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31.
J. P. Ranch L. Oglesby A. C. Zielinski R. B. Horsch 《In vitro cellular & developmental biology. Plant》1985,21(11):653-658
Summary Routine regeneration of fertile plants from a tissue culture of soybean has been achieved. Serially propagated embryogenic
cultures were initiated from immature embryos of many genotypes. Organized tissues developed only on the cotyledons of embryos.
Genotypic variation in the frequency of initiation of embryogenic tissue was noted. However, embryogenic tissue cultures were
generated from all genotypes tested. Embryogenic tissue was serially increased and underwent morphogenesis. Whole fertile
plants were recovered. Cultures have been maintained for two years without loss of morphogenic competency.
Editor's Statement This procedure for initiating embryogenic tissue cultures from commerical cultivars of soybean and the
subsequent development of fertile plants establishes a framework for studying the processes of embryogenesis and embryogenyin vitro as well as providing a system for tissue culture propagation and in vitro modification of soybean. Robert B. Horsch 相似文献
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Characterization of CGS 8515 as a selective 5-lipoxygenase inhibitor using in vitro and in vivo models 总被引:4,自引:0,他引:4
E C Ku A Raychaudhuri G Ghai E F Kimble W H Lee C Colombo R Dotson T D Oglesby J W Wasley 《Biochimica et biophysica acta》1988,959(3):332-342
CGS 8515 inhibited 5-hydroxyeicosatetraenoic acid (5-HETE) and leukotriene B4 synthesis in guinea pig leukocytes (IC50 = 0.1 microM). The compound did not appreciably affect cyclooxygenase (sheep seminal vesicles), 12-lipoxygenase (human platelets), 15-lipoxygenase (human leukocytes) and thromboxane synthetase (human platelets) at concentrations up to 100 microM. CGS 8515 inhibited A23187-induced formation of leukotriene products in whole blood (IC50 values of 0.8 and 4 microM, respectively, for human and rat) and in isolated rat lung (IC50 less than 1 microM) in vitro. The selectivity of the compound as a 5-lipoxygenase inhibitor was confirmed in rat whole blood by the 20-70-fold separation of inhibitory effects on the formation of leukotriene from prostaglandin products. Ex vivo and in vivo studies with rats showed that CGS 8515, at an oral dose of 2-50 mg/kg, significantly inhibited A23187-induced production of leukotrienes in whole blood and in the lung. The effect persisted for at least 6 h in the ex vivo whole blood model. CGS 8515, at oral doses as low as 5 mg/kg, significantly suppressed exudate volume and leukocyte migration in the carrageenan-induced pleurisy and sponge models in the rat. Inhibitory effects of the compound on inflammatory responses and leukotriene production in leukocytes and target organs are important parameters suggestive of its therapeutic potential in asthma, psoriasis and inflammatory conditions. 相似文献
34.
We have developed methods for the coculture of hepatocytes and mouse lymphoma cells and have shown that this system can be used for evaluating promutagens from several chemical classes (Brock et al., 1987). In the present study we investigated the use of hepatocytes isolated from rats pretreated with a cytochrome P-450 inducer (PB) or a P-448 inducer (BNF). CP-induced mutagenicity was higher in the presence of PB-induced hepatocytes than in control hepatocytes. Control and BNF-induced hepatocytes were evaluated with B(a)P, B(l)A, and BA. A dose-related positive response was observed with B(a)P and B(l)A both in the presence of control or induced hepatocytes; however, somewhat higher mutant frequencies were obtained in the presence of BNF-induced hepatocytes. BA induced a very weak positive response (approx. 2 X b.g.) in the presence of control hepatocytes and was weakly positive in the presence of BNF-induced hepatocytes. Benzene was tested using control and both PB- and BNF-induced hepatocytes. Neither of these approaches were successful in activating benzene to a mutagenic metabolite. These studies indicate that for some chemicals the mutagenic response of mouse lymphoma cells can be increased by inducing hepatocytes prior to isolation and cocultivation, and expands the use of hepatocytes for research evaluating chemicals requiring metabolic activation. 相似文献
35.
Shigella flexneri requires iron for survival, and the genes for iron uptake and homeostasis are regulated by the Fur protein. Microarrays were used to identify genes regulated by Fur and to study the physiological effects of iron availability in S. flexneri. These assays showed that the expression of genes involved in iron acquisition and acid response was induced by low-iron availability and by inactivation of fur. A fur null mutant was acid sensitive in media at pH 2.5, and acid sensitivity was also observed in the wild-type strain grown under iron-limiting conditions. Acid resistance of the fur mutant in minimal medium was restored by addition of glutamate during acid challenge, indicating that the glutamate-dependent acid resistance system was not defective. Inactivation of ryhB, a small regulatory RNA whose expression is repressed by Fur, restored acid resistance in the fur mutant, while overexpressing ryhB increased acid sensitivity in the wild-type strain. RyhB-regulated genes were identified by microarray analysis. The expression of one of the RyhB-repressed genes, ydeP, which encodes a putative oxidoreductase, suppressed acid sensitivity in the fur mutant. Furthermore, an S. flexneri ydeP mutant was defective for both glutamate-independent and glutamate-dependent acid resistance. The repression of ydeP by RyhB may be indirect, as real time polymerase chain reaction (PCR) experiments indicated that RyhB negatively regulates evgA, which encodes an activator of ydeP. These results demonstrate that the acid sensitivity defect of the S. flexneri fur mutant is due to repression of ydeP by RyhB, most likely via repression of evgA. 相似文献
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Christopher W. Benjamin Nancy K. Hopkins Thomas D. Oglesby Robert R. Gorman 《Biochemical and biophysical research communications》1983,117(3):780-787
Leukotriene C4 (LTC4) and, to a lesser extent, leukotriene D4 (LTD4) concentration dependently stimulate prostacyclin (PGI2) biosynthesis in cultured human umbilical vein endothelial cells. PGI2 biosynthesis was quantitated by radioimmunoassay and its structure confirmed by gas chromatography/mass spectrometry. Preincubation of endothelial cells with LTC4 resulted in desensitization to subsequent LTC4 stimulation. However, PGI2 biosynthesis in response to thrombin, PGH2 and arachidonic acid was not inhibited by preincubation with LTC4. The C-6-sulfidopeptide leukotriene receptor level antagonist FPL-55712 attenuates LTC4, but not thrombin-stimulated PGI2 biosynthesis. These data suggest that human umbilical vein endothelial cells have a C-6-sulfidopeptide leukotriene receptor, and that stimulation of this receptor results in PGI2 biosynthesis. 相似文献
38.
Identification and characterization of membrane cofactor protein of human spermatozoa. 总被引:6,自引:0,他引:6
F Cervoni T J Oglesby E M Adams C Milesifluet M Nickells P Fenichel J P Atkinson B L Hsi 《Journal of immunology (Baltimore, Md. : 1950)》1992,148(5):1431-1437
Membrane cofactor protein (MCP) regulates C activation by serving as a cofactor for the cleavage of C3b and C4b by the serine protease factor I. An MCP-like molecule on the inner acrosomal membrane of human spermatozoa has been characterized. Three mAb and a rabbit polyclonal antibody against MCP recognized the sperm protein. On SDS-PAGE, it migrated as a single band with a molecular mass of 38,000 and 44,000 Da under nonreducing or reducing conditions, respectively. The molecular mass was 10,000 to 20,000 Da less than the two forms of MCP expressed on others cells. The electrophoretic pattern, by one- and two-dimensional gel analysis, and the isoelectric point profile (4.5 to 5.0) of the sperm protein were similar among multiple individuals. In contrast to MCP of other cells, digestion with endoglycosidases did not alter either the m.w. or the pI of the protein, suggesting that it is a poorly or nonglycosylated form of MCP. The solubilized sperm protein bound C3 with broken thioester bond to Sepharose and possessed cofactor activity for factor I-mediated cleavage of C3 with the broken bond. A mAb that blocks the regulatory function of MCP inhibited the cofactor activity of the sperm lysate. Thus, the sperm protein is an antigenic and functional homologue of MCP but has the distinct structural features of a lower m.w. and an apparent lack of glycosylation. MCP may play an essential role in the survival of the acrosome-reacted spermatozoa by modulating C activation in the female genital tract. 相似文献