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31.
Summary Irradiation of Escherichia coli with UV light causes a transient inhibition of DNA replication. This effect is generally thought to be accounted for by blockage of the elongation of DNA replication by UV-induced lesions in the DNA (a cis effect). However, by introducing an unirradiated E. coli origin (oriC)-dependent replicon into UV-irradiated cells, we have been able to show that the environment of a UV-irradiated cell inhibits initiation of replication from oriC on a dimer-free replicon. We therefore conclude that UV-irradiation of E. coli leads to a trans-acting inhibition of initiation of replication. The inhibition is transient and does not appear to be an SOS function. 相似文献
32.
Action potentials initiated by single channels opening in a small neuron (rat olfactory receptor). 总被引:10,自引:0,他引:10
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Rat olfactory receptor neurons were enzymatically dissociated and studied with the cell-attached configuration of the patch-clamp technique. Biphasic current waveforms induced across the membrane patch by intracellular action potentials were observed in approximately 5% of cells studied. In one cell in particular, current injected by the opening of a single channel initiated an action potential in the remainder of the cell each time the channel opened. A conventional type of electrical model of the cell and patch allowed the accurate modeling of cell excitability. The same model was used to explain the shape of the action potential current waveforms induced across the patch. The analysis indicated that the whole cell resistance (Ro) was approximately 40 G omega and the membrane capacitance (Co) was close to the standard value of 1 microF.cm-2. In addition, the threshold potential change necessary to initiate an action potential (Vth) was approximately 13 mV and a minimum current injection of 1 pA was required to depolarize the cell to spike threshold. When the smaller size of mammalian receptors are taken into account, membrane electrical properties were found to be consistent with those of salamander cells investigated by others using whole-cell recording. The analysis also revealed possible errors in the determination of single-channel conductances and reversal potentials by cell-attached recording from small cells. 相似文献
33.
We investigated the inhibitory effects of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) on ATP-dependent H+ accumulation by membrane vesicles prepared from the turtle urinary bladder epithelium. NBD-Cl at 30 microM was found to completely inhibit the vanadate-insensitive component of H+ transport, with half-maximal inhibition occurring at 4.2 to 5.4 microM. In contrast, the vanadate-inhibitable component was unaffected by 30 microM NBD-Cl. At high concentrations (300 microM), both components were fully inhibited. The results confirm the presence of two distinct H+ transport processes in turtle bladder membranes and identify selective inhibitors, NBD-Cl and vanadate, for each process. 相似文献
34.
Water oxidation in photosystem II: from radical chemistry to multielectron chemistry 总被引:8,自引:0,他引:8
G T Babcock B A Barry R J Debus C W Hoganson M Atamian L McIntosh I Sithole C F Yocum 《Biochemistry》1989,28(25):9557-9565
35.
Characteristics of tripeptide transport in human jejunal brush-border membrane vesicles 总被引:3,自引:0,他引:3
These studies are aimed at characterizing the transport of the tripeptide, glycylglycyl-L-proline (GlyGlyPro) across human jejunal brush-border membrane vesicles. GlyGlyPro (0.65 mM) was hydrolyzed by brush-border membrane vesicles with the extent of hydrolysis per mg protein being 23% at 0.5 min, 57% at 1 min and complete hydrolysis at 60 min. Treatment of the membrane vesicles with gel-complexed papain (to remove membrane peptidases) resulted in minimal hydrolysis of GlyGlyPro up to 10 min of incubation. Measurement of GlyGlyPro influx with papain-treated vesicles in the presence of increasing medium osmolarity showed that uptake occurred into an osmotically reactive intravesicular space. Transport of GlyGlyPro with normal and papain-treated membrane vesicles was similar in the presence of an inward Na+ or K+ gradient. No overshoot phenomenon was observed in the presence of an inward proton gradient (extravesicular pH 5.5; intravesicular pH 7.5). An interior negative membrane potential induced by a K+ diffusion potential in the presence of valinomycin stimulated the uptake of the peptide. The effect of increasing concentrations on initial rates of GlyGlyPro uptake revealed the presence of a saturable component as well as a diffusional component. Preloading the membrane vesicles with 20 mM glycylsarcosylsarcosine stimulated uptake by 4-fold. Uptake of GlyGlyPro was inhibited greater than 50% by dipeptides and tripeptides and less than 15% by free amino acids. These results indicate that GlyGlyPro uptake in jejunal brush-border membrane vesicles is not energized by a Na+ or proton gradient and that transport occurs by carrier-mediated and diffusional processes. 相似文献
36.
Myocardial extracellular matrix is organized into a complex arrangement of intercellular and pericellular fibres and fibrils that serves as a supporting framework for contracting cells. Recent evidence suggests that changes in ventricular shape and function occurring after ischaemic injury may be related to alterations of this matrix. In this report we describe the rapid and extensive loss of collagen in myocardial infarction produced by ligating the left anterior descending coronary artery of the rat for 1-3 h. The total collagen content in the myocardial infarct zones after 1, 2 and 3 h of ligation was 75 +/- 8%, 65 +/- 7% and 50 +/- 10% respectively (mean +/- S.D.) of that of either the non-infarcted tissue controls or of the same regions in sex- and age-matched normal left ventricles. A marked decrease also occurred in the residual collagens which were not extractable with 6 M-guanidine hydrochloride, suggesting that rapid degradation of insoluble collagen fibres may also occur. The decreased collagen content in the 3 h myocardial infarct coincided with the appearance of several enzyme activities. Collagenase, other neutral proteinase and presumed lysosomal serine proteinase activities were increased by 3, 3 and 2 times the control values respectively. These results suggest that the increased activities of collagenase and other neutral proteinases may be responsible for the rapid degradation of extracellular matrix collagen in myocardial infarct. 相似文献
37.
38.
Genetic heterogeneity in tuberous sclerosis 总被引:8,自引:0,他引:8
L A Janssen L A Sandkuyl E C Merkens J A Maat-Kievit J R Sampson P Fleury R C Hennekam G C Grosveld D Lindhout D J Halley 《Genomics》1990,8(2):237-242
Tuberous sclerosis (TSC) is an autosomal dominant disorder characterized by widespread hamartosis. Preliminary evidence of linkage between the TSC locus and markers on chromosome 9q34 was established, but subsequently disputed. More recently, a putative TSC locus on chromosome 11 has been suggested and genetic heterogeneity seems likely. Here we describe an approach combining multipoint linkage analysis and heterogeneity tests that has enabled us to obtain significant evidence for locus heterogeneity after studying a relatively small number of families. Our results support a model with two different loci independently causing the disease. One locus (TSC1) maps in the vicinity of the Abelson oncogene at 9q34 and a second locus (TSC2) maps in the region of the anonymous DNA marker Lam L7 and the dopamine D2 receptor gene at 11q23. 相似文献
39.
Effect of Sorbinil on myo-Inositol Metabolism in Cultured Neuroblastoma Cells Exposed to Increased Glucose Levels 总被引:6,自引:6,他引:0
Neuroblastoma cells were used to determine the effect of sorbinil on myo-inositol metabolism in cells exposed to elevated levels of glucose in culture. Exposing cells to elevated levels of glucose led to an increase in levels of intracellular sorbitol. The increase in sorbitol levels was dependent on the extracellular glucose concentration. In contrast, the myo-inositol content of cells was decreased in the presence of increasing concentrations of extracellular glucose. Increasing the concentration of glucose in the culture medium caused a decrease in myo-inositol uptake and in the incorporation of extracellular myo-inositol into phospholipid. The effect of elevated glucose levels on myo-inositol metabolism and sorbitol accumulation was blocked by addition of 0.4 mM sorbinil. The ability of sorbinil to block the decrease in myo-inositol metabolism and sorbitol accumulation caused by 30 mM extracellular glucose was dependent on its concentration. Maximal effects were obtained with 0.4 mM sorbinil. However, there was some variation in the degree of effectiveness among batches of sorbinil. These results at the cellular level suggest that the intracellular accumulation of sorbitol is responsible for the alteration of myo-inositol metabolism observed in neuroblastoma cells exposed to elevated glucose concentrations. 相似文献
40.
Effect of Increased Glucose Levels on Na+ /K+ -Pump Activity in Cultured Neuroblastoma Cells 总被引:1,自引:0,他引:1
Neuroblastoma cells were used to analyze the effect of elevated glucose levels on myo-inositol metabolism and Na+/K+-pump activity. The activity of the Na+/K+ pump in neuroblastoma cells is almost totally sensitive to ouabain inhibition. Culturing neuroblastoma cells in 30 mM glucose caused a significant decrease in Na+/K+-pump activity, myo-inositol metabolism, and myo-inositol content, compared to cells grown in the presence of 30 mM fructose. Glucose supplementation also caused a large intracellular accumulation of sorbitol. The aldose reductase inhibitor sorbinil prevented the abnormalities in myo-inositol metabolism and partially restored Na+/K+-pump activity in neuroblastoma cells cultured in the presence of elevated glucose levels. These results suggest that the accumulation of sorbitol by neuroblastoma cells exposed to elevated concentrations of extracellular glucose causes a decrease in myo-inositol metabolism and these abnormalities are associated with a reduction in Na+/K+-pump activity. 相似文献