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41.
Ganglioside-Induced Neuritogenesis: Verification That Gangliosides Are the Active Agents, and Comparison of Molecular Species 总被引:11,自引:10,他引:1
Mary C. Byrne Robert W. Ledeen Fred J. Roisen † Glee Yorke † Joseph R. Sclafani 《Journal of neurochemistry》1983,41(5):1214-1222
Abstract: Gangliosides were previously reported to induce neuritogenesis in primary neuronal cultures and in some neurally derived cell lines. Because isolated gangliosides usually contain variable quantities of peptides, we investigated the possibility the neurite-stimulating activity could be caused by these contaminants. Ganglioside preparations from bovine brain and other sources were subjected to a three-step purification procedure that eliminated at least 95% of the contaminating peptides. These purfied preparations retained their capacity to induce extensive neurite growth in neuro-2A murine neuroblastoma. Proteolytic digestion and a number of additional procedures were used to reduce residual contamination further without loss of activity. Several crude ganglioside samples had negative effects on neurite development until freed of theri inhibitory factors, which were derived from the tissue and/or introduced during laboratory operations. This was particularly evident for bovine white matter gangliosides whose activity increased in proportion to peptide removal. When carefully purified, virtually all of 11 different gangliosides tested were highly active, with the possible exception of GM4, which demonstrated only moderate activity in a limited number of tests. All of the neutral glycolipids tested, as well as sulfatides and free sialic acid, were inactive. 相似文献
42.
Mary Taub Milton H. Saier Lorraine Chuman Sue Hiller 《Journal of cellular physiology》1983,114(2):153-161
Prostaglandin E1(PGE1), one of the components in the hormone-supplemented, serum-free medium for Madin Darby Canine Kidney (MDCK) cells (Medium K-1), is required for both long-term growth and for dome formation. Variant cells have been isolated from MDCK populations, which lack the PGE1, requirement for long-term growth in Medium K-1. These variants will be useful in identifying the molecular events initiated by PGE1 which are necessary for the growth response to be observed. The growth and functional properties of five independently isolated PGE1 independent clones have been examined. Normal MDCK cells grew at an equivalent rate in Medium K-1 and in serum-supplemented medium; the growth rate was lower in Medium K-1 lacking PGE1. In contrast, PGE1 independent clone 1 grew at an equivalent rate in Medium K-1 minus PGE1, and in serum-supplemented medium. When PGE1 was added to K-1 minus PGE1, less growth of PGE1 independent clone 1 was observed. A similar observation was made with one other PGE1 independent clone which was studied. A hormone deletion study indicated that PGE1 independent clone 1 still retained growth responses to the other four supplements in Medium K-1 (insulin, transferrin, T3, and hydrocortisone). The molecular alterations associated with loss of the PGE1 requirement for long-term growth were examined. At confluency, all of the PGE1 independent clones studied had higher intracellular cyclic AMP levels following PGE1 treatment, as compared with normal MDCK cells. The increased cyclic AMP levels in the variant cells could result from a number of different types of defects, including reduced cyclic adenylic acid (cyclic AMP) efflux, an increased affinity of PGE2 for the PGE1 receptor, or a defect in cyclic AMP metabolism. However, in all of the variant clones studied a decreased rate of cyclic AMP degradation by cyclic AMP phosphodiesterase was observed. Thus, the increased cyclic AMP levels in the PGE1 independent variants may result from alterations which affect cyclic AMP metabolism. The effect of PGE1 on dome formation by the variant cells was also examined. The frequency of dome formation by PGE1 independent clone 1 was enhanced in a dosage-dependent manner, like normal MDCK cells. This observation suggests that PGE1 affects MDCK cell growth and dome formation by different mechanisms. 相似文献
43.
The effect of continuous-wave ultrasound on the chromosomes of newborn infants has been investigated. Twenty-four women were studied during labour. The fetal heart was monitored by a Sonicaid FM2 monitor applied to the abdomen, and continuous monitoring undertaken for intervals varying from 1 hour 5 minutes to 9 hours 25 minutes. There was no increase in the number of chromosome aberrations in cultures of blood taken from the insonated babies when compared with controls. 相似文献
44.
Treatment of rabies virus with the nonionic detergent Nonidet P-40 resulted in solubilization of viral lipids and in a preferential release of the envelope glycoprotein. The other viral proteins and the viral ribonucleic acid remained associated in "core" particles sedimenting at a rate similar to that of intact virions. After fractionation of treated virus by velocity centrifugation in a sucrose density gradient, the amount of residual glycoprotein recovered in the "core" particle fraction and the extent of contamination of the glycoprotein fraction by other viral components were dependent on the ratio of detergent to viral protein used. 相似文献
45.
The Distribution of Crossovers along Unreplicated Lambda Bacteriophage Chromosomes 总被引:21,自引:4,他引:17 下载免费PDF全文
Franklin W. Stahl Kenneth D. McMilin Mary M. Stahl Jean M. Crasemann Stephen Lam 《Genetics》1974,77(3):395-408
The distribution of crossovers along unreplicated chromosomes of bacteriophage lambda has been examined by determining the density distributions and genotypes of particles in the progenies of crosses of density-labeled by ordinary parents in the presence of genetic blocks to replication. The Red and Rec systems combined produce crossovers primarily near the ends (especially the right end) of the chromosome. Removal of the generalized lambda recombination functions by red and gam mutations results in loss of these terminal crossovers; coupled with this loss is a disappearance of the differential dependence of recombination frequencies in terminal and central intervals on DNA synthesis. Removal of the bacterial system by a recA mutation results in severe depression of crossing over among unreplicated phage, with the few recombinants produced by the lambda system occurring near the right end. 相似文献
46.
Mary F. Lyon 《Genetics》1974,78(1):305-309
47.
48.
Mary J. Kurtz Peter Polgar Linda Taylor Alexander M. Rutenburg 《The Biochemical journal》1974,142(2):339-344
Inhibition of growth and DNA synthesis was observed in WI 38 cells incubated with 8-methylthioadenosine 3':5'-cyclic monophosphate or prostaglandin E(1). The effect of both compounds on cell growth was reversible. On removal of these compounds from culture media the cells initiated DNA synthesis and divided. In addition, prostaglandin E(1) stimulated cyclic AMP formation in these cells to over 40 times the normal basal value. The increase in cyclic AMP concentration in WI 38 cells after addition of prostaglandin E(1) showed a marked variation. Cells that had recently been treated with trypsin and plated at a lower cell density exhibited a smaller response to addition of prostaglandin E(1) than cells that had divided and reached confluence. 相似文献
49.
Comparison of Undefined Medium and Its Dialyzable Fraction for Growth of Mycoplasma 总被引:11,自引:3,他引:8 下载免费PDF全文
In examining the medium used in cultivation of Mycoplasma for deoxyribonucleic acid isolation, it was found that an aggregate was present which sedimented with the organisms and which was ethyl alcohol-precipitated during deoxyribonucleic acid purification. To eliminate the contaminating material, a method was devised to obtain only the dialyzable constituents of the medium. This report describes the preparation of a dialysate of soy peptone-yeast extract. The medium, obtained by immersion of the encased dehydrated ingredients in sodium chloride solution for 5.5 hr at approximately 80 C, has been employed as the basal medium for cultivation of a number of Mycoplasma species. Comparative growth curves of two saprophytic strains and two parasitic species indicated that multiplication in dialysate, with suitable supplement, followed the pattern typical of the common eubacteria. Thus, by elimination of the sediment which occurred in nondialyzed medium, Mycoplasma could be concentrated without concomitant accumulation of contaminating macromolecules. 相似文献
50.
The deoxyribonucleic acid homologies of Mycoplasma laidlawii type A and type B, M. pulmonis (#47 and #63), and M. hominis were determined by membrane methodology. The homology data revealed a difference in genome size between M. laidlawii type A and type B. This difference also held with stringent conditions of annealing (high temperature). Little or negligible homology was shown to exist between the M. laidlawii strains type A and type B and M. pulmonis strains 47 and 63 and M. hominis. M. hominis showed less than 10% homology to the M. pulmonis and M. laidlawii strains. Neither of the M. laidlawii strains showed more than 2% annealing to the M. pulmonis strains. Reaction rate studies are suggested as a means of demonstrating the phylogenetic relationship between the Mycoplasma and other microorganisms. 相似文献