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41.
Smart ML Panchal RG Bowser DN Williams DA Petrou S 《American journal of physiology. Cell physiology》2002,283(1):C77-C84
The present study examines whether changes in P2X7 purinergic receptor density precede formation of the cytolytic pore characteristic of this receptor. We fused P2X7 receptors with enhanced green fluorescent protein (EGFP) at the amino or carboxy termini (EGFP-P2X7 and P2X7-EGFP). Electrophysiological characterization in Xenopus oocytes revealed wild-type responses to ATP for GFP-tagged receptors. However, differences in sensitivity to ATP were apparent with the P2X7-EGFP receptor displaying a threefold reduction in ATP sensitivity compared with control. Ethidium ion uptake was used to measure cytolytic pore formation. Comparison of tagged receptors with wild type in HEK-293 and COS-7 cells showed there was no significant difference in ethidium ion uptake, suggesting that fusions with EGFP did not interfere with cytolytic pore formation. Confocal microscopy confirmed that tagged receptors localized to the plasmalemma. Simultaneous monitoring of EGFP and ethidium ion fluorescence revealed that changes in receptor distribution do not precede pore formation. We conclude that it is unlikely that large scale changes in P2X7 receptor density precede pore formation. 相似文献
42.
Kincer JF Uittenbogaard A Dressman J Guerin TM Febbraio M Guo L Smart EJ 《The Journal of biological chemistry》2002,277(26):23525-23533
Numerous studies have implicated either the presence or absence of CD36 in the development of hypertension. In addition, hypercholesterolemia is associated with the loss of nitric oxide-induced vasodilation and the subsequent increase in blood pressure. In the current study, we tested the hypothesis that diet-induced hypercholesterolemia promotes the disruption of agonist-stimulated nitric oxide generation and vasodilation in a CD36-dependent manner. To test this, C57BL/6, apoE null, CD36 null, and apoE/CD36 null mice were maintained on chow or high fat diets. In contrast to apoE null mice fed a chow diet, apoE null mice fed a high fat diet did not respond to acetylcholine with a decrease in blood pressure. Caveolae isolated from in vivo vessels did not contain endothelial nitric-oxide synthase and were depleted of cholesterol. Age-matched apoE/CD36 null mice fed a chow or high fat diet responded to acetylcholine with a decrease in blood pressure. The mechanism underlying the vascular dysfunction was reversible because vessels isolated from apoE null high fat-fed mice regained responsiveness to acetylcholine when incubated with plasma obtained from chow-fed mice. Further analysis demonstrated that the plasma low density lipoprotein fraction was responsible for depleting caveolae of cholesterol, removing endothelial nitric-oxide synthase from caveolae, and preventing nitric oxide production. In addition, the pharmacological removal of caveola cholesterol with cyclodextrin mimicked the effects caused by the low density lipoprotein fraction. We conclude that the ablation of CD36 prevented the negative impact of hypercholesterolemia on agonist-stimulated nitric oxide-mediated vasodilation in apoE null mice. These studies provide a direct link between CD36 and the early events that underlie hypercholesterolemia-mediated hypertension and mechanistic linkages between CD36 function, nitric-oxide synthase activation, caveolae integrity, and blood pressure regulation. 相似文献
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Analysis of the fecal microflora of human subjects consuming a probiotic product containing Lactobacillus rhamnosus DR20 总被引:13,自引:0,他引:13
Tannock GW Munro K Harmsen HJ Welling GW Smart J Gopal PK 《Applied and environmental microbiology》2000,66(6):2578-2588
The composition of the fecal microflora of 10 healthy subjects was monitored before (6-month control period), during (6-month test period), and after (3-month posttest period) the administration of a milk product containing Lactobacillus rhamnosus DR20 (daily dose, 1.6 x 10(9) lactobacilli). Monthly fecal samples were examined by a variety of methods, including bacteriological culture analysis, fluorescent in situ hybridization with group-specific DNA probes, denaturing gradient gel electrophoresis of the V2-V3 region of 16S rRNA genes amplified by PCR, gas-liquid chromatography, and bacterial enzyme activity analysis. The composition of the Lactobacillus population of each subject was analyzed by pulsed-field gel electrophoresis of bacterial DNA digests in order to differentiate between DR20 and other strains present in the samples. Representative isolates of lactobacilli were identified to the species level by sequencing the V2-V3 region of their 16S rRNA genes and comparing the sequences obtained (BLAST search) to sequences in the GenBank database. DR20 was detected in the feces of all of the subjects during the test period, but at different frequencies. The presence of DR20 among the numerically predominant strains was related to the presence or absence of a stable indigenous population of lactobacilli during the control period. Strain DR20 did not persist at levels of >10(2) cells per g in the feces of most of the subjects after consumption of the product ceased; the only exception was one subject in which this strain was detected for 2 months during the posttest period. We concluded that consumption of the DR20-containing milk product transiently altered the Lactobacillus and enterococcal contents of the feces of the majority of consumers without markedly affecting biochemical or other bacteriological factors. 相似文献
47.
Effects of sulfate on cadmium uptake by Swiss chard: II. Effects due to sulfate addition to soil 总被引:3,自引:0,他引:3
Sulfate complexation of Cd in nutrient solution has been shown to have little impact on Cd uptake by plants. This study examined the effect of sulfate added to soil on Cd concentrations in soil solution and Cd uptake by Swiss chard (Beta vulgaris L. cv. Fordhook Giant). Swiss chard was grown in soil which was wetted with complete nutrient solution containing equivalent salt concentrations of NaNO3 or Na2SO4. Plant growth was reduced by increasing both NO3 and SO4 concentrations in soil solution, with growth reductions similar for both salts. The Cd concentration in soil solution increased P< 0.05) more consistently with increasing concentrations of SO4 compared to NO3 in soil solution. Solution speciation, calculated with GEOCHEM-PC, showed significant increases of Cd2+ activities with increasing salt rates. Shoot Cd content in 19-day-old Swiss chard plants was marginally but significantly P <0.05) increased with increasing SO4 concentration but no effect was observed with increasing NO3 concentration. These results are compared with earlier work on the marked effect of Cl- salinity on Cd availability in Swiss chard. Possible mechanisms explaining the smaller effect of SO4 compared to Cl on Cd availability are proposed. 相似文献
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Structure of turnip crinkle virus. III. Identification of a unique coat protein dimer 总被引:1,自引:0,他引:1
P G Stockley A L Kirsh E P Chow J E Smart S C Harrison 《Journal of molecular biology》1986,191(4):721-725
The minor structural protein (p80), found in about one copy per virion in turnip crinkle virus (TCV), is shown by amino acid analysis and peptide mapping to be a covalent dimer of the major coat protein (p40). The covalent linkage occurs near the N termini of the crosslinked chains. These data suggest that TGV and related viruses contain 178 copies of p40 (89 non-covalent dimers) and one copy of p80 (covalent dimer of two additional p40 chains). The presence of p80 in the salt-stable RNA-protein complex formed when TCV dissociates, as described in an accompanying paper, indicates that the covalent modification affects binding to RNA. We suggest that p80 might be the final dimer to be incorporated into the shell and that it might also be the site for initiation of uncoating. 相似文献
50.
J. I. Smith N. J. Smart M. Misawa W. G. W. Kurz S. G. Tallevi F. DiCosmo 《Plant cell reports》1987,6(2):142-145
Vanadyl sulphate (10–500 mg/l), when added to cell suspension cultures of Catharanthus roseus stimulated increased intracellular accumulation of catharanthine and ajmalicine. This response was demonstrated in both flask and fermenter (30 litre) systems. The response varied, and depended upon cell line, concentration of vanadyl sulphate and the stage of the growth phase at which the cells were treated. This process has the potential to increase the yield and reduce the production time for commercially useful secondary plant metabolites.Abbreviations Ajm
ajmalicine
- Cath
catharanthine
- CAS
ceric ammonium sulphate
- VOSO4
vanadyl sulphate
- FW
fresh weight
- n.d.
not detected 相似文献