BACKGROUND: Effectiveness of SIT was well documented in many cases and published data. Selection of patients for SIT should be very serious and must include skin test and total and specific IgE measurement. How outcome of SIT correlate with changes of IgE, skin reactivity and overall symptoms reduction is aim of this study. MATERIAL AND METHODS: Skin testing, total and specific IgE measurements were performed before and after each year of treatment. Skin test assessment was performed according to recommendation of Manual of Laboratory immunology. IgE ws performed using ELISA method. Clinical outcome was assessed using AQLQ questionnaire. RESULTS: During five years period 58 asthmatic subjects with home dust and dermathophagoides allergy were treated by SIT. Bseline total IgE was 488,5 IU/ml (SD 78,9), mean specific IgE against dermatophagoides pteronissimus was 36,5 IU/ml (SD 15,2). Subcutaneous tests showed 15-20 mm weal in 43, and more than 21 mm in 15 cases. After 5 years mean total IgE was 227 IU/ml (SD 9,2) and mean specific IgE was 28,2 IU/ml (SD 8,9). Skin tests showed decrease diameter of weal. In 49 out of all patients clinical outcome were very well, and in 9 satisfied (according to AQLQ questionnaire). Using test of correlation, by linear regression, better correlation was shown between of skin testing and AQLQ than in total or specific IgE. So, in vivo skin tests were better predictor for success of SIT, than measurement of IgE. CONCLUSION: Results of skin tests in diagnostic assessment of allergy in asthmatic patients were better predictor of successful outcome of SIT than laboratory measurement of total and specific IgE.

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.

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 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.

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.

This is the first study performed in population from Bosnia & Herzegovina (BH), in which we analysed a significance of genetic variations in drug-metabolising enzyme, cytochrome P450 (CYP), in pathogenesis of Type 2 diabetes. We have determined allele frequencies for CYP2C9*2, CYP2C19*2, and CYP2D6*4 in diabetic patients and nondiabetic controls. Genomic DNA was extracted from blood samples collected from 37 diabetic and 44 nondiabetic subjects. A real-time polymerase chain reaction was used for the detection of specific CYP polymorphisms, with the application of the specific TaqMan® SNP genotyping tests (Applied Biosystems). Interestingly, results from this study have demonstrated that frequencies of CYP2C19*2 and CYP2D6*4 variants were in line, while frequency of CYP2C9*2 polymorphism seemed to be lower in this sample of BH population as compared to the Caucasians genotype data. Furthermore, no significant difference in allele frequencies for CYP2C9*2, CYP2C19*2, and CYP2D6*4 was demonstrated between diabetic and nondiabetic subjects. Thus, results form this study seem to indicate no relationship between CYP2C9, CYP2C19, and CYP2D6 genotype and diabetes susceptibility in Bosnian population. This in part may reflect a limited study population included in our study and would require larger cohorts to reveal potential relationships between analysed CYP genetic variants and diabetes risk. In addition, it would be pertinent to further explore possible effects of CYP genetic variations on therapeutic and adverse outcomes of oral antidiabetics, which might be the key in optimising therapy for individual patient with Type 2 diabetes.

Recent studies have introduced serum uric acid (UA) as a potential risk factor for developing diabetes, hypertension, stroke, and cardiovascular diseases. The value of elevated levels of UA in serum as a risk factor for diabetes development is still under scrutiny. Recent data suggest that clearance of UA is being reduced with increase in insulin resistance and UA as a marker of prediabetes period. However, conflicting data related to UA in serum of patients with Type 2 diabetes prompted us to study the urine/serum ratio of UA levels (USRUA) in these patients and healthy controls. All subjects included in the study were free of evidence of hepatitis B or C viral infection or active liver and kidney damage. Patients receiving drugs known to influence UA levels were also excluded from this study. Analysis of glucose and uric acid were performed on Dade Behring analyzer using standard IFCC protocols. Interestingly, our data demonstrated about 2.5 fold higher USRUA values in diabetic patients as compared to control subjects. Furthermore, there was a trend of correlation of USRUA value with the blood glucose levels in diabetic patients, which was more prominent in diabetic men than in women. With aging, levels of uric acid increased in serum of diabetic patients, and this effect was also more profound in male than in female diabetics. In conclusion, this study showed significantly elevated USRUA levels in patients with Type 2 diabetes, a negative USRUA correlation with the blood glucose levels in diabetic patients, and an effect of sex and age on the uric acid levels. Since literature data suggest a strong genetic effect on UA levels, it would be pertinent to perform further, possibly genetic studies, in order to clarify gender and ethnic differences in UA concentrations.

Th is is the fi rst study performed in population from Bosnia & Herzegovina (BH), in which we analysed a signifi cance of genetic variations in drug-metabolising enzyme, cytochrome P (CYP), in pathogenesis of Type  diabetes. We have determined allele frequencies for CYPC*, CYPC*, and CYPD* in diabetic patients and nondiabetic controls. Genomic DNA was extracted from blood samples collected from  diabetic and  nondiabetic subjects. A real-time polymerase chain reaction was used for the detection of specifi c CYP polymorphisms, with the application of the specifi c TaqMan® SNP genotyping tests (Applied Biosystems). Interestingly, results from this study have demonstrated that frequencies of CYPC* and CYPD* variants were in line, while frequency of CYPC* polymorphism seemed to be lower in this sample of BH population as compared to the Caucasians genotype data. Furthermore, no signifi cant diff erence in allele frequencies for CYPC*, CYPC*, and CYPD* was demonstrated between diabetic and nondiabetic subjects. Th us, results form this study seem to indicate no relationship between CYPC, CYPC, and CYPD genotype and diabetes susceptibility in Bosnian population. Th is in part may refl ect a limited study population included in our study and would require larger cohorts to reveal potential relationships between analysed CYP genetic variants and diabetes risk. In addition, it would be pertinent to further explore possible eff ects of CYP genetic variations on therapeutic and adverse outcomes of oral antidiabetics, which might be the key in optimising therapy for individual patient with Type  diabetes. ©  Association of Basic Medical Sciences of FBIH. All rights reserved

Numerous studies conducted on acetylsalicylic acid (ASA, aspirin) confirmed that ASA inhibits proliferation and induces apoptosis in various types of human cells. Therefore, it was of interest to examine possible effects of different concentrations of ASA on viability and proliferation of lymphocytes in the cell culture. After separation from blood, lymphocytes were suspended in RPMI 1640 medium and cultured at 37 degrees C. Solution of ASA was added to cultures after 24 h, in final concentrations of 1, 3 and 5 mmol/l. After 48 h, proliferative response was evaluated by WST-1 assay. Significant difference in viability between controls and cell cultures treated with ASA in three different concentrations was observed (p<0.01). Percents of viable cells in cultures after application of 1, 3 and 5 mmol/l ASA were 9.9%, 2.5% and 16.9% (compared to controls), respectively. To determine whether this cytotoxic effect was result of induction of apoptosis, DNA from cell cultures was isolated and subjected to agarose gel electrophoresis. Fragmentation of DNA was not detected, excluding apoptosis as possible cause of cytotoxic effects. Addition of ASA caused change of initial extracellular pH value for each treated culture. After addition of 1 mmol/l ASA, pH of culture was 7.19, after 3 mmol/L, 6.99 and after addition of 5 mmol/l solution, pH was 6.75. Decreased lymphocyte viability could be attributed to either the effects of the added substance or possible further acidification of cell cultures during three days of incubation.

Diabetes mellitus can be looked upon as an array of diseases, all of which exhibit common symptoms. While pathogenesis of IDDM (insulin dependant diabetes mellitus) is well understood, the same is not true for diabetes mellitus type II. In the latter case, relative contribution of the two factors (insulin resistance or decreased insulin secretion) varies individually, being highly increased in peripheral tissues and strictly dependant on insulin for glucose uptake. Moreover, in patients with diabetes mellitus type II, disbalance at the level of regulation of glucose metabolism as well as lipid metabolism has been noted in skeletal muscles. It is normal to assume that in this type of diabetes, these changes are reflected at the level of total activity of enzyme creatine kinase. This experimental work was performed on a group of 80 regular patients of Sarajevo General Hospital. Forty of those patients were classified as patients with diabetes type I and forty as patients with diabetes type II. Each group of patients was carefully chosen and constituted of equal number of males and females. The same was applied for adequate controls. Concentration of glucose was determined for each patient with GOD method, while activity of creatine kinase was determined with CK-NAC activated kit. Statistical analysis of the results was performed with SPSS software for Windows. Obtained results point out highly expressed differences in enzyme activity between two populations examined. Changes in enzyme activity are more expressed in patients with diabetes type II. Positive correlation between concentration of glucose and serum activity of the enzyme is seen in both categories of diabetic patients which is not the case for the patients in control group. At the same time, correlation between age and type of diabetes does exist . This is not followed at the level of enzyme activity or concentration of glucose.

lower HbA1c in normal glycaemia state but higher HbA1c in hyperglycaemia state (before pharmacological treatment was indicated in patients with type 2 diabetes); metformin, but not sulfonylurea treatment reverses the genetic impact on HbA1c