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Background: P-gp, MRP, LRP, Bcl-2, and Bax proteins play an important role in the multidrug resistance of leukemic cells. P-gp, MRP, and LRP proteins decrease the intracellular drug concentration, Bcl-2 and Bax proteins influence the apoptosis process. The overexpression of some of these proteins is associated with poor prognosis of leukemia. The aim of this study was to find the relationship between some multidrug resistance markers and some clinical and laboratory parameters. We compare also P-gp expression between acute myeloid leukemia (AML) FAB subgroups M0-M6., Methods and Results: With use of flow cytometry we measured the expression of P-gp, MRP, LRP, Bcl-2, and Bax proteins in acute myeloid leukemia patients cells. The study proved the association between blast percentage and P-gp protein expression (p=0.015) and the association between blast percentage and Bcl-2 protein expression (p=0.041). It was also shown the tendency of the LRP protein expression to associate with higher age of patients (p=0.062). Another correlation between MRP and Bax expression (p=0.006), as well as between LRP and Bax expression was found (p=0.034). In AML M0 FAB subgroup of patients the trend to higher P-gp protein expression was observed., Conclusions: The laboratory methodology to multidrug resistance markers detection was introduced. Some relations between multidrug resistance markers playing role in acute myeloid leukemia patients prognosis were suggested.

The apoptosis failure in cytostatic treatment of haemoblastosis is one of the means of chemoresistance. We were interested in the relationship of the after-doxorubicin-treatment-AML cells apoptosis and the immunophenotype, selected clinical and laboratory parameters, and also the P-gp, MRP, LRP, Bcl-2, Bax proteins expression. All analysis were evaluated with the flow cytometry method. To detect apoptotic cells in the sample, we used three methods: annexin test, TUNEL (TdT-mediated dUTP nick end labeling) test, and caspase 8 detection. After the cell cultivation the statisticaly important increase of apoptotic cells in the culture was apparent. The relation between the AML blast in vitro reaction and some clinical parameters such as the age of patient, white blood cell count, and blast percentage was also observed.

The publication provides an up-to-date review of the significance of cytogenetic abnormalities in chronic myelogenous leukemia (CML) and the development of a promising agent with specific molecular target against tyrosine kinase, product of the BCR-ABL fusion gene, namely imatinib mesylate (STI 571, Glivec). The publication summarizes the achieved results with this compound in the chronic phase CML (in patients resistant to interferon and in newly diagnosed patients) further in patients in the accelerated phase and in blast crisis and in patients in relapse after allogeneic stem cells transplantations for CML. The results in Ph+ acute lymphoblastic leukemia are also presented. The mechanisms of resistance to imatinib mesylate and the possibilities how to overcome or circumvent it are mentioned (escalation of the dosage, combination of imatinib with some other treatment modalities as immunotherapy, interferon or convention chemotherapy and development of new drugs).

The presented study compares the efficacy and the toxicity of idarubicine and mitoxantrone in combination with cytosar (3 + 7) in induction treatment of the patients with AML aged 55-75. 31 patients at the age of 55-75 (median 62) were evaluated in the arm with idarubicine and 29 patients at the age of 57-74 (median 64) in the arm with mithoxantrone. Complete haematological remission was achieved in 13 patients (41.9%) in the arm with idarubicine and 15 patients (51.7%) in the arm with mitoxantrone. The medians of overall survival time (OS) and disease free survival time (DFS) were 22 and 44 weeks in the idarubicine arm and 35 and 40 weeks in the mitoxantrone arm, respectively. Statistical analysis did not prove any significant difference in the complete remission rates, in the number of deaths during cytopenia, in the OS or DFS, in the duration of hospitalisation, severe neutropenia and thrombopenia, in the number of days with febrile neutropenia, or in the consumption of platelets and erythrocytes transfusion units between both arms. Despite the fact that these results are not statistically significant in favour of any treatment arm, which is probably influenced also by the small number of evaluated patients, more favourable results were achieved in the arm with mithoxantrone with the respect to the evaluated parameters. From the point of view of cost-effectiveness, the difference could be observed when considering the price of both intercalating cytostatics. The use of mitoxantrone (Refador, Lachema) is 15x times cheaper per course of treatment than the use of idarubicine (Zavedos, Pharmacia). Autologous peripheral blood stem cells transplantation (APBSC) was carried out only in 4 patients younger than 60. No one of them was cured by APBSC but the median of OS of these patients was longer than the median in the other patients of the group. The results achieved are comparable with those of other trials conducted by various foreign groups. The possible causes of our unfavourable treatment results in this high-risk category of aged patients and the ways how to individualize the treatment with the use of prognostic factors analysis and how to improve the quality of life of the patients has been discussed.

Several authors have tried to solve the problems in the classification of CMML. A fully suitable classification does not exist. The goal of our study was to determine common and different signs of MD and MP type of CMML and to observe frequency of shifts from MD to MP-CMML. Sixty nine CMML patients were divided according to FAB proposal into two groups: 31 patients into the MD group (WBC < or = 13 x 10(9)/l) and 38 patients into the MP group (WBC < or = 13 x 10(9)/l). Presenting features and the course of the disease in both groups were evaluated. The median age of patients was not different in both groups (71.5 and 74 years, respectively), male/female ratio was 1.1 and 2.4, respectively. The median follow-up time was 15.5 months (1-58.8) in MP group and 24 months (2-118) in MD group. In MP group splenomegaly, hepatomegaly, lymphadenopathy, abnormal karyotype and skin involvement were found more often than in MD group. Median LDH value was higher in MP group. Probability of survival was higher in the MD group than in MP group (median 30 and 11 months, respectively). Leukaemia transformation frequency was similar in both groups. In 12 out of 24 (50%) MD group patients WBC increased during the course of the disease over 13 x 10(9)/l. Oscillation of WBC values below and over 13 x 10(9)/l was observed in three patients. During the follow-up time number of patients with splenomegaly and/or immature granulocytes in the PB increased. After inclusion of 12 patients who shifted from MD to MP group a new CMML group resulted characterised by longer median survival (17 months) due to a higher number of patients in an earlier stage of the disease. Failure of evolution of myeloproliferative signs and lower frequency of AL in the remaining group might be explained by an early stage of CMML, untimely deaths due to unrelated causes and/or by patients suffering of RA with monocytosis rather than of CMML. In summary, our data suggest, that evolution from MD-CMML to MP-CMML is a frequent event and that MD-CMML could be the early stage of CMML in most of cases. The WBC at diagnosis as the single criterion for subclassification of CMML does not seem to be fully justified. We propose that CMML should not be divided in MD and MP types and that monitoring of patients and search for other signs of myeloproliferation such as PB immature granulocytes, splenomegaly, lymphadenopathy, skin involvement, pleural or peritoneal effusions, spontaneous growth of CFU-GM in vitro should be taken in consideration for a better classification of CMML, which would have an impact on the therapeutic approach.

Background: Detection of membrane molecules by immunophenotypisation is a routine counterpart of the laboratory tests which are needed for the complex diagnostic procedures of hematopoetic malignancies at present. The immunochemical detection of intracellular molecules is essential in such cases, where the expression of surface molecules on malignant cells is poor, or where the results of other diagnostic analyses are unequivocal (cases with ectopic expression of other lineage-specific molecules and cases of biphenotypic leukemias)., Methods and Results: We followed the procedure using 4% solution of paraformaldehyd in phosphate buffered saline for fixation of cells. Lysing solution G (Becton-Dickinson) was used for permeabilisation of cells to detect nuclear and cytoplasmatic molecules. The permeabilised cell suspension was than washed with 0.5% solution of Tween 20 in phosphate buffered saline. This simple and rapid procedure is readily available for the routine detection of intracellular molecules. This approach is important for the diagnosis of hematopoetic malignancies.

Background: An injury to the hemopoietic stem cell may lead to the aplasia of hemopoiesis, myelodysplasia and to an unregulated myeloproliferation. There is not a strict demarcation of them, so that mixed syndromes can develop as are hypoplastic syndromes on one side and mixed myelodysplastic and myeloproliferative syndromes (MDS-MPS) on the other side., Methods and Results: Among our 616 pts with MDS we looked for those cases, who had beside myelodysplasia signs of myeloproliferation with increased number of blood cells. They were examined in detail including bone-marrow histology, bone marrow cultivation, cytogenetics and bcr-abl gen. Signs of MDS-MPS were found in 22 patients at the first contact with the patient (13 patients had thrombocytemia and 9 patients had leukocytosis). Further 7 patients were diagnosed as MDS, proliferative syndrome developed after several months (MDS-MPS in evolution). The level of thrombocytemia was relatively stable, the number of leukocytes was progressive. All subtypes of MDS were found. All subjects had variable degree of anemia. Ring-sideroblasts and myelofibrosis were frequent finding in MDS-MPS. Men prevailed in patients with leukocytosis. Cytogenetic and cultivation findings were similar to MDS cases, deletion of long arm of chromosome 20 was present in 3 patients. Five patients transformed to acute myeloid leukemia., Conclusions: Sings of myelodysplasia and myeloproliferation were found in 4% of our MDS patients, designated as mixed myelodysplastic and myeloproliferative syndrome (MDS-MPS). In this syndrome beside evident signs of myelodysplasia thrombocythemia or leukocytosis with the release of bone marrow precursors are present. In only one case polycythemia was encountered.

A fully implantable venous port system with a cannula inserted into the central venous system was implanted to 91 patients. The system was used for the administration of different types of intravenous preparations, incl. transfusions of erythrocyte mass and parenteral nutrition. With in the framework of autologous transplantation via the port system haematopoietic progenitor cells from the peripheral blood stream were administered. Part of the introduced systems was used for monitoring of the central venous overpressure and for collecting blood samples. The system was introduced on an average for 289 days, with a median of 245 days, range 82-872 days. During the implantation seven complications developed, in three instances the port was removed prematurely. During treatment 16 complications were observed, which in six instances led to early explantation of the venous port. The number of complications in total amounted to 0.87 per 1000 treatment days. The system was well tolerated by the patients because of its functional effectiveness and minimal cosmetic changes, and because it is not demanding it was also accepted well by the nursing staff.

The effects of weekly intravenously administered a dithiol chelating agent-sodium 2,3-dimercaptopropane-sulphonate (DMPS)-in a single dose of 50 mg/kg/week for 10 weeks on biochemical and haematological parameters were studied in rabbits. DMPS was well tolerated, an increase in body weight was similar in the DMPS-treated and control animals. DMPS caused significant decrease in plasma calcium and vitamin E concentrations at the end of the experiment. No significant differences in haematological parameters between the DMPS and control groups were observed. A significant decrease in magnesium content in myocardial tissue was observed in the DMPS-treated rabbits. The above-mentioned biochemical changes should be taken into account in studies of possible chelating and radical scavenging effects of DMPS in various pathological conditions.

The effects of subchronical exposure to SO2 (400ppm, 3 hours daily, 28 days) on biochemical and hematological parameters were investigated in guinea pigs. Mostly no significant changes in the values of biochemical parameters and no significant changes in hematological parameters were found. The levels of investigated ions (K+, Na+, Cl-, Ca++, Mg++ and phosphates), proteins (albumines, globulines, total proteins), enzymes (LD, ALT, AST, CK) and other biochemical parameters (urea, creatinine, bilirubin) were not significantly different between groups, with the exception of a significantly higher ALP concentration in the exposed group as compared with controls (2.17 mukat and 1.85 mukat, respectively. It can be concluded that a subchronical exposure to sulphur dioxide mostly did not induce any definite changes in biochemical and hematological parameters in guinea pigs.

The authors describe a female patient suffering from hairy-cell leukaemia. Already at the onset of the disease, apart from marked splenomegaly, sonography revealed marked retroperitoneal lymphadenopathy. During the subsequent course skin changes developed such as vasculitis and leukaemic infiltrates of the cornea on both eyes. The patient was successfully treated with 2-chlorodeoxyadenosine (2-CdA, Leustatin).

The authors made immunophenotyping analyses of blood cells of 50 patients with the diagnosis of B-CLL. Malignant cells contained in all instances HLA DR and CD24 molecules. The superficial molecule CD5 was expressed on cells in 85.7% molecule CD6 on 66.7% cases. By statistical analysis the relationship between selected clinical parameters and expression of surface molecules CD5, CD6 and CD10 was sought. A statistically significantly higher number of leucocytes was found in patients with expression of molecule CD6.

Hairy cell leukemia is a chronic lymphoproliferative disorder of B-cell lineage. Malignant cells express the interleukin-2 receptor (IL-2R) which is released in vitro as well as in vivo. The sera of patients with hairy cell leukemia contain elevated levels of this soluble receptor (sIL-2R). Sera of 24 patients with hairy cell leukemia were tested for sIL-2R. In 9 patients treated with 2-chlorodeoxyadenosine an improved clinical status was associated with decreasing serum sIL-2R. The maximal rate of decrease of sIL-2R level was observed within the second and the third week after the therapy initiation. Patients with disease progression had an increase in serum sIL-2R level. Our results suggest that serial measurement of sIL-2R level can be used as a reliable, noninvasive means to assess the disease activity and its response to therapy.

The effects of the repeated i.v. administration of daunorubicin (50 mg/m2, once weekly, max. 9 weeks) were investigated in rabbits in vivo to analyze biochemical and hematological changes. Noninvasive polygraphic records were used to evaluate the function of the heart. The administration of daunorubicin induced changes especially in levels of protein (decrease in total protein and albumin) and of some ions (decrease in calcium, magnesium and phosphorus) as well as in hematological parameters (decrease in erythrocytes, leukocytes and thrombocytes). The results obtained correlate with data on mechanisms of daunorubicin toxicity.

The best treatment protocol for hairy cell leukaemia (HCL) has not yet been defined. Different chemotherapies failed to demonstrate sustained benefit. Splenectomy certainly has been beneficial for many patients but has no effect on bone marrow infiltration by leukaemic cells. The introduction of Interferon-alpha (IFN-alpha), 2-deoxycoformycin (DCF) and 2-chlorodeoxyadenosine (2-CdA) in the therapy of HCL dramatically improved treatment options during the past decade. IFN-alpha does not have curative potential in HCL. 2-CdA seems to be more promising than DCF. Serum IL-2R level can be used as non invasive means to assess disease response to therapy. The predictive reliability of this test remains to be evaluated. The definition of complete remission remains open being greatly influenced by the sensitivity of the methods applied to detect residual disease. The role of the spleen as the reservoir of malignant cells can not be excluded in respect to the relapses of the disease.

The authors present the results of immunophenotyping analysis of cells from 67 patients with the diagnosis of B-CLL from group of 393 adults examined by immunophenotyping because of a haematological malignity. Characteristic for B-CLL is the finding of surface molecules HLA DR, B lymphocytic superficial molecules and CD5. Aberrant membrane equipment of B-CLL cells is suggested by the rare absence of some pan B lymphocytic sign. Marked reduction of T lymphocyte ratios, their sub-populations and NK cells is associated with immunodeficiency manifestations which are common in patients with B-CLL.

Basic red cell ferritin was investigated in 28 patients with different phases of chronic granulocytic leukemia (CGL). Red cell ferritin was significantly decreased in remission after busulphan treatment and significantly elevated in the blast crisis as compared to healthy controls. Bone marrow stainable iron was decreased or absent in 86% of patients in the initial phase at the time of diagnosis and in 92% of those in remission. Red cell ferritin correlated with serum ferritin, however, serum ferritin level remained above normal range during all phases of the disease. A negative correlation between red cell ferritin and hemoglobin (Hb) (r = -0.605, p < 0.001) suggested that red cell ferritin level reflected the rate of iron utilization for heme synthesis. Decreased red cell iron stores observed in the remission may be explained by regression of dyserythropoiesis and by restoration of normal Hb synthesis after busulphan treatment. A progressive dyserythropoiesis in the blast crisis may lead to an increased red cell ferritin level.

Histological findings in the bone marrow and their changes in the course of the disease and therapy were evaluated with respect to the prognosis in a group of 32 patients with hairy cell leukemia (HCL). Two types of bone marrow infiltration by hairy cells (HCs), the diffuse type and the interstitial type, respectively were found. The diffuse type of infiltration and the minimal or absent residual hematopoiesis (RH) at presentation were found with statistical significance to be unfavorable prognostic findings when compared with interstitial infiltration and persisting RH (p < 0.01). Reticulin fibrosis is a characteristic finding in HCL and was found in all but 2 patients.

The authors describe a fatal haemorrhagic condition in a 53-year-old man which developed after assumed ingestion of so-called "superwarfarins". Symptoms of intoxication with substances of the warfarin type include the sudden development of haemorrhagic diathesis, marked prolongation of INR and APTT with persisting pathological values of these tests after the usual substitution doses of vitamin K1. The fluctuation of INR and APTT values during repeated examinations is also striking. In the event of such a coagulation finding it is important to consider possible superwarfarin intoxication, even if it is difficult to provide direct evidence, as it is not possible to assess the ratio of vitamin K1 and K 1, 2, 3 epoxide.

Enlargement of nodes in leukaemia with hairy cells (LVB) during the initial examination is rarely encountered. In the final stage of the disease it was found in six of our ten patients. A special situation is marked enlargement of retroperitoneal nodes observed in two patients. In one of them the nodes diminished after irradiation, in the second patient irradiation was not possible because of severe pancytopenia. Treatment with alpha-interferon failed previously in this patient. The question remains whether marked enlargement of the abdominal nodes is part of the picture of the disease or whether it is the manifestation of transformation of the disease into a more acute type. The authors recommend examination of the abdominal nodes by means of computed tomography (CT) already during the initial examination and during the subsequent course of the disease, in particular when there are signs of relapse.

Splenectomy as a therapeutic modality has been evaluated in a group of 24 consecutive patients. Before and after splenectomy only supportive therapy was applied with the exception of two patients in whom corticosteroids were administered because of vasculitis. Radiation therapy was applied in one patient because of massive retroperitoneal lymphadenopathy. One patient received Leukeran shortly before the fatal outcome because his response to splenectomy was poor. Out of 24 patients 14 patients died. The median survival for the partial responders and non-responders was 6 months only. Patients with a complete response did not reach median survival yet and 8 of them (33% of the whole group) have been alive for more than 9 years, one patient for more than 20 years. Two distinct groups of patients with respect to the outcome of splenectomy can be distinguished: The first group is characterized by a rapid fatal outcome mostly within two years after splenectomy, the second group has a more favorable course with a plateau and only occasional deaths after the fourth year.

In 193 patients with Ph-positive chronic myeloid leukaemia (CML) Sokal's calculation formulae prognosticating initial risk status were applied, using the computer program PHCML. The aim of this study was to the prognostic reproducibility of these prognostic staging systems in our patient population. Our results confirm the previous conclusions that prognostic discrimination is possible at the time of diagnosis of CML but in our patients with poorer prognosis the stratification seems to be more accurate. No significant advantage was achieved when we prognosticated our patients younger than 46 years using the formula recommended by Sokal et al. specifically for younger patients. We analyzed different criteria when segregating patients in groups with better or poorer prognosis. We found that the coefficient of the patient's relative risk as the boundary, was 1.9 our patient's group contrary to 1.2 in Sokal's studies.

On the example of a patients with eosinophil leukaemia, which at first was manifested as eosinophilia in the peripheral blood stream and bone marrow without involvement of other organs and only after three years acquired the character of malignant growth, the authors draw attention to difficulties in the differential diagnosis of hypereosinophil syndrome. At the same time the authors review briefly views on the origin of eosinophil leukaemia, morphological and cytogenetic findings considered useful as evidence of this rare type of leukaemia.