[The influence of some retarding agents NOS of dihydrothiazine-thiazoline rank on postradiational of recovery endogenous CFU-S-8 of mice].

In this work the attempt to estimate a nitric oxide (NO*) role in regulation of the number of pool haemopoietic stem cells at the irradiated mice was made. With this purpose the number of new compounds from dihydrothiazine-thiazoline line was synthesized, their NO-inhibiting activity was investigated in vivo by the method of ESR-spectroscopy of spin trap and their influence on an output endogenous spleen colonies (CFU-S-8) after the total sublethal y-irradiation of mice in a doze of 6 Gy was also investigated. Was shown, that the tested compounds reduced the contents of NO* in a liver tissue of mice which have received an injection of nitric oxide synthesis inductor - lipopolysaccharide, and also increased an output CFU-S-8 forming endogenous colonies in the spleen of the irradiated mice. Received data testify to perceptivity of search radioprotective agents among NO* synthesis inhibitors.

[Study of the impact of intramyocardial implantation of stem cells on myocardial perfusion and contractility in patients with coronary heart disease concurrent with postinfarct cardiosclerosis and chronic heart failure].

OBJECTIVE: to study of intramyocardial implantation of cultured bone marrow stem cells on myocardial perfusion and contractility in the surgical treatment of patients with coronary heart disease (CHD) and chronic heart failure (CHF), by synchronized single-photon emission computed tomography (SSPECT) of the myocardium. SUBJECTS AND METHODS: The study included 11 patients. Intramyocardial injection of cell injections into the myocardial periscarring areas was made at coronary bypass surgery. All the patients underwent 99mTc myocardial SSPECT MIBI before and 3, 6, 12 months after surgery. RESULTS AND CONCLUSIONS: Implantation of bone marrow stem cells into the left ventricular myocardium favorably affects left ventricular remodeling and contributes to the improvement of myocardial perfusion and contractility, as evidenced by 99mTc.

Immunology of apoptosis and necrosis.

A complex of reactions regulating the number of cells in organs and tissues under normal and pathologic conditions is one of the most important systems of multicellular organisms. In this system, which controls both cell proliferation and clearance, clearance has been given special attention during the last three decades. Some stages of the clearance are known (the choice of "unwanted" cells, their destruction not affecting the surrounding tissue, and, finally, removal of the corpses), and undeniable progress has been achieved in the understanding of the second stage mechanisms, whereas mechanisms of elimination per se of cells or their fragments still continue to be terra incognita. The clearance of such cells is mainly determined by different components of natural and adaptive immunity: phagocytes, complement, opsonins, antigen-presenting cells, etc. Recently specific "danger signals", such as hydrolases, DNA, heat shock proteins, and other potential immunogens released by cells during their elimination have been discovered. Entering the extracellular space, these signals induce inflammation and injury of the surrounding tissues, i.e., autoimmune reactions. Heat shock proteins, in addition to chaperon activity, act as signaling, costimulating, and antigen-carrying molecules in the interactions of dying cells and the immune system.

Agar cultures of human clonogenic hemopoietic precursor cells for early diagnosis of some myeloproliferative diseases.

Growth characteristics of human hemopoietic cells in erythremia and chronic myeloid leukemia were studied using agar cultures with and without hemopoietic growth factors. Agar cultures, similarly to cultures on other semisolid media (plasma clot, methylcellulose) can be used for early differential diagnosis of polycythemia vera (erythremia) and secondary erythrocytosis: erythremia, but not erythrocytosis, is characterized by spontaneous (erythropoietin-independent) formation of colonies from erythrocyte precursor cells. Spontaneous colony formation from granulocyte-macrophage precursor cells can serve as an important test for early diagnosis of chronic myeloid leukemia. The study of colony formation from granulocyte-macrophage precursors and of the capacity of bone marrow cells to form colonies from hemopoietic stromal precursor cells revealed new characteristics of the studied myeloproliferative diseases. Presumably, spontaneous colony formation from erythrocytic and myeloid precursors should be regarded as a sign of tumor transformation of the studied hemopoietic cells.

Effect of irradiation on colony-forming ability of stem cells from patients with ovarian cancer.

In patients with ovarian cancer, the colony-forming capacity and radiosensitivity of clonogenic tumor cells from the primary node and metastases (ascites) differed considerably.

Cellular alterations upon IR-laser (890 nm) exposures, in vivo.

Exposure of cultured cells and small animals to ionizing radiation as well as irradiation of cultured cells with He-Ne laser can cause changes in the functional condition of plasma membranes. The ionizing radiation-induced cell membrane alterations have been determined after either partial or local exposures. The aim of the present study was to reveal whether the local laser treatments cause a general, distant, so called abscopal" effect measured at cellular level, when the laser treatment is intended as a stimulatory procedure. The biological effect of infrared laser (mean power of 5 Watts, 150 Hz frequency, 890 nm wavelength) was demonstrated through 3H-concanavalin A binding by blood cells of daily irradiated (altogether 10 exposures) oncological and non-oncological patients as well as by changes in the proliferation of bone marrow cells of whole body gamma-irradiated (4 Gy) rats, partially laser-treated. The lectin binding of lymphocytes of oncological, as well as ischaemic heart disease patients was increased immediately after the first laser treatment. However, it was decreased after completion of the full course. In cases of inflammatory diseases the test parameters were either unchanged or decreased as compared to their self-control values. The platelets and erythrocytes did not react in any group. Gamma irradiation caused a deep inhibition of proliferation of rat bone marrow cells. The number of fibroblast colony-forming units (CFU-F) could be increased again if the animals were partially exposed to laser. Laser irradiation of one of the femurs led to some recovery of CFU-F values in the exposed as well as unexposed femur. Thus, local infrared laser treatment induces abscopal effects on the cell membrane and cell proliferation characteristics.

[New developments in understanding the compensatory hematological reactions and their outcomes in local fractionated therapeutic gamma irradiation]. / Novoe v ponimanii kompensatornykh gematologicheskikh reaktsii i ikh iskhodov pri lokal'nom fraktsionirovannom terapevticheskom gamma-obluchenii.

We have carried out a study of the bone marrow status in both irradiated and non-irradiated zones of 56 patients with stage I-II Hodgkin's disease in complete 9-12 (33 patients, group 1) and 18-23 (23 patients, group 2) year remission after therapeutic irradiation of the supradiaphragmatic lymphatic collectors at a dose of 40 Gy with irradiation of the spleen (33 patients) or splenectomy (23 patients). The total count of myelokaryocytes, myelogram, a relative and absolute content of lymphoid cells, immature granulocytes and elements of erythroid series were calculated in the aspirates from the exposed to radiotherapy sternum and non-irradiated upper portion of the ileum. The number of granulocyte-macrophage (CFU-GM) and stromal (CFU-F) precursor cells were defined using in vitro culture technique. There was a complete annihilation of the bone marrow in the irradiated zones, when the dose exceeded 35 Gy in 3-4 weeks. The concentration of myelokaryocytes, immature granulocytes, erythronormoblasts, CFU-GM, CFU-F in non-exposed bone marrow were significantly lower in all patients of group 2 than in normal subjects and in group 1 patients. Absolute lymphoid count in patients with 18-23 year remission was found to be normal but was considerably reduced in comparison to patients of group 1. These changes may be the result of the previous hyperactivity of the non-irradiated bone marrow which could be a cause of stem cell compartment depletion. The differential calculation of compact and diffuse subpopulations of CFU-F revealed a significant reduction of compact colony-forming CFU-F in both irradiated and unexposed bone marrow. Almost all the stromal precursor cells from irradiated zone formed diffuse colonies in cultures. These results confirm experimental data concerning greater radiosensitivity and proliferative potential of CFU-F, forming compact colonies versus diffuse colony-forming CFU-F. Aplasia of the irradiated bone marrow and hypoplasia of the non-irradiated bone marrow 18-23 years after radiotherapy completion coexisted with normal circulating CFU-GM and granulocyte blood count suggesting a compensatory mechanism involving a mitotic amplification between the progenitor cell and the final differentiated cell.

Differential sensitivity of two predominant stromal progenitor cell subpopulations in bone marrow to single and fractionated radiation doses.

The sensitivity of fibroblastoid precursor cells in rat bone marrow to single and fractionated doses of gamma rays delivered in vivo was measured. In vitro colonies were classified as being compact or diffuse, and the progenitor cells for both types were slowly cycling in vivo (survival levels after exposure to hydroxyurea were 90 +/- 6% and 93 +/- 11%, respectively). The progenitor cells forming diffuse colonies were more resistant (D0 = 1.39 Gy) than those forming compact colonies (D0 = 0.76 Gy). The fractionation sensitivities were characterized by an alpha/beta ratio of 12.7 +/- 5.5 Gy for diffuse colonies and 4.5 +/- 3.0 Gy for compact colonies, respectively. The progenitor cells forming diffuse colonies may contribute more to long-term regeneration after high doses in vivo.

[The effect of the gas hypoxic mixture GHM-8 on the capacity of the stromal clonogenic cells (CFU-F) in rat bone marrow for postradiation recovery]. / Vliianie gazovoi gipoksicheskoi smesi GGS-8 na sposobnost' stromal'nykh klonogennykh kletok (KOE-F) kostnogo mozga krys k postradiatsionnomu vosstanovleniiu.

The effect of gas hypoxic mixture, containing 8% of O2 (GHM-8), on the ability of cell precursors of haemopoietic stroma (which form colonies (clones) of fibroblasts (CFU-F) in a culture, and are present in the bone marrow of adult rats) to repair potentially lethal and sublethal radiation damages has been investigated. The recovery of CFU-F from potentially lethal damages, that was studied after their delayed survival in a culture following irradiation of animals, proceeds at nearly the same rate in cells irradiated both in the air and in hypoxic conditions (GHM-8). Fractionated irradiation reduces the radioprotective effect of GHM-8 for CFU-F, particularly for the radioresistant subpopulation; the ability of CFU-F to recover from sublethal radiation damages decreases.

[The capacity of rat clonogenic precursor cells of the hemopoietic stroma (CFU-F) to recover from radiation damages]. / Sposobnost' krysinykh klonogennykh kletok-predshestvennikov gemopoéticheskoi stromy (KOE-F) k vosstanovleniiu ot radiatsionnykh povrezhdenii.

The ability of cell precursors of the haemopoietic stroma (CFU-F), that are present in the bone marrow of adult rats, to recover from potentially lethal and sublethal radiation damages has been investigated. The highest reparability, with respect to potentially lethal damages, is displayed by the most radioresistant CFU-F population, that forms loose colonies (clones) in a culture; the slope of the dose-response curve, not the extrapolation number, changes, and heterogeneity of the CFU-F population is observed. The results obtained confirm the presence of heterogeneity in the population of CFU-F, that was revealed in studying their radiosensitivity by the formation of dense and loose fibroblast colonies in a culture.

[Radiosensitivity of cells-precursors of hemopoietic stroma (CFU-F) in the rat bone marrow under the effect of 60Co gamma irradiation in various conditions]. / Radiochuvstvitel'nost' kletok-predshestvennikov gemopoéticheskoi stromy (KOE-F), soderzhashchikhsia v kostnom mozge krys, pri deistvii gama-izlucheniia 60Co v razlichnykh usloviiakh.

Conditions have been developed for cloning cells-precursors of rat bone marrow haemopoietic stroma, that form in culture dense and sparse fibroblast colonies (CFU-F) at a plating efficiency of 10(-4). Radiosensitivity of rat bone marrow CFU-F, with 60Co-gamma-irradiation in vitro, is characterized by the values of Do and n of 1.87 Gy and 1.4 respectively for all clones; 0.65 Gy and 6.7 for dense clones, and 4.27 Gy and 1.0 for sparse clones. This confirms the observed heterogeneity of CFU-F population consisting of highly radiosensitive and radioresistant subpopulations. The parameters of rat bone marrow CFU-F are nearly the same with irradiation both in vivo and in vitro; with in situ irradiation, the oxygen effect comes into play in a radiosensitive subpopulation of CFU-F; the OER values are 1.6, 2.6 and 0.9 for all, dense and sparse clones respectively.

[The radioprotective effect of hypoxia on clonogenic cells of rat bone marrow stroma (KOE-F)]. / Radiozashchitnoe deistvie gipoksii na klonogennye kletki stromy kostnogo mozga (KOE-F) krys.

A comparative study was made on the survival rate of cell-precursors of haemopoietic stroma, that form, in a rat bone marrow culture, colonies (clones) of fibroblasts (CFU-F) after gamma-irradiation of animals in the air or in a gas hypoxic mixture, containing 8% of O2 (GHM-8). Irradiation in GHM-8 was shown to increase the survival rate of CFU-F by 1.7 times (as compared to exposure in the air) as estimated by the total number of colonies that are formed in a culture; the radioprotective effect of GHM-8 was more pronounced for CFU-F which form dense colonies: DMF for dense and loose clones was 2.4 and 1.6 respectively.

Age dependence of the number of the fibroblasts (CFUf) and haemopoietic stem cells (CFUs) in bone marrow and spleen of rats.

During maturation of rats the progenitor cells of fibroblasts (CFUf) increase in number in bone marrow, whereas they decrease in spleen. The numbers of the haemopoietic stem cells (CFUs) also exhibit similar changes.

[Patterns in the combined action of irradiation and glucose loading on tumor cells]. / Zakonomernosti kombinirovannogo deistviia oblucheniia i gliukoznoi nagruzki na opukholevye kletki.

It was shown that incubation of Ehrlich ascites tumor cells with glucose or in buffer solutions of low pH decreases their viability. The cell survival rate depends on pH values irrespective of the protoxidation method and oxygenation conditions used. At the same time, radiosensitivity of Ehrlich ascites tumor cells is practically invariable with pH being decreased from 7.0 to 5.0. The effects of glucose and radiation are additive in conditions simulating the effect of hyperglycaemia in vivo.

[Comparative radiosensitivity of clonogenic tumor cells of the transplantable Ehrlich carcinoma in the ascitic and solid forms of growth]. / Sravnitel'naia radiochuvstvitel'nost' klonogennykh opukholevykh kletok perevivaemogo raka Erlikha pri astsitnoi i solidnoi forme rosta.

Survival of clonogenic cells of solid Ehrlich ascites tumor (EAT) exposed to 60Co-gamma-radiation in vitro under the oxygenation conditions was investigated and the clonogenic capacity and radiosensitivity of these cells and cells of the previously studied EAT ascitic form and Lewis solid tumor comparatively studied to elucidate how the efficiency of colony formation (ECF) would affect their radiosensitivity. ECF for solid EAT cells was 2.6 +/- 0.3%, which was lower, by about an order of magnitude, than that for ascitic form of this tumor and was nearly the same as that for Lewis tumor cells. A median cell lethal dose (D0) was practically the same for all tumors under study. It is suggested that the differences in ECF do not substantially influence the radiosensitivity of clonogenic cells of the studied tumors.

[Thermosensitivity of clonogenic cells and the induction of thermal tolerance]. / Termochuvstvitel'nost' klonogennykh kletok i induktsiia v nikh termotolerantnosti.

Clonogenic cells of some normal tissues (CFU-GM of the bone marrow of mice, CFU-F of the bone marrow of rats, CFU-GM and CFU-F of the bone marrow of dormice) as well as tumor clonogenic cells of the ascitic variant of ascitic Ehrlich carcinoma were characterized by similar thermosensitivity during their heating under in vitro conditions whereas thermosensitivity in clonogenic cells of solid tumors (LLC, solid Ehrlich ascitic carcinoma) turned out twice as high. A possibility of inducing thermotolerance in these cells by various modes was shown.