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The article deals with indicators of cell composition of umbilical blood (fetus blood) in case of normal pregnancy (n = 38) and hypoferric anemia of light degree (n = 14)10 the hematologic analyzer of medium class was applied. The parameters of erythrocytes of fetus blood in case of hypoferric anemia of light degree were sufficiently stable and had no differences with normal values both in mean values and variability. The variety of adaptive index of fetus blood (Garkavi index) was significantly higher in case of hypoferric anemia as compared with normal values. This occurrence testifies higher stability of indicators of adaptation in case of moderate ferric deficiency. The study results testify the informativity of applied approach related to assessment of stability of cell composition of fetus blood.

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Human myeloid cells with Ph chromosome (Ph+ cells) from chronic myeloid leukemia (CML) in the course of proliferation and differentiation ex vivo are regulated under alternation of cell proliferation and neutrophil maturation stages by consecutive blocking and inducing apoptosis with of neutrophils participation as well bcr/abl, bax and bcl2 genes expression. Apoptosis regulation of three main Ph+ cells types from CML patients depends on alternation sequences of proliferation (1) and maturation (2) cell stages and realized by two ways. The first one is performed by consecutive blocking and inducing apoptosis under 2/1/2 stage alternation. The way is not described early. Neutrophils accumulation correlates with apoprosis blocking. Apoptosis level enhances under neutrophils exhausted. Apoptosis blockage allows cells to proliferate and, thus, to form new portion of neutrophils with consecutive regular their death as well a consequent alternation of apoptosis blocking and inducing. This way regulates proliferation efficiency indexes P/D that reflect Ph+ cells proliferating potential and performs cycle completion for proliferation and differentiation. The second way of apoptosis regulation starts from proliferation stage and performs for 1/2/1 alternations under diminished content of neutrophils and a little increase under next maturation. It leads to resistant depressed apoptosis levels that, at maximal points, are 3-8 times lower than those under alternation 2/1/2. Resistant apoptosis blocking is observed in the Ph+ cells with prolong proliferation or maturation stages, when blasts and myelosytes are accumulated under enhanced bcr/abl and bcl2 > box gene expression and remain under next maturation. Stable apoptosis blocking is accompanied by increasing amounts of blasts and myelocytes and enhancing bcr/abl and bcl 2 > bax expression. This is observed under CML progression. Ph+ cells cultivation may be useful for more distinct diagnostics of CML phases of individual CML patients and optimization of the treatment.

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The correlation between the erythrocyte sedimentation rate (ESR) and the electrophoretic mobility (EPM) of red blood cells in patients with hemophilia was studied due to the data available in the literature, showing an inverse relation. Anticoagulant-stabilized venous blood was a material to be tested. Patients with severe hemophilia A or B (factor VIII/IX < 1%; the normal values were 50-200%) were examined. The patients were divided into 2 groups: 1) those with an exacerbation of hemorrhagic syndrome and 2) those without an exacerbation. The patients with hemophilia with an exacerbation of the hemorrhagic syndrome showed a certain correlation between ESR and EPM, yielding two type of curves, which corresponded to two subgroups of patients. There was no correlation between EPM and ESR in the no-exacerbation group.

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Interaction of oligodeoxynucleotides (ODN), 18-mer, which included sequence of BCL2 mRNA translation start, with K562 cells has been studied. The kinetic curves of interaction showed that oligonucleotide total binding with the cells at 37 degrees C and low oligonucleotide concentration (< or = 30 nM), as well as under lipofection, were composed of two processes: 18-mer surface binding with cell membranes and its non-proportional internalization into the cells. The last, in turn, consisted of three consequent steps. The enhanced extent and rate of oligonucleotide internalization was diminished after first hour incubation and later they were increased again. This reflected rising additional binding sites that provided internalization. At chosen time-points the internalization of ODN into cells, been proceeded at 37 degrees C, were at most abruptly abrogated by cooling down. ODN to K562 cell membrane binding constants and specific number of binding sites have been determined. Time-intervals, providing equilibrium for each successive stage of multistep ODN bound/free determination, were maintained. It was established that receptor binding with increased binding constant (more than 2 x 10(9) M(-1)) promoted ODN internalization. Oligonucleotide binding and internalization with prolonged incubation were also up-regulated due to priming new binding sites of higher affinity. Lipofection enhanced ODN binding to cell membrane but conserved the main features of ligand-receptor interaction. During lipofection constants and ODN binding site numbers increased without changing the overall time-pattern of the process, observed for ODN without liposomes. Extent and rate of internalization of ODN in liposomal formulation did not differ substantially from ODN in solution.

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The following phenomena are observed in patients with late vascular complications of diabetes mellitus (DM): the smoking habit alters the quantitative and morpho-functional characteristics of the erythron cells; it inhibits the lipid peroxidation processes in the erythrocytes membranes and aggravates the clinical course of endotoxicosis, which has a pathological impact on the rheological blood properties and worsens the tissues' hypoxia. A correlation was found between the pronouncement degree of anemic syndrome in diabetic angiopathies, on the one hand, and smoking, on the other hand. Smoking was found in patients with diabetic angiopathies to be a factor enhancing the metabolic stress, early signs of vascular complications and a risk of developing the anemic syndrome.

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The count and size of erythrocytes are normally stable. In order to evaluate their ratio in health, some hematological diseases and during exposure to some ecological factors, peripheral blood erythrocytes were studied in 747 subjects (115 healthy controls aged 18-80 years, 56 subjects chronically exposed to ionizing radiation sources, 105 subjects working with aggressive liquids, 117 patients with anemias, and 81 with Hodgkin's disease). No relationship between erythrocyte count and MCV was detected in normal subjects and patients with anemia and Hodgkin's disease. It seems that this relationship is rather specific for hemopoiesis shifts associated with cell cycle delay, as it was detected in patients with B12 and folic deficiency anemias, in subjects chronically exposed to ionizing radiation sources, and in those handling nitrogen oxide in combination with heptyl.

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Diagnostic value of the NMR-relaxation method in the blood plasma was estimated in the patients with different pathologies. The time of hydrogen nuclei longitudinal relaxation (T1) in the health donors of the blood, in the patients with oncopathology (hemoblastoses) and in the cases with anemia and burning disease were investigated. The time of the longitudinal relaxation (T1) was measured by automated NMR-relaxometer "Palma" (Russia). The working frequency was equal to 35 MHz, the temperature was 45 +/- 0.1 degrees C. For the single measurement 0.2 ml of blood obtained from heparinized venous blood 1.5 hours after its taking was used. The time of the longitudinal relaxation (T1) was shown to be 1.78 +/- 0.02 in the health donors, 1.70 +/- 0.06 s in cases with anemia, 1.97 +/- 0.48 c in patients with leucosis, 2.40 +/- 0.12 s in patients with burns. The sensitivity and the specificity of diagnostics of leucosis based upon the results of the only single T1 measurement in blood plasma were concluded to be 75%. It proves the significant T1 change both in patients with anemia and burning disease of the II-III degree. However it is evidently insufficient for selective use of NMR-relaxation blood plasma (serum) in the diagnostics of anemia and leucosis. The data obtained prove also the possibility of use of NMR-relaxation blood plasma (serum) for control of the hemostasis state during treatment or remission.

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The paper presents the results of the studies of erythrocytic and iron metabolism parameters in the course of treatment with recombinant erythropoietin (RE) and iron preparations in 60 patients with chronic renal failure (CRF) on peritoneal dialysis (PD) aged 20 to 75 years. Pretreatment iron deficiency in CRF patients on PD persisted to the end of the third observation period. Hyperferritinemia registered in CRF patients on PD with frequent peritonitis may be considered as a result of active ferritin synthesis by the cells of the system of mononuclear phagocytes in response to inflammation. The only parameter indicating the presence of iron deficiency seems to be the percentage of hypochrome erythrocytes indirectly evidencing for insufficient supply and iron utilization in bone marrow. Pathogenetic mechanisms of anemia development in CRF patients on PD may be both low EP and high levels of proinflammatory cytokines as well as oversecretion of ferritin by activated by monocytes/macrophages ferrin, promoting apoptosis of late erythropoiesis precursors, development of ineffective erythropoiesis and having a suppressive effect on erythropoiesis cell precursors.

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AIM: To study proliferation of bone marrow cells in acute leukemia using flow DNA-cytometry. MATERIAL AND METHODS: Parameters of the cell cycle (S, G2 + M, G1/0) and myelogram were studied in 55 patients with acute myeloid leukemia (AML) and 46 patients with acute lymphoblastic leukemia (ALL) in remission (135 cases) and acute period (66 cases). Only cases with cell clones containing normal (diploid) number of DNA were included. RESULTS: In acute period, distribution of AML cases by the percentage of cells in S-phase of the cellular cycle was normal, in ALL--log-normal. In remission it was lognormal in both leukemias. Variability of cell cycle parameters in leukemia in some patients was much less than a mean sample one. The reestablishment of proliferation after chemotherapy (7 + 3) in AML was followed up. CONCLUSION: If proliferative activity of blast (tumor) cells in AML is subnormal (10%), proliferation of myelokaryocytes can be used as an additional test for assessment of remission completeness.

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AIM: Assessment of erythron parameters in subjects exposed occupationally to nitrogen oxides polluted air versus those who work in healthy environment. MATERIALS AND METHODS: Levels of red cells, reticulocytes, packed cell volume, resistance of red cell membranes to acid buffer were measured in 95 subjects working in polluted environment and 89 healthy controls. RESULTS: Long-term (10 years and longer) contact with nitrogases entails a tendency to lowering of red cell number in the exposed subjects, significant rise of packed cell volume, increased mean cell volume of red cells, anisocytosis and reticulocytosis, high acid resistance of red cells and their membrane permeability. The causes of these changes are discussed in terms of "uneffective erythropoiesis" conception. CONCLUSION: Thus, erythron parameters can serve for indication of toxic effects of some chemicals on human body.

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The proliferative activity of bone marrow cells was studied by flow cytofluorometry in 8 patients with aplastic anemia (AA), 6 with myelodyspastic syndrome (MDS), and 26 normal subjects aged 21-40 years. The myelokaryocyte proliferation was inhibited at the G0-1 stages of cell cycle, the number of cells in the S phase was decreased, and the coefficient of variations cV of DNA cytofluorometry values were increased in AA patients in comparison with the norm. By contrast, in MDS patients the number of cells in the G0-1 phase was decreased and that of cells in the S phase increased in comparison with the norm and with AA patients; cV was increased. These differences in the cell cycle characteristics are suggested to be used as a differential diagnostic criterion in AA and MDS.

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Studies of hemopoietic cell (myelokaryocyte) cycle and adequacy of applying flow DNA-cytometry for such analysis are discussed. Heterogeneity of the bone marrow, possibility of diluting a blood sample during puncture, reproducibility and stability of parameters, reliability of computer analysis of histograms demonstrating cell distribution, and correspondence of cell distribution by the cell cycle stages to the level of proliferative activity are the factors which may cause difficulties in the course of studies and while interpreting the results. Probable effects of these factors on the final results and efficacy of the method in clinical laboratory diagnosis are discussed.

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Reviews the history of flow cytometry, the philosophy of the method, main methods of cell investigation (conductometry, registration of light absorption and diffusion, cytofluorometry, study of fluorescence polarization, and analysis of a wave-shaped signal). Describes the equipment and methods for flow cytofluorometric analysis of cells, used in hematology. Characterizes the hematological analyzers with differentiated estimation of leukocytes.

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Proliferative activity of lymphocytes in a culture was assessed by flow cytometry and lymphocyte blast transformation test (LBTT) with the proliferative pool of cells (S+G2+M) from 150 patients with acute pneumonia, 30 of these with the mild form, 97 with medium-severe, and 27 with grave form; in 43 patients the disease duration was over 30 days (protracted pneumonia). In addition, levels of R.protein, homoreactant, immunoglobulins, complement components, T, B, and active T lymphocytes, phagocytosis, and granulocyte oxidative metabolism were assessed. LBTT values were in inverse relationship with the disease severity. Correlations between the studied immunity parameters were analyzed. The findings indicate that LBTT is an informative method for assessing the immune reactivity and may serve for predicting the disease course and for the differential diagnosis of pulmonary diseases.

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Cell cycle stage distribution of bone marrow cells was studied at flow cytometry in 137 hematological patients. They had acute lymphoblastic leukemia, acute myeloblastic leukemia, lymphosarcoma with leukemization, blast crisis of chronic myeloid leukemia, iron deficiency anemia, pernicious anemia, autoimmune hemolytic anemia, aplastic anemia (31, 34, 18, 12, 17, 8, 9, 8 cases, respectively). It was found that in leukemia, and to a lesser degree in anemia, stability of the cell cycle was impaired. In leukemia onset, recurrence and remission, the proportion S/(G2+M) appeared increased evidencing defective DNA synthesis by myelokaryocytes and ineffective hemopoiesis. Similar changes were seen in pernicious and aplastic anemia.

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The distribution of bone marrow cells by stages of cell cycle was studied by flow cytometry in 26 normal subjects and 107 adult patients (42 with anemias and 65 with acute lymphoblastic or myeloblastic leukemias). Normal myelokaryocyte cell cycle is rather stable, which manifests by low variability of cells. In anemias and, more so, in leukemias the proliferative status of cells varies within a wide range, which is explained by unstable and ineffective hemopoiesis.

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Bone marrow cellular DNA content was measured by flow cytometry in 107 adult patients with various types of acute leukemia. DNA-aneuploidy was detected in 28 of 107 patients (26%). In 18 patients with DNA-aneuploidy bone marrow was examined throughout 2-36 months. DNA-flow cytometry of bone marrow allows to determine DNA content in leukemia cells and distinguish hypodiploid, diploid, hyperdiploid and tetraploid DNA clones in patients with acute leukemia. In some cases this method allows monitoring of the minimal residual disease.

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