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111.
Norman R. Drinkwater Rebecca C. Corner J. Justin McCormick Veronica M. Maher 《Mutation research》1982,106(2):277-289
The sensitivity of diploid human fibroblasts to the cytotoxic effects of diphtheria toxin (DT) depended on the cell growth status. Exponentially growing cells treated with 10?3-1 lethal flocculating units (LF) of DT/ml for 4 days survived with a frequency of 4 × 10?4. However, the DT-resistant phenotype of colonies isolated under these conditions was not stable. When the growth of the cells had been arrested by confluence or deprivation of serum growth factors prior to treatment with DT (4 days, 10?3-0.6 LF/ml), the survival decreased to 2 × 10?6 and the resistance of isolated colonies was stable. An in situ assay for induced DT-resistant mutants was developed in order to avoid problems associated with the possible reduced viability of the mutants relative to that of wild-type cells. A reproducible and linear dose response was obtained for the induction of DT-resistant mutants by ethylnitrosourea. The mutants were induced with high frequency by this compound (e.g., 10?3 mutants/viable cell at a 37% survival dose); complete expression of the mutant phenotype occurred after 6 generations of growth under nonselective conditions. Isolated mutant colonies showed stable resistance to DT and were cross-resistant to Pseudomonas aeruginosa exotoxin A. 相似文献
112.
The ester groups of glycerophospholipids in tissue extracts can be cleaved in less than 10 min at room temperature if the lipids are extracted with hexane-isopropanol and the filtrate is treated with methanolic NaOH. The resulting mixture can be treated with aqueous Na-sulfate containing sulfuric acid and partitioned to remove the inorganic reagents and hydrophilic ester degradation products. When the procedure is applied to brain lipid extracts, the addition of alkali produces a second, lower phase that contains much of the hydroxycerebroside, virtually all of the sulfatide in the extract, and small amounts of other lipids. The sulfatide can be isolated from the lower phase by neutralizing it with HCl in aqueous methanol, and partitioning with chloroform to remove nonlipid components. The lower phase is evaporated to dryness and treated with periodic acid to convert the cerebroside to a less polar product. The lipids recovered from the reaction mixture are then fractionated with a Florisil column, which yields highly purified sulfatide. Starting with 300 g of pig brain one can obtain about 1.1 g of sulfatide in 4 working days, using conventional, compact equipment. Since the precipitation step is almost complete, and the procedure can be scaled down to very low levels, the method has promise for quantitation methods and isotopic studies of sulfatide metabolism. 相似文献
113.
Accessibility of sialo components in a murine tumor cell to extracellular glycohydrolase (sialidase)
Lipid-bound sialic acid in the murine melanoma cell is not totally inaccessible to an exogenous macromolecular probe, as formerly believed. Roughly 30% of the sialic acid bound to lipid, and an equal proportion of the sialic acid bound to protein is cleaved by the action of Clostridium perfringens glycohydrolase (neuraminidase, sialidase) when the purified enzyme is added to the suspension medium of intact murine melanoma cells freshly derived from the tumor. Cleavage of lipid-bound sialic acid is indifferent to the presence of Ca2+ in the medium. However, maximum release from protein requires a physiological concentration of this divalent cation. Variation in ionic strength has no effect on release of sialic acid. These findings show that a restricted portion of the bound sialic acid may be released from the intact murine melanama cell by the extracellularly supplied enzyme acting topographically. 相似文献
114.
Anthony W. Norman 《Steroids》1980,36(1):27-39
A heretofore unknown metabolite of vitamin D3 was isolated from the 1α,24,25-trihydroxyvitamln D3 fraction of lipid extracts obtained from plasma of rats which were given intravenous or oral doses of 100 pmol/100 g of either 1α-hydroxyvitamin D3 or 1α, 25-dihydroxy-vitamin D3. Doses of 25–250 pmoles of the new metabolite when given to a vitamin D deficient rat were completely inactive in terms of stimulating the classic vitamin D response of bone calcium mobilization. The nature of the metabolism of 1α-hydroxyvitamin D3 or 1α, 25-dihydroxy-vitamin D3 to the metabolite is not clear at the present time, but it is probable that neither of these steroids undergo side-chain cleavage to yield the new metabolite. 相似文献
115.
Early ventricular fibrillation occurs in approximately 5% of patients admitted for acute myocardial infarction. Although late ventricular fibrillation (> 48 hours postinfarction) may occur in stable patients, it occurs more commonly when severe left ventricular power failure is present. We have encountered late ventricular fibrillation in three of 42 (7%) patients treated with intraaortic balloon pumping (IABP) for profound cardiogenic shock secondary to myocardial infarction. These patients progressed to our hemodynamic Class A prior to weaning, and were thought to be stable prior to IABP removal. They were the only ones who expired after achieving Class A status. The episodes of late ventricular fibrillation occurred after the patients had been successfully weaned from IABP and were free of arrhythmias. This experience suggests that prolonged antiarrhythmic therapy may be indicated for postinfarction patients who have had ventricular dysrhythmias during IABP support. 相似文献
116.
Turner SA Bossart MI Klima T Leachman RD Cooley DA Norman JC 《Cardiovascular diseases》1980,7(3):272-277
Persistent atrial paralysis in a patient with complete heart block and mild mitral insufficiency is presented. Left atrial specimens obtained during implantation of a permanent cardiac pulse generator showed evidence of hypertrophy and fibrosis; subcellular degenerative changes ranged from near normal to irreversible, thus suggesting that atrial paralysis may be due to the replacement of normal atrial muscle with nonfunctional fibrous tissue. 相似文献
117.
Biology of Azospirillum-Sugarcane Association: Enhancement of Nitrogenase Activity 总被引:4,自引:2,他引:2
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R. Howard Berg Max E. Tyler Norman J. Novick Vimla Vasil Indra K. Vasil 《Applied microbiology》1980,39(3):642-649
Azospirillum brasilense was reisolated from associations with callus tissue cultures of sugarcane and compared with stock cultures of the inoculated bacterium and related strains. Although the reisolate had a growth rate similar to stock cultures, it exhibited a severalfold increase in maximum specific activity of nitrogenase. The reisolate and the parent culture had similar ultrastructure. The general ultrastructure of Azospirillum is described. The bacterium was capsulated when grown on nitrogen-free nutrient agar plates and on callus, but was not capsulated when growing in a subsurface zone in N-free semisolid nutrient agar, except rarely in aging cultures. Capsulation may be a protective mechanism against unfavorable pO2 under dinitrogen-fixing conditions. Pleomorphism occurred in capsulated forms, and the ultrastructure of these forms is described. 相似文献
118.
Renografin density gradient centrifugation and carboxymethyl (CM)-and diethylaminoethyl (DEAE)-cellulose ion-exchange chromatography were compared for purification of cells of phony peach, rickettsia-like bacterium (RLB) from plant extracts. Centrifugation on 30–40% linear gradients resulted in four bands. Cells of RLB sedimented into the third band located 4.9–5.4 cm below the meniscus. Phase-contrast microscopy, plant peroxidase activity, and viable bacterial assays showed the resulting cells ofRLB to be relatively free of host contaminants. A typical purification from 120 ml of extract containing 7.2×109
RLB cells resulted in the recovery of 7.0×107
RLB cells. Cells ofRLB were not successfully purified byCM andDEAE chromatography. Unitil a medium is available for cultivation of plantRLB, Renografin density gradient centrifugation should be a useful tool for obtaining cells ofRLB for biochemical and serological investigations. 相似文献
119.
Peter H. Burrill Isa Bernardini Hynda K. Kleinman Norman Kretchmer 《Journal of cellular biochemistry》1981,16(4):385-392
Rabbit intestinal epithelial cells, obtained after a limited hyaluronidase digestion, were incubated in medium with or without calf serum, on bacteriological plastic dishes. The dishes, either plain or coated with an air-dried type I collagen film, were pretreated with medium alone or with medium containing purified laminin or purified fibronectin. Cells did not attach in significant numbers to untreated bacteriological plastic, even in the presence of serum. Cells did attach to collagen-coated dishes, and were judged viable on the basis of their incorporation of radiolabeled leucine into cell protein. Cell adhesion to the collagen substrate increased in proportion to the concentration of serum in the medium, with maximal attachment at 5% serum or greater. Pretreatment of plain or collagen-coated dishes with increasing amounts of fibronectin enhanced cell adhesion in a concentration-dependent manner. Either serum, or fibronectin-free serum in the medium enhanced cell attachment to substrates pretreated with cither fibronectin or laminin. Thus, intestinal epithelial cells appear to possess surface receptors for both laminin and fibronectin. The evidence further suggests that calf serum may contain factors, other than fibronectin, capable of enhancing intestinal epithelial cell attachment to collagen substrates. 相似文献
120.
The rate of efflux of L-glutamate from renal brush-border membrane vesicles was enhanced by Na+ and by extravesicular L-glutamate, but not by D-glutamate nor analogs of L-glutamate that do not share the Na+-L-glutamate co-transport system. These results suggest that efflux was mediated by the Na+-L-glutamate carrier. The efflux of L-glutamate was increased by extravesicular K+ or Rb+ but not by Li+, choline+, or Tris+. These findings, together with previous results showing that intravesicular K+ or Rb+ increased L-glutamate uptake and that a K+ gradient energized the concentrative uptake of the acidic amino acid in the absence of other gradients, provide evidence consistent with the hypothesis that the co-transport of Na+-L-glutamate is coupled to the transmembrane flux of K+. 相似文献