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Introduction: In the present study we investigated the performance, precision, and recovery of three different automated methods in determining cancer antigen (CA) 15-3 levels.Methods: Serum samples were obtained from 60 hospitalized female patients. As controls, commercially available samples were used. Cancer antigen (CA) 15-3 levels were measured using ARCHITECT CA 15-3, Elecsys® CA 15-3, and Vitros CA 15-3 immunoassays. A comparison of the results between the three methods was conducted, and the precision and recovery were analyzed.Results: Coefficient of variations (CVs), determined with low- and high-level-CA 15-3 control samples, and reproducibility values were: 2.56-2.80% and 3.10-4. 20% for ARCHITECT i2000SR immunoassay analyzer; 3.50-5.55% and 4.88-6.47% for Cobas E 601 analyzer; 3.30-4.0% and 4.30-4.80% for VITROS 5600 Integrated System, respectively. The percent recoveries were 95-98% for Elecsys® CA 15-3 assay, 93-105% for Vitros CA 15-3 assay, and 92-95% for ARCHITECT CA 15-3 assay. Method comparison results demonstrated correlation coefficient (r) in range from 0.994 to 1. The average CA 15-3 concentrations measured by Vitros, ARCHITECT, and Elecsys® were 157.24 +/- 329.75 U/mL, 100.91 +/-213.75 U/mL, and 80.93 +/- 173.29 U/mL, respectively.Conclusions: Tumor marker CA 15-3 in individual patients should be monitored using the same immunoassay method, reagents, and analyzer. Different immunoassays tested on different analyzers, often show large discrepancies in reported values for individual patients. Different immunoassay technologies quantify analytes of clinical interest using monoclonal or polyclonal antibodies. Thus, the usage of antibodies with different specificities could explain the differences in CA 15-3 serum values between different methods.

A. Stanciu, N. Serdarević, A. Hurduc, M. Stanciu

Abstract Objective. To investigate the potential role of interleukin 4 (IL-4), interleukin 10 (IL-10) and high-sensitivity C-reactive protein (hs-CRP) as serum biomarkers of persistent/recurrent disease in papillary thyroid carcinoma (PTC) with/without Hashimoto's thyroiditis (HT). Methods. Eighty consecutive patients (64 F/16 M, 43.2 ± 12.7 years) with PTC and 40 (37 F/3 M, 40.6 ± 12.3 years) with papillary thyroid carcinoma associated with Hashimoto's thyroiditis (PTC + HT) were evaluated before radioiodine therapy. A control group of 20 patients with HT without thyroid cancer (18 F/2 M, 47.3 ± 2.8 years) was included in the study for the comparison of cytokine levels. Results. No meaningful differences were found in clinical outcomes between PTC and PTC + HT groups (47.5% vs. 45% persistent/recurrent disease). Serum IL-4, IL-10 and hs-CRP levels were higher in patients with persistent/recurrent disease compared to those without recurrence (p < 0.001). IL-4, IL-10 and hs-CRP were also found in substantially higher concentrations in PTC + HT patients with persistent/recurrent disease than in patients with HT or PTC (with or without recurrence) (p < 0.01). Positive correlations were observed between IL-4, IL-10, hs-CRP and thyroglobulin (Tg) (r between 0.48 and 0.56, p < 0.005) or antithyroglobulin antibodies (TgAb) (r between 0.63 and 0.80, p < 0.002) in PTC and PTC + HT patients with persistent/recurrent disease. Conclusions. Increased levels of serum IL-4, IL-10 and hs-CRP are associated with persistent/recurrent disease in PTC and PTC + HT patients. Our results suggest that these biomarkers might be used to improve patient stratification according to the risk of recurrence, especially in patients with PTC + HT, where Tg levels are not reliable due to presence of TgAb.

AIM To investigate effects of post-sampling analysis time, a type of blood samples and collection tubes on blood gas testing. METHODS This study included 100 patients at the Clinic for Pulmonary Diseases, Clinical Centre Sarajevo. The partial pressure of oxygen (pO2) and carbon dioxide (pCO2), and the oxygen saturation level of hemoglobin (sO2) were analyzed in the arterial blood samples (ABS) and capillary blood samples (CBS) by a potentiometric method using a blood gas analyzer ABL 555 (Radiometer, Copenhagen, Denmark). Paired measurements of ABS were performed within 15 minutes and after 60 minutes from sampling and compared. The results of CBS obtained within 15 minutes were compared with matching ABS results, as well as the results obtained from CBS within 15 minutes taken into glass and plastic tubes. RESULTS pO2 and sO2 values were significantly lower after 60 minutes compared to those within 15 minutes in ABS (9.20±1.89 vs. 9.51±1.95 and 91.25±5.03 vs. 92.40±4.5; p<0.01, respectively). Values of pO2 and sO2 in CBS were significantly lower than values obtained in ABS (8.92±2.07 vs. 9.51±1.95 and 91.25±4.86 vs. 92.40±4.50; p<0.01, respectively). Obtained pO2 and sO2 values in CBS in the plastic tubes were higher than those in the glass tubes (8.50±1.98 vs. 7.89±2.0 and 89.66±11.04 vs. 88.23±11.22, p<0.01 respectively). pCO2 blood values were not influenced significantly (p>0.05). CONCLUSION The length of post-sampling analysis time, a type of blood samples and collection tubes have significant impact on blood oxygen parameters. Analysis within 15 minutes after blood sampling is considered as appropriate.

Introduction: The diagnostic utility of B-type natriuretic peptide (BNP) has prompted interest in its use as an aid in the detection of early heart failure and assessment of diseases. The first objective of this study was measurement of BNP and troponin I (TnI) blood levels in patients with acute myocardial infarction (AMI) and unstable angina. The second objective of this study was to find a correlation between TnI and BNP in blood.Methods: The concentrations of BNP and TnI in 150 blood levels were determined using CMIA (chemiluminescent microparticle immunoassay) Architect and 2000 (Abbott diagnostics). The retrospective study included 100 patients who were hospitalized at the Department of Internal Medicine of the University Clinical Center Sarajevo and 50 healthy control. The reference blood range of BNP is 0-100 pg/mL and TnI is 0.00-0.4 ng/mL.Results: In the patients with AMI the mean value of BNP is 764.48 ± 639.52 pg/mL and TnI is 2.50 ± 2.28ng/mL. The patients with unstable angina have BNP 287.18 ± 593.20 pg/mL and TnI 0.10 ± 0.23 ng/mL. Our studies have shown that the correlation between BNP and TnI was statistically significant for p< 0.05 using Student t test with correlation coefficient r = 0.36. Conclusions: BNP and TnI levels can help to identify the patients with a high risk for cardiovascular diseases.

Acute kidney injury (AKI) is a complex, frequent and serious clinical problem with high rate of mortality. Therefore there is a serious need for early detection of AKI, with a tendency to detect early stage--RISK dut to start with therapy as soon as possible and prevent irreversible changes in renal function. Study's purposes were to explore the rhythm of urine neutrophile gelatinase associated lipocalin (urine NGAL) concentration changes before and after cardiovascular surgery and compare results of urine NGAL values with results of serum creatinine and creatinine clearance as main diagnostic indicators of renal function in order to define role of urine NGAL biomarker in early diagnosis of acute kidney injure. In the prospective clinical study 150 cardiovascular surgery patients were included. Basal value and concentration of urine NGAL were tested 3, 6 and 12 hours after cardiovascular surgery, and concentration of serum creatinine was tested once per day first three days after surgery. Also creatinine clearance value was calculated according to Cockrof-Goult formula. After above mentioned, rate of acute kidney injure was estimated according to RIFLE criteria. The study results showed that the value of urine NGAL was elevated above cutt-off after cardiac operation in a group of patients who developed AKI (defined according to RIFLE criteria). There were statisticaly significant difference between all four measerments (p < 0.05). There were also moderate positive correlation (0.500 and 0.502) between urine NGAL values and percentage difference of serum creatinine and creatinine clearence. All that indicate that higer values of urine NGAL are followed by higher percentage difference of serume creatinine and creatinine clearence. By using of automated urine NGAL test detection of acute kidney injure is posible 24-48 hours earlier comparing with actual results acquired by determination of serum creatinine concentration. The results of this study will indicate urine NGAL as a reliable biomarker of early acute kidney injure. A combination of early and late markers of kidney damage (urine NGAL, serum creatinine) can greatly contribute to better control the outcome of all those who are a risk group for the development of AKI.

The study covered 600 patients, aim of our study was to investigate serum concentration of homocysteine and lipids at patients with ischemic stroke and vascular dementia. We determined concentrations of homocysteine, creatinine and blood lipids in patients’ serum. Homocysteine concentration reading in serum was measured by AxSYM apparatus, lipids and creatinine were determined directly by DIMENSION LxR. Mean concentrations of homocysteine in patients was higher then in healthy subjects. Our results show that the concentration of HDL cholesterol was significantly lower in the patients group than in the control group. In the group with ischemic stroke, the concentration of LDL cholesterol was higher than 4.3 mmol/L in about 56% of patients, whereas in the group with vascular dementia, was higher than the upper reference value in 60 % of patients. The Pearson correlation coefficient have show significant negative difference between serum homocysteine with HDL cholesterol concentrations in patients groups for p<0.05. Our results showed that homocysteine concentration is inversely proportionate to HDL cholesterol concentration, i.e. as homocysteine concentration in serum increases, HDL concentration falls. Homocysteine concentration increases with LDL cholesterol concentration in serum of the patients with vascular dementia, which points to homocysteine impact in further atherosclerosis development.

N. Serdarević, A. Stanciu

BACKGROUND SCC (squamous cell carcinoma) antigen (SSCA), a tumor marker, is already used for the diagnosis and follow-up of carcinoma of the cervix and lungs. The aim of our study was the determination of SCCA concentration in 96 serum samples using CMIA (chemiluminesecent microparticle immnoassay) on an Architect i2000 (Abbott Diagnostics) and MEIA (microparticle enzyme immnoassay) on an IMx Chemistry Analyzer (Abbott Diagnostics). METHODS In our investigation we used a CMIA (chemiluminesecent microparticle immnoassay) Architect i2000 and MEIA (microparticle enzyme immnoassay) Imx Chemistry Analyzer for SCCA determination. RESULTS All patients were hospitalized at the Department of Gynecologic Oncology and Department for Oncology at the University Clinics Center of Sarajevo. The normal serum range of SCCA should be < 0.1 microg/L. The quality control, precision and accuracy of the Architect i2000 were assessed. The quality control was done using quality control serums for low (X = 2.17 microg/L), medium (X = 10.23 microg/L), and high (X = 48.99 microg/L) range. The precision for the Architect i2000 has a CV of 3.44% to 4.44%. We established that the main difference between Architect i2000 and IMx was statistically significant at p < 0.05 according to Student's t-test. The correlation coefficient was r = 0.990. The some samples have higher concentrations on the Architect than on the IMx because the sensitivity of the Architect assay is high (< 0.1 microg/L). CONCLUSIONS The CMIA Architect technology is an applicable method significant in diagnostic of SCCA.

N. Serdarević, Lejla Zunic

Background: Cyclosporine has been shown effective drug in suppressing acute rejection in recipients of allograft organ transplants. Methods: The cyclosporine concentration of 96 blood samples was determined using CMIA (chemiluminesecent microparticle immnoassay) Architect i 2000 and FPIA (fluorescence polarization immunoassay) AxSYM Abbott diagnostic. All patients have transplantation of kidneys and were hospitalized at Department of Nephrology at the Clinical center of University of Sarajevo. The reference serum range of cyclosporine for kidney organ transplantation for maintenance lies between 50 and 150 ng/mL. The quality control, precision and accurancy of Architect i 2000 were assessed. Results: The quality control was done using quality control serums for low (= 91 ng/mL), medium (= 328 ng/mL) and high (= 829 ng/mL). We have used commercial BIORAD controls and got reproducibility CV 5.83 % to 13 % for Architect i 2000. It was established that the main difference between Architect i 2000 and AxSYM and it was statistically significant for P < 0.05 according to Student t-test. Correlation coefficient was r = 0.903. Conclusion: The CMIA Architect assay has significant reduced cyclosporine metabolite interference relative to other immunoassay and is a convenient and sensitive automated method to measure cyclosporine in whole blood.

Introduction: Serum CA15-3 has been one of the most reliable tumor markers used in monitoring of breast cancer patients. To increase its sensitivity, the combined measurement of other tumor markers (CEA and ferritin) with CA15-3 was investigated. The aim of this study was determination of CA 15-3, CEA and ferritin in female patients with breast cancer, lung cancer and mastitisMethods: 300 patients with carcinoma, hospitalized at Department of Gynecologic Oncology and Department for Oncology at the University Clinics Center of Sarajevo and 200 healthy subjects were compared.Results: In patients with breast cancer the mean value of tumor markers were CEA 155.61 ng/mL, CA 15-3 106.38 U/mL and ferritin 197.03 ng/mL. In patients with lung cancer CEA was 58.97 ng/ml, CA 15-3 40.62 U/mL and ferritin 544.16 ng/mL. Patients with mastitis had CEA 5.17 ng/mL, CA 15-3 112.67 U/mL and ferritin 174.92 ng/mL. The control group had values of tumor markers CEA 1.62 ng/mL, CA 15-3 11.72 U/mL and ferritin 85.35 ng/mL. We found good correlation between CA 15-3 and CEA correlation coeffi cient was r = 0.750. There was a low correlation between CA 15-3 and ferritin with correlation coeffi cient r = 0.274.Conclusions: The CA 15-3 and CEA are useful markers in patients with confi rmed diagnosis of breast and lung cancers. The ferritin concentration has not increased in patients with breast cancer but it increased inlung patients. The future study has to make investigations of tumor markers and ferritin in different stage of breast cancer.

Introduction: The aim of this study was determination of troponin I at serum using Architect (Abbott) and AxSYM System (Abbott). Troponin is regulatory subunit of the troponin complex associate with actin filament within muscle cells and it is a marker for diagnosis of myocardial damage.Methods: We used Architect STAT chemiluminescent microparticle immunoassay (CMIA) and AxSYM microparticle Enzyme Immunoassay (MEIA), techniques for quantitative determination of cardiac TnI in human serum or plasma. At our study we have proved precision, reproducibility and accuracy from both methods. The investigation included patients (n=119) who have myocardial infarction or ischemic heart damage and were treated at cardiology, emergency, internal medicine and neurology unit in Clinical Center University in Sarajevo.Results: The precision for three controls using Architect STAT TnI asssay technology were 3.6 – 5.2 % and reproducibility was 3.7 to 5.6 %. The AxSYM STAT TnI has precision for three controls 4.3–6.6 % and reproducibilitywas from 4.8 to 7.8 %. We have got very good correlation between Architect and AxSYM technology r = 0.999 in the investigation of troponin I in serum.Conclusions: We can conclude that chemiluminescent troponin assay I (Architect) showed good analytical performance and gave new possibility at troponin I determination.

Introduction: The aim of this study is to examine whether moderate hiperhomocysteinemia is an independent risk factor for cerebral infarction.Methods: We have measured homocysteine levels in 50 patients with ischemic stroke during acute phase and postacute phase, 50 patients diagnosed with vascular dementia and healthy group of 50 subjects. Homocysteine concentration in serum was measured, on the basis of fluorescent polarisation measuring.Results: The study demonstrated that homocysteine concentration was 16.93 µmol/L in the patient group with ischemic stroke, and in the group of patients with vascular dementia was 20.39 µmol/L. Homocysteine increases during the postacute phase of ischemic stroke after 7 days for 1.54 µmol/L and 14 days for 3.66 µmol/L compared to the concentration of homocysteine after the first hours of hospitalization. Using Wilcoxon signed ranks and Mann-Whitney (P < 0.05) tests we got significant difference between homocysteine concentration at acute phase and post-acute phase of ischemic stroke and it was significant difference between concentrations of homocysteine in the acute and post-acute phase of ischemic stroke and vascular dementia. The Spearman correlation test was found signifiant correlation between the number of strokes and the concentration of homocysteine in serum of patients with vascular dementia.Conclusions: The homocysteine concentration rises significantly during of acute phase of ischemic brain stroke, and it is significantly increased during post-acute phase, which is a predictor factor for further development of vascular dementia, or a new ischemic brain stroke.

N. Serdarević, F. Kozjek, Ivan Malešič

Th e lithium ions concentration in human serum and saliva was determinated using dry-slide technology Vitros  Analyser (Ortho Clinical Diagnostic) and atomic absorption spectrometry Perkin Elmer  (AAS). We analyzed lithium ions in  serum and saliva specimens of patients after oral administration of lithium carbonate (x mg) Jadran, Galen Laboratory Rijeka. Saliva and blood were taken  and  hours after the last dose. At the same time lithium ions at samples of blood and saliva were determined with bouth methods which showed high level of correlation. Th e mean diff erence of lithium ions between saliva and serum was statistically signifi cant for p<, using t student test. At saliva we got constant of elimination Kel = ,-h and elimination half life (t/) was t/=, h. For serum was t/=  h what means that lithium ions elimination is slower from saliva then from serum. Th at is the reason why probably concentration at saliva is higer then at serum. Lithium elimination is two compartment pharmacokinetic model where important part of compartment are saliva and salivary glands. At a certain point in medical treatment it could be expected to use controlled determination of lithium ions in saliva with serum as control.

N. Serdarević, F. Kozjek, Aleš Goropevšek

The lithium ions concentration in human serum and saliva was determined using dry-slide technology Vitros 250 Analyser (Ortho Clinical Diagnostic) and atomic absorption spectrometry Perkin Elmer 403 (AAS). We analyzed lithium ions in 100 serum and saliva specimens of patients after oral administration of lithium carbonate (3 x 300 mg) Jadran, Galen Laboratory Rijeka. Saliva and blood were taken 2 and 12 hours after the last dose. At the same time lithium ions at samples of blood and saliva were determined with both methods which showed high level of correlation. The mean difference of lithium ions between saliva and serum was statistically significant for p<0.05 using t student test. At saliva we got constant of elimination Kel = 0.02(-1)h and elimination half life (t(1/2)) was t(1/2)=34.6 h. For serum was t(1/2)= 24 h what means that lithium ions elimination is slower from saliva then from serum. That is the reason why probably concentration at saliva is higher then at serum. Lithium elimination is two compartment pharmacokinetic model where important part of compartment are saliva and salivary glands. At a certain point in medical treatment it could be expected to use controlled determination of lithium ions in saliva with serum as control.

N. Serdarević, Ivan Malesić, F. Kozjek

The lithium ions concentration in human serum was determined using Dry-slide technology Vitros 250 Analyser (Ortho Clinical Diagnostic), atomic absorption spectrometry (AAS) method Perkin Elmer 403 and ion-selective electrode (ISE) potentiometry AVL 9181. We compared lithium ions results in sample sera between these methods. Our reference method was AAS. We analyzed lithium ions concentration in 23 sera samples of patients after oral administration of lithium carbonate (3x 300mg) Jadran, Galen Laboratory Rijeka, by dry-slide technology, AAS and ISE methods. The quality control, precision, reproducibility and accuracy for Vitros dry slide technology were assessed. We established that the main difference between AAS method and dry slide technology was not statistically significant at p< 0.05 according to Student t-test. Therefore, the dry slide technology may be a useful alternative or it may even replace other methods, such as AAS. The main difference between dry slide technology and ISE methods was statistically significant at p<0.05 using Student t-test. By ISE method, we obtained considerably higher results, which may be explained by the presence of electrolytes or medicaments interfering with lithium ions.

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