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Introduction. Poor graft function is one of the most severe complications after allogeneic hematopoietic stem cell transplantation, which manifests as pancytopenia/cytopenia in the blood count, with the presence of complete or incomplete donor chimerism. There are three entities of graft weakness: 1. poor graft function: pancytopenia with complete donor chimerism, 2. graft failure: pancytopenia with incomplete, i.e., mixed donor chimerism and 3. graft rejection: progressive decline of donor chimerism. Definition. Poor graft function is diagnosed as pancytopenia (hemoglobin < 70 g/L, absolute neutrophil count < 0.5 x 109/L, platelets < 20 x 109/L) for 3 consecutive days from D+28, excluding the presence of severe graft versus host disease and relapse, with complete donor chimerism in poor graft function, and incomplete in graft failure. Risk factors and therapeutic principles. The most common risk factors for poor graft function are a small dose of CD34+ hematopoietic stem cells in the transplant, graft versus host disease, cytomegalovirus infection, the presence of donor-specific antibodies, high serum ferritin, i.e., iron overload, as well as splenomegaly. Pathogenetic mechanisms in the development of poor graft function are still not fully elucidated. The role of the microenvironment of the patient?s bone marrow is also important, as well as disorders of the immune system Therapeutic options for overcoming this complication include using selected ?stem cell boost?, mesenchymal stem cells, and newer medical agents (N-acetyl cysteine, atorvastatin, thrombopoietin receptor agonists). Conclusion. The type of poor function of the graft is defined in relation to the percentage of donor chimerism, and is necessary for planning further treatment strategy.

Conventional hematopoietic stem cell transplantation is a well-known treatment method for numerous acquired and congenital hematopoietic disorders, disorders of the immune system, as well as certain metabolic disorders. Stem cells (SCs) can be defined as cells capable of self-renewal with a high proliferative capacity and the potential to differentiate into functionally competent mature cells. Stem cells can be divided into embryonic SCs (ESCs) and tissue-specific or adult SCs - such as bone marrow (BM) stem cells, peripheral blood (PB) stem cells, and SCs derived from umbilical cord blood (UCB), as well as other non-hematopoietic or somatic SCs. SCs in adults are characteristically considered to be restricted in their regenerative and differentiative potential, while embryonic stem cells are 'true' totipotent/pluripotent cells, due to their ability to develop into endoderm, ectoderm, or mesoderm - all three embryonic tissue types in the human body. They are the most promising, but also the most controversial type of potentially transplantable SCs. Immature hematopoietic SCs have the potential of differentiating, not only into all blood cells, but also into some somatic cell types (SC plasticity). In different clinical settings, the transplantation of immature stem cells leads to the repopulation of recipient bone marrow, with subsequent complete, stable, and long-term reconstitution of hematopoiesis. Given that immature stem cells are also capable of homing to different tissues, autologous stem cell implantation into a damaged and/or ischemic area induces their colonizing and consecutive transdifferentiating into cell lineages of the host organ, including neovascularization. Thus, they are clinically applicable in the field of regenerative medicine for the treatment of myocardial, brain, vascular, liver, pancreatic, and other tissue damage. The purpose of this overview is to recapitulate the key developments in the rapidly evolving area of stem cell research, as well as to review the use of SCs in conventional transplantations and in regenerative medicine. Additionally, a brief critical evaluation of our own stem cell research will be summarized.

Background/Aim. After the introduction of a careful selection procedure for blood donors and the implementation of highly sensitive screening tests for transfusion-transmitted infections (TTIs), blood has become a very safe product concerning TTIs. However, due to the existence of a ?window? period during which these ?markers? cannot be detected, as well as the emergence of new pathogens, the risk is still present. Implementation of pathogen reduction technology (PRT) provides a proactive approach to improving blood safety. By damaging nucleic acids, PRT selectively inactivates pathogens and leucocytes. Nevertheless, during the process, plasma proteins are also damaged to some extent. The aim of this study was to conclude whether there is a difference in the effect of PRT on protein S (PS) and alpha 2-antiplasmin (?2AP) regarding the time of inactivation: inactivation immediately after plasma separation from whole blood (before freezing) vs. inactivation after freezing/thawing. Methods. The voluntary donors? blood is taken into a quadruple bag system, centrifuged, and separated into blood products. Control group plasma was first inactivated by the Mirasol? PRT system and then frozen. Experimental group plasma was immediately frozen and, after four months, thawed and inactivated. PS and ?2AP activity was examined in samples after separation, inactivation, and thawing. Results. Analyzing PS and ?2AP activity, no statistically significant difference was found between the initial samples. The trend of protein activity reduction after inactivation and freezing/thawing was present in both groups but without a statistically significant intergroup difference. Conclusion. No statistically significant difference was found between the activity values of PS and ?2AP after immediate inactivation, before freezing, and after freezing/thawing, making stored plasma units suitable for safe and efficient inactivation directly before clinical use and according to the patient?s blood type.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a new human disease. December 31, 2019 marked the day the World Health Organization (WHO) first became aware of an infectious outbreak in the Hubei province in China. Until January 2021, more than two million people died from COVID-19. The use of convalescent plasma (CP) has been widely used in different outbreaks as the first therapeutic option given the lack of effective medications or vaccines, and often as a last chance or experimental treatment. CP is a strategy of passive immunisation. Possible mechanisms of CP-COVID-19 action are antiviral and immunomodulatory. The established protocol for CP-COVID-19 collection defines activities and criteria related to recruiting and informing potential CP donors, clinical and laboratory examination, plasma collection, labelling and storage. Plasma is collected by apheresis/plasmapheresis. Administration of plasma is performed at the request of clinicians, according to the strict indications based on the severity of clinical picture, expressed by precisely determined "scoring" of symptoms. The risks transfusion recipients are likely to be exposed to do not differ from those of standard plasma recipients. At the Blood Transfusion Institute of Serbia, the first plasmapheresis from the recovered patient-donor was performed on 11 April 2020 and so far, collection and distribution of CP-COVID-19 have been performed continuously. During the observation period, preliminary results of the effect of CP transfusion, along with other applied therapy, indicate its favourable effect, both worldwide and in Serbia. CP-COVID-19 should be used as early as possible in the course of infection in order to achieve the best outcomes.

Background/Aim. The treatment of chronic myeloid leukemia (CML) has changed dramatically with the advent of targeted therapies. This study aimed to assess the efficacy of generic imatinib in CML patients treated in our center. Methods. The study was retrospective. It included 101 patients diagnosed with CML ? chronic phase (CP). The patients were divided into two groups. Group 1 included 55 patients initially treated with branded imatinib and then switched to generic imatinib. Group 2 consisted of 46 newly diagnosed patients who received only generic imatinib from the beginning of therapy. Results. The patients were treated with branded imatinib for the mean of 42 months (range 6?132 months) before switching to generic imatinib. Treatment with generic imatinib lasted for 25 months on average (range 3?66 months). A quarter of the patients from the group 1 lost their cytogenetic reponse after being switched to generic imatinib, but with-out signs of transformation to acute leukemia. The patients treated with branded imatinib had a significantly longer event-free survival (EFS) and failure-free survival (FFS) (log-rank p = 0.01 and p = 0.03, respectively). These results could have been influenced by frequent changes of the brand and dosage formulation of generic imatinib. Conclusions. Our study showed a significantly longer EFS and FFS in the patients who were initially treated with branded imatinib, compared to those treated with generic imatinib only. These results provide useful information, but have to be interpreted within the context of the crossover study.

Stem cells (SC) are the unique and "key-cells" in the human body "working" as a source of producing a large number (proliferation) of mature (differentiation) cells inside different tissues ("cytopoiesis") - while at the same time maintaining the ability to "reproduce" themselves (self-renewal). These events are balanced by interactive signals from the extracellular matrix, as well as microenvironment provided by stromal cells. On the other hand, SC plasticity (so-called "inter-systemic plasticity") is the ability of the most "primitive" (immature) adult SCs to switch to novel identities. The phrase SC plasticity also involves phenotypic potential of these cells, broader than spectrum of phenotypes of differentiated cells in their original tissues. Recent increasing clinical use of cell-mediated therapeutic approaches has resulted in enlarged needs for both, higher quantity of SCs and improved operating procedures during extracorporeal manipulations. The aim of harvesting procedures is to obtain the best SC yield and viability. The goal of optimised cryopreservation is to minimise cellular thermal damages during freeze/thaw process (cryoinjury). Despite the fact that different SC collection, purification and cryopreservation protocols are already in routine use - a lot of problems related to the optimal SC extracorporeal manipulations are still unresolved. The objective of this paper is to provide an integral review of early haemobiological and cryobiological research in the unlimited "SC-field" with emphasis on their entities, recent cell-concepts, extracorporeal manipulative and "graft-engineering" systems. Their therapeutic relevance and efficacy in "conventional" SC transplants or regenerative medicine will be briefly summarised. Finally, in this paper original results will not be pointed out - related to neither SC transplants nor regenerative medicine - but a light will be shed on some of them.

Background/Aim. Despite the introduction of new oral anticoagulants (dabigatran, rivoroxaban, apixaban), vitamin K antagonists (VKA), such as warfarin and acenocoumarol are still the most widely used oral anticoagulants for the treatment of non-valvular atrial fibrillation (NVAF). Time in therapeutic range (TTR) represents a measure of the quality of the anticoagulant effect of these drugs, and it is considered that the lower value of TTR is associated with the adverse effects of therapy. The study aim was to evaluate of the effectiveness of VKA therapy in patients with NVAF and to identify factors affecting the anticoagulation efficacy. Methods. A retrospective study was conducted on a population of 725 outpatients with NVAF, treated with VKA and followed in Blood Transfusion Institute of Nis from January to December 2017. Laboratory control of the INR was done from capillary blood of patients on Thrombotrack Solo (Axis Shield, Norway) and Thrombostat (Behnk Elektronik, Germany). Targeted therapeutic INR was between 2.0 and 3.0. For each patient we evaluated all available INR values to calculate the individual TTR according to the Rosendaal method. Results. The study included a total of 725 patients with NVAF which had 6105 INR measurements, what is 8.13±2.47 INR measurements per patient. The mean value of TTR and was 60.15±17.52%, but 49.72% of patients had a TTR less than 60%. Patients were at high risk of thrombosis in 6.15% of time (INR 4.5). The most significant independent factors affecting the quality of VKA therapy are gender, arterial hypertension, diabetes mellitus and the use of amiodarone and antiplatelet drugs (aspirin, clopidogrel). Conclusion. The TTR is undoubtedly useful indicator of the effectiveness of VKA treatment. The most important predictors of poorer efficacy of VKA therapy are arterial hypertension, diabetes mellitus, patints' gender and the use of amiodarone and antiplatelet (aspirin, clopidogrel) drugs. To improve the quality of VKA therapy, education of patient and better collaboration with them, as well as a successful teamwork with clinicians are also imperative.

Using intraoperative salvage of blood/erythrocytes (ISB/E) in the practice, rationalized application of allogeneic blood products and hemostatic-active drugs - based on "point-of-care" (POC) monitoring findings - it is possible to improve diagnosis and evaluate of transfusion hazards and increase efficacy of therapy of patients with excessive bleedings, with reduction of perioperative morbidity and mortality in cardiac surgery. This study was performed as a prospective analysis of platelet function using "multiple platelet function analyzer" (MEA; by Multiplate) system and examination of hemostasis by rotational thromboelastometry (ROTEM) during and immediately following myocardial revascularization and surgical treatment of valves in the Institute of Cardiovascular Diseases "Dedinje" for six years period for therapy of 1021 random selected patients. The study aim was to evaluate the influence of ISB/E and hemostatic drugs - indicated based on the results of platelet count and function (Multiplate) and hemostasis monitoring (ROTEM) in compared to allogeneic transfusions - on the incidence of bleedings, treatment efficiency and overall clinical outcome. In the perioperative period, a total of 617 (60.4%) patients were treated with ISB/E reinfusion only. Other patients (404; 39.6%) received allogeneic blood components too. Total 391 of them (38.3%) were treated (together by ISB/E) with transfusion of one to three units of allogeneic red blood cells (RBCs). There were only 13 (1.3%) polytransfunded (typically 10 - 15 units) patients. The rate of cardio-surgical reinterventions due to bleeding was only 2.5%. In conclusion, the application of the ISB/E strategy represents an effective and safe (reduced immune-mediated complications and risk of disease transmission) therapeutic approach. By monitoring MEA/ROTEM and implementation the algorithm of current transfusion therapy, it is possible to reduce significantly of allogeneic blood component therapy. The use of allogeneic RBCs is justified only when the possibilities of autologous transfusion and pharmacological hemostatic therapy have been exhausted.

There are four stem cell (SC) transplant centers in Serbia that carry out their activities within the European Group for Blood and Marrow Transplantation (EBMT). They are positioned in the Clinical Center of Serbia, Military Medical Academy, Institute for Mother and Child Healthcare "Dr. Vukan Čupić" and in the Clinical Center of Vojvodina. The aim of this paper is to analyze the basic approaches and aspects, as well as main parameters and guidelines - related to the optimization of the functioning of SC transplantation centers. The existence of a competent and motivated professional employees of the transplant center, as well as an adequate operational locality/area and up to date equipments are imperative for the successful performance of this specific and complex procedure. The current tendency - and in the near future an obligation or requirement - is for each transplant center within the EBMT to be fully staffed and structurally equipped and ready to be included in the accreditation process by the Joint Accreditation Committee International. Begining the accreditation process implies, in addition to the implementation of education and training of employees, the establishment and implementation of specific quality control standards (ISO 9001: 2015, ISO 13485, ISO 13485: 2016, ISO 15189: 2007, ISO 17025, ISO / IEC 7025: 2005). The activity indicators of every transplant center have to be transparent and presented by: the number of treated patients, the rate of transplant mortality, the duration and quality of patient's lives. Also important parameters are various methods of economic analyses - different specific cost-benefit analyses. In conclusion, in order to achieve consistent/comparable therapeutic effects and outcomes for patients globally - it is imperative that transplant centers and related organizations work together on the quality system assurance, using a continuous and adequate "educational and organizational-structural models".

The spread of the COVID-19 virus has a strong influence on blood collection, maintaining a stable supply of all blood components and the safety of the transfusion itself. SARS-CoV-2 has a long incubation period (1-14 days, on average 5-6 days, longest reported 24 days) and causes asymptomatic infection in a large number of patients, which is a great challenge in a recruitment of blood donors and achieving a safe transfusion. Precise recommendations and precautions have been adopted regarding the criteria for temporary refusal of blood donors during the COVID-19 pandemic, organization of mobile teams and collection sites, disposal of medical waste, examination of potential donors and mandatory body temperature measurement. Although transmission of COVID-19 via blood and blood components has not been demonstrated, some countries have also introduced mandatory NAT testing for SARS-CoV-2 as a part of blood screening testing. Also, proactive measures have been taken, such as temporary storage of blood in quarantine for 14 days after collection, while special attention is paid to efficient management of blood component stocks and development of a collection plan, in order to avoid shortage of certain blood components or their expiration. The first step in this regard is to revise the measures which have the aim for improving the usability of blood components, ie reducing waste of stocks, which primarily refers to the temporary extension of the shelf life of blood components. Extending the shelf life of erythrocytes (longer than 35 to 49 days, which is defined at the national level) should be considered as early as possible, because once a shortage of erythrocytes occurs, they will be issued long before the expiration date. Previous studies have not shown significant side effects of erythrocyte transfusion with extended shelf life, so it is possible to consider the flexibility of blood processing and erythrocyte storage conditions with mandatory internal process validation and component quality control. The shelf life of platelet concentrate should be extended from 5 days to 7 or even 8 days, with mandatory bacteriological testing or pathogen inactivation. Another option to increase the platelet supply for prophylactic purposes is to reduce the platelet dose by dividing the existing components. Frozen fresh plasma has the longest shelf life (up to 3 years), so maintaining stable reserves is much safer than for cellular components. Liquid plasma (never previously frozen) has a shelf life of 7-40 days, and can be used in conditions of reduced freezer capacity, shortage of staff working on blood processing or for the production of convalescent plasma. Pathogen inactivation of plasma and platelets allows 3-6 log reduction of SARS-CoV-2 and MERS-CoV. The decision to introduce some of the methods of pathogen inactivation should be made taking into account the costs and resources required for implementation. For countries that do not have pathogenic inactivation already in routine practice, its rapid introduction is a big task. For now, the risk of SARS-CoV-2 transmission through the blood appears to be very low, although our understanding of the virus and behavior during a pandemic will improve over time. In this regard, pathogen inactivation of convalescent plasma should also be considered.

Hematopoietic stem cells (SCs) are responsible for the production and replacement (proliferation) of an extensive quantity of functionally competent blood cells (differentiation) during the entire life, while simultaneously maintaining the ability to reproduce themselves (self-renewal). A complex network of interactive substances and factors organize and protect the survival, maturation and multiplication of SCs. Hemobiological events in the bone marrow (BM) are synchronized and balanced by the extracellular matrix and microenvironment provided by stromal cells. These cells-including macrophages, fibroblasts, dendritic, endothelial and other cells-stimulate SCs by producing specific hematopoietic growth factors. Other cytokines secreted by stromal cells regulate the adhesion molecules positioned on SCs, allowing them to remain in the BM or migrate to an area where the respective cell type is needed. Thus, hematopoietic SCs could be defined as cells with high proliferative capacity and extensive potential to differentiate into all blood cells or some somatic cell types (SC plasticity)-such as cardiomyocytes, myocytes, osteocytes, chondrocytes, hepatocytes, and even endothelial cells. Recent increasing clinical use of cell-mediated therapeutic approaches has resulted in increased needs for SCs, but in superior operating procedures during their ex vivo manipulations. The aim of cell harvestings is to obtain a higher SC yield and improved viability or clonogenicity. The goal of optimized cryoinvestigation protocols is to get a minimized cell damages (cryoinjury). Despite the fact that different SC collection protocols and cell freezing practice are already in routine use, a lot of questions related to the optimal SC ex vivo manipulations are still unresolved. This review summarizes fundamental knowledge and methodological approaches, and recapitulates data enabling progress on constantly evolving research frontiers in the SC area. The studies (including also our investigations) that evaluated the efficiency and safety of SC-treatment (transplants and regenerative medicine) will be also concisely presented.

Chronic granulocytic leukemia (CGL), Ph + is a chronic myeloproliferative disorder, which, due to its specificity for many decades, has attracted the attention of researchers of specialties. This disease was among the first malignant haemopathies that received their “ID card” since molecular precisely defined the basic pathological substrate, that origin of the disease. Over the past decades we have witnessed the evolution of the therapeutic approach in the treatment of from oral cystostatic therapy, radiotherapy, through recombinant interferon alpha (IFN-alpha), haematopoeza stem cell transplantation, to the targeted molecular therapy of the tyrosine inhibitor (TKI). In this report we present patient with a diagnosis of Ph + in the early chronic phase at the age of 13 all relevant analyzes. During the past 26 years, all modalities of CGL treatment were applied in patients, according to the time period, concluding with the second generation resulting in complete remission of diseases with excellent quality of life and identical expectations for the future, as well as in healthy population.

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The patient was admitted for urgent coronary angiography following an acute anterior ST segment elevation myocardial reinfarction (STEMI) caused by acute stent thrombosis. A stent had been implanted 10 days prior to the reinfarction for an acute anterior STEMI. However, the patient had stopped taking ticagrelor post-discharge. Primary percutaneous coronary intervention of the left anterior descending artery was performed. Subsequently, due to a high C-reactive protein (CRP) level, 3 CRP apheresis sessions were performed, with the first session starting 12 h after the onset of symptoms. A significant drop in CRP was noted after each apheresis. The post-procedural course was uneventful.

Background/Aim. Intravenous drug users (IDUs) are still a high risk-group for cross-reacting blood-borne infections, for vertical pathogen transmission as well as for potentially blood/plasma donation (especially as ?paid? donors). The aim of our study was to establish the profile of opiate addict and prevalence of blood-borne pathogens ? Hepatitis B virus (HBV), Hepatitis C virus (HCV) and Human Immunodeficiency Virus (HIV) among 99 patients on substitution therapy with methadone and buprenorphine from Sumadia District. Methods. The Treatment Demand Indicator (TDI) of Pompidou-questionnaire was used to assess the history of drug abuse and risk behavior. All blood samples were tested for HBV surface antigen (HBsAg), anti-HCV antibody (anti-HCV) and HIV antigen/antibody (HIVAg/ Ab) by Enzyme-Linked ImmunoSorbent Assay (ELISA) or Chemiluminescent Immuno-Assay (CIA). Investigations were also performed for HBV, HCV and HIV by molecular testing ? Polymerase Chain Reaction (PCR) method. Results. The majority of patients were males (81.8%), median age 32 (19?57) years, lived in a city (99%), unemployed (58.6%), with finished secondary school (67.7%), unsafe injecting practices (34.3%) and never previously tested for HBV (39.4%), HCV (36.4%) nor HIV (28.3%); only 4% of them previously got HBV-vaccine. The complementary testing resulted with following results: HBV ELISA/CIA and PCR negativity for 66 patients and positive results (by ELISA/CIA and PCR) for 19 patients. However, a difference was observed in the ELISA/CIAnegative/ PCR-positive result for 12 and ELISA/CIApositive/ PCR-negative for two patients respectively. Further, the negative results for HCV (ELISA/CIA and PCR testing) were found in 15 IDUs and positive results (using both methods) were found in 58 patients. Different results for ELISA/CIA-negative / PCR-positive results were found in 11 IDUs and ELISA/CIA-positive/PCR ? negative results were found in 15 patients. All investigated IDUs were negative for HIV (ELISA/CIA and PCR testing) and for pathogens of opportunistic infection (Cryptococcus neoformans; Pneumocystis carinii; PCR testing), as well as for West Nile Virus (PCR testing). Just one IDU was positive for syphilis (ELISA and confirmatory testing). Conclusion. Our study demonstrated that the positivity for HBV and HCV is still very high (33.4% and 84.8%, respectively) in IDUs. Thus, we suggest that drug users have to be periodically screened using a complementary serological/molecular testing ? concerning differences/discrepancies in the results obtained using these methods.

Introduction. Risk stratification is an important aspect of COVID-19 management, especially in patients admitted to ICU as it can provide more useful consumption of health resources, as well as prioritize critical care services in situations of overwhelming number of patients. Materials and Methods. A multivariable predictive model for mortality was developed using data solely from a derivation cohort of 160 COVID-19 patients with moderate to severe ARDS admitted to ICU. The regression coefficients from the final multivariate model of the derivation study were used to assign points for the risk model, consisted of all significant variables from the multivariate analysis and age as a known risk factor for COVID-19 patient mortality. The newly developed AIDA score was arrived at by assigning 5 points for serum albumin and 1 point for IL-6, D dimer, and age. The score was further validated on a cohort of 304 patients admitted to ICU due to the severe form of COVID-19. Results. The study population included 160 COVID-19 patients admitted to ICU in the derivation and 304 in the validation cohort. The mean patient age was 66.7 years (range, 20–93 years), with 68.1% men and 31.9% women. Most patients (76.8%) had comorbidities with hypertension (67.7%), diabetes (31.7), and coronary artery disease (19.3) as the most frequent. A total of 316 patients (68.3%) were treated with mechanical ventilation. Ninety-six (60.0%) in the derivation cohort and 221 (72.7%) patients in the validation cohort had a lethal outcome. The population was divided into the following risk categories for mortality based on the risk model score: low risk (score 0–1) and at-risk ( score > 1 ). In addition, patients were considered at high risk with a risk score > 2 . By applying the risk model to the validation cohort ( n = 304 ), the positive predictive value was 78.8% (95% CI 75.5% to 81.8%); the negative predictive value was 46.6% (95% CI 37.3% to 56.2%); the sensitivity was 82.4% (95% CI 76.7% to 87.1%), and the specificity was 41.0% (95% CI 30.3% to 52.3%). The C statistic was 0.863 (95% CI 0.805-0.921) and 0.665 (95% CI 0.598-0.732) in the derivation and validation cohorts, respectively, indicating a high discriminative value of the proposed score. Conclusion. In the present study, AIDA score showed a valuable significance in estimating the mortality risk in patients with the severe form of COVID-19 disease at admission to ICU. Further external validation on a larger group of patients is needed to provide more insights into the utility of this score in everyday practice.

Introduction. Mortality among critically ill COVID-19 patients remains relatively high despite different potential therapeutic modalities being introduced recently. The treatment of critically ill patients is a challenging task, without identified credible predictors of mortality. Methods. We performed an analysis of 160 consecutive patients with confirmed COVID-19 infection admitted to the Respiratory Intensive Care Unit between June 23, 2020, and October 2, 2020, in University Hospital Center Bezanijska kosa, Belgrade, Serbia. Patients on invasive, noninvasive ventilation and high flow oxygen therapy with moderate to severe ARDS, according to the Berlin definition of ARDS, were selected for the study. Demographic data, past medical history, laboratory values, and CT severity score were analyzed to identify predictors of mortality. Univariate and multivariate logistic regression models were used to assess potential predictors of mortality in critically ill COVID-19 patients. Results. The mean patient age was 65.6 years (range, 29–92 years), predominantly men, 68.8%. 107 (66.9%) patients were on invasive mechanical ventilation, 31 (19.3%) on noninvasive, and 22 (13.8%) on high flow oxygen therapy machine. The median total number of ICU days was 10 (25th to 75th percentile: 6–18), while the median total number of hospital stay was 18 (25th to 75th percentile: 12–28). The mortality rate was 60% (96/160). Univariate logistic regression analysis confirmed the significance of age, CRP, and lymphocytes at admission to hospital, serum albumin, D-dimer, and IL-6 at admission to ICU, and CT score. Serum albumin, D-dimer, and IL-6 at admission to ICU were independently associated with mortality in the final multivariate analysis. Conclusion. In the present study of 160 consecutive critically ill COVID-19 patients with moderate to severe ARDS, IL-6, serum albumin, and D-dimer at admission to ICU, accompanied by chest CT severity score, were marked as independent predictors of mortality.

The objective of this chapter is to provide a systematic overview of current knowledge regarding therapeutic apheresis—primarily therapeutic plateletapheresis (TP)—and to summarize evidence-based practical approaches related to cytapheresis treatment of “hyperthrombocytosis” or “extreme thrombocytosis” (ETC). Our results of platelet (Plt) quantitative/qualitative analyses and evaluation of efficacy of apheresis systems/devices—on the basis of Plt removal and in vivo Plt depletion—will be presented. Our preclinical researches confirmed that in Plt concentrates, the initial ratio of discoid shapes was 70%, spherical 20%, and less valuable (dendritic/balloonized) shapes 10%—with morphological score of platelets (MSP = 300–400). After storage, the ratio of discoid and spherical shapes was decreased, while the less valuable ones progressively increased (MSP = 200). Electron microscopy has shown discoid shapes with typical ultrastructural properties. Spherical shapes with reduced electron density and peripheral location of granules/organelles were detected. Also, dendritic shapes with cytoskeletal “rearrangement,” membrane system integrity damages, and pseudopodia formations were documented. Our clinical study demonstrated that TP was useful in ETC treatment and should help prevention of “thrombo-hemorrhagic” events—until chemotherapy, antiplatelet drugs, and other medication take effect. During TP treatment, Plt count and morphology/ultrastructure were examined. Plt functions by multiplate analyzer were evaluated. We concluded that intensive TP was an effective, safe, and rapid cytoreductive treatment for ET.

Background/Aim. Autologous stem cell transplants (ASCTs) improve the rate of overall survival (OS) in patients with hematological malignancies such as multiple myeloma (MM) after induction chemotherapy, aggressive non-Hodgkin's lymphomas (NHL), and relapsed, chemotherapy- sensitive Hodgkin's lymphoma (HL). The study aim was to evaluate influence of applied CD34+ cell quantity on clinical outcome, as well as early post-transplant and overall survival (OS) of HL and MM patients following ASCT. Methods. This study included a total of 210 patients (90 HL/120 MM) who underwent ASCT. Stem cell (SC) mobilization was accomplished by granulocyte-colony stimulating factor (G-CSF) 10–16 μg/kg body mass (bm) following chemotherapy. For proven poor mobilizers, mobilization with G-CSF (16 μg/kgbm) and Plerixafor (24 or 48 mg) was performed. To our best knowledge, it was the first usage of the Plerixafor in our country in the ASCT-setting. Harvesting was initiated merely at "cut-off-value" of CD34+ cells ≥ 20 × 106/L in peripheral blood with "target-dose" of CD34+ cells ≥ 5 × 106/kgbm in harvest. The CD34+ cell count and viability was determined using flow cytometry. Results. The majority of HL patients (76.7%) were infused with > 5.0 × 106/kgbm CD34+ cells, while 68.3% of MM patients were treated by approximately 4.0–5.4 × 106/kgbm CD34+ dose, respectively. Beneficial response (complete/ partial remission) was achieved in 83.3% (HL) and 94.2% (MM) patients. Among parameters that influenced survival of HL patients with positive response to the therapy, multivariate analysis (pre-ASCT performance status, CD34+ cell quantity applied, rapid hematopoietic, i.e. lymphocyte and platelet recovery) indicated that higher CD34+ cell dose used, along with pre-ASCT performance status correlated with superior event-free survival (EFS) and OS following ASCT. In MM patients, multivariate analysis (renal impairment, infused CD34+ cell quantity, early platelet recovery) indicated that the number of CD34+ cells infused was the most important parameter that influenced both EFS and OS after ASCT. Conclusion. Data obtained in this study undoubtedly confirmed that CD34+ cell dose applied is an independent factor that may contribute to superior clinical outcome and OS of HL and MM patients following ASCT. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 41004 and Grant no. III 41030]

Background/Aim. Stem cells (SCs) guarantee complete/longterm bone marrow (BM) repopulation after SC-transplants. The aim of the study was to evaluate absolute count of total SCs (determined by ISHAGE-sequential-gating protocol – SCish) and relative frequency of immature CD34+/CD90+ (CD90+SCish) subset in peripheral blood (PB) as predictive factors of mobilization and apheresis product (AP) quality. Methods. Mobilization included chemotherapy and granulocytegrowth- factor (G-CSF). Harvesting was performed by Spectra- Optia-IDL-system. The SCsish were determined as a constitutional part of CD34+ cells in the “stem-cell-region” using FC- 500 flow-cytometer. In this study, the original ISHAGEsequential- gating protocol was modified by introduction of anti-CD90-PE monoclonal-antibody into the analysis of CD90 expression on SCish (CD90+SCish). The results were presented as a percentage of SCish per nucleated-cell count, absolute SCish count in μL of the PB or the AP, percentage of the CD90+SCish expressed to SCish and absolute CD90+SCish count in μL of the PB or the AP. Results. The absolute count of total SCish and CD90+SCish was significantly higher (p = 0.0007 and p = 0.0266, respectively) in the AP than in the PB samples. The CD90+SCish/total SCish indexes from PB were higher than indexes from the AP (p = 0.039). The relative frequency of CD90+SCish showed a highly significant inverse correlation with the absolute count of total SCish in both, the PB and AP (p = 0.0003 and p = 0.0013 respectively). The relative frequency of CD90+SCish from the PB also showed a significant (p = 0.0002) inverse relationship with total SCish count in the AP. Patients with less than 10% CD90+SCish in the PB had evidently higher (p = 0.0025) total SCish count in the AP. Conclusion. We speculate that lower CD90+SCish yield in the AP is not a consequence of an inferior collection efficacy, but most likely a result of several still not fully resolved immature SC cytomorphological/ biophysical features. Therefore, following the mobilization by chemotherapy G-CSF, some logical questions appear – whether we should follow the absolute count of total SCish, or, whether we should test for relative frequency of CD90+SCish prior to harvesting. To reach the final conclusions, it is essential to conduct further controlled and larger investigations concerning the correlation of circulating and harvested SCs with patients' hematopoietic recovery. [Project of the Ministry of Defence of the RS, Project MF/VMA 9/17-19 and Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 41031]

Introduction. Bence-Jones myeloma multiplex is a progressive disease characterized by excessive numbers of abnormal plasma cells in the bone marrow and overproduction of incomplete immunoglobulins, containing only the light chain portion of the immunoglobulins. This type of myeloma occurs 15-20%. The median overall survival is approximately 4 years. Aim of this study was to define prognostic factors affecting overall survival in Bence-Jones multiple myeloma patients. Material and Methods. From 1995 to 2015, we treated 75 patients (49 men and 26 female), average age 55.9 years (range 31-85). Results. Conventional chemotherapy introductory clinical response was achieved in 54 patients (72%), while in 21 patients (28%) the established disease was resistant. Transplantation was done in 45 patients (60%), while 30 patients (40%) were treated with conventional chemotherapy. In the group of patients with transplantation done, tandem was carried out in 11 patients and secondary stem cell transplantation was done in 5 relapsed patients. With 1 patient with tandem stem cell transplantation allogenic (singen) stem cell transplantation was done. Transplant related mortality is 1.5%. The transplanted patients had significantly longer PFS (mediana 13 months vs 7 months, p

Introduction. Autologous stem cell transplantation combined with high dose chemotherapy is an effective and safe approach in the treatment of different hematological malignancies. Nowadays, autologous stem cell transplantation represents a standard therapeutic option in the treatment of multiple myeloma, lymphomas and other hematological malignancies. Aim is to analyze the available medical data of patients with multiple myeloma, Hodgkin’s lymphoma, non-Hodgkin’s lymphoma acute myeloid and lymphoblastic leukemia, who underwent autologous stem cell transplantation, and to compare the results with published data from other similar studies. Material and Methods. A retrospective study included 90 patients with multiple myeloma, acute myeloid and lymphoblastic leukemia, non-Hodgkin’s and Hodgkin’s lymphoma who underwent autologous stem cell transplantation in the period from 2004 to August 2017. Results. In relation to the underlying disease, the distribution of the respondents was as follows: 39 patients had multiple myeloma, 25 non-Hodgkin’s lymphoma, 20 Hodgkin’s lymphoma and 6 had acute leukemia. 75 patients (89.3%) had the large volume apheresis procedure, while 9 patients (10.7%) had the conventional two-day apheresis procedure. The average number of the mononuclear cells in the apheresis product was 7,8x108/kg, and the number of the CD34 + cells was about 12,11x106 kg. After applying the conditioning regimen, depending on the underlying disease, neutrophils engraftment mainly occurred on the 11th while the platelet engraftment occurred on the 14th post-transplant day. Transplant-related mortality was low, and the mortality rate was 3.57%. Conclusion. Autologous stem cell transplantation is an efficient method of treatment for patients with he­matological malignancies. It is associated with a low rate of complications as well as low rate of transplant-related mortality.

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Autologous stem cell transplant with high-dose chemotherapy is considered to be an effective treatment strategy for outcome improvement in lymphoma patients, especially those with refractory and relapsed disease. Despite the feasibility and efficacy of autologous stem cell transplant, patients with lymphoma still face the risk for relapse, mostly patients who have adverse prognostic features. Salvage chemotherapy followed by high-dose chemotherapy and autologous stem cell transplant is recognized as the most effective strategy for relapsed or refractory Hodgkin lymphoma or aggressive non-Hodgkin lymphoma improving their response rate. First line therapy is ABVD (Hodgkin lymphoma patients), R-CHOP or R-CHOP-like regimens while salvage regimens such as DHAP, ESHAP, ICE represent the standard of care for relapsed/refractory lymphoma patients. Before autologous stem cell transplant, standard condition regimens are BEAM or CBV.

Stem cells could be defined as cells capable for self-renewal with high proliferative capacity and extensive potential to differentiate into blood cells or some somatic cell types - ?plasticity? due to ?trans-differentiation? - such as osteocytes, chondrocytes, hepatocytes, myocytes, cardiomyocytes and even endothelial cells. Recent increasing clinical use of various cell-mediated therapeutic approaches has resulted in amplified needs for both stem cells and operating procedures to get a minimized cell damages during collection, purification and cryopreservation. The aim of cell harvesting procedures is to obtain the best stem cells yield, high purity and good viability/clonogenicity. The goal of optimized cryoinvestigation protocols is to minimize cell injuries during the freeze/thaw process (cryoinjury). Despite the fact that different stem cells collection protocols and cell freezing practice are already in routine use, a lot of questions related to the optimal blood stem cells harvesting, purification and cryopreservation are still unresolved.

Background: In patients with ST-elevation myocardial infarction (STEMI), C-reactive protein (CRP) levels are associated with larger infarct size, transmural extent, and poor function of left ventricle and independently predict 30-day mortality. CRP-apheresis following STEMI showed to be feasible, safe, and has significant beneficial effect both on myocardial infarction size and wall motion. To the best of our knowledge, this is only the second published clinical evaluation of the efficacy and safety of selective CRP-apheresis in the STEMI treatment using Spectra-Optia and Pentrasorb CRP-adsorber systems. Case Report: A 53-year-old female was referred with anterior STEMI. After percutaneous coronary intervention, patient received standard post-STEMI therapy according to current guidelines. Selective therapeutic plasma exchange (TPE) was performed using Spectra-Optia (Terumo BCT; USA) and Pentrasorb CRP-adsorber (Pentracor GmbH; Germany) systems. Antecubital veins were used for vascular access and acid-citrate-dextrose solution (ACD formula A; total volume = 1,026 mL) was utilized as anticoagulant. The volume of processed blood was 15,600 mL. The removed “natural” plasma (total volume = 8,329 mL) was replaced with CRP-depleted autologous plasma (total volume = 8,085 mL). This intensive TPE-treatment was well tolerated, without adverse effects, or complications. The CRP plasma levels were: initial = 4.2 mg/L 6 h after acute myocardial infarction (AMI), pre-apheresis = 16.4 mg/L, and post-apheresis = 4.59 mg/L (CRP-depletion = 72%). There were neither significant changes observed in biochemistry nor any alterations in plasma hemostatic activity investigated before and after CRP-adsorption performed. Conclusion: Early performed CRP-apheresis is a promising innovative therapeutic approach for STEMI treatment that could provide a reduced size of infarction zone – with inferior occurrence of heart failure after AMI. However, precise and complete evaluation of the efficacy and safety of this treatment requires further multicenter randomized and larger clinical studies.

The common signs of canine babesiosis caused by an infection with Babesia canis are fever, anorexia, lethargy, pulse alterations, anemia, and occasionally mild icterus. Dogs with these clinical signs can be divided into two groups: those with acute-phase reaction and those with systemic inflammatory response syndrome (SIRS). Factors associated with the occurrence of SIRS in canine babesiosis have not been thoroughly researched. This article outlines a cross-sectional study of 54 client-owned dogs with an acute B. canis infection, and evaluates the differences in age, gender, laboratory findings, parasite load, and seroreactivity against B. canis between the SIRS and the SIRS-free dogs. We have analyzed a complete blood count, serum biochemistry, serum amyloid A, ceruloplasmin, paraoxonase-1, serology, and PCR testing using standard methodologies. The frequency of SIRS among the investigated dogs reached 0.59. Male dogs and those seronegative against B. canis, were more frequent in the SIRS group, whilst age and parasite load could not be associated with the presence of SIRS. Dogs with SIRS had a lower count of total leukocytes, neutrophils, lymphocytes, and monocytes, and a lower concentration of iron and bilirubin compared with SIRS-free dogs. No significant differences in the concentration of acute-phase proteins have been noticed to exist between the groups of dogs. Further, the seronegative dogs had a lower count of lymphocytes and monocytes and a higher parasite load than the seroreactive dogs. Multivariate logistic regression analysis has identified leukopenia (

Prognostic significance of immune microenvironment has been emphasized using the most advanced analysis, with consecutive attempts to reveal prognostic impact of this findings. The aim of this study was to compare and define prognostic significance of clinical parameters, microvessel density (MVD) in tumour tissue and expression of CD44s as adhesive molecule on tumour cells in diffuse large B cell lymphoma-DLBCL, primary central nervous system DLBCL-CNS DLBCL and follicular lymphoma-FL. A total of 202 histopathological samples (115 DLBCL/65 FL/22 CNS DLBCL) were evaluated. Overall response (complete/partial remission) was achieved in 81.3 % DLBCL patients, 81.8 % primary CNS DLBCL and 92.3 % FL. Absolute lymphocyte count-ALC/Absolute monocyte count-AMC2.6 in DLBCL and ALC/AMC ≥ 4.7 in FL were associated with better event-free survival (EFS) and overall survival (OS) (p 0.05). In DLBCL, MVD 42 blood vessels/0.36 mm(2) correlated with primary resistant disease (p 0.0001), poorer EFS and OS (p = 0.014). High CD44s expression in FL correlated with inferior EFS and OS (p 0.01). In DLBCL, multivariate Cox regression analysis showed that ALC/AMC was independent parameter that affected OS (HR 3.27, 95 % CI 1.51-7.09, p = 0.003) along with the NCCN-IPI (HR 1.39, 95 % CI 1.08-1.79, p = 0.01). Furthermore, in FL, ALC/AMC mostly influenced OS (HR 5.21, 95 % CI 1.17-23.21, p = 0.03), followed with the FLIPI (HR 3.98, 95 % CI 1.06-14.95, p = 0.041). In DLBCL and FL, ALC/AMC is simple and robust tool that is, with current prognostic scores, able to define long-term survival and identify patients with inferior outcome. The introduction of immunochemotherapy might altered the prognostic significance of microenvionmental biomarkers (MVD and CD44s).

Introduction. Multiple sclerosis (MS) is an immunemediated disease of the central nervous system that affects young individuals and leads to severe disability. High dose immunoablation followed by autologous hemopoietic stem cell transplantation (AHSCT) has been considered in the last 15 years as potentialy effective therapeutic approach for agressive MS. The most recent long-time follow-up results suggest that AHSCT is not only effective for highly aggressive MS, but for relapsing-remitting MS as well, providing long-term remission, or maybe even cure. We presented a 10- year follow-up of the first MS patient being treated by immunoablation therapy and AHSCT. Case report. A 27-year-old male experienced the first symptoms - intermitent numbness and paresthesia of arms and legs of what was treated for two years by psychiatrist as anxiety disorder. After he developed severe paraparesis he was admitted to the Neurology Clinic and diagnosed with MS. Our patient developed aggressive MS with frequent relapses, rapid disability progression and transition to secondary progressive form 6 years after MS onset [the Expanded Disability Status Scale (EDSS) 7.0 Ambulation Index (AI) 7]. AHSCT was performed, cyclophosphamide was used for hemopoietic stem cell mobilization and the BEAM protocol was used as conditionig regimen. No major adverse events followed the AHSCT. Neurological impairment improved, EDSS 6.5, AI 6 and during a 10-year followup remained unchanged. Brain MRI follow-up showed the absence of gadolinium enhancing lesions and a mild progression of brain atrophy. Conclusion. The patient with rapidly evolving, aggressive, noninflammatory MS initialy improved and remained stable, without disability progression for 10 years, after AHSCT. This kind of treatment should be considered in aggressive MS, or in disease modifying treatment nonresponsive MS patients, since appropriately timed AHSCT treatment may not only prevent disability progression but reduce the achieved level of disability, as well.

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Background/Aim. Intramyocardial bone marrow mononuclear cells (BMMNC) implantation concomitant to coronary artery bypass grafting (CABG) surgery as an option for regenerative therapy in chronic ischemic heart failure was tested in a very few number of studies, with not consistent conclusions regarding improvement in left ventricular function, and with a follow-up period between 6 months and 1 year. This study was focused on testing of the hypothesis that intramyocardial BMMNC implantation, concomitant to CABG surgery in ischemic cardiomyopathy patients, leads to better postoperative long-term results regarding the primary endpoint of conditional status-functional capacity and the secondary endpoint of mortality than CABG surgery alone in a median follow-up period of 5 years. Methods. A total of 30 patients with ischemic cardiomyopathy and the median left ventricular ejection fraction (LVEF) of 35.9 ± 4.7% were prospectively and randomly enrolled in a single center interventional, open labeled clinical trial as two groups: group I of 15 patients designated as the study group to receive CABG surgery and intramyocardial implantation of BMMNC and group II of 15 patients as the control group to receive only the CABG procedure. All the patients in both groups received the average of 3.4 ± 0.7 implanted coronary grafts, and all of them received the left internal mammary artery (LIMA) to the left anterior descending (LAD) and autovenous to other coronaries. Results. The group with BMMNC and CABG had the average of 17.5 ± 3.8 injections of BMMNC suspension with the average number of injected bone marrow mononuclear cells of 70.7 ± 32.4 × 106 in the total average volume of 5.7 ± 1.5 mL. In this volume the average count of CD34+ and CD133+ cells was 3.96 ± 2.77 × 106 and 2.65 ± 1.71 × 106, respectively. All the patients were followed up in 2.5 to 7.5 years (median, 5 years). At the end of the follow-up period, significantly more patients from the group that received BMMNC were in the functional class I compared to the CABG only group (14/15 vs 5/15; p = 0.002). After 6 months the results on 6-minute walk test (6-MWT) were significantly different between the groups (435 m in the BMMNC and CABG group and 315 m in the CABG only group; p = 0.001), and continued to be preserved and improved on the final follow-up (520 m in the BMMNC and CABG group vs 343 m in the CABG only group; p < 0.001). Cardiovascular mortality was also significantly reduced in the BMMNC and CABG group (p = 0.049). Conclusion. Implanatation of BMMNC concomitant to CABG is a safe and feasible procedure that demonstates not only the improved functional capacity but also a reduced cardiac mortality in a 5-year follow-up in patients with ischemic cardiomyopathy scheduled for CABG surgery.

Worldwide, the syndromes of paraganglioma (PGL), somatostatinoma (SOM) and early childhood polycythemia are described in only a few patients with somatic mutations in the hypoxia-inducible factor 2 alpha (HIF2A). This study provides detailed information about the clinical aspects and course of 7 patients with this syndrome and brings into perspective these experiences with the pertinent literature. Six females and one male presented at a median age of 28 years (range 11–46). Two were found to haveHIF2Asomatic mosaicism. No relatives were affected. All patients were diagnosed with polycythemia before age 8 and before PGL/SOM developed. PGLs were found at a median age of 17 years (range 8–38) and SOMs at 29 years (range 22–38). PGLs were multiple, recurrent and metastatic in 100, 100 and 29% of all cases, and SOMs in 40, 40 and 60%, respectively. All PGLs were primarily norepinephrine-producing. All patients had abnormal ophthalmologic findings and those with SOMs had gallbladder disease. Computed tomography (CT) and magnetic resonance imaging revealed cystic lesions at multiple sites and hemangiomas in 4 patients (57%), previously thought to be pathognomonic for von Hippel–Lindau disease. The most accurate radiopharmaceutical to detect PGL appeared to be [18F]-fluorodihydroxyphenylalanine ([18F]-FDOPA). Therefore, [18F]-FDOPA PET/CT, not [68Ga]-(DOTA)-[Tyr3]-octreotate ([68Ga]-DOTATATE) PET/CT is recommended for tumor localization and aftercare in this syndrome. The long-term prognosis of the syndrome is unknown. However, to date no deaths occurred after 6 years follow-up. Physicians should be aware of this unique syndrome and its diagnostic and therapeutic challenges.

Background/Aim. Introducing tyrosine kinase inhibitors (TKIs) has essentially changed curative approach, to be precise, clearly improved treatment efficacy for chronic myeloid leukemia (CML). Thus, the place and usage of allogeneic stem cell transplant (SCT) in CML treatment - as a former "nearly monopolistic" therapeutic manner - is nowadays controversial. The objective of this retrospective study was to evaluate the results obtained in the treatment of CML patients, with a particular attempt to define parameters critical for clinical benefit and superior overall outcome following allogeneic SCT. Methods. A total of 32 CML patients (27 in chronic phase and 5 with advanced disease), with female/male ratio 11/21, aged from 9 to 54 (32 in average) years, underwent allogeneic SCTs (1993 to 2009). The initial treatment for 25 patients was interferon alpha (IFN-α) with or without ARA-C, and additional 7 patients with no response to imatinib mesylat (IM). The time from diagnosis to SCT was approximately 12 (range 3- 37) months. The patient were categorized according to the risk for the disease, transplant-related mortality (TRM) scoring system, and stem cell (SC) source. The basic conditioning regimen was a combination of busulphan and cyclophosphamide (BuCy-2). Graft-versus-host disease (GvHD) was typically prevented with cyclosporine-A (CsA) and methotrexate (MTX). Results. Engraftment was observed in 26 (84.4%) patients, with polymorphonuclear (PMNs) and platelet (Plt) recovery on the 15th (range 10-22) and 19th (range 11-29) posttranspalnt days, respectively. Acute GvHD (aGvHD) had 13/26 (50%), and chronic GvHD (cGvHD) 10/21 (47.1%) patients. The incidence of overall TRM was 46.8% (15/32), while early death was noticed in 4 (12.5%) patients. A cause of death in 9 (28.1%) patients was cGvHD, in 2 (6.25%) patients infection, and in 3 (9.35%) cases disease-relapse was occurred. Fourteen (43.7%) of the patients are still alive, 9 from the low-risk group for TRM, with long-term survival from 1 to 16 years. Patients who received SCs from peripheral blood (PB) vs bone marrow (BM) had significantly faster engraftment (p < 0.05), lower oropharingeal mucositis rate (25% vs 70%; p < 0.05), but more frequent cGvHD (83.3% vs 30.3%; p < 0.05). A significantly improved (log-rank = 2.39; p < 0.01) overall survival (OS) was obtained in BM-setting. Conclusion. The results obtained in this study are in accordance with data from analogous clinical trials. Exactly, in the era of the new target therapy (TKI application), allogeneic SCT can be still a convenient therapeutic approach for well-selected CMLpatients, especially for those with initial high-risk disease and lower probability of TRM.

The aim of this study was to evaluate the prognostic significance of international prognostic index (IPI), mantle cell lymphoma IPI (MIPI), simplified MIPI (sMIPI), and MIPI biological (MIPIb), as well as their correlation with immunophenotype, clinical characteristics, and overall survival (OS), in a selected group of 54 patients with advanced-stage mantle cell lymphoma (MCL), treated uniformly with CHOP. Seventeen patients had IV clinical stage (CS), while other 37 had leukemic phase at presentation. Diffuse type of marrow infiltration was verified in 68.5% and nodular in remainder patients. Extranodal localization (25.9%) included bowel (20.4%), pleural effusion, sinus, and palpebral infiltration. All of analyzed patients expressed typical MCL immunophenotypic profile: CD19(+)CD20(+)CD22(+)CD5(+)Cyclin-D1(+)FMC7(+)CD79b(+)smIg(+)CD38(+/-)CD23(-)CD10(-). Median OS of the whole group was 23 months, without significant differences between IV CS and leukemic phase patients. Thirty-two patients (59.3%) responded to initial treatment, 9 (16.7%) with complete and 23 (42.6%) with partial remission. Negative prognostic influence on OS had high IPI (P0.01), high sMIPI (P0.001), MIPI (P0.01), MIPIb (P0.01), extranodal localization (P0.01), and diffuse marrow infiltration (P0.01). Testing between randomly selected groups showed that patients with lower proportion of CD5(+) cells (80%) correlated with cytological blastoid variant and had shorter survival comparing with the group with higher proportion of CD5(+) cells (80%) (P0.01). Using univariate Cox regression, we proved that IPI, sMIPI, MIPI, and MIPIb had an independent predictive importance (P0.01) for OS in uniformly treated advanced MCL patients, although sMIPI prognostic significance was the highest (P0.001).

Background/Aim. Peripheral blood (PB) is used more frequently as a source of stem cells (SCs) for allogeneic transplantation. However, the influence of cell source on the clinical outcome of SC transplantation is not yet well established. The aim of this study was to compare the results of PBSC transplantation (PBSCT) with bone marrow transplantation (BMT) on the basis of engraftment, frequency and severity of immediate (mucositis, acute Graft versus Host Disease - aGvHD) and delayed (chronic GvHD - cGvHD) complications, as well as transplant-related mortality (TRM), transfusion needs, relapses and overall survival (OS). Methods. We analyzed 158 patients, women/men ratio 64/94 median age 29 (range 9-57), who underwent allogeneic SC transplantation between 1989 and 2009. All included patients had diseases as follows: acute myeloid leukemia (AML) - 39, acute lymphoblastic leukemia (ALL) - 47, chronic myeloid leukemia (CML) - 32, myelodysplastic syndrome (MDS) - 10, Hodgkin?s lymphoma (HL) - 2, multiple myeloma (MM) - 3, granulocytic sarcoma (GrSa) - 3, severe aplastic anemia (sAA) - 22. The patients underwent transplantations were divided into two groups: BMT group (74 patients) and PBSCT group (84 patients). Each recipient had HLA identical sibling donor. SCs from bone marrow were collected by multiple aspirations of iliac bone and from PB by one ?Large Volume Leukapheresis? (after recombinant human granulocyte colony stimulating factor, rHuG-CSF) application (5-12 ?g/kgbm, 5 days). Conditioning regimens were applied according to primary disease, GvHD prophylaxis consisted of combination of a cyclosporine A and methotrexate. Results. Engraftment, according to the count of polymorphonuclear and platelets, were significantly (p < 0.001) faster in the PBSCT vs BMT group. The needs for transfusion support were significantly (p < 0.01) higher in the BMT group. Those patients had more frequently oropharingeal mucositis grade 3/4 (33.3% vs 10.0%, p < 0.05). There were no significant differences in the incidence of aGvHD and cGvHD between the two groups. The patients who underwent PBSCT had more frequently extensive cGvHD in comparison with the BMT group (29.1% vs 11.29%, p < 0.05). SC source (SCS) had no significant influence on the TRM (21.62% vs 23.8%, p = 0.64) and the incidence of relapses (21.6% vs 29.7%, p = 0.32). Finally, the patients treated by BMT had a significantly better OS (logrank 2.33, p < 0.05). Conclusion. SCs harvesting from PB resulted in improved cell yield, faster engraftment, as well as in a decrease of immediate transplantation related complications with a reduced treatment cost. Allogeneic PBSCT were associated with more frequent extensive cGvHD, while the influence of SCS in TRM and relapses was not observed. Finally, the longterm OS was better in the patients treated by BMT. To verify impact of SC source on transplantation (PBSCT vs BMT) overall efficacy, more larger randomized clinical studies are needed.

Therapeutical concept for myeloma mulitplex (MM) patients has been significantly improved over the previous years. As in other haematological malignancies, in MM also, induction treatment goal is to achieve complete remission (CR) of the disease. The importance of CR on overall survival (OS) in MM was first estimated in the high dose chemotherapy (CT) with autologous stem cell transplantation (ASCT). Induction treatment with 3-6 cycles of combined CT based on Dexamethason (VAD) following with one or two courses of high doses of Melphalan with ASCT leads to 20-40% CR and 40-55% CR/VGPR with median survival of 4-5 years. These are significantly better results compared to previous conventional therapy, and considering such findings, ASCT became the treatment of choice for MM patients aged below 65 years. The introduction of novel agents, such as immunomodulatory drugs (Thalidomid, Lenalidomid) and proteasome inhibitors (Bortezomib) into induction therapy have improved treatment of MM patients even more. The combination of Dexamethason with Thalidomid, Bortezomib or Lenalidomid, on the average in 3-4 cycles, arises the level of CR/ VGPR (very good partial response) even to 60-75%. Bortezomib based regimens are recommended in the treatment of ?high risk? group of patients with MM. Recommended conditioning regimen is 200 mg/m2 of Melphalan. If either CR or VGPR is not achieved, recommendation is to perform the second ASCT as a part of ?tandem?concept. Thalidomid applied as a ?maintenance therapy? after ASCT may prolong OS. Until now, allogeneic ASCT, as the first line treatment for MM patients aged below 65 years, has been applied only in clinical trials.

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.

Recently, a guanosine analog, 7-allyl-7,8-dihydro-8-oxo-guanosine (loxoribine), has been identified as a selective Toll-like receptor (TLR)7 agonist. Bearing in mind the controversy regarding the expression of TLR7 by human myeloid dendritic cells (DCs) and its significance for functions of these cells, the goal of this study was to investigate the effect of loxoribine on differentiation, maturation and functions of human monocyte-derived (Mo)DCs. Immature MoDCs were obtained by cultivation of monocytes for 6 days with recombinant granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4. These cells were stimulated with loxoribine (250 μM) for an additional 48 h. Phenotypic properties of MoDCs were determined by flow cytometry, cytokine production was assayed by ELISA, whereas their allostimulatory capability was tested using a mixed leukocyte reaction. We showed that loxoribine up-regulated the expression of TLR7, CD40, CD54, CD80, CD83 and CCR7 and stimulated the production of IL-12, IL-23, IL-27 and IL-10 by MoDCs, whereas the level of interferon (IFN)-β was not modulated. Allogeneic CD4(+)T cells in co-culture with loxoribine-treated MoDCs proliferated more strongly, at lower DC/CD4(+)T-cell ratio (1:80), and secreted significantly higher levels of IL-17 and IFN-γ compared to the cultures with control MoDCs. The stimulatory effect of loxoribine on T helper (Th)1 polarization capability of MoDCs was further potentiated by ligation of CD40. In conclusion, our results show that loxoribine stimulated differentiation, maturation, allostimulatory as well as Th1 and Th17 polarization capability of human MoDCs and suggests that these effects might be associated with up-regulation of TLR7 expression, but not increased IFN-β production.

Langerhans' cells (LCs) represent a specific subset of dendritic cells (DCs) which are important for detecting and processing pathogens that penetrate the skin and epithelial barriers. The aim of our study was to explain what makes their in vitro counterparts - monocyte-derived Langerhans'-like cells (MoLCs) - unique compared with monocyte-derived dendritic cells (MoDCs). Immature MoDCs were generated by incubating peripheral blood monocytes with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. The addition of transforming growth factor-β (TGF-β) to this cytokine cocktail resulted in the generation of MoLCs. MoLCs showed a lower expression of CD83, CD86, HLA-DR and CCR7 compared with MoDCs, regardless of their maturational status. Both immature and mature MoLCs secreted higher quantities of IL-23 compared with MoDCs and this finding correlated with a higher secretion of IL-17 in co-culture of MoLCs with allogeneic CD4(+) T cells. Mature MoLCs, which produced higher levels of IL-12 and lower levels of IL-10 compared with mature MoDCs, were more potent at inducing interferon-γ (IFN-γ) production by CD4(+) T cells in the co-culture system. In conclusion, the finding that mature MoLCs stimulate stronger T-helper 1 and T-helper 17 immune responses than mature MoDCs, makes them better candidates for use in the preparation of anti-tumour DC vaccines.