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Biomedical subjects

Bela Balint

Publications and source records attributed to Bela Balint.

16 recordsLinked to original sources

Cytapheresis in the treatment of cell-affected blood disorders and abnormalities.

This report presents our experience with cytaphereses performed in treatment of 476 patients. Leukapheresis was used in management of 68 patients with hyperleukocytosis leukostasis (WBC > or = 150 x 10(9)L(-1)). Average decrease in cell count after treatment was 73.3%. Plateletapheresis for 32 patients (platelets > or = 1500 x 10(9)L(-1)) was applied in order to prevent the thrombotic-hemorrhagic syndrome and resulted in a moderate platelet count reduction (84.3%). Erythrocytaphereses performed in treatment of 376 patients by manual or automated technique resulted in a rapid blood viscosity drop (42.4+/-7.1%). Patients with red blood cell exchanges (severe malaria and autoimmune hemolytic crisis) were in life-threatening situations and resulted in a prompt reduction of parasitized or antibody-coated RBCs and anemia correction. This study indicates that "conventional" TCs resulted in considerable cytoreduction only in patients with especially high cell count. This effect was not associated with bone marrow remission. The best clinical effect and long-term benefits were obtained using RBCX and antimalarial drugs in malaria patients who have had high-level parasitized-RBCs with multiorgan dysfunction.

Adolescent↗

Controlled-rate versus uncontrolled-rate freezing as predictors for platelet cryopreservation efficacy.

BACKGROUND: Cryobiologic variables responsible for cell injuries and freezing techniques applicable in medical cryopractice should be revised and/or reengineered for minimizing cryoinjuries and maximizing cell recovery. In this study, the efficacy of different cryopreservation protocols based on platelet (PLT) recovery was evaluated. STUDY DESIGN AND METHODS: PLTs (n = 33) were prepared from whole-blood units. Cell count and viability, PLT morphologic score (PMS), and hypotonic shock response were determined. PLT surface antigens were measured by flow cytometry. Controlled-rate (with compensated fusion heat) and uncontrolled-rate freezing methods combined with 6 percent dimethyl sulfoxide were used. RESULTS: PLT recovery was superior in the controlled-rate setting (91.0 +/- 5.5 vs. 86.0 +/- 6.5; p < 0.05). PMS was significantly better in controlled-rate freezing (p < 0.01). GPIb/CD42b expression was reduced in both freezing groups versus control. GP140/CD62p expression was significantly (p < 0.05) lower in the controlled-rate group and in both frozen groups was significantly higher than in the control groups. CONCLUSION: The use of strictly equalized (1 degrees C/min) controlled-rate freezing, combined with an intensified cooling rate (2 degrees C/min) during the liquid-to-solid-phase transition period, allows advanced quantitative and qualitative PLT recovery, even though the minor intergroup differences for some variables were observed.

Adult↗

Microprocessor-controlled vs. "dump-freezing"platelet and lymphocyte cryopreservation: a quantitative and qualitative comparative study.

BACKGROUND/AIM: Thermodynamical and cryobiological parameters responsible for cell damages during cryopreservation (cryoinjuries) have not yet been completely explained. Thus, freezing procedures should be revised, exactly optimized to obtain an enhanced structural and functional recovery of frozen-thawed cells. The aim of this study was to compare microprocessor-controlled (controlled-rate) with the compensation of the released fusion heat and "dump-freezing" (uncontrolled-rate) of the platelet and lymphocyte cryopreservation efficacy. METHODS: Platelet quantitative recovery (post-thaw vs. unfrozen cell count), viability (using hypotonic shock response--HSR), morphological score (PMS), ultrastructural (electron microscopy) properties and expression of different surface antigens were investigated. In lymphocyte setting, cell recovery and viability (using trypan blue exclusion test) as well as functionality (by plant mitogens) were determined. Controlled-rate freezing and uncontrolled-rate cryopreservation were combined with 6% (platelets) and 10% (lymphocytes) dimethyl sulfoxide (DMSO). RESULTS: Platelet recovery and functionality were superior in the controlled-rate system. The majority of surface antigen expression was reduced in both freezing groups vs. unfrozen cells, but GP140/CD62p was significantly higher in controlled-rate vs. uncontrolled-rate setting. Controlled-rate freezing resulted with better lymphocyte recovery and viability (trypan blue-negative cell percentage). In mitogen-induced lymphocyte proliferative response no significant intergroup difference (controlled-rate vs. uncontrolled-rate) were found. CONCLUSION: The data obtained in this study showned the dependence of cell response on the cryopreservation type. Controlled-rate freezing provided a superior platelet quantitative and functional recovery. Lymphocyte recovery and viability were better in the controlled-rate group, although only a minor intergroup difference for cell proliferative response was obtained.

Blood Platelets↗

Apheresis in donor and therapeutic settings: recruitments vs. possibilities--a multicenter study.

In our country, the first apheresis was performed in the late 1960s (by manual technique), and the first cell separator was used in 1979. The number of blood component collections performed from 1994 to 2004 was: 11,170 (total), i.e., 8540 (NBTI), 1180 (IT-MMA), 1050 (BTI of Novi Sad) and 400 (BTI Nis). The number of PBSC harvests during 1996-2004 was 386 for treatment of 272 patients. For treatment of myocardial infarction, "cell-therapy" by autologous stem cells was introduced in 2004 at the MMA. The results of PE treatments performed (7632 sessions) by our group for various immune-mediated and other disorders were generally beneficial, but the effect is not associated with bone marrow remission. TC procedures (total number=1279) resulted in a significant fall in the blood cell counts and hemorheological improvement, as well as the removal and replacement of abnormal red blood cells. Greater standardization of different apheresis protocols is required.

Cell- and Tissue-Based Therapy↗

Possibilities and limitations of fibrin glue usage in nephron-sparing surgery: experimental study.

INTRODUCTION: The possibilities and limitations of fibrin glue (FG) usage in nephron-sparing surgery were studied. MATERIALS AND METHODS: A prospective experimental study was carried out in 50 pigs: 30 with polar resection, and 20 with mediorenal wedge resection of the kidney. Hemostatic sutures, FG, and FG with a muscle 'cup' in animals with polar resection of the kidney were compared. FG and sutures in animals with the wedge resection of the kidney were studied as well. Bleeding, hot ischemia time, complication rate, and additional scarring were also analyzed. RESULTS: Suture hemostasis is safe but with significant adverse effects in both polar and wedge resection of kidney. FG was not efficient as a sole hemostatic agent for polar resection. It was as efficient as hemostatic suture for wedge resection of the kidney. FG with a muscle 'cup' on a pole of the kidney achieved good results in animals with polar resection of the kidney. Histological analysis confirmed better results with FG because of both the less intense and smaller area of additional scarring. CONCLUSION: FG is a reliable and efficient hemostatic agent for nephron-sparing surgery whenever both sided gluing is possible.

Administration, Topical↗

[Significance of iron reduction for the therapy of chronic hepatitis C].

BACKGROUND: It has been established that many patients with chronic hepatitis C have elevated serum iron, feritin levels and iron deposites in the liver. Therefore, the liver damage due to hepatitis C virus may be aggravated with iron overload. In many studies higher levels of iron in the blood and the liver were connected with the decreased response to interferon-alfa therapy for chronic viral hepatitis C. Recent introduction of pegylated interferons plus ribavirin has improved the therapeutic response, so it is now possible to cure more than 50% of the patients. CASE REPORT: Three patients with chronic hepatitis C and iron overload were presented. Iron reduction therapy using phlebotomy or eritrocytapheresis with plasmapheresis was done at different times in regard to specific antiviral therapy or as a sole therapy. CONCLUSION: It has been shown that iron reduction, sole or combined with antiviral therapy, led to the deacreased aminotransferase serum activity and might have slown down the evolution of chronic hepatitis C viral infection.

Adult↗

Multicenter evaluation of a whole-blood filter that saves platelets.

BACKGROUND: Whole-blood (WB) leukoreduction filters in current use retain the majority of PLTs. A new whole-blood filter, which retains significantly fewer of the PLTs (or saves PLTs [WB-SP]), has been developed. The performance characteristics of the WB-SP filter have been evaluated in a multicenter study. STUDY DESIGN AND METHODS: A total of 617 units of WB was collected into quadruple bag sets with an integrated WB-SP filter, leukoreduced, and processed into leukoreduced RBCs (LR-RBC), plasma (LR-PL), and buffy coats (LR-BC) from which, pooled, leukoreduced, PLT concentrates (LR-PCs) were produced. Recovery, yield, and residual WBCs were assessed in prepared blood components. RESULTS: The median residual WBC number in the LR-RBCs was 0.05 x 10(6) (range, <0.05-3.8), exceeding 1 x 10(6) in 0.6 percent of the units. Median Hb content in LR-RBC was 50 g (range, 34-72), reflecting a final RBC recovery of 81 +/- 6 percent. The median WBC content of the LR-PC was 0.05 x 10(6) (range, <0.05-0.28), with none exceeding 1 x 10(6). The median PLT content of the LR-PC, per individual donation, was 6.4 x 10(10) (range, 4.1-10.7), representing a final recovery of 62 +/- 10 percent. The mean FVIII activity was 104 +/- 25 percent and 83 +/- 11 percent in plasma separated from fresh or overnight stored WB, respectively. CONCLUSION: Use of the WB-SP filter makes it possible to obtain three leukoreduced blood components with only one filtration step. The WB-SP filter showed good leukoreduction performance and recovery of all blood components including PLTs.

Blood Component Removal↗

The use of single-donor fibrin glue prepared by recycled cryoprecipitation in experimental liver surgery.

The purpose of the study was to evaluate the hemostatic effectiveness of fibrin glue (FG) prepared by a modification of cryoprecipitation technique in experimental rat liver surgery. FG component 1 was prepared by triple or 'recycled' cryoprecipitation method from single-donor plasma. Rats subjected to liver incision, partial and total lobectomy were treated with FG on the surgical cut surface or underwent standard surgical technique. The efficacy of FG-treatment was evaluated on the basis of the 24-hour survival ratio and peripheral blood hematological parameters. The mean values of fibrinogen, FXIII, fibronectin and horizontal tensile strength of FG were 54.2 +/- 19.9 g/l, 13.5 +/- 3.6 IU/ml, 3103.1 +/- 148.9 mg/l, and 1.076 +/- 0.18 N/cm2, respectively. The survival of FG-treated rats subjected to partial and total lobectomy was significantly higher in comparison to the FG-nontreated animals, accompanied with higher values of red blood cell counts, hemoglobin concentration and hematocrit. When liver incision was performed, although there were no differences in survival rate, FG-treated animals had significantly higher values of the tested hematological parameters. The presented results demonstrated that by using 'recycled' cryoprecipitation it is possible to obtain high quality single-donor FG with successful hemostatic therapeutical effects, as confirmed in the experimental rat model of liver surgery.

Animals↗

Quantitative, functional, morphological and ultrastructural recovery of platelets as predictor for cryopreservation.

Cryopreservation of platelets is of great interest, since it could extend the shelf life of therapeutic platelet concentrates and facilitate stockpiling and inventory control in blood banking. Despite the use of many cryopreservation procedures the optimal cryopreservation procedure is not defined yet. We have compared the cryopreservation of human platelets by various protocols employing controlled-rate and non-controlled-rate freezing procedures in combination with different concentrations of DMSO (6% and 10%) or 5% DMSO + 6% HES combination. After storage for 1 to 3 months, samples were thawed and analyzed. Measurements included cell recovery, platelet viability according to hypotonic shock response (HSR), platelet aggregation with ADP, morphological and ultrastructural properties of defrozen platelets. Our findings show that the application of our original procedure for controlled-rate freezing consisting of six cooling steps (cooling rate 1 degree C/min) with compensation of released heat of fusion (cooling rate 2 degrees C/min) has significantly influenced the quality of thawed platelets. At the same time, a concentration of 6% DMSO proved to be the most effective. In summary, cryopreservation of human platelets using controlled-rate freezing procedure in combination with lower (6%) DMSO concentration resulted in less damage from freezing and higher recovered function of platelets.

Blood Platelets↗

Differentiation of human dendritic cells from monocytes in vitro using granulocyte-macrophage colony stimulating factor and low concentration of interleukin-4.

Several laboratories have developed culture systems that allow the generation of large numbers of human dendritic cells (DC) from monocytes using granulocyte-macrophage colony stimulating factor (GM-CSF), and interleukin-4 (IL-4). In this work we provided evidence that GM-CSF (100 ng/ml) in combination with a low concentration of IL-4 (5 ng/ml) was efficient in the generation of immature, non-adherent, monocyte-derived DC as the same concentration of GM-CSF, and ten times higher concentration of IL-4 (50 ng/ml). This conclusion was based on the similar phenotype profile of DC, such as the expression of CD1a, CD80, CD86, and HLA-DR, down-regulation of CD14, and the absence of CD83, as well as on their similar allostimulatory activity for T cells. A higher number of cells remained adherent in cultures with lower concentrations of IL-4 than in cultures with higher concentrations of the cytokine. However, most of these adherent cells down-regulated CD14 and stimulated the proliferation of alloreactive T cells. In contrast, adherent cells cultivated with GM-CSF alone were predominantly macrophages, as judged by the expression of CD14 and the inefficiency to stimulate alloreactive T cells. DC generated in the presence of lower concentrations of IL-4 had higher proapoptotic potential for the Jurkat cell line than DC differentiated with higher concentrations of IL-4, suggesting their stronger cytotoxic, anti-tumor effect.

Apoptosis↗

Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro.

BACKGROUND: Dendritic cells (DC) have been used for immunotherapy of malignant tumors, different kinds of infections, and other clinical conditions. For that purpose, optimal conditions for the generation of functionally mature DC in vitro are required. Two different protocols for the induction of maturation of monocyte-derived DC (MDDC) were compared in this study. METHODS: MDDC were generated in vitro by cultivating adherent monocytes of healthy volunteers with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) during 6-days period. The immature DC thus prepared were induced to mature using two protocols. DC were stimulated for 2 days with lipopolysaccharide (LPS), or with a cocktail of proinflammatory mediators (PM) containing IL-1beta, IL-6, tumor necrosis factor alpha (TNFalpha), and prostaglandin E2 (PGE2), respectively. Phenotypic characteristics of MDDC and their endocytic activity were studied by flow cytometry. Allostimulatory activity of these cells was tested in the mixed leukocyte reaction (MLR), whereas the production of cytokines was determined by ELISA kits. RESULTS: MDDC matured with PM (PM-DC) were predominantly non-adherent cells, while about 30% of LPS-matured DC were adherent cells. In comparison with LPS-DC, PM-DC expressed higher levels of CD86 and CD83, had lower endocytic activity, produced higher levels of IL-10 and lower levels of IL-12, and more strongly stimulated proliferation of allogeneic lymphocytes. CONCLUSION: The protocol based on the combination of proinflammatory cytokines and PGE2 is better for the induction of maturation of human MDDC in vitro than the protocol using LPS alone.

Cell Culture Techniques↗

Autologous bone marrow-derived progenitor cell transplantation for myocardial regeneration after acute infarction.

BACKGROUND: Experimental and first clinical studies suggest that the transplantation of bone marrow derived, or circulating blood progenitor cells, may beneficially affect postinfarction remodelling processes after acute myocardial infarction. AIM: This pilot trial reports investigation of safety and feasibility of autologous bone marrow-derived progenitor cell therapy for faster regeneration of the myocardium after infarction. METHODS AND RESULTS: Four male patients (age range 47-68 years) with the first extensive anterior, ST elevation, acute myocardial infarction (AMI), were treated by primary angioplasty. Bone marrow mononuclear cells were administered by intracoronary infusion 3-5 days after the infarction. Bone marrow was harvested by multiple aspirations from posterior cristae iliacae under general anesthesia, and under aseptic conditions. After that, cells were filtered through stainless steel mesh, centrifuged and resuspended in serum-free culture medium, and 3 hours later infused through the catheter into the infarct-related artery in 8 equal boluses of 20 ml. Myocardial viability in the infarcted area was confirmed by dobutamine stress echocardiography testing and single-photon emission computed tomography (SPECT) 10-14 days after infarction. One patient had early stent thrombosis immediately before cell transplantation, and was treated successfully with second angioplasty. Single average ECG revealed one positive finding at discharge, and 24-hour Holter ECG showed only isolated ventricular ectopic beats during the follow-up period. Early findings in two patients showed significant improvement of left ventricular systolic function 3 months after the infarction. There were no major cardiac events after the transplantation during further follow-up period (30-120 days after infarction). Control SPECT for the detection of ischemia showed significant improvement in myocardial perfusion in two patients 4 months after the infarction. Echocardiographic assessment in these two patients also showed significant improvement of systolic function three months after the infarction. CONCLUSION: Preliminary results of the study showed that the transplantation of bone marrow-derived progenitor cells into the infarcted area was safe, and feasible, and might improve myocardial function. Further follow-up will show if this treatment is effective in preventing negative remodeling of the left ventricle and reveal potential late adverse events (arrhythmogenicity and propensity for restenosis).

Aged↗