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Cationic current (Icat) and inhibition of the voltage-dependent Ca2+ current (ICa) evoked by muscarinic receptor activation with carbachol were studied using whole-cell patch clamp technique in smooth muscle cells isolated from longitudinal muscle of guinea pig small intestine. With low buffering of [Ca2+]i (0.1 mM BAPTA [1,2-bis-(2-aminophenoxy)-ethane-N,N, N', N'-tetraacetic acid] in pipette solution) Icat and ICa inhibitory responses had a rapid onset to an initial peak followed by a sustained phase. The sustained phase of ICa suppression was bigger than in the case when [Ca2+]i was clamped to 100 nM, but decreased with repeated stimulation. Upon repeated stimulation with 50 microM carbachol in cells where [Ca2+]i was clamped to 100 nM and when GTP was absent, Icat amplitude decreased strongly and more substantially compared to ICa inhibition, but both responses declined only slightly when 1 mM GTP was present in the pipette solution. GDP-betaS (1 or 5 mM) in pipette solution or pre-treatment of cells with pertussis toxin (6 microg/ml, for 4 h or longer) blocked Icat more than ICa suppression by carbachol, whereas L-NAME (N-omega-nitro-L-arginine methyl ester hydrochloride) (100 microM in pipette solution) affected neither of them significantly. We conclude that the cationic current and the suppression of the voltage-dependent Ca2+ current evoked by muscarinic receptor activation are mediated by pertussis toxin-sensitive G-protein(s) but the latter response was less sensitive to blockade by GDP-betaS and to GTP deficiency in the cell. 相似文献
574.
A microplate assay, for use with a variety of glycohydrolase enzymes, was developed to aid the screening of Chinese medicinal herb extracts for the presence of potential anti-viral and anti-lymphoma compounds. The microplate assay method described offers greater convenience, speed and reproducibility over existing methods. The enzymes tested were alpha-glucosidase, beta-glucosidase and beta-glucuronidase. The assay can be easily adapted for use with other glycohydrolase enzymes. Of the 12 herb extracts examined four did not inhibit any of the enzymes (< 50% inhibition), one inhibited alpha-glucosidase only (> 50% inhibition), six inhibited beta-glucuronidase only, and one inhibited both alpha-glucosidase and beta-glucuronidase. None of the extracts were capable of inhibiting beta-glucosidase to any significant extent. 相似文献
575.
DA Taira DG Safran TB Seto WH Rogers AR Tarlov 《Canadian Metallurgical Quarterly》1997,278(17):1412-1417
CONTEXT: The US Preventive Services Task Force recommends that physicians assess patients' health risk behaviors, addressing those needing modification. OBJECTIVE: To examine the relationship between patient income, health risk behaviors, the prevalence of physician discussion of these behaviors, and the receptiveness of patients to their physicians' advice. DESIGN: Employee survey. PARTICIPANTS: A random sample of 6549 Massachusetts state employees in 12 health plans. MAIN OUTCOME MEASURES: Data were obtained using a patient-completed mail survey. Trend tests were used to discern differences in the prevalence of health risk behaviors, physician discussion of these behaviors, and patient receptiveness to discussions by patient income. RESULTS: Although unhealthy behaviors were common among all income groups, physician discussion of health risk behaviors fell far short of the universal risk assessment recommended by the US Preventive Services Task Force. Low-income patients were more likely to be obese and smoke than high-income patients and were less likely to wear seat belts and exercise. In contrast, stress and alcohol consumption increased with income, while the proportion of heavy drinkers did not vary significantly. Physicians were more likely to discuss diet and exercise with high-income patients in need of these discussions than with low-income patients, but were more likely to discuss smoking with low-income patients who smoked than with high-income patients who smoked. Among patients with whom discussions occurred, low-income patients were much more likely to report attempting to change their behavior based on physician advice. CONCLUSIONS: Physician counseling of patients regarding health risk behaviors should be greatly improved if the US Preventive Services Task Force recommendations are to be fulfilled. Improvement is especially needed in regard to alcohol consumption, safe sex, and seat belt use. Physicians also need to be more vigilant in properly identifying and counseling low-income patients at risk in regard to diet and exercise and high-income patients who smoke. 相似文献
576.
BACKGROUND: Replication-deficient adenovirus, one of the most efficient vectors in gene therapy, has been limited by transient transgene expression due to its episomal location and loss during cell division, as well as a host immune response against viral proteins. METHODS: Murine pancreatic islets were infected ex vivo with ad5-cytomegalovirus (CMV)-beta-galactosidase and transplanted into diabetic recipients with normalization of glucose metabolism. RESULTS: High levels of beta-galactosidase activity were detectable histologically for at least 20 weeks after transplant, and beta-galactosidase and viral mRNA were also present that long. Sera from transplanted animals did not significantly inhibit ad5-CMV-interleukin-2-Ig infection of HeLa cells in vitro, whereas sera from intravenously delivered ad5-CMV-beta-galactosidase drastically diminished HeLa cell infection, suggesting the presence of reduced levels of antibodies in transplanted animals as compared with intravenously infected animals. Immunofluorescent staining of islet isografts infected with ad5-CMV-beta-galactosidase revealed the presence of CD8+ and CD4+ T lymphocytes at all time points, however, no islet destruction was seen. CONCLUSIONS: Treatment of islet isografts ex vivo with ad5-CMV-beta-galactosidase results in prolonged transgene expression, possibly due to an attenuated immune response against adenovirus. 相似文献
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578.
TB Levine AB Levine AD Goldberg MC Tobes B Narins S Goldstein M Lesch 《Canadian Metallurgical Quarterly》1997,79(1):105-106
For transplant wait-list patients with end-stage congestive heart failure, reversibility of pulmonary hypertension tested with acute administration of vasodilators is a prerequisite to listing for transplantation. We have shown that the magnitude of the initial pulmonary vasodilatory response to nitroprusside predicts neither the extent of the long-term hemodynamic response nor the subsequent need for transplantation versus clinical improvement and removal from transplant consideration. 相似文献
579.
Chitosan microspheres containing oxytetracycline (OTC), an antibiotic agent, were prepared by spray hardening and interfacial acylation methods. The object of this study was to prepare oxytetracycline-containing microspheres for oral administration and injection using different molecular weight (Mw 70,000 approximately 2,000,000) of chitosan. By the spray hardening method, microspheres with particle sizes between 5 and 30 microns could be obtained and might be suitable for intramuscular injection. On the other hand, chitosan microspheres with the ability to extend the dissolution period of oxytetracycline in low pH medium were also prepared by the interfacial acylation method. The result indicated that the releasing of oxytetracycline from various acylated chitosan microspheres was decreased with increasing the molecular weight of chitosan and would show well sustained-release property. Besides, the morphology of various microspheres and crystalline form transition of oxytetracycline were also studied using electron scanning microscope and X-ray analysis. 相似文献
580.