[Morphological evaluation of dysmyelopoiesis in decitabine-treated patients with myelodysplastic syndromes].

AIM: To estimate a change in myelodysplasia in decitabine-treated patients with myelodysplastic syndromes (MDS). SUBJECTS AND METHODS: Thirteen MDS patients from a high-risk group were examined; 75 bone marrow puncture specimens and 67 bone marrow trepanobiopsy specimens from these patients were analyzed before and after decitabine treatment. Dysplastic changes in the hematopoietic cells were monitored during the treatment. RESULTS: The dysplastic changes in the hematopoietic cells are a morphological portrayal of the ineffective hematopoiesis in patients with MDS. The study has indicated that the use of the hypomethylating agent decitabine promotes the restoration of cell differentiation to mature forms, causing hematopoiesis to be more effective. The incidence of myelodysplasias (including mixed double- and triple-lineage ones) was statistically significantly reduced by decitabine treatment, which was associated with a positive response to treatment as a whole. The count of cells with dysplastic features remained unchanged in patients with therapy resistance or further disease progression. CONCLUSION: Analysis of myelodysplastic manifestations in different hematopoietic lineages in patients with MDS should be based on the comprehensive dynamic assessment of cytological and histological parameters at both the primary diagnosis of the disease and different stages of treatment. With a response to decitabine therapy (as shown by the results of aspiration and trepanobiopsy), all cell lines displayed reduced myelodysplastic changes, indirectly indicating a decrease of the abnormal clone itself in high-risk MDS patients.

[Trisomy of chromosome 8 in Ph-negative cells of the bone marrow in patients with chronic myeloid leukemia treated with inhibitors of BCR-ABL tyrosine kinases].

AIM: To analyse clinical implications of chromosome 8 trisomy in Ph-negative cells of the bone marrow in patients with chronic myeloid leukemia (CML) treated with inhibitors of tyrosinkinases (ITK). MATERIAL AND METHODS: A total of 386 patients with CML (chronic phase--288, acceleration phase--77) received imatinib (400-800 mg/day). Because of resistance and/or intolerance some patients were switched to ITK II (nilotinib, dasatinib, bozutinib). This study included 8 CML patients (7 in a chronic phase, 1 in acceleration phase) treated with BCR-ABL ITK inhibitors of the first (imatinib) and the second line (ITK-II). The standard cytogenetic examination, on demand--investigation of the interphase nuclei with FISH, in some cases morphological, cytochemical and histological examinations of the bone marrow were made. RESULTS: The existence of a Ph-negative clone with trisomy of chromosome 8 had no negative effect on the course of the disease. The patients showed a stable hematological and cytogenetic response and no need in changing treatment policy. In long-term follow-up Ph-negative clone with trisomy of the chromosome 8 persisted without a clear trend to rise in most patients. CONCLUSION: Detection of a Ph-negative clone with chromosome 8 trisomy at early stages suggests parallel existence of Ph-positive and Ph-negative clones. None of the patients had myelodisplasia.

[Hairy cell leukemia in young patients].

AIM: To characterize clinical symptoms, course, immediate and long-term treatment results in young patients with hair cell leukemia (HCL). MATERIAL AND METHODS: The data on 41 HCL patients were analysed. The diagnosis was made by standard diagnostic protocol for HCL detection. RESULTS: The analysis of the age of 160 HCL patients studied demonstrated high (26%) incidence of HCL at young age. Young patients with HCL had special clinical manifestations and specific long-term outcomes of treatment with standard schemes. CONCLUSION: Differences in occurrence of recurrences after standard therapy make it necessary to consider young HCL patients as a separate group who need adjuvant treatment to prolong remission.

[Principles and potentialities of the standardization of morphocytochemical diagnosis of acute leukemias].

Under the present conditions, the competitive capacity of a health care facility is provided by the high level and timeliness of diagnosis of disease. The diagnosis of the types of acute leukemia (AL) may be accomplished immunologically, by using a 33-marker panel and without consideration of the morphocytochemical parameters of blast cells. But such an approach complicates and prolongs the examination of patients with AL. Moreover, morphocytochemical data more exactly define the stages of blast cell differentiation than does the immunological phenotype. Their preparation methods are simple and cost-effective. Only M0, M6, and M7 forms of leukemia require compulsory blast cell phenotyping, particularly in the differential diagnosis of acute myeloid leukemia and acute lymphoblastic leukemia. Search for new markers of leukemia cells, including lesions at the chromosomal or molecular levels, is under way. Some of them are only of theoretical value while other markers have been already used by hematologists to diagnose leukemia. Standardization in this essence is the self-assessment of a facility and reference comparison, which are based on the principles of its orientation to the patient, the adjusted system for controlling the quality of health care that is up to the world standards rather than the compliance with the state-regulated standard. The present paper discusses the ways of establishing the uniform rates and requirements for the morphocytochemical diagnosis of acute leukemias.

[Clinical significance of erythrocyte ferritin in refractory anemia and chronic myeloid leukemia]. / Klinicheskoe znachenie ferritina éritrotsitov pri refrakternykh anemiiakh i khronicheskom mieloleikoze.

AIM: To find out if the RBC ferritin elevation can serve as an additional criterion of inefficient erythropoiesis during progression of chronic myeloid leukemia (CML) and in various types of refractory anemia. MATERIAL AND METHODS: The study group consisted of 56 MDS patients and 73 patients at various stages of CML. 20 healthy donors and 105 patients with verified inefficient erythropoiesis (20--with B12 deficiency before and after the treatment, 85--with beta-thalassemia) were the controls. A ferritin level was measured by radioimmunoassay in RBC hemolysates. RESULTS: The RBC ferritin level in all types of refractory anemia was elevated throughout the disease course, increasing with the development of transfusion dependency. The CML progression was also accompanied by RBC ferritin level elevation associated with abnormal erythroid cell accumulation and elevation of intracellular PAS-positive substance (p < 0.05). CONCLUSION: RBC ferritin level elevation can be considered as an additional biochemical criterion of inefficient erythropoiesis that may be useful in differentiation of anemias, adequate therapy selection and follow-up of erythropoiesis.

[Effectiveness of cyclosporin A in the treatment of adult patients with aplastic anemia]. / Effektivnost' tsiklosporina A v lechenii vzroslykh bol'nykh aplasticheskoi anemiei.

AIM: To evaluate the efficacy of cyclosporin A (CyA) at different stages of immunosuppressive therapy (IST) in patients with aplastic anemia (AA). MATERIALS AND METHODS: The efficacy of CyA was studied in 56 patients with AA. The agent was orally given in an initial dose of 10 mg/kg as solution or capsules. Its daily dose during a treatment course varies with the serum CyA levels and clinical tolerance. CyA was used in 17 patients at the first stage of treatment, in 8 with recurrent AA, and in 31 after ineffective previous therapy (antilymphocytic globulin therapy--ALGT, splenectomy). Erythropoiesis was evaluated by the count of erythrokaryocytes and by relative erythroid hyperplasia of the bone marrow and by using erythrokaryocytic PAS reaction, by calculating the total count of sideroblasts and ringed sideroblasts. RESULTS: A positive response was obtained in 41% of the patients with AA. Its pattern depended on the severity of AA, on CyA use regimens, and treatment duration: when treatment with CyA lasted 6-12 months, its efficacy considerably increased (positive responses in severe AA and mild AA being in 64 and 94%, respectively). It has been found that high (over 6%) baseline bone marrow ringed sideroblasts in patients with AA may be regarded as a poor predictor in the context of the efficacy of this agent. CONCLUSION: CyA is recommended for combined IST in patients with AA at the second stage of treatment (after antilymphocytic globulin administration) in order to perform long-term (12-month) immunosuppression by choosing the optimum dose on an individual basis and by continuously monitoring the quality of a response.

[Quantitative morphology of periferal erythrocytes and bone marrow erythrokaryocytes in acute leukemia and hemopoietic depressions]. / Kolichestvennaia morfologiia éritritsitov perifericheskoi krovi i éritrokariotsitov kostnogo mozga pri ostrykh leikozakh i depressiiakh krovetvoreniia.

Morphology of peripheral blood erythrocytes was studied in patients with acute leukemia and aplastic anemia by a Russian cytoanalyzer Mekos-C. Twenty-eight patients with acute myeloblastic leukemia, 15 with acute lymphoblastic leukemia, and 11 with aplastic anemia were examined. Erythrocytes (n = 500) were examined in fixed non-stained blood smears. Hemoglobin content and morphometric parameters of each cell were studied and automatic classification of cells was carried out. The data of computer morphodensitometry are compatible with the data of cytochemical studies of the bone marrow erythroid cells (PAS reaction after McManus). The results indicate circulation of erythrocyte subpopulations differing by shape and other signs (hemoglobin content, section area, shape factor) in the blood of patients with acute leukemia and aplastic anemia. The share of pathological erythrocytes in the peripheral blood reflects failure of erythropoiesis.

[Physiological (programmed) cell death in hemopoiesis]. / Fiziologicheskaia (zaprogrammirovannaia) gibel' kletok pri gemopoéze.

According to current concepts, pluripotent cells proliferate and differentiate into "committed" precursors. These committed precursors divide, mature, and give rise to red cells, granulocytes, monocytes, and platelets of the blood. The life span of mature circulating cells being short, and their populations in the blood very stable, a constant and strict regulation of hematopoiesis is needed. The regulation of hematopoiesis is believed to be mediated through precursor cell interaction with specific molecules (glycoproteins) in their microenvironment. Soluble forms of these molecules are termed "hematopoietic growth factors" and include erythropoietin, colony-stimulating factors, interleukins, and stem cell factors. Hematopoietic growth factors not only stimulate the proliferation of precursor cells, but activate the differentiation program and maintain the viability of these cells as well. The normal fate of precursor cells devoid of these factors is programmed suicide. The morphology of such cell death is usually that of apoptosis, rather than of necrosis. The concept of apoptosis was proposed 22 years ago. Apoptosis is a widespread and morphologically distinct process of cell death. The significance of apoptosis stems from its active nature and its ability of controlling biological systems. The present review of published and authors' own data describes apoptosis morphology and presents evidence of the participation of cell reactions of this type in hematopoiesis regulation. The major point in the review is the balance of three normal processes: proliferation, differentiation, and apoptosis, which maintains the homeostasis of hematopoiesis, similarly as of any other cell system; it is well illustrated by recent findings in experimental and clinical hematology.

[A "primitive" variant of the blast crisis in chronic myeloleukemia]. / "Primitivnyi" variant blastnogo kriza khronicheskogo mieloleikoza.

While immunotyping blast cells from 45 patients with CML blast crisis, we detected 5 cases with immunologically primitive blast cells. The immunological phenotype of these cells corresponded to that of primitive stem cells which are characterized by expression of CD34 and HLA-DR antigens in the absence of other immunological markers. We suggest that blast cells from these patients may undergo differentiation similar to that of primitive stem cells that implies the existence of a new immunological variant of CML blast crisis, a primitive variant. Morphologically, blast cells in 3 cases could be classified as myeloid, in 2 cases precise identification was impossible. Cytochemically, this type of cells can be defined as mixed. The patients with CD34+ phenotype do not differ clinically or hematologically from those with CML blast crisis. Blast cells with membrane marker CD34 are likely to arise in any CML phase either as a component of overall leukemic population or predominant, single subclone.

[Possible relationship between the cytologic characteristics of mouse leukemia P-388 cells and their sensitivity to rubomycin]. / Vozmozhnaia sviaz' mezhdu tsitologicheskimi kharakteristikami opukholevykh kletok leikoza P-388 myshei i ikh chuvstvitel'nost'iu k rubomitsinu.

In search for markers of tumor cell resistance to cytostatic drugs, cytologic characteristics were compared of resistant and sensitive to rubomycin variants of mouse P-388 leukemia. Morphocytochemical, morphometrical and cytogenetic investigations were performed as well as measurements of intracellular pH. As shown by the fluorescent test, pH levels in rubomycin-sensitive and resistant subpopulations differed. This fact evidences an important role of intracellular pH in inducing the resistance and revealed some mechanisms of rubomycin-resistant tumor cell selection.

[Morphocytochemical characteristics of hemopoietic cells in sideroblastic anemia]. / Morfotsitokhimicheskaia kharakteristika gemopoéticheskikh kletok pri sideroblastnoi anemii.

The findings of morphocytochemical and cytogenetic examinations of hemopoietic cells in 8 patients with sideroblastic anemia are analyzed. The authors consider that the final diagnosis of sideroblastic anemia can be made if the count of ring-shaped sideroblasts surpasses 30%. The leukemic nature of this condition is discussed.