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101.
Lambert-Eaton myasthenic syndrome (LEMS) is an autoimmune neuromuscular disease in which impairment of Ca2+ entry into the nerve ending and consequent impaired release of acetylcholine (ACh) results in muscle weakness. The identity of the primary antigenic target molecule(s) of the autoantibodies is uncertain. Electrophysiological studies and 45Ca2+ uptake studies implicate a direct effect on the Ca2+ channel complex at the motor nerve terminal. Some recent studies, however, suggest a more indirect interference caused by binding of autoantibodies to synaptotagmin or syntaxin, molecules presumed to be involved in docking and/or coupling the synaptic vesicles to the Ca2+ channels in the active zone for vesicle exocytosis and transmitter release. Western blot analyses of rat and human brain membrane proteins and pure recombinant synaptotagmin and syntaxin were used to examine directly the targets of LEMS autoantibodies and determine specifically whether or not synaptotagmin and/or syntaxin were general targets in LEMS. IgG from 14 patients with LEMS was used to probe western blots of gels containing synaptotagmin, syntaxin, rat synaptosomal proteins, and human brain membrane proteins. Several similar immunoreactive bands were observed using both rat and human brain membranes. These include high-molecular-weight protein bands whose size would be consistent with being components of Ca2+ channels. No reactive component was observed against either syntaxin or synaptotagmin in IgG of the 14 LEMS patients. However, both human and rat brain membranes contain proteins recognized by antibodies directed against synaptotagmin or syntaxin, indicating their immunologic relatedness and evolutionary conservation. These results suggest that large-molecular-weight proteins consistent with being Ca2+ channel subunits rather than syntaxin and synaptotagmin are general targets of LEMS autoantibodies. 相似文献
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J Whittaker WD McFadyen G Wickham LP Wakelin V Murray 《Canadian Metallurgical Quarterly》1998,26(17):3933-3939
Cisplatin analogues were synthesised that consisted of platinum(II) diamine complexes tethered via a polymethylene chain ( n = 3, 5, 8 and 10) to a phenanthridinium cation. Both chloro and iodo leaving groups were examined. DNA adduct formation was quantitatively analysed using a linear amplification system with the plasmid pGEM-3Zf(+). This system utilised Taq DNA polymerase to extend from an oligonucleotide primer to the damage site. This damage site inhibited the extension of the DNA polymerase. The products were electrophoresed on a DNA sequencing gel enabling adduct formation to be determined at base pair resolution. The damage intensity at each site was determined by densitometry. The platinum phenanthridinium complexes were shown to damage DNA at shorter incubation times than cisplatin. To produce similar levels of damage, an 18 h incubation was required for cisplatin compared to 30 min for the n = 3 platinum phenanthridinium complexes; this indicates that the intercalating chromophore causes a large increase in the rate of platination. A reaction mechanism involving direct displacement of the chloride by the N-7 of guanine may account for the rate increase. These results indicate that further development of these compounds could lead to more effective cancer chemotherapeutic agents. 相似文献
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Studies of isolated cell membranes and animal brain extracts have shown that ethanol (EtOH) partitions into cell membranes. We tested the hypothesis that EtOH in the living brain after EtOH administration exists in two or more pools: a free, mobile pool of EtOH and one or more EtOH pools that are restricted in their molecular mobility, possibly because of association with membranes. In vivo brain proton magnetic resonance spectroscopy (1H MRS) routinely detects the methyl protons of the mobile EtOH pool but does not detect motionally restricted EtOH. We used in vivo brain 1H MRS in rat brain (n = 11) after intraperitoneal EtOH administration to measure the signal intensity of methyl EtOH protons in the presence and absence of off-resonance saturation. Off-resonance saturation resulted in a 33 +/- 4% decrease of the EtOH methyl proton signal. We interpret this signal reduction as a magnetization transfer effect. It is consistent with the existence of an MRS-invisible EtOH pool with restricted molecular mobility, which is in exchange with the free EtOH pool. Off-resonance saturation at the water frequency resulted in an even larger decrease of the EtOH methyl signal, consistent with water molecules being in close proximity to EtOH molecules at the restricted motion site(s). These results provide support for the hypothesis that partial MRS-invisibility of brain EtOH is at least to some extent caused by the presence of a (MRS-invisible) pool of motionally restricted EtOH. They also strongly suggest that water suppression, routinely used in in vivo 1H MRS, may reduce the observable EtOH methyl signal intensity through a magnetization transfer mechanism. These studies may provide both a mechanism of, and a means to investigate the alterations of EtOH MRS visibility observed in heavy drinkers. 相似文献
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DL Broyles RG Nielsen EM Bussett WD Lu IA Mizrahi PA Nunnelly TA Ngo J Noell RH Christenson BC Kress 《Canadian Metallurgical Quarterly》1998,44(10):2139-2147
The performance characteristics of the Tandem-MP Ostase assay, a new microplate immunoassay for bone-specific alkaline phosphatase (bone ALP; EC 3.1.3.1) in human sera, are described. Bone ALP is bound to streptavidin-coated microwells by a single biotinylated anti-bone ALP monoclonal antibody. Antigen is detected by the addition of p-nitrophenyl phosphate. The assay is performed at room temperature in <90 min. Imprecision was 2.3-6.1% with a detection limit of 0.6 microg/L. Method comparison of bone ALP measurements with the Tandem-MP Ostase assay and the mass-based Tandem-R Ostase assay (n = 285) indicated regression statistics of Tandem-MP Ostase = 1.03 Tandem-R Ostase + 0.22 microg/L, S(y/x) = 4.0 microg/L, r = 0.97. Serum bone ALP values in apparently healthy men and in pre- and postmenopausal women were also similar between the two Ostase assay formats. Liver ALP reactivity determined using the slope and heat inactivation methods was similar in both Ostase assays. Liver ALP reactivity ranged from 3 microg/L (heat inactivation) to 6 microg/L (slope method) per 100 U/L of liver ALP activity, whereas bone ALP reactivity was 37 microg/L per 100 U/L of bone ALP activity, indicating a liver ALP relative reactivity of 8.1-16.2%. Similar results were obtained with the Alkphase-B bone ALP immunoassay. The Tandem-MP Ostase bone ALP assay demonstrated increased concentrations of serum bone ALP in conditions where bone metabolism is increased and showed a rapid, temporal decrease in serum bone ALP in Paget disease patients on bisphosphonate therapy. In conclusion, the Tandem-MP Ostase assay for serum bone ALP is a rapid, simple, robust nonisotopic alternative to the Tandem-R Ostase immunoradiometric assay that provides an accurate and sensitive assessment of bone turnover. 相似文献
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WD van Marken Lichtenbelt GA Heidendal KR Westerterp 《Canadian Metallurgical Quarterly》1997,51(12):826-830
OBJECTIVES: To assess sleeping metabolic rate (SMR), average daily metabolic rate (ADMR), and total bone mineral density (TBMD) in women with anorexia nervosa, and to evaluate the effect of daily physical activity on TBMD. DESIGN: We compared women with anorexia nervosa and controls using measurements on body composition, and energy expenditure. Relations between these measurements were investigated. SETTING: Daily living environments in The Netherlands, and body composition and energy expenditure laboratory of the Department of Human Biology. SUBJECTS: Twelve adult, non-hospitalized women with anorexia nervosa, and sixteen adult normal weight women. INTERVENTIONS: Average daily metabolic rate was measured with the doubly labeled water method and sleeping metabolic rate in a respiration chamber. TBMD was measured by dual energy X-ray absorptiometry, and percentage body fat was calculated combining the results from underwater weighing and deuterium dilution. RESULTS: TBMD was significantly lower in anorexia than in controls (0.989 +/- 0.081 vs 1.144 +/- 0.054 g/cm2). Also ADMR and SMR were reduced in anorexia. The physical activity index (PAI = ADMR/SMR) was not significantly different from PAI in controls. In anorexia, TBMD was related to the PAI (R2 = 0.35, P < 0.05). Finally, stepwise multiple regression revealed that PAI together with the study groups as dummy variables could explain 69% of the variation in TBMD. CONCLUSION: These findings show that in anorexia TBMD is reduced, but that nonetheless physical activity has a significant positive effect on bone density. 相似文献