Separation of resting and proliferating granulocytic precursors

We have obtained cells in various stages of granulocytic development by a combination of isopycnic separation and electronic cell sorting. Not only were immature cells (blast cells, promyelocytes and myelocytes) separated from mature cells (bands and polys), but the immature cells were separated into proliferating (S + G2 + M) and resting (Go/G1) compartments of the cell cycle. This permits the study of the morphological and biochemical changes associated with development apart from those changes associated with proliferation.     

The influence of histamine on precursors of granulocytic leukocytes in murine bone marrow

The effect of histamine on granulocytic progenitor cells in murine bone marrow was studied in vitro. When bone marrow cells were cultured for three days with the drug, 10(-8) M to 10(-5) M of histamine stimulated differentiation and proliferation of myeloid precursor cells. Subsequently, the number of descendant cells, such as metamyelocytes and neutrophils, increased dose-dependently. Co-existence of equimolar H2 blockers such as cimetidine and ranitidine completely suppressed this effect of histamine, though this was not the case with an H1 blocker/histamine combination. Significant increase in 3H-thymidine incorporation was observed almost exclusively in myeloblasts, promyelocytes and myelocytes after exposure to histamine at concentrations higher than 10(-8) M. Also, selective incorporation of 3H-histamine into bone marrow cells was observed in myeloblasts and promyelocytes, but histamine incorporation was not influenced by the presence of either of histamine agonists or antagonists. While histamine, via H2 receptors, selectively increased the number of granulocytic colony forming units in culture (CFU-C), it had no such effect on macrophage colonies. Considering these findings, it was concluded that histamine promotes proliferation and differentiation of granulocytic myeloid cells via 1) H2 receptors in the CFU-C stage and 2) histamine receptors which are neither H1 nor H2 in the stages of myeloblast and promyelocyte differentiation.     

孓遗植物桫椤RAPD分析中主要反应参数的影响

为得到桫椤RAPD扩增的最佳条件,对影响桫椤RAPD扩增的模板纯化方法、模板含量、酶浓度、Mg²⁺浓度、dNTP浓度和引物浓度及扩增程序等多种因素进行了探讨.结果表明,用玻璃奶和蛋白酶K纯化后获得的DNA作为模板进行RAPD反应,其扩增带比用未纯化的和仅用无水乙醇再沉淀的DNA为模板更亮,更清晰,重复性更好;模板浓度、酶浓度、Mg²⁺及dNIT浓度、引物浓度及退火温度这些因素都会对RAPD产生影响.经过本实验的探索,发现桫椤RAPD扩增的最佳体系是(2.5×10^-2mL反应体积):模板浓度为70ng,Mg²⁺浓度为2.5mmol·L^-1,dNTP为0.2mmol·L^-1,引物3×10^-4mL;最佳扩增程序为:94℃200s,一个循环;94℃60s,36℃60s,72℃120s,40个循环:72℃600s,一个循环:4℃保存.     

Thein vitro andin vivo effects of carrageenin on humoral and cellular factors of natural resistance

The inhibitory effect of carrageenin, a sulfated algal polygalactose, on humoral factors of natural resistance is discussed. In dependence on dose and time, the influence of non-specific and specific bacteriolysis, on thein vitro andin vivo opsonic activity and on the course of infection was studied.     

To the intranucleolar translocation of AgNORs in leukemic early granulocytic and plasmacytic precursors

Early leukemic granulocytic and plasmacytic precursors were studied in vitro and in vivo to provide an information on the intranucleolar distribution of AgNORs (silver stained nucleolus organizer regions). In most of these cells AgNORs appeared as clusters of silver stained particles distributed in the whole nucleolar body. On the other hand, in some leukemic early granulocytic precursors, i.e., in myeloblasts and promyelocytes enlarged AgNORs were translocated in the nucleolar peripheral part. In addition, the number of translocated AgNORs at the nucleolar periphery was significantly smaller. Such translocation of a reduced number of AgNORs was easily produced by experimental aging, i.e., starving of cultured leukemic early granulocytic precursors (HL-60 and K562 cells) in vitro and seems to be reversible. Similar translocation of a reduced number of AgNORs was also produced by aging of leukemic plasmacytic precursors. Thus, the translocation of the reduced number of AgNORs to the nucleolar periphery in some blastic leukemic hematopoietic cells might be an useful marker of their aging at the single cell level. However, more studies in this direction are required in the future.     

Investigating serum factors promoting erythrocytic growth of Plasmodium falciparum

The elucidation of factors inducing the growth of Plasmodium falciparum can provide critical information about the developmental mechanisms of this parasite and open the way to search for novel targets for malaria chemotherapy. The ability of components of a growth-promoting factor derived from bovine serum and various related substances to sustain growth of P. falciparum was characterized. A simple total lipid fraction (GFS-C) containing non-esterified fatty acids (NEFAs) as essential factors was noted to promote the parasite's growth. Various proteins from a variety of animals were tested, indicating the importance not only of GFS-C, but also of specific proteins, such as bovine and human albumin, in the parasite growth. Several combinations of the NEFAs tested sustained low parasite growth. Among various phospholipids and lysophospholipids tested, lysophosphatidylcholine containing C-18 unsaturated fatty acids was found to sustain the complete development of the parasite in the presence of bovine albumin. Several other lysophospholipids can partially support growth of P. falciparum.     

Radiosensitivity of clonogenic granulocytic macrophage cell precursors in the bone marrow and spleen of tumor-bearing mice

Clonogenic granulocytic macrophagal cells-precursors (CFU-DC) of bone marrow and spleen of intact C57Bl/6 mice and those inoculated subcutaneously with LLC tumor cells do not substantially differ in their radiosensitivity; the concentration of CFU-DC in the spleen markedly varies as tumor grows. The values of Do and extrapolation number n for CFU-DC of the bone marrow are 0.9-1.4 and 1.5-3.0 Gy, and of the spleen, 0.8-1.6 and 1.0-2.6 Gy, respectively.     

Plasmodium falciparum and Plasmodium berghei: effects of ornithine decarboxylase inhibitors on erythrocytic schizogony

Five ornithine decarboxylase inhibitors: alpha-difluoromethylornithine (DFMO) (eflornithine); alpha-monofluoromethyl-3,4-dehydroornithine; alpha-monofluoromethyl-3,4-dehydroornithine methyl ester; alpha-monofluoromethyl-3,4-dehydroornithine ethyl ester; and (2R,5R)-delta-methyl-alpha-acetylenic putrescine were shown to inhibit erythrocytic schizogony of Plasmodium falciparum in vitro and reduced spermidine levels in infected erthrocytes. Only DFMO was effective at limiting erythrocytic schizogony of P. berghei in vivo. Administration of DFMO as a 2% solution in the drinking water for 4 days reduced parasitemia in mice by 50% in a 4-day suppression test but did not increase survival time of infected mice. This is the first demonstration of an effect of DFMO on plasmodial erythrocytic schizogony in vivo and suggests that interference with polyamine biosynthesis may, in fact, be a viable chemotherapeutic target in erythrocytic malaria.     

A short note on the nucleolar size and density in apoptotic leukemic granulocytic precursors (HL-60 cells)

The present study was undertaken to provide more information on the nucleolar size and density in mononuclear blastic granulocytic precursors represented by HL-60 cells the proliferation of which was blocked by photodynamic treatment (PDT) which induced apoptotic process without preceding terminal maturation. Both the nucleolar size and density did not change in apoptotic cells in comparison with controls. Thus, large and dense nucleoli in apoptotic cells are not necessarily related to the nucleolar biosynthetic or cell proliferation activity.     

Cellular and humoral immunity factors as regulators of regenerative morphogenesis

The published and author's data concerning changes in the immune system during regeneration of various organs are summarized. During the first few hours after partial removal of organs possessing high regeneration capacity, lymphocytes stimulate proliferation of nonlymphoid cells of an organ identical to the operated one; the production of antibodies against thymus-dependent antigen also increases. At the following stages of regeneration, the lymphocytes suppress the cell proliferation and decrease the antigen production. The level of these changes correlates with the level of post operational deficiency of the organ being maximal after total removal of the organ. The functional properties of splenocytes at different stages of regeneration suggest that the high T-helper and T-suppressor activities correlate with stimulation and suppression of non-lymphoid cells proliferation respectively. Culture medium supernatant after cultivation of these lymphocytes also changes the proliferation of hepatocytes. The author considers the impairment of natural immune tolerance caused by deficiency of organ auto-antigens that normally suppress lymphocyte proliferation to be the cause of the changes in lymphocyte activity.     

Comparative immunological analysis of echinoderm cellular and humoral defense factors

Coelomocyte are found in the fluid filling coelomic cavity of echinoderms and depending on species can be a mixture of several morphologically different types. There are among them: granular and agranular amoebocytes, morula cells, vibratile and lymphocyte-like cells. All these cells take part in cellular response to immune challenges through phagocytosis, clotting, encapsulation of foreign particles, cytotoxicity, and the production of antimicrobial agents, such as reactive oxygen and nitric oxide. The data are given on a variety of humoral factors found in the coelomic fluid, including different types of lectines, agglutinins, hemolysins, acute phase proteins and antimicrobial factors. The discussion on cooperation between cellular and humoral arms of defense reactions during inflammation reveals the crucial role of coelomocytes in immune response. It is suggested that the sea urchin complement system (that is homologous to the alternative pathway in vertebrates) is appeared initially in echinoderms as a protein cascade that points to opsonization of foreign cells and particles, augmenting their phagocytosis and subsequent destruction by coelomocytes. So the identification of a simple complement system as a part of the echinoderm immune response shows that these animals as well as all invertebrate deuterostomes share innate immune system homologies with vertebrates. Studying the simpler immune response demonstrated by echinoderms is important for understanding the ancestral deuterostome defense system and reconstructing the evolution of immune system in higher vertebrates.