BACKGROUND Clinical Chemistry is the backbone of medical treatment, diagnostics, and prevention. The laborato-ries are trying to improve the quality and to reduce diagnostic errors and processing time and safeguard trace-ability of all laboratory procedures to ensure patient safety. Six sigma belongs to statistical quality control and provides a new methodology for measuring and improving process performance in laboratory. METHODS Activities of AST, ALT, CK, LDH, Amy, and γ-GT were determined by standard kinetic methods on a Vitros 5600 biochemistry analyzer. Two daily quality controls (Verifier I and Verifier II) were run over 60 days. Total percent CV was calculated from routine daily QC. Between-instrument bias was also calculated from daily QC. RESULTS The calculated sigma metrics for AST were 6.9 and 3.8; for ALT 9.3 and 5.6; for CK 6.6 and 5.3; LDH 5.2 and 5.2; for γ-GT 4.9 and 2.7; and for amylase 8.7 and 7.1. Analytical performance for AST, ALT, CK, LDH, and Amylase is world class. On the other hand, γ-GT analytical performance is poor. CONCLUSIONS Six Sigma benefits from earlier quality management approaches that creates new challenges for medical laboratories.

Summary Background HIV infection is characterized by progressive depletion of CD4+ T cells due to their reduced synthesis and increased destruction followed by marked activation and expansion of CD8+ T lymphocytes. CD4/CD8 ratio was traditionally described as a marker of immune system ageing in the general population, but it increasingly appears as a marker of different outcomes in the HIV-infected population. The main objective of this study is to examine the power of CD4/CD8 ratio in predicting the occurrence of metabolic syndrome (MetS) in HIV-positive patients receiving cART therapy. Methods 80 HIV/AIDS subjects were included in a retrospective case-control study. Flow cytometry was used to determine the percentage of CD4+ and CD8+ cells in peripheral blood of these patients. The values of biochemical parameters (triglycerides, HDL, blood sugar, blood counts), immunological parameters (CD4/CD8, PCR), anthropometric measurements and type of cART therapy were evaluated in this study. Results After six months of cART therapy 19 (23.8%) subjects had all the elements necessary for making the diagnosis of MetS. Using multivariate analysis CD4/CD8 ratio was statistically significant (p < 0.05) and had the largest effect on development of MetS (Wald = 9.01; OR = 0.45), followed by cART (Wald = 7.87; OR = 0.10) and triglycerides (Wald = 5.27; OR = 1.7). On the other hand, body weight and waist circumference showed no statistically significant effect on the development of MetS after six months of cART, p > 0.05. Conclusions CD4/CD8 ratio proved to be a significant marker for prediction of metabolic syndrome in HIV/AIDS patients.

Introduction: Diabetes mellitus type 2 has become a global health-care problem of modern society due to a pronounced increase of prevalence to pandemic proportions and vascular complications. At present, glycated hemoglobin (HbA1c) is widely accepted as a measure of glycemic control in established diabetes. The aim of this study was to analyze the lipid profile in serum of patients with diabetes mellitus type 2, and its relationship with HbA1c levels. Methods: The observational cross-sectional study included 60 diabetic patients, 30 men, and 30 women, age 32–94 years. Patients were assigned into two groups based on HbA1c values; Group 1: HbA1c ≤ 7% (good glycemic control) and Group 2: HbA1c > 7% (poor glycemic control). We analyzed the concentration of glucose, HbA1c, and lipid profile including total cholesterol levels, triglycerides (TAG), low-density lipoproteins (LDL), and high-density lipoproteins (HDL). Results: Significantly lower values of glucose concentration, TAG and the ratio TAG/HDLc were obtained in the group of patients with good glycemic control. (p < 0.0005) Patients with good glycemic control had lower values of Castelli 1 and Castelli 2 index, and atherogenic index of plasma, compared to patients with poor glycemic control, but this difference was not significant. (p > 0.005) Our study revealed a significant positive correlation between HbA1c and triglyceride level (r = 0.375; p = 0.003) and HbA1c and ratio triglyceride/HDLc (r = 0.335; p = 0.009). Conclusion: HbA1c can also be used as a predictor of dyslipidemia in type 2 diabetics in addition to as a glycemic control parameter.

Summary Background: There is still no reliable, specific biomarker for precision diagnosis and clinical monitoring of systemic lupus erythematosus. The aim of this study was to investigate the importance of the determination of immunofenotypic profiles (T, B lymphocytes and NK cells) and serum cytokine concentrations (IL-17 and IFN-alpha) as potential biomarkers for this disease. Methods: The study included 55 patients with SLE and 25 healthy controls. The proportion of T, B, NK cells were assessed in peripheral blood using flow cytometric assays while the serum cytokine concentration (IL-17 and IFNalpha) was determined by ELISA test. Results: ROC curve analysis showed good accuracy to distinguish between patients and healthy individuals for activated T cells (AUC=0.798; p<0.001), Treg (AUC= 0.651; p=0.036), and memory B cells (AUC=0.285; p=0.002). We found statistically significant difference (p=0.036) in the levels of serum IL-17 between patients with SLE (IL-17=49.27 pg/mL) and controls (IL-17= 28.64 pg/mL). Conclusions: Significant increase in the relative number of Treg lymphocytes, and decrease in memory B cells, as well as decrease level of IL-17, in SLE patients may be implicated in the pathogenesis of the disease. These parameters, as biomarkers, could distinguish SLE patients and no-SLE patients. Monitoring subpopulations of immune cells in peripheral blood using flow cytometry provides insight into abnormal T and B cell function in SLE. Progress in understanding the immunity at SLE, results in concrete benefits for the SLE patients, which include new clinical management and therapeutic strategies.

BACKGROUND Metastasis-Associated in Colon Cancer-1(MACC1) was first identified as a transcriptional activator of the HGF/MET pathway. Deregulation of HGF/MET signaling is reported as a prognostic marker for tumorigenesis, early stage invasion, and metastasis which is associated with poor clinical outcome in breast cancer patients. The aim of the present study was to further investigate the prognostic or predictive value of MACC1 expression in breast cancer. MATERIALS AND METHODS We analyzed the MACC1 expression in 105 primary breast cancer samples by Western-Blot analysis and immunohistochemistry. RESULTS A significant correlation of high MACC1 expression with shorter disease-free survival was found within the group of lymph-node-negative patients. Additionally, an association of high MACC1 expression and shorter disease-free survival was observed within estrogen receptor positive tumors and patients without adjuvant chemotherapy. CONCLUSION Our results support a biologic role and potentially open the perspective for the use of MACC1 as a prognostic marker for treatment decision in breast cancer patients.

Summary Background: The production of erythrocytes is regulated by the hormone erythropoietin (EPO), which maintains the blood haemoglobin (Hb) levels constant under normal conditions. Human EPO is a glycoprotein hormone and its synthesis is controlled by the hypoxia-inducible transcription factor. The aim of this study was to establish EPO and Hb levels in patients with chronic kidney disease (CKD), as well as in control subjects, and to investigate the relationship between these parameters. Methods: This cross-sectional, observational study included 356 subjects with CKD divided into 4 subgroups according to their glomerular filtration rate (GFR). The control group consisted of 206 age and sex matched healthy subjects with GFR rate ≥90 mL/min/1.73 m2. EPO, Hb and serum creatinine levels were determined by using immunochemical and spectrophotometric methods. GFR was determined using the MDRD formula. Results: The CKD patients had significantly lower levels of haemoglobin (p<0.0005) and hematocrit (p<0.0005) compared to control group. Our results showed that Hb levels decreased, whereas serum creatinine increased with the increasing renal failure. The CKD patients in all four groups had significantly lower (p<0.0005) Hb levels, and significantly higher (p<0.0005) creatinine levels compared to the control group. The median EPO in group I and II were significantly higher (p=0.002; p=0.018), while median EPO in group III and IV were significantly lower (p=0.03; p=0.011) compared to the control group. Conclusions: In patients with CKD, GFR positively correlated with Hb and EPO, while the correlation between GFR and serum creatinine was negative.

Introduction: Transfusion is an activity that assures sufficient supply of blood and blood components to treat the sick and injured. In transfusion departments is necessary to timely provide adequate amounts of blood and blood products for various surgical procedures. Material and methods: To determine the total amount of preoperative requirements (BT/AB and BT/AB/MT) for blood and blood products in surgical departments of General Hospital “Prim. Dr. Abdulah Nakas” in the period from June 1, 2014 – December 31, 2014 and analyze the requirements for blood in relation to surgical procedures, surgical discipline, period, age and gender of patients. To determine the maximum consumption levels surgeries. Results: The total amount of preoperative requirements for blood and blood products in surgical departments amounted to 927. Almost the same number of requests with a slightly higher percentage was in December and October and the lowest in June. The average age of patients was 52.2±20.1 years with the youngest patient aged 9 and the oldest at the age of 97 years. Women were more prevalent with 686 or 74% of the time compared to men. The largest number of requests for surgery elective cesarean section 208 (22.5%) with delivery wards, then for surgery or hip replacement 98 (10.6%). Maximum consumption for surgical operations Hysterectomy totalis abdominals 15 doses of blood. Conclusion: The largest number of requests were for elective cesarean section 208 (22.5%) with delivery wards, then for surgery or hip replacement with 98 (10.6%). Maximum consumed doses had gynecology surgery at Hysterectomy totalis abdominals 15 doses of blood, then orthopedics surgery at Primary hip prosthesis 11 doses.

Introduction: The hospital blood bank (HBB) need to timely provide adequate amounts of blood and blood products for surgeries. For various surgical programs are performed assessments of the average number of blood doses needed for surgery. By using two types of requisitions BT/AB (blood type/antibody) and BT/AB/MT (blood type/antibody/match test) for pretransfusion immunohaematological testing in General Hospital “Prim. Dr. Abdulah Nakas” is achieved more rational consumption of blood and blood derivatives and financial savings through reduced number of matching tests (MT). Goal: To determine the total amount of pre-operative requisitions (BT/AB and BT/AB/MT) for blood and blood products at surgical departments of the General Hospital “Prim. Dr. Abdulah Nakas” in the period from June 1, 2014 – December 31, 2014 and analyze the consumption/return of blood in reserve in relation to the surgical disciplines, the total number of savings in MT. Conduct assessments MSBOS (Maximum Surgical Blood Ordering Schedule). Results: The total amount of preoperative requisitions for blood and blood products in surgical wards was 927 requests from which 623 demands or 67.2% is tested by BT/MT, while 304 or 32.8% was tested by BT/AB/MT. Transfused in total was 617 units of blood and blood products, 275 units were not transfused. Probability of transfusions for surgery was 51.3, the highest in the case of surgical intensive care 70.4 and the lowest for the department of general surgery 37.2%. Assessment of indicators of efficient resource management indicates they are the best at the delivery ward 0.89, while a total for surgical wards is 0.69. In total for surgery on the average were required 2.1 units of blood. By using two types of requisitions for pretransfusion immunohaematological testing (BT/AB and CG/AB/MT) is achieved more rational use of MT. In 623 requests for BT/AB only 61 MT were performed. Average of blood units issued in accordance with these requirements is 0.08 and the savings in the number of MT amounts to 562.

Introduction: Anti GAD (antibodies on glutamic acid decarboxylase) and anti-IA2 antibodies (against tyrosine phosphatase), today, have their place and importance in diagnosis and prognosis of Type 1 diabetes. Huge number of patients with diabetes mellitus type 1 have these antibodies. Insulin antibodies are of critical importance in diagnosis of diabetes mellitus type 1 for pediatric population. Materials and methods: During 2014, the samples of 80 patients from Clinical Center University Sarajevo (CCUS) Pediatrics clinic’s, Endocrinology department were analyzed on anti-GAD and IA2 antibodies. The samples of serums of all patients were analyzed with ELISA tests using Anti GAD ELISA (IgG) kites from EUROIMMUN company. These are quantitative in vitro tests for human antibodies against decarboxylase of glutamine acid (GAD) and IA2, in serum or EDTA plasm. Results: During the period of one year, in CCUS’s Organizational unit, Institute for Clinical Immunology, 80 samples of patients with anti GAD and IA2 antibodies were analyzed. Out of total number of samples, 41 were male patients, or 51% and 39 female, or 49%. The youngest patient was born in 2012, and the oldest in 1993. Age average was represented by the patients born in 2001. Share of positive results for IA2 antibodies and GAD antibodies was 37% for IA2 antibodies, and 63% for GAD antibodies. Discussion: During an autoimmune – mediated Diabetes mellitus type 1 leads to T-cell mediated destruction of beta cells of pancreatic islets, reduced production of insulin and glucose metabolism. Studies have shown that these bodies are the most intense single marker for identifying persons with increased risk for diabetes development.

Summary Background: High-density lipoproteins (HDL) have atheroprotective biological properties: antioxidative, anti-apoptotic, anti-inflammatory, and they have the efflux capacity of cellular cholesterol. Plasma mRNA analysis can be used to investigate statin pleiotropy in vivo as a new analytical tool for non-invasive assessment of gene expression in vascular beds. The aim of this study was to assess the pleiotropic effects of atorvastatin in stable angina patients with highrisk values (group A) as compared with patients who had borderline and desirable HDL-cholesterol (HDL-C) values (group B). Methods: The atorvastatin therapy (20 mg/day) was given to forty-three patients with stable angina for 10 weeks. We investigated three statin pleiotropy-targeted genes: intercellular adhesion molecule-1, chemokine (C-C motif) ligand 2 and cathepsin S and assessed by gel electrophoresis gradient the effects of atorvastatin on HDL size and subclasses. Results: In group A, after therapy, HDL-C concentration was significantly increased but not in group B. Atorvastatin lowered plasma chemokine (C-C motif) ligand 2 and intercellular adhesion molecule-1 mRNA levels in both groups, but did not change the plasma cathepsin S mRNA levels. In group A only, baseline total bilirubin showed negative cor relations with the genes of cathepsin S (r=-0.506; p=0.023) and significantly increased after therapy. Conclusion: HDL-C and bilirubin can be promising therapeutic targets in the treatment of cardiovascular diseases. Analysis of cell-free mRNA in plasma might become a useful tool for estimating statin pleiotropy Kratak sodržaj Uvod: Lipoproteini velike gustine (HDL) imaju ateroprotektivne biološke osobine: antioksidativne, antiapoptotičke, antiinflamatorne kao i kapacitet da izvlače holesterol iz ćelija. Analiza plazmatske mRNA može da se koristi za ispitivanje plejotropnih efekata statina in vivo kao novo analitičko sredstvo za neinvazivnu procenu ekspresije gena u zidu krvnog suda. Cilj ove studije je bio da se procene plejotropni efekti atorvastatina kod pacijenata sa stabilnom anginom sa visokorizičnim vrednostima (grupa A) u odnosu na pacijente sa graničnim i poželjnim vrednostima HDL holesterola (HDL- C) (grupa B). Metode: Četrdesettri pacijenta sa stabilnom anginom su primala terapiju atorvastatinom (20 mg/dan) 10 nedelja. Mi smo ispitivali tri gena značajna za plejotropno delovanje statina: intracelularni adhezioni molekul-1, hemokin (C-C motiv) ligand 2 i katepsin S i procenjivali smo efekte atorvastatina na veliCinu i raspodelu HDL subfrakcija promoću elektroforeze na poliakrilamidnom gradijent gelu. Rezuttati: U grupi A, posle terapije, HDL-C koncentracija se značajno povećala, ali ne i u grupi B. Atoivastatin je snizio plazmatski nivo hemokin (C-C motiv) liganda 2 i intracelularnog adhezionog molekula-1 mRNA u obe grupe, ali nije promenio plazmatski nivo gena za katepsin S. Samo u grupi A, ukupni bilirubin je pokazao negativnu korelaciju sa genom za katepsin S (r=-0,506; p=0,023) pre zapotinjanja terapije i značajni porast nakon terapije atorvastatinom. Zaključak: HDL-C i bilirubin mogu biti obećavajući terapijski ciljevi u lečenju kardiovaskularnih bolesti. Analiza slobodne mRNA (eng. cell-free mRNA) u plazmi može postati korisno sredstvo za procenu plejotropnog delovanja statina

Introduction: Hypoxia is a basic stimulant in production of erythropoietin (EPO). The primary function of erythrocytes is the transport of oxygen to tissues. Erythropoietin stimulates erythropoiesis which leads to increased production of erythrocytes- their total mass. This increases the capacity of the blood to carry oxygen, reduces the hypoxic stimulus and provides a negative feedback of stopping EPO production. The aim of this study was to establish a quantitative relationship between the concentration of erythropoietin, hemoglobin and hematocrit in different values of renal insufficiency. Material and methods: The survey was conducted on 562 subjects divided into two groups: with and without renal insufficiency. EPO, hemoglobin, hematocrit, serum creatinine and additional parameters iron, vitamin B12, and folic acid were determined by using immunochemical and spectrophotometric methods and glomerular filtration rate (GFR) was calculated as well. Results: EPO values (median) grow to the first degree of renal insufficiency, as compared to EPO values of healthy subjects, this increase is statistically significant, p=0.002. With further deterioration of renal function the values of EPO between all pathological groups are decreasing, and this decrease is statistically significant between first and second degree of renal insufficiency (RI) p<0.001. In the group of healthy subjects EPO is correlated rho = -0.532, p <0.0005 with hematocrit. The correlations are negative and strong and can be predicted by regression line (EP0 = 41.375- Hct * .649; EPO = 61.41–Hb * 0.355). In the group of subjects with the first degree of renal insufficiency EPO is in correlation with hematocrit rho=-0.574, p<0, 0005. It is also correlated with hemoglobin rho=-0.580, p< 0.0005. The correlation is negative (EP0= 42.168- Hct * 0.678). In the group of subjects with the third degree of renal insufficiency EPO is in correlation with hemoglobin rho=0.257, p=0.028. The correlation is medium strong and positive. In the group of subjects with third and fourth degree of renal insufficiency EPO is not in correlation with hemoglobin and hematocrit p>0.05. Conclusion: Renal dysfunction, depending on the level of RI effects differently on the biosynthesis of EPO in a diseased kidney, and consequently it also has a different effect on biosynthesis of HB in bone marrow and its content in the blood.

Background: The serum prostate specific antigen for the early detection and screening for prostate cancer are very common used among physicians as the best screening tool for prostate cancer. The result of prostate specific antigen levels discriminates whether or not a prostate biopsy should be performed. The lack of specificity is a limitation of PSA as tumor marker, increased PSA concentrations are found not only in patients with prostate cancer but also in patients with benign prostatic disease. The object of this study was to improve the specificity and sensitivity of prostatic cancer detection. We evaluated total PSA levels, free PSA levels and the prostate volume in asymptomatic patients which came for routine check without medical history of prostate cancer. Methods: We received medical record of 90 patients between 50-60 years. Total and free PSA in serum was measured with the analyzer Architeckt i2000 SR. Prostate volume was determined by transrectal ultrasound. Results : The ratio of total and free PSA levels to prostate volume was significantly (p < 0.001) between all three groups. It was observed that increased prostate volume correlates with increased level of total and free PSA in serum. Conclusion: Early studies have demonstrated the advantage of measuring prostate volume with PSA total and free levels in serum as a useful tool for early diagnosis of prostate cancer. Data from this study on 90 patients with total PSA in the range from 0,22-7,0 ng/ml confirmed the well known correlation. All three parameters total PSA, free PSA and prostate volume showed significant correlation and a useful tool in prediction of prostate cancer for Bosnia and Herzegovina men.

Introduction: Glycated hemoglobin (HbA1c) is formed by non-enzymatic binding of glucose to the free amino group of the N-terminal end of the s-chain of hemoglobin A. HbA1c is representative of the mean blood glucose level over three months. The aim of the study was to evaluate the Hemoglobin A1c immunoturbidimetric assay performance on two different commercial systems.Methods: We evaluated the precision and trueness for determination of HbA1c in whole blood. Concentrations of total hemoglobin and HbA1c were evaluated on Dimension Xpand (Siemens) and Cobas 501 (Roche) analyzers. HbA1c was measured in a latex agglutination inhibition test. Commercial controls Liquichek Diabetes Control Level 1 and Liquichek Diabetes Control Level 2 (Bio Rad) at two levels were used for quality control. Analytical validation of HbA1c included: within-run imprecision, between-day imprecision, inaccuracy and comparison determination on the human samples on 2 systems: Dimension Xpand and Cobas 501 analyzers. Results: Within-run imprecision on the commercially controls for Level 1 is 4.5% and Level 2 is 3.2% between-day imprecision on commercially controls is 6.1% Level 1 and 5.1% Level 2 for respectively inac- curacy on commercially controls for Level 1 is 1.8% and Level 2 is 4.8%. Method comparison on human samples shows the correlation coefficient of 0.99.Conclusion: The presented results of the analytical evaluation methods for the determination of HbA1c showed an acceptable accuracy and precision.

ABSTRACT The study aim was to explore the pattern of the changes in haematological and iron status parameters of acute coronary syndrome patients through period 1-7 day of hospital admission in order to define the type of anaemia. Forty-one patients (15 female and 26 male patients, aged 36-81years) of the Clinic for Heart Disease and Rheumatism, University Clinical Center Sarajevo have been included in the cross-sectional study. Haematological and serum iron status parameters have measured on days 1 and 7 of hospital admission. A decrease in haemoglobin levels to <13g/dl in men and <12g/dl in women was notified as anaemia. A significant reduction in red blood cells count, haemoglobin, haematocrit (p<0.01), iron, total iron binding capacity (p<0.05) and significant ferritin elevation (p<0.05) within period the 1-7 day were noted. Percent of anemic patients on day 1 was 17.07 % with increase of number on day 7 (36.36%). Serum ferritin has been elevated with reduction of red blood cells count, mean cell volume, mean corpuscular haemoglobin (p<0.05); haemoglobin, haematocrit (p<0.01) at first 24 hours of admission in anemic versus non-anemic patients. Anemic patients had significantly lower values of percent transferrin saturation (p<0.05), red blood cells count, haemoglobin, haematocrit (p<0.01) compared to non anemic on day 7. A statistically significant negative correlations were obtained between serum iron and C-reactive protein; cardiac troponin I and total iron binding capacity (rho=-0.389, p<0.05; rho=-0.331; p<0.05, respectively). Observed changes in laboratory parameters through period 1-7 day indicate inflammatory type of anaemia in acute coronary syndrome. Key words: anaemia, acute coronary syndrome, haematological, iron status, parameters

Aim: The main aim of this research was to determine the influence of socioeconomic status and residence/living conditions on the status of oral health (e.g. health of mouth and teeth) in primary school students residing in Canton Central Bosnia. Methods: The study was designed as a cross-sectional study. Our research included two-phased stratified random sample of 804 participants. The quantitative research method and newly designed survey instrument were utilized in order to provide data on the oral health of the examined children. The alternate hypothesis foresaw that “there were significant statistical differences between the levels of incidence of dental caries in comparison to the incidence in children of different socioeconomic status. Results: The Chi square () of 22.814, degree of freedom (Df) = 8, coefficient of contingency of 0.163 and T-test (Stat) of–0.18334 showed that there were no significant statistical differences at p < 0.05 level between the primary school children from urban and rural areas. The obtained results showed that the caries indexes in elementary schools in Central Bosnia Canton were fairly uniform. Research showed that there were a difference in the attitudes towards a regular dental visits, which correlated with social-educational structure of the children's’ families. Conclusion: According to the results, we can see that the socioeconomic status of patients had an effect on the occurrence of dental caries and oral hygiene in patients in relation to the rural and urban areas, because we can see that by the number of respondents, the greater unemployment of parents in both, rural and urban areas, caused a host of other factors, which were, either, directly or indirectly connected with the development of caries.