Abstract The enzymes of the cytochrome P450 superfamily play a critical role in phase I drug metabolism. Among them, CYP2C9 and CYP2C19 are clinically important, as they can mediate severe toxicity, therapy failure, and increased susceptibility to cancer and other diseases caused by chemicals. The aim of this study was to determine the prevalence of pharmacologically most important allelic variants of the CYP2C9 and CYP2C19 genes in the general population of the Republic of Srpska (Bosnia and Herzegovina) and to compare them with other populations. For this purpose we determined the genotype profile and allele frequency of 216 randomly selected healthy volunteers using real-time polymerase chain reaction (RT-PCR). The prevalence of the CYP2C9 *2 and *3 alleles was 13.6 and 7.4 %, respectively. Based on these frequencies, of the 216 participants four (1.86 %) were predicted to be poor metabolisers, 78 (36.11 %) intermediate, and the remaining 134 (62.03 %) normal metabolisers. Based on the prevalence of CYP2C19 *2 and *17 variants – 16.2 and 20.4 %, respectively – nine (4.17 %) were predicted to be poor, 57 (26.39 %) rapid, and nine (4.17 %) ultra-rapid metabolisers. We found no significant differences in allele frequencies in our population and populations from other European countries. These findings suggest that genetically determined phenotypes of CYP2C9 and CYP2C19 should be taken into consideration to minimise individual risk and improve benefits of drug therapy in the Republic of Srpska.

Introduction: Oganophosphorus compounds (OP) bind to acetylcholinesterase (AChE) and inactivate it. In the synaptic cleft, undestroyed and accumulated acetylcholine produce the acute cholinergic effects. The aim of this study was to determine the frequency, speed of onset and intensity of certain signs of paraoxon poisoning depending on dose and outcome of poisoning. Methods: The study was conducted in adult Wistar rats. The median lethal dose (LD50) of paraoxon as well as protective ratio (PR) of atropine (10 mg/kg intramuscularly) was determined. Clinical signs of poisoning were observed: fasciculations, tremor, seizures, ataxia, piloerection, lacrimation, exophthalmos, bizzare/stereotypic behaviour and dyspnoea. The time from paraoxon injection to the first appearance of the sign of poisoning was recorded as well as the intensity of poisoning with evaluation at 10 time intervals throughout the 4 h observational period. Results: The LD50 of paraoxon was 0.33 mg/kg (subcutaneously) and PR of atropine was 2.73. Dose-dependent, piloerection occurred more often (p = 0.009) and at higher intensity (p = 0.016) at higher doses. Fasciculations, tremor, seizures and ataxia occurred significantly earlier at higher doses of paraoxon (p = 0.015, 0.002, 0.021 and 0.016, respectively), as well as the intensity of seizure, tremor and fasciculation. Piloerection (p = 0.002) and seizures occurred more frequently (p = 0.009) in non-survivors. Fasciculations, tremor, seizures and ataxia occurred significantly earlier and at higher intensity in non-survivors (p < 0.001, for all parameters), as well as dyspnoea (p = 0.009 and p = 0.048). In atropine-protected rats, nicotinic effects persevered, so they were the prognostic parameter of the severity of the poisoning. Conclusion: Seizures and fasciculations followed by tremor were strong prognostic parameters of the probability of lethal outcome of paraoxon poisoning. Also, the mentioned poisoning signs were with their intensity and speed of occurrence in a clear positive correlation with the administered dose of paraoxon. Even at high doses of paraoxon, atropine blocked the muscarinic (but not nicotinic) effects and somewhat mitigated the CNS toxic effects.

Background/Aim: The prevention of cardiovascular risk factors and cardiovascular disease management contributes to the cardiovascular mortality reduction. The effects of these activities have been measured by quality indicators. The aim of this study was to determine the effects of family medicine team training workshop and implementation of clinical guidelines on the cardiovascular risk factors and diseases management in primary health care in the Republic of Srpska/Bosnia and Herzegovina. Methods: The "CardioVascular Risk Assessment and Management" study included a sample of 373 teams from 41 primary health care centres trained to provide adequate services and to compare the quality of cardiovascular risk management before and after the training workshop and implementation of clinical guidelines. The comparison was based on nine project defined performance indicators related to hypertension, type 2 diabetes mellitus, hyperlipidaemia, tobacco smoking and obesity. Results: Significant improvements were observed in six indicators after the training workshop and implementation of guidelines. Target values for blood pressure and HbA1c were achieved in over 80 % of patients (82.12 ± 15.81 vs 84.49 ± 12.71 and 84.49 ± 12.71 vs 85.49 ± 24.55; before and after the training workshop, respectively), while the target values for LDL cholesterol were achieved in 54.98 % ± 20.33 before and 57.64 % ± 16.66 after the training workshop. The number of teams that had less than 20 % of recorded data significantly decreased after the training workshop and guidelines implementation, and adequate recording of all indicators was improved. Conclusion: The training workshop of family medicine teams and implementation of clinical guidelines resulted in significant quality improvement of cardiovascular diseases management in primary health care.

Increasing evidence suggests that apoptosis of tubular cells and renal inflammation mainly determine the outcome of sepsis-associated acute kidney injury (AKI). The study aim was to investigate the molecular mechanism involved in the renoprotective effects of simvastatin in endotoxin (lipopolysaccharide, LSP)-induced AKI. A sepsis model was established by intraperitoneal injection of a single non-lethal LPS dose after short-term simvastatin pretreatment. The severity of the inflammatory injury was expressed as renal damage scores (RDS). Apoptosis of tubular cells was detected by Terminal deoxynucleotidyl transferase-mediated dUTP Nick End Labeling (TUNEL assay) (apoptotic DNA fragmentation, expressed as an apoptotic index, AI) and immunohistochemical staining for cleaved caspase-3, cytochrome C, and anti-apoptotic Bcl-xL and survivin. We found that endotoxin induced severe renal inflammatory injury (RDS = 3.58 ± 0.50), whereas simvastatin dose-dependently prevented structural changes induced by LPS. Furthermore, simvastatin 40 mg/kg most profoundly attenuated tubular apoptosis, determined as a decrease of cytochrome C, caspase-3 expression, and AIs (p  <  0.01 vs. LPS). Conversely, simvastatin induced a significant increase of Bcl-XL and survivin, both in the strong inverse correlations with cleaved caspase-3 and cytochrome C. Our study indicates that simvastatin has cytoprotective effects against LPS-induced tubular apoptosis, seemingly mediated by upregulation of cell-survival molecules, such as Bcl-XL and survivin, and inhibition of the mitochondrial cytochrome C and downstream caspase-3 activation.

Centre for Biomedical Research, Faculty of Medicine, University of Banja Luka, Banja Luka, the Republic of Srpska, Bosnia and Herzegovina. Public Health Institute of the Republic of Srpska, Banja Luka, the Republic of Srpska, Bosnia and Herzegovina. Agency for Certification, Accreditation and Healthcare Quality Improvement of the Republic of Srpska (ASKVA), Banja Luka, the Republic of Srpska, Bosnia and Herzegovina.

Background/Aim: There have been different experimental conditions for in vitro studies on human umbilical arteries (HUA) in tissue bath system. This diversity was mainly reflected in variables such as stretching tension, incubation period and initial constriction challenging with potassium (KCl). The aim of the study was to establish optimal experimental conditions which will provide better responsiveness of HUA preparations, as well as to examine the impact of 24 h cold storage on viability and responsiveness of HUA to KCl and serotonin. Methods: The KCl-induced constrictions at different stretching tensions (0.5 g, 1.0 g, 2.0 g, 4.0 g), incubation times (30 min, 60 min, 120 min), and after multiple initial constriction challenging were compared. Dose response curves for serotonin were obtained under different conditions (1.0 g and 60 min vs. 2.0 g and 120 min). The influence of 24 h cold storage on KCland serotonininduced vasoconstriction of HUA preparations was examined as well. Results: The strongest constrictions induced by serotonin or KCl were obtained when preparations were adjusted at 2.0 g and incubated for 120 min. The KCl-induced constrictions observed after 120 min were statistically higher (p < 0.05) when preparations were challenged three times (30 min, 60 min, 120 min), compared to those challenged only once. The preparations that were stored at 4 0C for 24 h showed significantly stronger serotonin-induced constrictions (p < 0.01). The cold storage had no influence on KCl-induced constriction. Conclusion: For performing in vitro studies on HUA preparations in tissue bath, we propose stretching tension of 2.0 g, incubation period of 120 min and multiple initial constriction challenging with KCl as optimal experimental condition. We also showed that HUA preparations retained functional viability even after 24 h of cold storage.

Department of Hygiene, Faculty of Medicine, University of Banja Luka, Banja Luka, the Republic of Srpska, Bosnia and Herzegovina. Department of Hygiene, Public Health Institute of Republic of Srpska, Banja Luka, the Republic of Srpska, Bosnia and Herzegovina. Department of Pharmacology, Toxicology and Clinical Pharmacology, Faculty of Medicine, University of Banja Luka, Banja Luka, the Republic of Srpska, Bosnia and Herzegovina. Institute for Medicinal Plant Research "Dr Josif Pančić", Belgrade, Serbia. Institute of Hygiene and Medical Ecology, Faculty of Medicine, University of Belgrade, Belgrade, Serbia.

Background: The manipulation of gut microflora composition and activity by probiotics could modify the enzymatic activity of intestinal bacteria. In this study, we sought to investigate the influence of probiotic treatment on sulphasalazine (SSZ) excretion in inflammatory bowel disease (IBD) patients. Methods: Newly diagnosed IBD patients were randomised in two groups; half of subjects were treated with SSZ and other half were treated with combination of SSZ and probiotics. At the each visit, patients were assesed clinically andfecal samples and total volume of 24 h urine was measured and noted. Urine samples were collected and analized by liquid chromatography-mass spectrometry/mass spectrometryfor determination of SSZ and its metabolites. The enzymatic activity of azoreductase by intestinal bacteria in the fecal contents was determined spectrophotometrically. Results: Urinary levels of SSZ and its metabolites showed no statistically significant changes after probiotic administration. Azoreductasa activities, in both experimental groups, decreased comparing with pretreatment values in both cultivation conditions. Transient colonization with Bifidobacterium BB12 was confirmed in 22% of samples. Lactobacillus rhamnosus LGG did not show transient colonisation of the digestive tract. Conclusions: Co-administration of probiotics in patients treated with SSZ did not change the amounts of execreted SSZ and its metabolites