension, diabetes, gout, dyslipoproteinemia, genes for premature atherosclerosis, etc. because of the possible direct impact on a patient, an implication for life-insurance, employers and commercial exploitation.

Conflicts of interest for the clinician(physician)-researcher are not limited only to direct and clear financial support by manufacturers of the pharmaceutical and medical device industry, but rather include delicate indirect monetary and research support. Today professionals face an inevitable choice between two opposing moral orders, one based in the primacy of ethical obligations to the sick, the other in the primacy of self-interest and the marketplace. Some medical ethicists urge, reshape ethical codes to conform to the ethos of the marketplace, which legitimates self-interest over beneficence and makes vices out of most of traditional virtues. Second opinion represents the ethicists who recommend a firm stand in belief that being a physician imposes certain specific obligations. Medicine is at heart a moral enterprise and those who practice it are de facto members of a moral community. The market introduces an alien-till this time unknown-set of economic values into an institution (medicine) whose inherent ends are altruistic, but in countries under health care reform it brings a complex of special ethical issues in connection with deficient legislation and not firm ethical rules adopted. (Ref. 17.)

BACKGROUND:Linolenic, linoleic, and arachidonic acids as well as other polyunsaturated fatty acids are necessary for health as the precursors of eicosanoids and for the structure of developing membranes.OBJECTIVES:The aim of the present study was the determination of the level of 11 individual free fatty acids (FFA) in the milk and in the blood of mothers and newborns during the perinatal period.METHODS:In 21 women the FFA was determined in their colostrum as well as in the venous blood at the delivery in the hospital, and then again 5 days later at leaving the hospital. Simultaneously, the blood of newborns was collected as umbilical samples at birth and as venous blood on the 5th day. The study was performed on health term infants and mothers with normal gestational age.RESULTS:The results show a marked increase in total milk FFA as well as in most, but not all, individual FFAs during the followed period of the first 5 days. The values in the milk were always remarkably higher (the increase more than 2 orders) than in the blood. We have found no significant statistical correlation between values in the blood and those in the milk. The concentrations of all very important omega-3 FFAs (which are present in fish oil and in foods of marine fish origin) were always lower in all blood and milk samples in comparison with the levels of omega-6 FFAs (which are prevailing in lipids of our usual nutrition as are margarines and most of commercial oils).CONCLUSIONS:Our findings seem to be very important for preventive medicine and need to study further the relationship of low intake of O3FA to increased incidence of various allergies and other pathological syndromes in children. The very large range of a variance in the values of FFAs in the milk suggests the need of more profound study of the role of the food composition probably during the whole period of pregnancy, mainly as to the type of lipid composition. (Tab. 1, Fig. 6, Ref. 24.)

In young rats of different age the effect of short-term administration of thyroxine (T4) on the level of 6 FFA was determined. The concentration of FFA was expressed in mmol/l as well as the molar ratio to serum albumin. The results show that the level of the total FFA and the ratio of saturated to unsaturated (S/U) FFA are high in control animals till the age of 21 days in comparison with adult animals. Administration of T4 caused marked increase in the total FFA in the youngest age group (8-day-old) and the effect increased with age. Differences in the trend of changes in individual FFA were found. In the concentration of myristic acid (C 14:0) a significant increase was found only till the age of 16 days; in the case of palmit-oleic acid the increase was significant in all age groups. T4 caused no significant changes in the values of stearic acid (18:0). When the data were expressed in terms of the molar ratio FFA/albumin, the changes were more pronounced. The peak values found in 16-day-old animals were 33.9 +/- 0.75; they represent a 2-times higher supply of FFA to tissues in comparison with control animals. In this study it was also found that the concentration of serum albumin in newborn animals is low and represents just 57% of adult values. The short-term application of T4 caused no changes in the concentration of serum albumin. The mechanism of T4 action on FFA during ontogenesis was not explained in this study.(ABSTRACT TRUNCATED AT 250 WORDS)

The authors presented their own experience with treatment of cystine lithiasis in 7 patients, which has been performed during the course the past of ten years. Identification of cystine was performed by concrement submitted examination, using a polarizating microscope, and x-ray difractometric structural analysis. After stone extraction the patients were treated by a high fluid intake (diuresis over 2.5 l), alcalizating treatment involving citrates (Alkalit Spofa), and Penicilamin (Spofa), or Thiola (Santen) which are currently proved to be the most optimal approach. Recurrence of stones requiring percutaneous treatment, was observed in one of the patients during five years metaphylaxy. Another patient of the same group had three times stated cystine stone recurrence which passed spontaneously.