[Kinetics of proliferation, differentiation and transcription of genes regulating apoptosis in cultured human BCR/ABL+ Ph+-cells].

Ph+, bcr/abl+ cells arise due to t(9,22) chromosome translocation and Ph+ chromosome formation in hematopoietic stem cells. The cells show appreciable apoptosis suppression but retain their ability to differentiate and maturate. Ph chromosome, bcr/abl oncogene and Ph+, bcr/abl+ cells themselves are the hallmark of chronic myeloid leukemia. Under leukemia progression differentiating Ph+, bcr/abl+ cells transform into leukemic malignant cells with differentiation block. It is assumed to be a result of subsequent mutations or activation of proliferation of long silent Ph+ cells arisen previously in the stem cells because of the translocation. Real mechanism underlying the cell transformation remains unknown. This work was performed to develop a proper cell model allowing us to study functioning of differentiating Ph+, bcr/abl+ cells and their real transformation into malignant cells with block of differentiation. For this purpose we have investigated kinetics of Ph+, bcr/abl+ cells proliferation, differentiation, cell death and transcription of antiapoptotic genes in cultured 14-day of Ph+ mononuclear cells isolated from peripheral blood of a patient in chronic phase of chronic myeloid leukemia before treatment. The results obtained revealed that Ph+ cell differentiation proceeded in accord with characteristic scheme of chronic myeloid leukemia in vivo. Myeloid cells of hematopoietic cell lineage amounted to 3/4 of live Ph+ mononuclear cells undergoing accumulation and subsequent consumption in the course of differentiation. 95% myeloid cells were differentiating Ph+ granulocytes. The most deal of differentiating Ph+ cells was myelocytes. The rate ratio of myelocyte accumulation to its subsequent consumption showed that the rate of transformation into metamyelocytes was significantly decreased at this differentiation stage. Ph+ cells cultivation curves characterized cell death at different differentiation stages. There were observed the cell death of proliferating Ph+ cells and Ph+ myelocytes, and intensive death of mature cells as well. P/D index, that is ratio of immature Ph+ granulocytes differentiated by cell dividing (blasts, promyelocytes and myelocytes) to the cells differentiated without dividing (metamyelocytes and mature neutrofiles), revealed active of proliferation at the beginning of cultivation and unexpected new proliferative activity at the end of cultivation in the presence of growth factor. The peaks of antiapoptotic bcr/abl gene transcription activity coincided with the observed active proliferation at the beginning and at the end of cultivation. Cell proliferation, differentiation and apoptosis were noticeably accelerated by growth factor treatment. Thus, the study of the Ph+ cells cultivation kinetics is rather informative approach to investigation of continuous regulation of cellular and molecular processes in vitro in the case of chronic myeloid leukemia and allows more complete consideration of Ph+, bcr/abl+ cells hematopoiesis.

[Possibilities of early diagnosis of tumors].

AIM: To study diagnostic significance of blood serum cancer antigens levels in patients who subsequently develop pulmonary cancer (PC) and gastrointestinal cancer (GIC). MATERIAL AND METHODS: ELISA was used to study expression of tumor antigens (CEA, NSE, CA19-9, CA242, AFP) in the blood serum of 27 PC and 31 GIC patients; 22 patients with lymphatic tumors and 32 patients with pulmonary and gastrointestinal inflammation served control. After removal of the tumor the same antigens including cytokeratines (CK) and differentiated leukocytic markers were studied immunocytochemically in the tumor cells with relevant monoclonal antibodies. RESULTS: Sera of patients with verified afterwards cancer contained elevated concentrations of the antigens: NSE and CEA in PC, CA19-9, CA242 in GIC. The expression of these antigens including CK was found also in tumor cells of these patients. Atypical cells of lymphatic tumors had hemopoietic markers in the absence of CK. In inflammation and in lymphatic tumors, tumor antigens levels remained normal. CONCLUSION: The test for tumor antigens levels in the serum may be used for early (preoperative) diagnosis of cancer, especially in tumors with difficult access or if they are asymptomatic.

[Differential diagnosis of complicated cases of acute leukemia by morphocytochemical and immunological methods]. / Differentsial'naia diagnostika slozhnykh sluchaev ostrogo leikoza s pomoshch'iu kompleksa morfotsitokhimicheskikh i immunologicheskikh metodov.

Twelve intricate cases with different variants of acute leukemia were diagnosed by morphocytochemical and immunological methods. In six patients with initially diagnosed acute lymphoblastic leukemia (ALL) comprehensive studies revealed acute myelomonoblastic leukemia (AMML), in four ALL patients AML(M1,M0) and AMML were diagnosed, and in two patients with AML B-lymphoblastic lymphoma/leukemia was detected. These data indicate that immunophenotyping methods in complex with morphological and cytochemical studies helped correctly diagnose the disease variants, particularly in cases difficult to diagnose.

[The immunodiagnosis of breast cancer metastases to the bone marrow]. / Immunodiagnostika metastazov raka molochnoi zhelezy v kostnyi mozg.

Thirty-eight patients with stages II-III breast cancer after mastectomy, two patients with other tumors, and six patients with lymphosarcoma were examined. For detecting cancer metastases to the bone marrow, morphological analysis of puncture biopsy specimens was carried out in all patients with breast cancer; in patients with other diseases, puncture biopsy specimens of the tumor or lymph node were examined. Immunodiagnosis was carried out by the immunocytochemical method with detection of tumor antigens--monoclonal antibodies to cytokeratins, to breast cancer (ICO-25 and ICO-103), and to hemopoietic cells. All patients with breast cancer were divided into two groups: 1) 9 patients with cancer metastases to the bone marrow, confirmed by x-ray, histological, and morphological methods and 2) 29 patients without documented metastases to the bone marrow. Immunocytochemical methods detect isolated tumor cells in patients with suspected metastases to the bone marrow, lymph nodes, and other organs in cases when these cells are undetectable by routine morphological and histological methods.

[Mathematical model of light diffraction on red blood cells]. / Matematicheskoe modelirovanie difraktsii sveta na kletkakh krasnoi krovi.

Mathematical model diffraction of laser radiation on red blood smear for population erythrocytes with their normal distribution according to sizes is presented. It was shown during normal distribution of erythrocytes according to sizes that the first maximal pattern diffraction case does not correspond to the medium diameters, as it is given in literature. It was established that the depth of the first maximum is changed depending on the dispersion value according to sizes.