[The multiabberant cells in groups of people exposed to radiation due to different situations and their possible biological part].

The results of the analysis of multiaberrant cells (MAC) obtained in the course of long-term investigation of cytogenetic effects in human peripheral blood lymphocytes are presented. MAC were discovered in different groups of the population exposed to the radiation factor. No such cells were found in the control groups. The greatest number of MAC "carriers" was registered among employees of radiochemical plants who had contacts with plutonium salts. The highest frequency of MAC (2.49 +/- 0.59 per 1000 cells) was also revealed in the same group. It exceeded by an order of magnitude the analogous values in other examined groups. In the groups of radiochemical workers, cosmonauts, and miners from Tselinograd the frequency of dicentrics and centric rings was also the highest as compared to that in other groups. The character of chromosome aberrations observed in MAC suggests that they are formed under the action of the radiation factor, and their frequency among different groups of people exposed to radiation makes it possible to assume that the formation of MAC is a result of the action on lymphocytes of alpha-particles emitted by radionuclides incorporated in the organism. Classical MAC was observed in routine studies (FPG staining) are only an extreme manifestation of cell damage. To elucidate the true picture of chromosome rearrangements induced by radiation and the role of MAC in the tumor process, it is necessary to use methodical potentialities of modern molecular cytogenetics, including the FISH method.

[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.

Origination of stromal precursor cells replenishing the population of these cells in heterotopic mouse bone marrow transplants after curettage in recipients.

Curettage of bone marrow cavities of two bones (femoral and crural) in recipient mice causes a drastic (more than 7-fold) increase in the count of stromal precursor cells in heterotopic bone marrow transplants. Stromal colonies in cell cultures from these transplants consist of fibroblasts with an appreciable admixture of macrophages. All y chromosome-typed colonies from cultures of female donor bone marrow transplants in recipient males (intact and subjected to curettage) contained cells carrying and not carrying y chromosome. Quantitative results of Y chromosome typing of cells from colonies corresponded to the fibroblast/macrophage ratio in colonies and the predominant localization of the label corresponded to predominant localization of macrophages (at the periphery of colonies). The results indicate that the pool of bone marrow stromal precursor cells under conditions of increased demands originates from local sources, which confirms ample data on inability of these cells to migration.

[Molecular-cytogenetic monitoring of different regimens of treatment in patients with chronic myeloid leukemia]. / Molekuliarno-tsitogeneticheskii monitoring razlichnykh rezhimov terapii u bol'nykh khronicheskim mieloleikozom.

AIM: To conduct molecular-cytogenetic monitoring of bone marrow cells in different regimens of chronic myeloid leukemia (CML) treatment. MATERIAL AND METHODS: A total of 651 samples of bone marrow from 319 CML patients were studied. 229 patients received polychemotherapy and 90 patients--interferon-alpha. Primary examination and monitoring of the treatment efficacy were performed using G-differential chromosome staining. Fluorescent in situ hybridization (FISH) was made in 75% cases. RESULTS: Interferon therapy resulted in a significant increase in the number of complete and significant cytogenetic responses. With aggravation of the disease the above responses occurred less frequently while minor and no response are encountered more often. Treatment with interferon-alpha in combination with chemotherapy is much more effective than monotherapy with interferon. CONCLUSION: G-differential chromosome staining karyotypes metaphases and detects clonal chromosome restructuring. Molecular-cytogenetic methods study chromosome restructuring at DNA level. FISH detects chimeric gene bcr/abl in cases when Ph-chromosome is not detectable by standard cytogenetic methods.

[Synchronous programmed intensive therapy of patients with severe rheumatoid arthritis]. / Sinkhronnaia programmnaia intensivnaia terapiia bol'nylh revmatoidnym artritom tiazhelogo techeniia.

AIM: To evaluate efficiency and safety of intensive treatment program (synchroneous plasmapheresis, large-dose methotrexate and methypred) for patients with severe rheumatoid arthritis (RA). MATERIAL AND METHODS: 45 patients with highly active and progressive RA, systemic symptoms, corticosteroid dependence who had intolerance to standard therapy or had not responded to it were divided into 2 comparable groups. 25 patients of group 1 for a month got 6 plasmapheresis procedures with synchroneous intravenous injection of 40 mg of methotrexate and 250 mg of methypred. 20 patients of group 2 received pulse therapy with methypred (3 g) and methotrexate (200 mg). The intensive therapy was followed in all the patients with methotrexate in a dose 10-20 mg/week. RESULTS: One, six and twelve months after treatment patients of group 1 demonstrated a decrease in RA clinical activity and inflammation. In a year remission by ACR criteria was achieved in one-third of the patients. CONCLUSION: The sychroneous program of intensive therapy is highly effective in RA patients with vasculitis, ineffective standard therapy and corticosteroid dependence.

[Induced leukemias and their connection with radiation exposure]. / Indutsirovannye leikozy, ikh sviaz' s vozdeistviem radiatsii.

The cytogenetic study of bone marrow cells was performed in 40 patients with secondary leukemias which have arisen after application of cytostatic and/or radiotherapy for primary tumours (Hodgkin's disease, lymphomas, acute lymphoblastic leukemias, breast cancer and other solid tumours). The comparative analysis of results has shown, that the leukemias after irradiating or application of alkylating agents and irradiation, have the quite particular clinico-morphological and cytogenetic characteristics. In 70% of cases these diseases develops as smouldering leukemias with subsequent transformation in M-4, M-6, and rarely M-2 cytochemical variants. Primary cytogenetic events in 60% of researched karyotypes are the losses of long arms or whole chromosomes 5 and 7. In 20% of the researched cases normal karyotype was found, in the left 20%, the changes of a karyotype not including anomalies 5 and 7 chromosomes were detected. The obtained outcomes allow to consider the discharged complex of tags as reference for leukemias, induced by irradiating or chemical agents with similar mechanism of action (alkylating agents, benzene and its derivates). This complex of tags is typical for induced leukemias, and in a combination with definition of a level of stable aberrations in peripheral blood lymphocytes, can be utilised for abjection of radiation-induced leukemias from common mass of cases detected in regions, polluted by radionuclides. In this study in 60% of cases only specific for secondary leukemias chromosomal aberrations, including monosomy 5 and 7, rearrangements of 11q23 were found. On the base of the obtained data the differences in concepts of "secondary" and "induced" leukemias are considered.

Stable and unstable aberrations in lymphocytes of Chernobyl accident clearance workers carrying rogue cells.

Cells with multiple chromosomal aberrations, the so-called rogue cells, were found in blood samples from more than 100 Chernobyl accident clearance workers. A comparative analysis of frequencies of stable and unstable chromosomal aberrations in two worker groups--those with or without rogue cells was made. A higher level of unstable aberrations in persons carrying rogue cells was observed. No difference in the level of stable aberrations between the groups was seen. The possibility of low dose alpha irradiation causing the chromosomal damage is raised.

Cytogenetic features of leukaemias diagnosed in residents of areas contaminated after the Chernobyl nuclear accident.

A comparison of chromosomal abnormalities in bone marrow leukaemic cells and of stable and unstable aberrations in lymphocytes of patients with hematological malignancies who live in areas with or without contamination by the Chernobyl nuclear accident has been made using FISH and G-banding. Healthy residents of these areas comprised the control group. No systematic cytogenetic differences of leukaemic cells between patients from contaminated and uncontaminated areas were observed. Lymphocyte aberrations, however, were generally higher in all subjects from contaminated areas. Comparison has been made with specific cytogenetic features of leukaemic cells and a high level of stable aberrations in lymphocytes of patients with secondary leukaemias that had developed after chemo- and/or radio-therapy.

[The first case of Hb Camden carriage in Russia]. / Pervyi sluchai nositel'stva Hb Camden v Rossii.

Structural hemoglobinopathy due to carriage of Hb Camden was diagnosed in a male patient and his mother. Anomalous Hb was detected after a detailed analysis of the clinical evidence, blood smears microscopy, marrow examination, data obtained by a complex of methods to identify unstable Hb. It was defined as Hb Camden beta 131 GLN-->GLU. The carriage of this variant brings no clinical manifestations, but leads to impaired red cells morphology, decline of their life span, enhanced hemolysis. These properties are typical for mutant Hb lacking stability. Hb Camden carriage is considered as a compensatory hemolytic process.

Plasma desorption mass spectrometry of haemoglobin tryptic peptides for the characterization of a Hungarian alpha-chain variant.

S-Aminoethylated-alpha A and -beta A globin tryptic peptides separated by reversed-phase high-performance liquid chromatography have been analysed by plasma desorption mass spectrometry. Almost all the expected alpha A and beta A tryptic fragments were tentatively assigned relative to the known globin chain sequences based on the molecular weight obtained by plasma desorption mass spectrometric analysis of the purified peptides. The application of plasma desorption mass spectrometry for structure elucidation of a haemoglobin alpha-chain variant revealed the first case of Hb Hasharon in Hungary.

[The second family with hemoglobin Dagestan (alpha 60 Lys----Glu)]. / Vtoraia sem'ia s gemoglobinom Dagestan (alpha 60 Liz----Glu).

Data have been presented on a new hemoglobin detected in a family of the Lezghin population in the Daghestan ASSR. The structural analysis of this hemoglobin has shown its correspondence to hemoglobin "Daghestan" that was previously detected among the population living in the Tabasaransk region. A detailed genetic analysis has shown a possibility of the gene drift. It was found that the proband's grandmother (his father's mother) had come from the Tabasaransk population of the Republic.

[A new strategy for structural identification of abnormal human hemoglobins]. / Novaia strategiia strukturnoi identifikatsii anomal'nykh gemoglobinov cheloveka.

A new strategy for structural identification of abnormal human hemoglobins is proposed. It is based on micropreparative modification of electrophoretic separation of globins on Cellogel strips with subsequent quantitative isolation of a pure, desalted globin chain, in a form suitable for its subsequent structural investigation. Among the major advantages of the new strategy age possibility to use small blood samples (0.1-0.2 ml), short analysis time, relative simplicity and low cost.