Daria Pašalić
Department of Medical Chemistry, Biochemistry and Clinical Chemistry
Zagreb University School of Medicine
Šalata ul 2.
10 000 Zagreb, Croatia
Phone +385 (1) 4590 205; +385 (1) 4566 940
E-mail: dariapasalic [at] gmail [dot] com

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Pavić M, Bronić A, Milevoj Kopčinović L.P14-1: Procalcitonin in systemic and local bacterial infection. Biochemia Medica 2009;19(Suppl 1):S173-S174.
University Hospital of Traumatology, Zagreb, Croatia
Corresponding author:marpavic [at] yahoo [dot] com
Background: Procalcitonin (PCT) has been proposed as a marker of infection in critically ill patients. The aim of the study was to compare the serum concentrations of PCT in patients with systemic and local bacterial infection.
Materials and methods: According to clinical and microbiologic findings we divided 25 patients into two groups: group A-patients with systemic bacterial infection (positive blood cultures) and group B-local bacterial infection (other positive cultures). We determined in all patients C-reactive protein (CRP), PCT, leukocyte, non-segmented neutrophyle and platelet count. PCT was measured by enzyme-linked fluorescent immunoassay (miniVidas) and CRP by immunoturbidimetric method (Dimension Xpand). Leukocyte and platelet count were determined on a XT-1800i (Sysmex). Differential blood count was determined by microscopic method in stained blood peripheral smear. Data were analyzed using the Medcalc software. The level of significance was set at P < 0.05.
Results: The median concentrations of PCT were 1.32 (range: 0.13-7.37) ng/mL in group A and 0.21 (range: 0.05-9.07) ng/mL in group B and differences between groups were statistically significant (P = 0.038). A significantly higher median platelet count (P = 0.012) was found in group B (327, range: 40-325 x 109/L) as compared to group A (140, range: 40-325 x 109/L). In contrast, median values of CRP, leukocyte and non-segment neutrophyle were not significantly different between groups (P = 0.071; P = 0.189; P = 0.239, respectively).
Conclusion: According to obtained results, higher PCT concentrations combined with lower platelet counts in patients with systemic bacterial infection contribute to an improved stratification of patients at risk to develop major complications.
Milić M1, Berecki I2, Šerić V1.P14-2: Case report: usefulness of procalcitonin in infant sepsis complicated with liver failure. Biochemia Medica 2009;19(Suppl 1):S174.
1Department for Clinical Laboratory Diagnostics, Osijek University Hospital, Osijek, Croatia
2University Department of Pediatrics, Osijek University Hospital, Osijek, Croatia
Corresponding author:milic [dot] marija [at] kbo [dot] hr
Introduction: Procalcitonin is a precursor of hormone calcitonin produced by the C-cells of the thyroid gland. Bacterial infections increase its expression in all cell types and circulating levels of procalcitonin increase up to several thousand-fold. CRP is an acute-phase protein released by the liver. Procalcitonin is more sensitive and specific marker than CRP for severe bacterial infection.
Materials and methods: Procalcitonin was determined by Vidas Biomerieux. CRP, AST, ALT was determined by biochemistry analyzer Olympus AU 400. Complete blood count was determined by hematology analyzer Sysmex SF3000 and coagulation parameters by coagulation analyzer BCT Siemens and Siemens reagents.
Results: Two months old infant was hospitalized with clinical and laboratory signs of septic shock, disseminated intravascular coagulation, acute liver failure and hemorrhage in the gastrointestinal tract with consequent anemia. Results of laboratory analysis were: WBC 40.3 x 109/L, CRP 0.7 mg/L, procalcitonin 17.0 ng/mL, D-dimeri 1627 ug/L, fibrinogen 0.5 g/L, PT 0.14, aPTT 3.48, AST 475 U/L, ALT 68 U/L.
Conclusion: Procalcitonin proved as useful and valuable parameter as it correlated with septic condition, while CRP was within reference interval because of liver failure.
Petrovečki M1,2, Bilić-Zulle L2,3, Šimundić AM2,4, Nikolac N2,4.P14-3: Evaluation of “Statistics” Curriculum at the Zagreb University School of Pharmacy and Medical Biochemistry. Biochemia Medica 2009;19(Suppl 1):S175.
1Dubrava Clinical Hospital, Zagreb, Croatia
2Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia
3Rijeka Clinical Hospital Center, Rijeka, Croatia
4Sestre Milosrdnice University Hospital, Zagreb, Croatia
Corresponding author:mp [at] kbd [dot] hr
Introduction: “Statistics” reformed curriculum was introduced to first year students of the Zagreb University School of Pharmacy and Medical Biochemistry (PMB) during the 2008/09 academic year. The content of curriculum and final exam follows textbook Essential Statistics for the Pharmaceutical Sciences by Philip Rowe (Wiley, 2007).
Methods: Evaluation of curriculum was estimated using anonymous questionnaire filled out by students after the last class. Questionnaire consisted of questions regarding organization and content of curriculum (marked 1-5) and comprehensive questions. Final exam marks were collected. Out of 183 students, 135 filled out the questionnaire and 142 took the exam. Results are presented as mean and standard deviation.
Results: Students enrolled PBM had high marks at high school (4.90  0.31). They expected statistically lower exam mark (3.67  0.83, N = 135) comparing to their real achievement at the exam (4.19  1.07, N = 145, P < 0.001). Organization of curriculum was highly appreciated between students (4.48  0.66) with significantly higher grade than organization of all other curricula (2.39  0.78, P < 0.001). The interest for curriculum contents was also graded higher for Statistics (3.89  0.94) than for all other curricula (3.26  0.75, P < 0.001). Student's knowledge gained through classes was self evaluated as 3.18  0.69, but when answering four multiple choice comprehensive questions the rate of correct answers were as follows: 52% (regarding standard deviation), 90% (regarding sampling), 99% (regarding correlation) and 100% (regarding randomization). Similar question, asked at the first class remained unanswered, i.e., before Statistics classes students didn't recognize basic statistical terms.
Conclusion: Results revealed good organization and achievement of proper interest for curriculum content among students.
Bilić K, Fumić K, Čvorišćec D. P14-4: Enzyme assay on dried blood filter paper samples for detection of Pompe disease. Biochemia Medica 2009;19(Suppl 1):S176.
Clinical Institute of Laboratory Diagnosis, Zagreb University School of Medicine and Clinical Hospital Center, Zagreb, Croatia
Corresponding author:karmen [dot] bilic [at] zg [dot] t-com [dot] hr
Introduction: Pompe disease is an autosomal recessive disorder of glycogen metabolism caused by a deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA). Abnormal storage of glycogen in different tissues leads to cellular injury, with enlargement and dysfunction of the involved organs. Broad clinical spectrum varies with respect to age at one set, rate of disease progression and extent of organ involvement. Three types of Pompe disease are recognised: infantile, juvenile and adult-one set. Progressive hypertrophic cardiomyopathy and marked muscle weakness are typical for the infantile type. The slowly progressive adult type and juvenile type present with skeletal muscle weakness and respiratory failure. As enzyme replacement therapy (recombinant human alpha-glucosidase) has recently become available, screening for this disorder and early intervention may significantly improve the outcome. Enzymatic diagnosis in blood is not usually used due to the presence of GAA isoenzyme maltase-glucoamylase which shows a significant overlap in activity within the acidic pH range. Recently, novel enzymatic methods have been described that measure GAA activity in dried blood spot (DBS) and eliminate the maltase-glucoamylase activity by the use of the specific inhibitor acarbose.
Methods: We tested a fluorometric enzyme assay using 8 mM acarbose at pH 3.8, 10 mM substrate 4-MU-alpha-D-glucopiranoside and DBS samples.The activity of a reference enzyme was also measured to confirm the quality of the sample. In suspected cases of Pompe disease, a second sample was requested (new DBS and lymphocytes).
Results: As a result, patients with infantile and adult types of Pompe disease were discriminated from healthy controls.
Conclusion: In summary, the fluorometric DBS enzyme assay using inhibitor acarbose can be successfully used for screening.
Flegar-Meštrić Z, Perkov S, Kardum Paro MM, Ožvald I, Šimonović B, Sikirica M. P14-5: Harmonization of reference intervals for serum creatinine concentrations. Biochemia Medica 2009;19(Suppl 1):S177.
Institute of Clinical Chemistry, Merkur University Hospital, Zagreb, Croatia
Corresponding author:zlata [dot] mestric [at] zg [dot] t-com [dot] hr
Background: Reference intervals for the serum creatinine concentrations were determined on the representative reference sample group of Croatian population (2246 adults and 998 children, age 8-70 years) using Jaffe kinetic method. In order to harmonize the laboratory results the Croatian Chamber of Medical Biochemists recommended obligatory use of the obtained reference intervals for all laboratories using the same analytical method since January, 2005. A review of long-term results collected on national external quality assessment surveys has shown that serum creatinine concentration in almost 200 medical-biochemistry laboratories were determined with average CV between of 4.3% to 6.2%, but in 2002, according to the International Measurement Evaluation program (IMEP-17) our national results for the creatinin concentrations showed significant positive bias to the IDMS certified reference materials.
Aim: Due to the growing requirement for accurate and specific diagnostic tests and their traceability to reference methods we evaluate the reference intervals for creatinine concentrations using accurate and specific enzymatic method.
Methods: The reference sample group consisted of 240 a priori selected healhy adults. Serum creatinine concentrations were measured by the kinetic Jaffe method and by an enzymatic assay traceable to the IDMS method on the Olympus AU 600 Analyzer. The manufacturer´s calibrator traceable to the IDMS method and NIST SRM 967 was used.
Results: The reference intervals (central 95th percentiles) obtained with Creatinine Enzymatic assay ranged from 54 to 107 µmol/L for males and from 50 to 93 µmol/L for females vs. IFCC recommended reference intervals: 64-104 and 49-90 µmol/L for males and females, respectively.
Conclusions: In accordance with an ongoing activity for worlwide harmonization based on traceability in laboratory methods evaluation of the reference intervals using enzymatic creatinine determinations traceable to the IDMS method in our study showed that recommended reference intervals for global application as proposed by the IFCC Committee on Reference Intervals, and Decision Limits, could be adopted for the Croatian population.
Garilović J, Škorvaga S, Šimić-Vojak S. P14-6: Values of procalcitonin and „classic“ markers in neonatal sepsis: Case report. Biochemia Medica 2009;19(Suppl 1):S178.
Požega General Hospital, Požega, Croatia
Corresponding author:jasna [dot] garilovic [at] po [dot] t-com [dot] hr
Introduction: Procalcitonin (PCT) is the newest biomarker for early recognition and diagnosis of sepsis. Determination of PCT helps in making clinical decisions for appropriate antibiotic therapy. “Classic” markers include determination of these parameters: complete blood count, immature to mature ratio (I/M ratio), percentage of band cells and C-reactive protein (CRP).
Materials and methods: PCT concentration was determined in serum on Elecsys 2010 Roche analyzer, method of electrochemiluminescence. CRP was determined in serum on Olympus AU 400 analyzer (high sensitive application). Complete blood cell count was determined on automated blood counter Abbott Cell-Dyn 1700 and deferential blood count was determined by microscopic method in stained peripheral blood smear. We measured concentration of CRP, PCT, WBC, I/M ratio and percentage of band cells on days 2, 5, 6 and 11.
Results: During the first measuring (on day 2) concentration of PCT (2.69 ng/mL) was higher, while the values of CRP, WBY, I/M ratio and % band cells were in range. On day 5 of newborns life for all study parameters were found increased values: PCT (21.0 ng/mL), CRP (30.4 mg/L), WBC (29.3 x 109/L), I/M ratio (0.17), % band cells (12). Next measurements (on days 6 and 11) recorded decrease in values of investigated markers and their return to normal. Results show that PCT has the highest diagnostic accuracy among studying biomarkers. PCT levels rise rapidly after infections insult and in sepsis.
Conclusion: Using appropriate antibiotic therapy values of all biomarkers return to normal