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PURPOSE: Possible transmission of SARS-CoV-2 from donors to recipients via cornea grafts is still a concern of the transplantation community. Current recommendations are to avoid corneal transplants from donors with ongoing SARS-CoV-2 infection or those recently exposed to it. During pandemic period in Croatia 21/1113; (1,9%) corneas were procured from donors positive for SARS-CoV-2 by postmortem nasopharyngeal swab tests. That tissue was discarded. Due to the lack of knowledge about the infectivity of such corneas, we started prospective study of SARS-CoV-2 presence in cornea tissue. Here we show our first results. METHODS: In the study period we had four corneas procured from two post-mortem SARS-CoV-2 positive donors. For the presence of SARS-CoV-2, analysis is performed on donor serum, hypothermic storage medium and cornea tissue lysate. Corneas were stored in hypothermic condition for 8 to 10 days, after which tissue was macerated and washed with PBS. The intracellular content was released by incubation with lysis buffer, followed by centrifugation. Next, tissue lysate, serum and hypothermic storage medium were in parallel subjected to fully automated nucleic acid isolation and RNA expression was analyzed by qRT-PCR. During isolation, RNasaP was used as internal control for successful nucleic acids isolation. RESULTS: No SARS-CoV-2 RNA was detected in the donors serum, storage medium and cornea tissue from donors who were SARS-CoV-2-positive upon tissue procurement. In nasopharyngeal swabs of post mortem positive donors cycle threshold values of viral copies were high (CT>34), indicating that there was small number of viral particles in infected donors that could have impact on negative results in tested tissue. CONCLUSION: Our data suggested that corneas may not be SARS-CoV-2 permissive if the donor was postmortem positive. Further research is required to gain more coherent insight into SARS-CoV-2 transmission via corneal transplantation.

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PURPOSE: Croatian Tissue and Cell Bank (CTCB) regularly monitors the effectiveness of cornea donation program on the national level. All hospitals are required to have designated tissue donation coordinators in charge of detection, family interview and tissue procurement. If hospital has cornea donation program only from donors after brain death (DBD), tissue donation coordinator can be the same as for organs. Five collection centres have cornea donation program for donors after circulatory death (DCD) with designated cornea donation coordinators. METHODS: We retrospectively analyzed all monthly reports from tissue donation coordinators in the period from May 2019 to September 2022. Additional data was collected from national organ and tissue database Croatian National Transplantation Network (NTM). RESULTS: During the analyzed period, 25.753 deaths were recorded, from which 38,6% to 54,7% of DCD and 0,6% to 1,1% of DBD donors were considered for cornea donation, depending on the hospital. Out of all deceased, 2,4% to 5,2% of patients were realized as cornea donors, 0,4 to 0,5% of which were DBD and 2 to 4,7% were DCD. Cornea donations were realized in 18,2% to 38,9% cases of all DBD donors. As SARS-CoV-2 pandemic has strated in March 2020, the cumulative number of donations declined for 26,1% in 2020 and 12,1% in 2021, compared to the pre-pandemic 2019. Moreover, CTCB received 30,5% less DCD in 2020 and 21,9% less in 2021. Despite that, we recorded increase in DBD during 2020 and 2021, for 13,3% and 44,7%, respectively. The same trend continued throughout 2022, where only until September 16,1% more DBD were received than in the whole 2019. CONCLUSION: Hospitals involved in cornea donation program record high number of deaths, however only a small proportion of which are realized for cornea donation. This is particularly pronounced in DBD donors. SARS-CoV-2 pandemic left significant impact on donation program. However, CTCB recorded higher number of DBD donors during that period. The current situation leaves plenty of room for improvement of CTCB and corresponding donation hospitals, to increase disproportionately low rate of cornea procurement in respect to the total rate of deaths and considered donors.

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BACKGROUND: Corneas procured post mortem are at risk of microbiology contamination, therefore decontamination procedures before storage, aseptic techniques during processing and antimicrobials used in the storage medium are routinely used. Despite that, corneas are discarded due to microbiology contamination. According to professional guidelines, corneas can be procured preferably within 24 hours after cardiac arrest but up to 48 hours. Our goal was to evaluate the risk of contamination depending on the post mortem time and the spectrum of microbes isolated. METHODS: Corneas were decontaminated before procurement using 0,5% povidone iodine and tobramycin, stored in the organ culture medium and microbiologically tested after four to seven days of storage. Ten millilitres of cornea preservation medium were inoculated in two blood bottles (aerobic, anaerobic/fungi, Biomeriex) and incubated for seven days.Microbiology testing results in the period of four years (2016-2020) were retrospectively analysed. Corneas were divided in four groups depending on the duration of post mortem interval: group A post mortem interval < 8 h, group B post mortem ranging from 8 to 16 h, group C post mortem ranging from 16 to 24 h and group D post mortem > 24 h. Contamination rate and spectrum of isolated microorganisms in all four groups were analysed. RESULTS: 1426 of 2019 procured corneas were stored in organ culture and microbiologically tested. 65/1426 of tested corneas were contaminated (4,6%). In total, 28 strains of bacteria and fungi were isolated.Contamination rate of post mortem groups are as following: group A 3,1% (14/455), group B 4,1% (23/561), group C 6,7% (27/402) and group D 12,5% (1/8).In the group A bacteria family Staphylococcaceae, Moraxellaceae, Morganellaceae were predominately isolated (64,3%). In the group B fungi Saccharomycetaceae, bacteria Moraxellacea, Staphylococcaceae, Morganellaceae and Enterococcaceae were predominately isolated (78,1%). In the group C, bacteria family Enterococcaceae, Moraxellaceae and fungi Saccharomycetaceae are most often isolated (70,3%). In the group D bacteria family Enterobacteriaceae was isolated (100%). CONCLUSION: Organ culture allows detection and discard of microbiology contaminated corneas. Our results show higher microbiology contamination rate for corneas with longer post mortem intervals, suggesting these contaminations can be rather related to donor post mortem changes and contamination than previous infection. In order to keep the best quality and safety of the donor cornea, all efforts should be directed in disinfection of the cornea and keeping post mortem interval shorter.

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BACKGROUND: The aim of this study was to evaluate the incidence and clinical features of non-melanoma tumors of the head and neck, as well as the validity of surgical therapy in their treatment. SUBJECTS AND METHODS: The study included 530 patients who were operated in the Otorhinolaryngology department of the Livno County Hospital. RESULTS: In 295 cases (65.1%), it was basal cell carcinoma of the skin and was followed by squamous cell carcinoma of the skin, in 119 cases (29.9%) while the remaining 5% of cases referred to other non-melanoma skin carcinomas. Statistically significant, the most common non-melanoma skin carcinoma was basal cell carcinoma (χ2=625,67; df=4; p<0.01). The most co mmon localization was the skin of the nose (24.2%), which proved to be statistically significant (χ2=290.824; df=5; p=0.00). All patients underwent classic surgery, and in 358 cases (89.5%) the tumor was completely removed, while in 40 cases (10.5%) the tumor was partially removed which proved to be statistically significant (χ2=254,08; df=1; p=0.00). CONCLUSIONS: The results of the study fully confirm the assertion that classical surgery is the method of choice in the treatment of non-melanoma skin cancers and in the vast majority it is proven to be sufficient.

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OBJECTIVE: To evaluate the performance of the epoc hand-held analyzer against the RAPIDPoint 500 blood gas analyzer and laboratory analyzers where applicable. METHODS: Venous or arterial whole blood samples collected in balanced heparinized syringes were obtained from 69 patients (35 females, 34 males) predominantly (77%) from the surgical unit and intensive care unit (ICU). Method comparison was performed for all analytes on the epoc System against the RAPIDPoint 500 Blood gas analyzer or laboratory analyzers where applicable. Results: Mean bias was <5% for blood gases, electrolytes, lactate and glucose. Hematocrit showed a bias of -6.76% (95% CI â€‹= â€‹-8.91, - 4.61) compared to the HemataSTAT-II method, whereas calculated total hemoglobin showed a bias of 1.51% (95% CI â€‹= â€‹-1.04, 4.06) against the Sysmex XN-10 hematology analyzer. Creatinine showed the largest bias relative to laboratory analyzers, Abbott Architect c8000 Jaffe method (13.54%, 95% CI â€‹= â€‹5.43, 21.65) and Roche Cobas c702 enzymatic method (30.01%, 95% CI â€‹= â€‹12.64, 47.38). Conclusions: The epoc system is fit for use in the surgical and ICU setting for the measurement of all analytes except for creatinine.

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BACKGROUND AND OBJECTIVES: The number of CD34+ cells collected in apheresis procedures depends mainly on the collection efficiency of the device and the blood volume processed. Large volume leukapheresis (LVL) can improve CD34+ cell yield and has previously been investigated using the COBE Spectra device (Terumo BCT, USA). MATERIALS AND METHODS: This was a retrospective analysis of LVL performance in patients undergoing continuous mononuclear cell collection (CMNC) using the new Spectra Optia apheresis system (Terumo BCT, USA) at the University Hospital Center, Zagreb, from March 2016 to September 2016. CD34+ cell yield predictability, determined using a customized algorithm, was also assessed. RESULTS: In total, 67 procedures performed in 46 adults and 14 performed in 11 children were included in the analysis. In adults, 30 (65.2%) patients successfully reached their target preapheresis CD34+ cell count on day 1, with a median (interquartile range [IQR]) CD34+ collected cell dose of 4.8 × 106 /kg (2.3-10.6 × 106 /kg). In the pediatric group, 81.8% successfully collected the target CD34+ cell dose on the first day, with a median (IQR) CD34+ collected cell dose of 11.1 × 106 /kg (3.2-16.3 × 106 /kg). The customized algorithm showed a strong and significant linear correlation with actual CD34+ cell dose (P < 0.0001). CONCLUSION: The results of this study support the use of LVL and the customized prediction algorithm in apheresis procedures. The ability to tailor the procedure to meet the needs of the individual patient may help to minimize the blood volume processed, shorten the duration, reduce the volume of infused anticoagulants, and improve patient comfort.

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BACKGROUND: Acute and delayed haemolysis, alloimmunisation and pure red cell aplasia (PRCA) are potential complications after ABO incompatible haematopoietic stem cell transplantation (HSCT). The aims of this study were to investigate acute and delayed red blood cell (RBC) antibody-associated complications, including haemolysis, PRCA and alloimmunisation in major and bidirectional ABO incompatible HSCT. MATERIALS AND METHODS: We retrospectively examined the transplant courses of 36 recipients of bone marrow or peripheral blood stem cells from ABO incompatible donors and evaluated the current practice of performing plasmapheresis in patients with higher isoagglutinin titres. We investigated the role of ABO incompatibility in haematopoietic recovery, transfusion requirements, alloimmunisation and PRCA. RESULTS: Laboratory signs of acute haemolysis were noted in five (14%) patients, one (3%) of whom had clinically overt haemolysis. Patients with haemolysis had IgM titres ≥1:8 and received >16 mL of RBC in the HSCT. In patients with higher titres, plasmapheresis performed prior to the transplant prevented acute haemolysis. Delayed haemolysis was not recorded in the follow up. Haematopoietic recovery and transfusion requirements did not differ notably between patients with and without haemolysis. De novo RBC antibodies were detected in two (5.5%) patients after HSCT, and PRCA was noted in one (3%) patient. DISCUSSION: Carried out with adequate graft processing, plasmapheresis and blood component support, haemolysis is not a common complication after HSCT. Our results confirm that the occurrence of haemolysis depends on larger RBC volumes and higher isoagglutinin titres. Despite the reduction of patients' isoagglutinin titres by plasmapheresis, we still noted a critical combination for the development of laboratory signs of haemolysis (IgM titre ≥1:8 and RBC volume >16 mL). De novo immunisation to RBC antigens and PRCA are rare events following ABO incompatible HSCT.

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OBJECTIVE: To estimate time trends in prevalence of symptoms and reported diagnosis related to asthma, allergic rhinitis/conjunctivitis and eczema among school children in the north-west coastal part of Croatia. METHODS: Results of two identical cross-sectional surveys conducted on the same area 8 years apart (school years 2001/02 versus 2009/10) in complete adherence to the protocol of the International Study of Asthma and Allergies in Childhood were compared. Surveyed population comprised two age groups: 6-7 years (n = 1634 versus n = 1052) and 13-14 years (n = 2194 versus 1181). RESULTS: Significant (p < 0.001) increases in prevalence (%) of symptoms related to asthma (8.4 versus 14.0), allergic rhinitis (17.5 versus 25.6), allergic rhinoconjunctivitis (6.7 versus 15.3) and eczema (3.4 versus 5.9) were observed in the 13-14-year-olds. In the 6-7-year-olds there were observed significant (p < 0.001) increases in prevalence of symptoms of eczema (5.4 versus 8.7) and allergic rhinitis (16.9 versus 22.1) whereas prevalence of symptoms related to asthma (9.7 versus 9.4; p = 0.398) and allergic rhinoconjunctivitis (5.6 versus 6.8; p = 0.102) showed to be stable. Significant increases in prevalence of reported diagnosis were observed for asthma (5.2 versus 6.9; p = 0.02) and hay fever (10.5 versus 14.6; p < 0.001) in the older, and for eczema (10.6 versus 14.1; p < 0.001) in the younger age group. CONCLUSION: Prevalence of asthma and allergic diseases among the school children living on the surveyed area showed a rising trend.

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