Mirasol PRT system inactivation efficacy evaluated in platelet concentrates by bacteria-contamination model

Background/Aim. Bacterial contamination of blood components, primarily platelet concentrates (PCs), has been identified as one of the most frequent infectious complications in transfusion practice. PC units have a high risk for bacterial growth/multiplication due to their storage at ambient temperature (20 ± 2°C). Consequences of blood contamination could be effectively prevented or reduced by pathogen inactivation systems. The aim of this study was to determine the Mirasol pathogen reduction technology (PRT) system efficacy in PCs using an artificial bacteria-contamination model. Methods. According to the ABO blood groups, PC units (n = 216) were pooled into 54 pools (PC-Ps). PC-Ps were divided into three equal groups, with 18 units in each, designed for an artificial bacteria-contamination. Briefly, PC-Ps were contaminated by Staphylococcus epidermidis, Staphylococcus aureus or Escherichia coli in concentrations 102 to 107 colony forming units (CFU) per unit. Afterward, PC-Ps were underwent to inactivation by Mirasol PRT system, using UV (l = 265-370 nm) activated riboflavin (RB). All PC-Ps were assayed by BacT/Alert Microbial Detection System for CFU quantification before and after the Mirasol treatment. Samples from non-inactivated PC-P units were tested after preparation and immediately following bacterial contamination. Samples from Mirasol treated units were quantified for CFUs one hour, 3 days and 5 days after inactivation. Results. A complete inactivation of all bacteria species was obtained at CFU concentrations of 102 and 103 per PC-P unit through storage/ investigation period. The most effective inactivation (105 CFU per PC-P unit) was obtained in Escherichia coli setting. Contrary, inactivation of all the three tested bacteria species was unworkable in concentrations of ≥ 106 CFU per PC-P unit. Conclusion. Efficient inactivation of investigated bacteria types with a significant CFU depletion in PC-P units was obtained - 3 Log for all three tested species, and 5 Log for Escherichia coli. The safety of blood component therapy, primarily the clinical use of PCs can be improved using the Mirasol PRT system

HIV/AIDS and other sexually transmitted infections among military members of the armed forces of Serbia

Background/Aim. Military personnel is a population group at special risk of exposure to sexually transmitted diseases (STD). In peacetime, STD infection rates among service members are generally 2 to 5 times higher than among civilian population. In time of conflict, the differences can be 50 or more times greater. This study describes sexual behavior as a risk factor for STD in the Armed Forces of Serbia. Methods. The sample of 5 617 voluntary blood donors from the Armed Forces of Serbia gave blood and filled World Health Organization Questionnaire about sexual behavior within January 2007 - December 2008 period. The mandatory testing of voluntary blood donors was performed in the Institute of Transfusiology Military Medical Academy in Belgrade, by the specific immunoenzyme tests and polymerasa chain reaction tests for HIV, hepatitis B, C and syphilis. Statistical analysis of data was done using State for Windows 93, USA, 1996. Results. We identified 36 soldiers with some form of STDs. This study showed that 1 668 (29.7%) tested soldiers reported always using condoms, 1 725 (30.72%) almost always, 1 238 (20.04%) sometimes, 495 (8.81%) almost never and 490 (8.73%) never. Among the sample, 449 (7.99%) soldiers reported sexual contacts with partners with high risk of sexual behavior, whilst 22 (0.37%) of them reported homosexual and bisexual contacts. Conclusion. This study reported STDs found in voluntary blood donors among the service members of the Armed Forces of Serbia, but none of them was identified to be HIV positive. Soldiers with the most frequent risk behavior were reported to be those with inconsistent condom use. In the future, the STD Control and Prevention Program should be more intensively conducted among the members of the Armed Forces of Serbia

Stem cell harvesting protocol research in autologous transplantation setting: Large volume vs. conventional cytapheresis

Background/Aim. The use of peripheral blood as a source of hematopoietic stem cells (SCs) is progressively increasing and has nearly supplanted bone marrow transplantation. Interpatient variability in the degree and kinetics of SC mobilization into peripheral blood is an expected event after conventional chemotherapy-based treatment, followed by sequential administration of recombinant granulocyte-colony- stimulating factor (rHu-CSF). In this study, specific factors associated with the application of two different SC-harvesting approaches, including the use of large volume leukapheresis (LVL) vs. repetitive conventional apheresis (RCA), were analyzed. The basic goal of the study was to evaluate the influence of apheresis protocol (collection timing, processed blood volume and cell yield) upon the clinical outcome of transplantation. Methods. Results obtained by LVL (76 pts) and RCA (20 pts - control group) were compared. The SC mobilizing regimen used was cyclophosphamide (4-7 g/m2) or polychemotherapy and rHuG-CSF 10-16 μg/kg of body mess (bm) per day. Cell harvesting was performed using COBE-Spectra (Caridian-BCT, USA). The volume of processed blood in LVL setting was ≥ 3.5 - fold of the patient's circulating blood quantity (ranged from 12.7 to 37.8 l). All patients tolerated well the use of intensive treatment, without any side or adverse effects. Our original controlled-rate cryopreservation was carried out with 10% dimethyl sulfoxide (DMSO) using Planer R203/200R or Planer 560-16 equipments (Planer Products Ltd, UK). Total nucleated cell (NC) and mononuclear cell (MNC) counts were examined by flow cytometry (Advia-2120 Bayer, Germany; Technicon H-3 System, USA). The CD34+ cell surface antigen was investigated by the EPICS XL-MCL device (Coulter, Germany). Results. Performing LVL-apheresis, high-level MNC and CD34+ cell yields (7.6±4.6 × 108/kg bm and 11.8±6.5 × 106/kg bm, respectively) were obtained. As a result, rapid hematopoietic reconstitution ("graft-healing") - on the 9.4th and 12.4th day for granulocytes and platelets, respectively was achieved. Using repetitive conventional apheresis (2-3 procedures), the total MNC count was high (8.2±7.0 × 108/kg bm), but the total CD34+ yield was lower 10.8±9.9 due to inferior CD34+ vs. MNC ratio. Conclusion. The results obtained suggest that well-timed LVL-apheresis increased SC-yield in cell harvest, resulting in faster bone marrow repopulation and hematological reconstitution, as well as better overall clinical outcome of transplantation. These results necessitate additional examinations of CD34+ subsets ratio in cell harvest

Ispitivanje prinosa i indukcuske sposobnosti humanih leukocita pripremljenih iz 'buffy coat-a', namenjenih za proizvodnju interferona - alfa

Jedan od osnovnih ciljeva ovog rada je ispitivanje mogućnosti uvođenja novih postupaka u pripremi, skladištenju i daljoj obradi prečišćene suspenzije leukocita - "huffy coat-a" (BC-α) namenjene za proizvodnju humanog interferona-alfa (MFN-α). Naša ispitivanja su pokazala da su srednje vrednosti (SV) broja lukocita (Le) u jedinicama cele krvi (JCK) upotrebljenim za pripremu BC-α, za sve ispitane serije približno iste (oko 2,4 x 109). Prosečno je oko 90% tih Le izdvojeno u svakom BC-u. Tokom skladištenja broj Leje smanjen za oko 9% u BC-u skladištenom na konstantnoj temperaturi (22 °C) u eksperimentalnim serijama BC-α (ESBC-I i ESBC-II), a znatno vise u kontrolnoj seriji BC-α (KS-BC) skladištenoj na ambijentalnoj temperaturi. Indukciona sposobnost Le ispitivana je proračunom srednjih vrednosti titra MFN-α. Srednje vrednosti titra hlFN-aiz uzoraka KSBC su manje za 18% od srednjih vrednosti titra MFN-α u uzorcima ESBC-II, a za oko 5% man-je od srednjih vrednosti titra MFN-α u uzorcima ESBC-I

Living unrelated donor kidney transplantation: A fourteen-year experience

Background. In countries without a national organization for retrieval and distribution of organs of the deceased donors, problem of organ shortage is still not resolved. In order to increase the number of kidney transplantations we started with the program of living unrelated - spousal donors. The aim of this study was to compare treatment outcome and renal graft function in patients receiving the graft from spousal and those receiving ghe graft from living related donors. Method. We retrospectively identified 14 patients who received renal allograft from spousal donors between 1996 and 2009 (group I). The control group consisted of 14 patients who got graft from related donor retrieved from the database and matched than with respect to sex, age, kidney disease, immunological and viral pretransplant status, the initial method of the end stage renal disease treatment and ABO compatibility. In the follow-up period of 41 ± 38 months we recorded immunosuppressive therapy, surgical complications, episodes of acute rejection, CMV infection and graft function, assessed by serum creatinine levels at the beginning and in the end of the follow-up period. All patients had pretransplant negative cross-match. In ABO incompatible patients pretransplant isoagglutinine titer was zero. Results. The patients with a spousal donor had worse HLA matching. There were no significant differences between the groups in surgical, infective, immunological complications and graft function. Two patients from the group I returned to hemodialysis after 82 and 22 months due to serious comorbidities. Conclusion. In spite of the worse HLA matching, graft survival and function of renal grafts from spousal donors were as good as those retrieved from related donors