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The aim of this study was to define risk factors for jaundice and anemia in newborns with a positive direct antiglobulin test (DAT) and/or with an incompatible crossmatch due to ABO incompatibility between mother and newborn. ABO incompatibility has become a more significant cause of hemolytic disease of the fetus and newborn since the introduction of effective anti-D prophylaxis. The condition is common and, if clinically significant at all, causes only mild jaundice, which can be treated with phototherapy (PT). However, rare and serious presentations, requiring transfusion therapy, have been noted. Clinical, laboratory, and immunohematologic data were collected retrospectively from medical records of ABO-incompatible newborns and their mothers over a 5-year period (2016–2020) from University Hospital Centre Zagreb. Two groups of newborns were compared: those who needed medical intervention because of hyperbilirubinemia or anemia and those who did not. Within the group of newborns requiring intervention, we also compared those with A and B blood groups. Over the 5-year period, 72 of 184 (39%) newborns required treatment. The treatment was PT in 71 (38%) newborns and erythrocyte transfusion in 2 (1%). In 112 (61%) newborns, ABO incompatibility was an accidental finding while performing blood group typing ; these newborns did not require any therapy. In conclusion, we found a statistical, but not clinically significant, difference between the groups of treated and untreated newborns, related to the mode of delivery and DAT positivity within hours of delivery. There were no statistically significant differences in characteristics between the groups of treated newborns, except for two newborns with blood group A who received erythrocyte transfusions.

Objective Foundation project for the first human milk banks (HMB) in Croatia was launched in 2017 as the result of a collaboration between the Ministry of Health, University Hospital Centre (UHC) Zagreb, UNICEF and the Croatian Association of Breastfeeding Support Groups. The HMB was opened in UHC Zagreb on November 15, 2019as a part of the Croatian Tissue and Cell Bank, a division of the Department of Transfusion Medicine and Transplantation Biology. The specific technical side is based on the PATH standards and Guide to the quality and safety of tissues and cells for human application, Council of Europe. On January 10, 2020, HMB passed a Competent Authority (CA) inspection, conducted in accordance with the requirements of national tissue legislation that has been adapted to EU Directive 2004/23/EC on setting standards of quality and safety for the donation, procurement, testing, processing, preservation, storage and distribution of human tissues and cells. The specific technical part is based on the PATH standards and Guide to the quality and safety of tissues and cells for human application, Council of Europe. The aim of this study was to review the very first experience in human milk banking in Croatia. Methods We retrospectively analysed the data of the first 28 human milk donors. Results From the opening to mid-February, 135 women showed interest in donating human milk, of which 28 became donors. In February 2020, we had 24 still active donors. Four women stopped donating, with a median donation period of 2 months. All donors were tested for blood borne viruses by serology and NAT and were negative. In total, we received 79 L of donated human milk.We started pasteurizing the milk after obtaining a license. Of the 30 controlled pre-pasteurization milk pools, 10% was over the allowed microbial contamination. Microbiological controls were performed for each pasteurization cycle. They were all sterile. The nutritional values of milk were all within the expected range. Conclusions Human milk is recognised as the optimal feeding for all newborn infants. When mothers’ own milk is not available, donor human milk provided by HMB is the second-best choice, especially for premature or sick infants. The opening of the HMB in Croatia is highly important in helping to provide the best possible medical care for prematurely born babies and infants with a serious medical condition when they cannot receive their mother’s milk.

Validacija i provedba periodičke provjere metode za određivanje CD34+ stanica na protočnom citometru

Darovano humano mlijeko za svako novorođenče u Hrvatskoj

Testiranje antiheparinskih protutijela u bolesnika na izvantjelesnoj membranskoj oksigenaciji.

Prikupljanje i krioprezervacija autolognih mononuklearnih stanica iz periferne krvi za potrebe proizvodnje car-t stanica

Značaj imunohematološkog ispitivanja u ABO nepodudarnosti majki i novorođenčadi

Učestalost antitrombocitnih aloprotutijela i genotipizacija 12 specifičnih trombocitnih antigena (HPA) u trudnica

Darovano humano mlijeko je najbolja zamjena za majčino mlijeko u okolnostima kad ona ne može hraniti svoje dijete. Sigurno i kvalitetno darovano humano mlijeko osiguravaju banke humanog mlijeka. S tim ciljem u Hrvatskoj banci tkiva i stanica Kliničkog bolničkog centra Zagreb osnovana je Banka humanog mlijeka u siječnju 2020. Rad Banke u skladu je sa Zakonom o primjeni ljudskih tkiva i stanica. U ovom radu prikazujemo rezultate njenog rada od otvaranja do lipnja 2020. godine. Iz logističkih razloga uzrokovanih epidemijom COVID-19 i potresom u Zagrebu Banka humanog mlijeka nije prikupljala mlijeko 43 dana. Mlijeko je darovala 31 majka. Medijan dobi bio je 31 godina a 81% ih je bilo visokoobrazovanih. U 52% slučajeva majke su počele darivati mlijeko tri mjeseca nakon porođaja. Najviše darivateljica darovalo je mlijeko samo jedan put (45%). Medijan razdoblja darivanja bio je 46 dana. Većina (52%) darivateljica rodilo je prvi put, u očekivanom terminu porođaja (94%), djecu porođajne mase >2 500 gr. Samo troje od njihove djece (9%) bilo je na intenzivnom liječenju. Ukupno je prikupljeno 175, 5 L mlijeka (prosječno 5, 7 L/darivateljici), od kojih je 151, 5 L zadovoljilo zahtjeve ulazne kontrole kakvoće, a 141 L je bila i pasterizirana. Kritičan broj vijabilnih, aerobnih i fakultativnih bakterija imalo je 32, 6% mješavina mlijeka pripremljenih za pasterizaciju, a nakon nje 8, 9%. Za kliničku primjenu izdano je 78, 7 L u tri jedinice intenzivnog liječenja novorođenčadi. Već u prvim mjesecima rada Banka humanog mlijeka pokazala je važnost svog djelovanja. Kako bi mogli zadovoljiti potrebe za darovanim humanim mlijekom na nacionalnoj razini, potrebno je trajno podučavati/educirati majke o važnosti humanog mlijeka i promicati njegovo darivanje.

U današnje vrijeme svjedoci smo neprekidnog i sve većeg interesa u otkrivanju novih biokemijskih biljega. Takvo što i ne čudi s obzirom da novootkriven, ispravno validiran biokemijski biljeg može poslužiti kao vrijedan prognostički i/ili dijagnostički pokazatelj pojave bolesti, progresije ili oboje. Prije uvođenja, svaki novi potencijalni biokemijski biljeg potrebno je evaluirati, odnosno provesti opsežnu validaciju, kako bi se potvrdilo da će biti koristan za kliničku prim jenu.

Introduction: Autovalidation is a part of the laboratory information system whereby laboratory results are released without manual human intervention through defined rules and algorithms. Autovalidation rules may include: analytical measurement range, interference indices (hemolysis, icterus, lipemia), critical values or delta checks. Rules must be set for laboratory testing and patient population for which autovalidation will be applied. The aims of the study were to examine the credibility of the rules in the algorithm for autovalidation and determine the percentage of samples that were autovalidated in comparison to the total number of samples included in validation, as well as rules that stop autovalidation. Materials and Methods: The validation results included 9805 samples of biochemical tests analyzed in the Department of Laboratory Diagnostics, University Hospital Centre Zagreb from May to July 2014. Validation was performed in such a way that, before starting the system of autovalidation, the results were reviewed by a medical biochemist who recorded samples that should meet the rules set in the algorithm. Statistical analysis was made using the software MedCalc (version 9.3.2.0). Results: 78.3% (7677) of the samples included in validation were autovalidated. The highest percentage of samples (54.9%) was not autovalidated due to set rules for analytical measurement ranges, while the lowest percentage of non-validated samples was recorded in the rules for interference indices for icterus (0.6%). The correspondence of autovalidation and the person who carried it out was 99.5%, i.e. the mismatch is observed only in 0.5% of samples (38). An analysis of Χ2-test data showed no statistically significant difference (P=0.523) in the number of samples that were autovalidated (7677) in relation to those validated by medical biochemist (7639). Conclusion: The analysis of results showed that set rules in the algorithm are credible, and that system autovalidation can be implemented in routine laboratory use.

Prije primjene novih reagensa u svakodnevnom radu potrebno je, prema dobroj laboratorijskoj praksi, napraviti njihovu analitičku procjenu. Cilj rada bio je verifikacija koagulacijskih reagensa Sclavo Diagnostics International S.r.I.(Sovicille, Italija) za određivanje protrombinskog vremena (PV), aktiviranog parcijalnog tromboplastinskog vremena (APTV), funkcionalne koncentracije fibrinogena (Fbg) i aktivnosti faktora V (FV).

Background: Efficient managing of laboratory test utilization is required as overutilization of diagnostic laboratories is constantly increas-ing. The implementation of hospital information system has improved the communication between laboratory and clinic, but at the same time has enabled uncontrolled placing of electronic laboratory orders even for highly differentiated coagulation tests. The aim of this study was to determine whether testing for fibrinogen (Fbg) and antithrombin (AT) antigen concentration is ordered appropriately. Materials and methods: A two-step procedure was employed in the study. First we performed the analysis of all requests for the determination of Fbg and AT antigen in a 1-year period by searching through the laboratory information system. The appropriateness of each request was evaluated in the second step by analyzing if determination of functional activity was required prior to requesting the determination of specific antigen concentration. Results: In the investigated period, a total of 48 requests for Fbg:Ag and 72 requests for AT:Ag were identified. Among them, 27/48 requests for Fbg:Ag and 10/72 requests for AT:Ag were inappropriate as functional activities were not performed in those patients. The determination of Fbg:Ag was not performed in 7/48 patients and of AT:Ag in 35/72 patients because functional activities were within the reference intervals. We provided this comment with each report so that physicians may understand why the requested test was not performed. Finally, due to decreased functional activities, Fbg:Ag and AT:Ag were determined in 14/48 and 27/72 patients, respectively, and were clinically indicated. Conclusions: According to obtained results, the majority of requests were inappropriate, especially for Fbg:Ag as physicians were actually requesting the determination of fibrinogen functional activity. Probably this was the consequence of the availability of requesting functional activity and antigen concentration for Fbg and AT in the same computerized laboratory order with no restriction.

Background: Ristocetin cofactor activity of von Willebrand factor (VWF:RCo) and VWF antigen concentration are still essential laboratory assays for the diagnosis of von Willebrand’s disease (VWD). As VWF:RCo has poor reproducibility and sensitivity to low levels of VWF, new generation assays for measurement of VWF activity (VWF:Ac) are developed. They are based on binding VWF to recombinant GPIb which in turn binds to particle-fixed anti-GPIb antibodies, and do not contain ristocetin. Aims: The aim was to validate the new VWF:Ac assay and compare obtained results with VWF:RCo assay. Methods: We have validated the performance of a new automated latex- immunoturbidimetric assay (INNOVANCE VWF Ac) on Sysmex CS2000i and on BCS analyzer (Siemens Healthcare Diagnostics, Germany). Within-run and between-run imprecision were estimated in normal and pathological range using Control plasma N (CPN) and Control plasma P (CPP). Low-range calibration curve was tested using CPP prediluted 1/3 on Sysmex CS2000i. Method comparison was performed in normal and clinical samples with VWF:RCo (BC von Will-brand Reagent) using Passing and Bablok regression. Results: Low coefficients of variations (CVs) for within-run and between-run imprecision were obtained on both analyzers using CPN (BCS within-run CV 1.5%, between-run CV 3.0% ; CS2000i within-run CV 2.0%, between-run CV 3.8%) and CPP (BCS within-run CV2.2%, between- run CV 4.4% ; CS2000i within-run CV 3.7%, between-run CV 4.4%). Even for CPP 1/3, within- run and between- run imprecision CVs were 3.6% and 3.5%, respectively. Normal and clinical samples demonstrated good correlation between VWF:RCo and VWF:Ac on both analyzers (BCS y = 0.88x + 5.77, r = 0.968, P < 0.001 ; CS2000i y = 1.05x–0.04, r = 0.987, P < 0.001). Conclusion: Considering low CV values, VWF:Ac assay had satisfactory accuracy and precision on both analyzers (even for low VWF activities) and good correlation with VWF:RCo. The performance of VWF:Ac assay makes it suitable for the laboratory diagnosis of VWD.

Background: Accurate coagulation test results depend on all phases of laboratory process. Besides sampling, transport and centrifugation, sample stability following blood collection is an important part of preanalytical phase, but it is still a matter of concern. Although samples for coagulation testing are routinely processed according to current guidelines, sometimes there are requests for performing additional tests in already analyzed fresh plasma samples, especially in critical care patients. The aim of this study was to examine whether fresh plasma samples stored at room temperature for up to 24-h after blood collection can still be accepted for second- line routine coagulation testing. Materials and methods: We have measured prothrombin time (PT), activated partial tromboplastin time (aPTT), fibrinogen (Fbg) and antithrombin (AT) in citrated plasma samples in primary tubes within 4-h of blood collection and after storage at 25 °C for up to 24-h. Additionally, plasma aliquots were separated from primary tubes and stored at 25 °C for up to 24-h. Stored plasma samples in primary tubes and aliquots were processed the next day under the same conditions. For each parameter, samples from 20–30 patients were investigated covering a wide range of normal and pathological results. Measurements were performed on the coagulation analyzer BCS XP by using reagents from Siemens Healthcare Diagnostics. Results: The highest variation of results in stored samples was observed for aPTT (primary tubes: mean 11.8%, range 3.4–32.8% ; aliquots: mean 15.5%, range 5.0–30.4%). For all other parameters the mean percentage change was below 10% either in primary tubes (ranging from 1.3– 5.7%) or in aliquots (ranging from 1.1–3.6%). Conclusion: No advantage of separating plasma from cells was observed. Samples stored in primary tubes for up to 24-h can be used for second-line routine coagulation testing, even for aPTT as changes of results were not clinically relevant.

BACKGROUND: According to the guidelines of the European Society of Cardiology (ECS), the diagnosis of acute myocardial infarction (AMI) is made according to criteria that include an increase in cardiac markers (troponin) with the presence of characteristic clinical symptoms or electrocardiogram changes or imaging of myocardial damage. At the Department of Emergency Medicine, University Hospital Centre Zagreb, the use of the ECS guidelines was introduced in year 2013. In an emergency laboratory high-sensitive troponin (hs-TnT) is used for determination of troponin (TnT). The guidelines define dynamic determination of TnT, i. e. at patient admission and after three hours. The decision cutoff point for definition of AMI is the 99th percentile in the general population with the coefficient of variation of less than 10%. The 99th percentile for the hs- TnT test that we routinely used is 0.014 µg/L and the lower detection limit is 0.003 µg/L. To facilitate physicians' understanding, we have provided the comment with each TnT report: "The values exceeding 0.014 µg/L indicate the presence of cardiac damage. In patients with clinically suspected AMI, changes in value ≥ 0.007 µg/L after 3 hours may assist in diagnosis confirmation“. The aim of this study was to investigate how the application of new guidelines affected the number of requests for determining TnT concentration. METHODS: We compared the number of patients for whom the TnT analysis was requested during three months in 2012 and 2013 and the number of released TnT reports in these periods. Data were taken from the laboratory information system. For statistical analysis, the chi- square test was used. RESULTS: The number of patients with requested TnT analysis in 2012 was 1109/1091/1046 and in 2013 it was 1150/1152/1134 (October/November/December, respectively). The number of released TnT reports in 2012 was 1267/1194/1186 and in 2013 it was 1402/1414/1407. Statistical analysis showed significant increase in the number of released TnT reports (p0.05). CONCLUSIONS: During the period of observation in 2013 there was a significant increase in the number of released TnT reports, which is attributed to the application of ECS guidelines.