Abstract Common variants in MTNR1B, encoding melatonin receptor 1B, have been recently associated with impaired glucose homeostasis and an increased risk for developing Type 2 diabetes (T2D). In this study we investigated the association of MTNR1B variant rs10830963 with T2D and related quantitative traits in a population from Bosnia and Herzegovina (BH). A total number of 268 subjects were recruited in the study (162 T2D patients and 106 nondiabetic controls). Subjects were genotyped for MTNR1B rs10830963 SNP by using hydrolysis probes. Our data showed that the prevalence of the MTNR1B rs10830963 risk G-allele in BH population was 26%. Furthermore, we demonstrated a significant association of MTNR1B rs10830963 variant with fasting plasma glucose (FPG) levels in nondiabetic subjects. Under the additive genetic model, each variant G-allele was associated with an increased FPG levels of 0.29 mmol/L (95% CI 0.12, 0.46, p=0.001). Strikingly, our results also showed a significant association of this MTNR1B polymorphism with increased glycated hemoglobin (HbA1c) levels in nondiabetic subjects (p=0.040, additive genetic model). An association of the MTNR1B variant rs10830963 with T2D risk was not detected in our cohort. In conclusion, here we have demonstrated the association between the common MTNR1B rs10830963 variation and fasting plasma glucose levels in BH population. Furthermore, the influence of this polymorphism on the HbA1c levels was also shown in this study, further strengthening its role in blood glucose control.

Introduction: Oxidative stress represents a pathophysiological mechanism lying behind occurrence of different acute and chronic diseases. Pregnancy, mainly due to placenta rich with mitochondria, is also being associated with the state of oxidative stress. Numerous markers have been proposed in order to test oxidative stress in pregnancy state, one of them being 8-isoprostane. The aim of this study was to analyse serum concentrations of 8-isoprostane as a possible oxidative stress marker in pregnancy.Methods: Serum concentrations of 8-isoprostane were measured in overall population of 84 woman, between 20 and 30 years of age. Tested population was divided in 2 groups: 42 pregnant woman were classifi ed as being either in the fi rst or second trimester of pregnancy. In the control group healthy nonpregnant women were included. Concentration of 8-isoprostane in serum was determined by commercial 8-isoprostane EIA test kit of Cayman Chemical Company, USA.Results: The 8-isoprostane levels were signifi cantly increased in pregnant women in relation to healthy non pregnant women (p<0.05). The 8-isoprostane levels were signifi cantly increased in second and third trimester of pregnancy (p<0.05).Conclusions: According to the obtained results, 8-isoprostane might be used as a possible marker of oxidative stress in pregnancy state, but not as a biomarker for the risk of complications development in pregnancy in analysed population.

Type 2 diabetes mellitus (T2DM) is a worldwide epidemic with considerable health and economic consequences. T2DM patients are often treated with more than one drug, including oral antidiabetic drugs (OAD) and drugs used to treat diabetic complications, such as dyslipidemia and hypertension. If genetic testing could be employed to predict treatment outcome, appropriate measures could be taken to treat T2DM more efficiently. Here we provide a review of pharmacogenetic studies focused on OAD and a role of common drug-metabolizing enzymes (DME) and drug-transporters (DT) variants in therapy outcomes. For example, genetic variations of several membrane transporters, including SLC22A1/2 and SLC47A1/2 genes, are implicated in the highly variable glycemic response to metformin, a first-line drug used to treat newly diagnosed T2DM. Furthermore, cytochrome P450 (CYP) enzymes are implicated in variation of sulphonylurea and meglitinide metabolism. Additional variants related to drug target and diabetes risk genes have been also linked to interindividual differences in the efficacy and toxicity of OAD. Thus, in addition to promoting safe and cost-effective individualized diabetes treatment, pharmacogenomics has a great potential to complement current efforts to optimize treatment of diabetes and lead towards its effective and personalized care.

Introduction: Several decades of basic science and animal research provided considerable support for significant role of plasma free fatty acids (FFAs) in etiology of Type 2 diabetes mellitus (T2DM). Contradicting data related to signifi cance of elevated FFAs in plasma of patients with Type 2 diabetes prompted us to study concentrations of palmitic acid, stearic acid, and linoleic acid, in patients and healthy controls in an attempt to possibly use them as potential biomarkers in progression of the disease. Since aging is associated withincreased plasma glucose and insulin levels that are consistent with an insulin resistant state, in this study,age differences in the concentration of the above mentioned acids were tested.Methods: Progressive changes in their concentrations were followed through a period 6 months. All subjects included in the study were free of evidence of hepatitis B or C viral infection or active liver and kidney damage. Analysis of glucose and glycated hemoglobin levels were performed on BT PLUS 2000 analyzer using standard IFCC protocols, while concentrations of FFAs were analyzed by gas chromatography.Results: Our data demonstrated signifi cantly higher FFA values in plasma of diabetic patients as compared to healthy controls. There was a trend of correlation of FFAs levels with the blood glucose levels in diabetic patients, which was more prominent in diabetic men than in women.Conclusion: With aging, levels of free fatty acids signifi cantly increased in plasma of diabetic patients, and this effect was also more profound in male than in female diabetics.

Introduction: The enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) catalyzes the conversion of the hormonally inactive cortisone to active cortisol, thus facilitating glucocorticoid receptor activation in target tissues. Increased expression of 11β-HSD1 in adipose tissue has been associated with obesity and insulin resistance. In this study, we investigated the association of two 11β-HSD1 gene (HSD11B1) polymorphisms with the metabolic syndrome (MetS) and its characteristics in the Bosnian population. Materials and methods: The study included 86 participants: 43 patients diagnosed with MetS and 43 healthy controls. Subjects were genotyped for two HSD11B1 gene polymorphisms: rs846910: G>A and rs45487298: insA, by the high resolution melting curve analysis. Genotype distribution and an influence of genotypes on clinical and biochemical parameters were assessed. Results: There was no significant difference in the mutated allele frequencies for the two HSD11B1 gene polymorphisms between MetS patients and controls. In MetS patients, no significant associations between disease-associated traits and rs45487298: insA were found. Regarding rs846910: G>A variant, heterozygous patients (G/A) had significantly lower systolic (P = 0.017) and diastolic blood pressure (P = 0.015), lower HOMA-IR index (P = 0.011) and higher LDL-cholesterol levels (P = 0.049), compared to the wild-type homozygotes. In the control group, rs45487298: insA polymorphism was associated with lower fasting plasma insulin levels (P = 0.041), lower homeostasis model assessment insulin resistance (HOMA-IR) index (P = 0.041) and lower diastolic blood pressure (P = 0.048). Significant differences between rs846910: G>A genotypes in controls were not detected. Haplotype analysis confirmed the association of rs45487298: insA with markers of insulin resistance in the control subjects. Conclusions: Our results indicate that a common rs45487298: insA polymorphism in HSD11B1 gene may have a protective effect against insulin resistance.

Aim: To study the influence of C-reactive protein (CRP) level in the blood, fibrinogen level and general inflammatory syndrome as the predictors of development of secondary fibrosis in patients with pulmonary tuberculosis (TB). Methods: Concentration of CRP, fibrinogen level was measured using immunoturbidimetric methodIncluding criteria was presentation of TB process in both lungs, as the sign of widespread TB process. Results: We examined 85 patients treated in one year. Mean CRP level was 22,6 mg/mL, range 5-245 mg/mL; normal level (up to 8 mg/mL) was measured in 23,4% patients, medium level (9-20 mg/mL) was measured in 31,3% patients, high level (21-50 mg/mL) were measured in 26,2% patients, and in 23,7% patients CRP were higher than 50 mg/mL. Average fibrinogen level in whole group was 6,9 g/L (SD 5,8). Normal level of fibrinogen (up to 4 g/L) were measured in 6,4% of patients; 4,1-1,0 g/L were measured in 24,6% patients, 10,1-20 g/L were measured in 31,1% patient and level more than 20 g/L were measured in 37,9% patients. Using statistic method of partial correlation statistical significane at level p<0,05 was shown between them. Correlation of CRP and fibrinogen level with appearance of fibrosis on X-ray of the lung was shown. Thereafter, closer correlation was shown with fibrinogen and fibrosis than with CRP and fibrosis. Conclusion: Predicted value of CRP and fibrinogen for pulmonary fibrosis was shown in TB patients. So, attenuation of fibrosis development, possible with antifibroblastic activity of pentoxyphyllin, should be taken in consideration, for prevention of widespread development of lung fibrosis in these patients.

Aim: To study correlation of IgE level and C-reactive protein (CRP) for exacerbation of the disease in asthmatic patients. Methods: Asthmatic subject were examined for achieving of asthma control according to GINA recommendation. Numbers of exacerbation of asthma during one month were analyzed. The patients were followed in six month period (since first January to 30th of Jun. Average monthly days of exacerbations was calculated. IgE level in the blood was measured using Enzyme-linked Immunoassay (ELISA), and CRP was measured by immunotubidimetry. Assessment of asthma control was considered using Asthma Quality of Life Questionnaire (AQLQ). Results: The study includes 63 patients with asthma. Average level of IgE was 674 IU/mL (SD 167), range 56-3785 IU/mL, 1 IU=3,2 ng; average level of CRP was 16,4 mg/mL (SD 6,3), range 5-48; Average number of days in exacerbation during one month was 3,6 (SD 2,4), and varied from zero, patients with no exacerbation, to 21. Using test of multiple correlation it was shown statistical significant correlation (level p Conclusion: In this study CRP was shown as stronger predictor of asthma exacerbation and worse quality of life than total IgE level in asthmatic subjects.

In this paper we would like to briefly introduce readers to the situation in the field of laboratory medicine in Bosnia and Herzegovina, with a focus on training in the field of medical biochemistry. As in some of neighboring countries, term Medical biochemist is the usual name for the Clinical biochemist or Clinical chemist in Bosnia and Herzegovina. Despite the difficult period through which the profession had passed in the last two decades, laboratory work, particularly clinical biochemistry, has managed to retain the necessary quality and keep pace with the developed world. In post war period, Society of Medical Biochemists of Bosnia and Herzegovina held regular meetings each year as a part of "life long learning" process, where both scientific and vocational lecturers presented their work. A single law on the state level would provide us with more defined and precise answers, such as: who can get a specialization, how long should last the training for medical biochemistry specialists (duration in years). This law should be in consent with the program described in EC4 or other documents given by the EFCC (European Federation of Clinical Chemistry and Laboratory Medicine) and IFCC (International Federation of Clinical Chemistry and Laboratory Medicine).

AIM Lipin 1 is a recently discovered multifunctional protein involved in the metabolism of lipids, while PPARgamma is involved in adipocyte differentiation, and regulation of lipid metabolism. Up to now, LPIN1 and PPARG gene polymorphisms have been associated with type 2 diabetes, metabolic syndrome, and central obesity. In this study, we hypothesized that genetic variants within LPIN1 and PPARG genes were associated with traits of metabolic syndrome. Correlation between biochemical parameters (including but not limited to, glucose, HbA1c, insulin levels, HDL and LDL cholesterol, triglycerides, serum proteins, liver enzymes) and frequency of polymorphisms in LPIN1 (rs11693809 and rs2716610) and PPARG gene (rs10865710, rs3856806 and rs1801282), was tested in this study. METHODS The study included 70 patients diagnosed with metabolic syndrome and type 2 diabetes. Two polymorphisms of LPIN1 gene (rs11693809 and rs2716610), and three polymorphisms of PPARG gene (rs10865710, rs385806 and rs1801282) were analyzed by real time PCR and conventional PCR-RFLP methods. RESULTS Our analysis revealed correlation between insulin levels and rs11693809 LPIN1 polymorphism in diabetic patients. Also the results of this study showed an association of rs10865710 and rs385806 polymorphism of PPARG with HDL cholesterol and LDL plus total cholesterol levels, respectively. CONCLUSION These data reflect an association of analyzed PPARG and LPIN1 gene polymorphisms with values of insulin, HDL, LDL and total cholesterol witch indicates an important role of these genes in lipid metabolism and pathogenesis of type 2 diabetes and metabolic syndrome.

AIM Differences in the frequency of distribution of the cytochrome P450 (CYP) allelic variants have been demonstrated between distinct ethnic groups, contributing to observed interindividual variation in drug response. In this study we determined, for the first time, prevalence of the common allelic variants of the polymorphic CYP enzymes, CYP3A4*1B and CYP3A5*3, in the population of Bosnia and Herzegovina (BH). METHODS Genomic DNA was extracted from blood samples collected from 140 unrelated subjects. A real-time PCR was used for the detection of CYP polymorphisms, with the application of the specific TaqMan SNP Genotyping Assay (Applied Biosystems) for CYP3A5*3, while CYP3A4*1B was genotyped by high-resolution melting analysis. RESULTS Our results have shown that the distribution of CYP3A4*1B and CYP3A5*3 alleles was in line with the data reported in European Caucasians. We confirmed that CYP3A4*1B mutant allele is rare in Caucasians, being present in only 5.1% individuals. However, CYP3A5*3 polymorphism was found to be predominant in the Bosnian population with an incidence of 94%, similarly to other European populations tested so far. Interestingly, we have demonstrated a strong linkage disequilibrium between CYP3A5*3 and CYP3A4*1B alleles. No significant difference in allele frequencies for CYP3A4*1B and CYP3A5*3 has been shown between male and female subjects participating in our study. CONCLUSION Our data demonstrated the high prevalence of CYP3A5*3 allele in Bosnian population, indicating significance of analysis of CYP3A5 and CYP3A4 polymorphisms and corresponding allele frequencies in specific ethnic groups. Importantly, results of this study may lead to translation of pharmacogenetics and individualized therapeutic approach in current clinical practices in BH.

AIM To analyze usefulness of measurement amino-terminal pro-B type natriuretic peptide of (NT pro-BNP) as the one of parameters of water overload in patients with chronic kidney diseases. METHODS A total number of 277 patients with chronic kidney diseases (CKD) were followed up in the period often years between January 2000 and July 2010. Patients with creatinine clearance of 60 ml/min or less were included in the study. Changes of creatinine clearance, and in last five years changes of NT pro-BNP were followed. Water overload was analyzed using chest x-ray in relation with concentration of NT pro-BNP in the blood. RESULTS Decrease of clearance of creatinine ranged from average 54.7 ml/min in the first year to 14.6 ml/min in the fifth year of the monitoring. Average NT pro-BNP level in patients without any sign of water overload was 94 pg/ml (SD 21), mean value in those with Kerley lines was 231 pg/ml/L (SD 64), in those with clear signs of water overload but without pleural effusion it was 525 pg/ml (SD 223), and in those with water retention including pleural effusion it was 1606 pg/ml (SD 1134). Using test of multiple correlation a statistically significant correlation between X-ray signs of water overload and NT pro-BNP concentration was shown, p < 0.05. CONCLUSION Measurement of NT pro-BNP was increased in the beginning of water overload in patients with CKD. Increased value of NT pro-BNP may be found earlier than any other signs of water overload. NT pro-BNP was a useful parameter in estimation of water overload in these patients.

BACKGROUND Polymorphisms in HSD11B1, the gene encoding 11β-hydroxysteroid dehydrogenase type 1 enzyme, have been associated with obesity, metabolic syndrome, and type 2 diabetes. In this study, we present an optimized high-resolution melting (HRM) method for genotyping two common polymorphisms of the HSD11B1 gene: rs846910: G>A and rs45487298: insA. METHODS One hundred DNA samples from patients with polycystic ovary syndrome and healthy controls were genotyped by HRM. The results were compared with those obtained with classic polymerase chain reaction followed by restriction fragment length polymorphism analysis. RESULTS Various approaches were used during HRM specificity optimization. With the optimized method, genotyping accuracy of 100% was achieved. CONCLUSIONS HRM analysis is a fast, simple, and cost-effective method compared with the alternative genotyping approaches. The work required for optimizing the method (improvement of specificity) is minor compared to the advantages.

Problem: Laboratory medicine, medical-biochemical diagnosis in primary health care is much represented. By organization of family medicine medical-biochemical diagnosis is defined as a branch of diagnostic services in primary health care. For these actions is necessary in the morning prior to admission of users and their demands that all jobs are properly prepared. On previous day should be provided and prepared: accessories, reagents and machines. Morning daily routine work of preceding control and calibration equipment, methods and process quality control of work in the laboratory. Only after the fulfillment of the procedures followed overview of search control of samples. After validating the results of daily quality control and after they met the criteria can be analytically examined samples from users. These procedures are not sufficiently familiar to users and doctors, for that are very often necessary the direct telephone communication between them. To make the results of laboratory tests needed are huge material resources. This is evident in the economic analysis where laboratory tests are valued with a score of: search by type and material resources expended for analytical examination. These technical and financial performances of laboratory medicine are not appropriately classified as blatant as that in other industries, technology and other primary health care (PHC) and family medicine (FM). Goal: The overall objective of the research is to define a model of efficiency (or effectiveness) of medical-biochemical diagnosis for users with the requirements of units of family medicine (FM), in a representative sample of patients in the unit for the laboratory diagnosis of the Primary Health Care Center Gracanica. Confirm what is the usefulness of the application of laboratory diagnosis in family medicine. Determine the frequency of the need for laboratory tests in the therapeutic treatment of major diseases. Evaluate the need for using laboratory diagnostics to try to prevent major diseases. Material and methods: The study included a total of 1000 respondents. All subjects were users of primary health care in Primary Health Care Center Gracanica (Tuzla Canton) in primary health care units have received requests for laboratory diagnosis. This paper is an analysis of the representation requirements for the laboratory diagnosis by doctors in primary health care and the most frequent diseases in primary care. An analysis is made of laboratory test results, based on requests for laboratory diagnosis by doctors and illnesses in primary care. Made is analysis of the presence of normal and pathological laboratory test results from the request for the laboratory diagnosis by doctors in primary health care. Made is an analysis of the most common laboratory tests requests, and based on requests for laboratory diagnosis by doctors in primary health care and the most frequent diseases in primary health care. Incorporated is the economic analysis of labor