A mini extracorporeal photopheresis (mini‐ECP) “off line” technique has been developed for use in the treatment of small children and patients with apheresis contraindications. Until now various ...methods have been used for buffy coat separation from whole blood. In this report we describe a protocol for mini buffy coat preparation using the automated Sepax laboratory separator for “off line” ECP treatment in a low body weight child with graft‐vs‐host‐disease. According to our results this alternative method has been proven feasible and tolerable.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Background
Patients with hematological diseases are polytransfused and often immunocompromised, therefore susceptible to transfusion reactions (TR). This study aims to document the incidence of TRs ...in adult hematological patients and assess the effect of changes in the production of blood components and transfusion practice on their occurrence.
Study design and methods
Retrospective observational analysis of TRs reported from 1993 to 2019 was performed. For the analysis of the effect of changes on the incidence of TRs, the evaluated time was divided into two periods: the 1st period before the introduction of changes in production, when leukoreduced blood components were used only selectively, and the 2nd period, when semi‐automated method of production and universal leukoreduction was introduced.
Results
The decrease in the incidence of TRs was observed for both red blood cell (RBC) and platelet concentrate (PC) transfusions in the 2nd period. Since platelet additive solution has been used, a further decrease in the incidence was reported. The decrease in incidence was also observed for delayed hemolytic/serological transfusion reactions and for transfusion‐transmitted bacterial infections. Four cases of incorrect blood transfusions were uniquely related to the hematological patients, caused by antigen loss and transfusion ordering after ABO‐incompatible hematopoietic stem cell transplantation.
Discussion
Our results provided evidence that the introduction of tools offered by modern transfusion medicine: universal leukodepletion, plasma replacement with additive solutions, sensitive laboratory techniques, prophylactic antigen matching policy, informatization, and automatization, decreased the incidence of TRs and improved transfusion safety.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Due to their ability to induce immunological tolerance in the recipient, mesenchymal stromal cells (MSCs) have been utilized in the treatment of various hematological and immune- and ...inflammation-mediated diseases. The clinical application of MSCs implies prior in vitro expansion that usually includes the use of fetal bovine serum (FBS). The present study evaluated the effect of different platelet lysate (PL) media content on the biological properties of MSCs. MSCs were isolated from the bone marrow of 13 healthy individuals and subsequently expanded in three different culture conditions (10% PL, 5% PL, 10% FBS) during 4 passages. The cells cultured in different conditions had comparable immunophenotype, clonogenic potential, and differentiation capacity. However, MSC growth was significantly enhanced in the presence of PL. Cultures supplemented with 10% PL had a higher number of cumulative population doublings in all passages when compared to the 5% PL condition (p < 0.03). Such a difference was also observed when 10% PL and 10% FBS conditions were compared (p < 0.005). A statistically significant difference in population doubling time was determined only between the 10% PL and 10% FBS conditions (p < 0.005). Furthermore, MSCs cultured in 10% PL were able to cause a 66.9% reduction of mitogen-induced lymphocyte proliferation. Three chromosome aberrations were detected in PL conditions. Since two changes occurred in the same do nor, it is possible they were donor dependent rather than caused by the culture condition. These findings demonstrate that a 10% PL condition enables a higher yield of MSCs within a shorter time without altering MSC properties, and should be favored over the 5% PL condition.
BACKGROUND
Oral chronic graft‐versus‐host disease (cGvHD) impairs oral function and patients' quality of life. Some lesions are refractory to local and systemic immunosuppressive therapy, and new ...therapeutic modalities are required. The aim of the study was to assess the efficacy and safety of topical application of autologous platelet gel (PG) in patients with oral cGvHD.
STUDY DESIGN AND METHODS
PG was prepared from autologous blood and applied on ulcerous lesions using an automated system. The oral cGvHD was assessed using the 273‐point Oral Mucositis Rating Scale (OMRS) prior and after completion of the PG treatment. The overall response to treatment of particular topography expressed as the total score on OMRS was compared to total score on National Institutes of Health cGvHD Oral Mucosal Score (NIH OMS). The pain intensity was measured by the Numeric Pain Rating Scale (NRS).
RESULTS
In five patients, 12 autologous blood collections were performed; median 3 (range 1‐3) per patient, and 26 PG applications were performed; median 6 (range 2‐8) per patient. PG applications reduced lesions in oral cGvHD: median OMRS total score was reduced for 43.2% (range 9.6%‐47.3%), and median NIH OMS total score for 27.3% (range 20.0%‐50.0%) from baseline values. Median of pain intensity reduction on NRS scale was 57.1% (range 50%‐100%). No side effects were observed.
CONCLUSION
Application of autologous PG in oral cGvHD showed as an efficient and safe treatment option for patients who do not respond to standard local treatment.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
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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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Alloimmunization is a known risk of transfusion therapy caused by exposure to foreign RBC antigens. However, alloimmunization is not observed in all transfused patients. Human leukocyte antigen (HLA) ...molecules may contribute to the recognition and presentation of foreign antigens and to the potency of immune responses that result in the production of antibodies. The aim of this study was to determine the association of HLA-DR and HLA-DQ polymorphisms with alloimunization to Fy
antigen in Croatian patients.
The study was conducted on 70 alloimmunized patients to Fy
antigen and two control groups: 165 healthy Croatian individuals (Control 1) and 45 Fy
antigen-negative nonimmunized patients exposed to Fy
antigen (Control 2). Phenotype frequencies for HLA-DRB1 and HLA-DQB1 alleles were compared between the cases and control groups.
Statistically significant differences in phenotype frequencies between cases and controls were found for DRB1*04 (odds ratios ORs, 10.5 and 18.7 for Control 1 and Control 2, respectively), DRB1*15 (ORs, 8.0 and 6.9), and DQB1*02 alleles (ORs, 0.2 and 0.03); and DRB1*04-DQB1*03:01 (ORs, 7.9 and 17.6), DRB1*04-DQB1*03:02 (ORs, 5.5 and 7.6), DRB1*15-DQB1*06:02 (ORs, 7.3 and 5.5), DRB1*03-DQB1*02:01 (OR, 0.1), and DRB1*07-DQB1*02:02 (OR, 0.3) haplotypes.
Several HLA-DRB1 and HLA-DQB1 alleles and haplotypes were proved to contribute to and protect from alloimmunization to Fy
antigens. Alleles DRB1*04 and DRB1*15, as well as haplotypes DRB1*04-DQB1*03:02 and DRB1*15-DQB1*06:02 can be considered as risk factors, while allele DQB1*02 and haplotype DRB1*03-DQB1*02:01 have a protective role in Fy
alloimmunization.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
BackgroundCorneas 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.MethodsCorneas 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.Results1426 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%).ConclusionOrgan 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.
This study provides an overview of tissue banking activities at the Croatian Cardiovascular Tissue Bank (CTB) during past ten years and presents the outcomes of cryopreserved heart valve allografts ...(CHAs) use in different patient groups. From June 2011 until December 2021, 75 heart donations were referred to CTB: 41 recipient of heart transplant (RHT), 32 donors after brain death (DBD) and 2 donors after circulatory death (DCD) donations. Processing resulted in 103 valves of which 65 met quality requirements for clinical use. Overall tissue discard rate was 37%. The most frequent reasons for discard were inadequate morphology (12%) in RHT donations and microbiological contamination (19%) in DBD donations. Altogether, 38 CHAs were transplanted to 36 patients. Recipients were divided in three groups; infective endocarditis (IE), non-infectious heart disease and congenital heart disease group. In the IE group, the 30-day, 1-year and 3-year survival was 71%, 53% and 47%, respectively. Freedom from re-operation due to all graft-related causes was 76% and due to structural valve deterioration 88%. There were no cases of graft reinfection. In the congenital heart disease group CHAs were predominantly (94%) used for right ventricular outflow tract reconstruction and 88% of patients recovered without graft-related complications. At present, the number of demands for CHAs at CTB considerably outweighs their availability.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
An estimated 20% of allogeneic blood transfusions in the United States are associated with cardiac surgery. It is estimated that 11% of red cell resources were used for transfusion support of ...patients undergoing coronary artery bypass grafting (CABG) with a documented wide variability in transfusion rate (7.8 to 92.8%). To address the issue of unnecessary transfusions within the CABG population, we developed a model to predict which patients are at low risk of bleeding for whom transfusion treatment might be considered unnecessary. Herein we present our "SHOULD-NOT-BLEED-SCORE" application developed for the Windows® software platform which is based on our previous research.
This study is aimed to develop a user-friendly application that stratifies patients with respect to bleeding risk. The statistical model we used in our previous research was focused on detection of CABG patients at low risk of bleeding. The rationale behind such an approach was to identify a CABG patient subgroup at low risk of bleeding. By identifying patients at low risk of bleeding we can define a subgroup of patients for whom transfusion treatment might be considered unnecessary. We developed a Windows platform application based on risk modelling which we previously calculated for 1426 patients undergoing elective CABG from January 2010 to January 2018.
The SHOULD-NOT-BLEED-SCORE risk score is developed for the Windows software platform. A mathematical model that is based on multivariate analysis was used for app development. The variables that entered the scoring system were: Age; Body Mass Index; Chronic Renal Failure; Preoperative Clopidogrel Exposure; Preoperative Red Blood Cells Count; Preoperative Fibrinogen Level; Preoperative Multiplate ASPI test area under the curve (AUC) units. The SHOULD-NOT-BLEED-SCORE identifies/predicts patients without a risk for excessive bleeding with strong discriminatory performance (Receiver Operating Curve (ROC) analysis AUC 72.3%, p < 0.001).
The SHOULD-NOT-BLEED risk scoring application may be useful in the preoperative risk screening process. The clinical and economic burden associated with unnecessary transfusions may be adequately addressed by a preoperative scoring system detecting patients at low risk of bleeding for whom transfusion treatment might be considered unnecessary.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Fetal/neonatal alloimmune thrombocytopenia (FNAIT) is a rare life‐threatening disorder, leading to severe thrombocytopenia and potentially bleeding, with intracranial haemorrhage (ICH) being the most ...serious complication. We report on a FNAIT case with fourth‐degree ICH that arose due to antibodies against human platelet antigen (HPA)‐1b. The male infant, born to an otherwise healthy mother, presented with severe signs of ICH soon after delivery. Since only moderate thrombocytopenia was noted and there were no active signs of bleeding, the infant did not receive intravenous immunoglobulins (IVIg) or platelet transfusion. Spontaneous recovery of platelets was noted on the eighth day of life, but permanent neurological impairment remained as a consequence of ICH. We report the results of HPA and human leukocyte antigen (HLA) antibodies in the mother's and the infant's sera, the family's HPA genotype and the mother's HLA genotype, and summarise previously described cases of FNAIT due to anti‐HPA‐1b antibodies in the literature. FNAIT with severe ICH due to anti‐HPA‐1b antibodies is rarely diagnosed. An association between HLA genes and sensitization to HPA‐1b antibodies was not demonstrated. The severity of FNAIT and the occurrence of ICH is often difficult to predict. In this case, the infant presented with moderate thrombocytopenia and ICH, with subsequent permanent consequences.
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DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, SIK, UILJ, UKNU, UL, UM, UPUK
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