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141.
    
Fluorogenic peptide substrates designed to encompass the reported-secretory and amyloidogenic cleavage sites of the amyloid- precursor protein (PP) were used to analyze proteinase activities in brain extracts from control patients and those with Alzheimer's disease (AD). Activity against the secretory substrate atpH 7.5 in control and AD brains produced a major endopeptidase cleavage at the Lys687-Leu688 bond (PP770 numbering), consistent with thePP secretase cleavage. Activity in control brains against the amyloidogenic substrate atpH 7.5 produced one cleavage at the Ala673-Glu674 bond, two residues C-terminal to the amyloidogenic Met-Asp site. However, in three of four AD brains, the major cleavage was at the Asp-Ala bond, one residue from the amyloidogenic site. Both endopeptidase and carboxypeptidase activities in AD brains were lower than in control brains. Proteinase activities against the secretory substrate had a major optimum atpH 3.0–4.0 and another atpH 6.0–7.5. Proteinase activities against the amyloidogenic substrate had a major optimum at or belowpH 3.0 and another atpH 6.0. Using both substrates, activities at lowpH were higher in AD brains than in controls, while atpH above 6.5, activities in control brains were higher than in AD. These results indicate that the levels of proteolytic enzymes in AD brains are altered relative to controls.Abbreviations A Amyloid- - ACN acetonitrile - AD Alzheimer's disease - PP amyloid- precursor protein - DABCYL 4-(4-dimethylaminophenylazo)-benzoic acid - EDANS 5-{(2-aminoethyl)amino}napthalene-1-sulfonic acid - MES morpholinoethane sulfonic acid - MOPS morpholino-propane sulfonic acid - RP-HPLC reverse-phase high-performance liquid chromatography - SDS-PAGE sodium do-decyl sulfate-polyacrylamide gel electrophoresis - TFA tri-fluoroacetic acid - Tris tris(hydroxyethyl)aminomethane  相似文献   
142.
葱兰黄化病病原类菌原体的研究   总被引:3,自引:0,他引:3  
葱兰为石蒜科(Amaryllidaceae)草本植物,是一种在我国各地普遍栽培的多年生观赏花卉.葱兰黄化病是1992年在杭州发现的一种新病害,该病对葱兰的生长和观赏价值有严重影响.罹病株(丛)最初表现为深红色斑点,逐渐扩大为鲜红色条斑或斑块并转为亮黄色,最后整株叶片大部分转为鲜艳的黄色,呈现典型的系统侵染.发病1—2月后整株(丛)地上部分枯死.发病一般始于早春(3月上旬),春末夏初有一个发病高峰期;秋末有另一个发病期,但病状较轻;感染株有恢复现象,即在下一个生长季能从地下部分重新生长出叶片,但病株叶片明显细小,有丛簇现象,病株开花少且花期短.作者从1992年春起,对该病害进行了病原分离、生物学测定和病组织超薄切片电镜检查,发现该病害可能与类菌原体有关,现将研究结果报告如下.1 材料和方法1.1 样品感病植株分别于1992年4月采自浙江农业大学校园和1992年6月采自杭州花圃,用盆栽保存于防虫温室中,同时采取健株作为对照.1.2 用病株分离病原和接种  相似文献   
143.
Strawberry tissues infected with Phytophthora cactorum were comminuted and plated in a selective antibiotic agar medium to determine levels of tissue colonisation as indicated by the number of colony forming units (CFU) recovered per gramme of infected tissue. The number of CFU recovered per gramme of tissue increased logarithmically with the amount of necrosis in infected crown, leaf and petiole tissues. Under the conditions of enhanced susceptibility to infection and colonisation caused by cold storage treatments, this relationship between colonisation and necrosis was not significantly altered in the susceptible cv. Tamella. A recovery index was used to determine the effect of infected tissues on the recovery of CFU. This indicated that increasing levels of host colonisation stimulated CFU recovery and may partly explain the large increase in CFU g-1 with larger amounts of necrosis. The amount of tissue colonisation was greater in inoculated plants of the susceptible cv. Tamella than in less susceptible cv. Cambridge Favourite, although the necrotic tissues of the latter contained more CFU g-1, indicating a greater level of tolerance to colonisation. In cv. Tamella small amounts of colonisation were capable of causing wilt symptoms, although no wilted plants contained less than 200 CFU g-1. Conversely, plants containing more than 1000 CFU g-1 always wilted. In the early stages of infection, low levels of colonisation could be detected in strawberry crowns in the absence of symptoms. Dormant strawberry plants of cv. Tamella were readily infected by P. cactorum zoospore inoculations but, unlike actively growing plants, the majority of infections remained latent. These latent infections exhibited little or no symptoms and CFU recoveries from infected tissues were always below 100 CFU g-1.  相似文献   
144.
Cerebrospinal Fluid Nitrite/Nitrate Levels in Neurologic Diseases   总被引:5,自引:0,他引:5  
Abstract: Nitric oxide has been proposed to mediate cytotoxic effects in inflammatory diseases. To investigate the possibility that overproduction of nitric oxide might play a role in the neuropathology of inflammatory and noninflammatory neurological diseases, we compared levels of the markers of nitric oxide, nitrite plus nitrate, in the CSF of controls with those in patients with various neurologic diseases, including Huntington's and Alzheimer's disease, amyotrophic lateral sclerosis, and HIV infection. We found that there were no significant increases in the CSF levels of these nitric oxide metabolites, even in patients infected with HIV or in monkeys infected with poliovirus, both of which have significantly elevated levels of the neurotoxin quinolinic acid and the marker of macrophage activation, neopterin. However, CSF quinolinic acid, neopterin, and nitrite/nitrate levels were significantly increased in a small group of patients with bacterial and viral meningitis.  相似文献   
145.
Cortical Cytochrome Oxidase Activity Is Reduced in Alzheimer's Disease   总被引:21,自引:5,他引:16  
Abstract: A defect in energy metabolism may play a role in the pathogenesis of neurodegenerative diseases, such as Alzheimer's disease. In the present study, we examined the activities of the enzymes that catalyze oxidative phosphorylation in frontal, temporal, parietal, and occipital cortex from Alzheimer's disease patients and age-matched controls. Complex I and complex II–III activities showed a small decrease in occipital cortex, but were unaffected in the other cortical areas. The most consistent change was a significant decrease of cytochrome oxidase (complex IV) activity of 25–30% in the four cortical regions examined. These results provide further evidence of a cytochrome oxidase defect in Alzheimer's disease postmortem brain tissue. A deficiency in this key energy-metabolizing enzyme could lead to a reduction in energy stores and thereby contribute to the neurodegenerative process.  相似文献   
146.
Abstract: Ganglioside analysis and quantitative Golgi studies of the cerebral cortex of cats with ganglioside and nonganglioside lysosomal storage diseases reveal a correlation between the amount of accumulated GM2 ganglioside and the extent of ectopic dendrite growth on cortical pyramidal neurons. This correlation was not observed with any of the other gangliosides assayed for, including GM1 ganglioside. These results suggest a specific role for GM2 ganglioside in the initiation of ectopic neurites on pyramidal cells in vivo and are consistent with the developing hypothesis that different gangliosides have specific roles in different cell types dependent upon the receptor or other effector molecules with which they may interact.  相似文献   
147.
Abstract: ADP-ribosylation is an essential process in the metabolism of brain neuronal proteins, including the regulation of assembly and disassembly of biological polymers. Here, we examine the effect of HgCl2 exposure on the ADP-ribosylation of tubulin and actin, both cytoskeletal proteins also found in neurons, and B-50/43-kDa growth-associated protein (B-50/GAP-43), a neuronal tissue-specific phosphoprotein. In rats we demonstrate, with both in vitro and in vivo experiments, that HgCl2 markedly inhibits the ADP-ribosylation of tubulin and actin. This is direct quantitative evidence that HgCl2, a toxic xenobiotic, alters specific neurochemical reactions involved in maintaining brain neuron structure.  相似文献   
148.
Antioxidant Properties of Bromocriptine, a Dopamine Agonist   总被引:3,自引:1,他引:2  
Abstract: It has been suggested that free radicals may adversely influence the pathogenesis of Parkinson's disease. We conducted this study to determine whether bro-mocriptine, an agent widely used for treating parkinsonism, possesses antioxidant effects. Bromocriptine scavenged superoxide produced from a superoxide generating system (hypoxanthine-xanthine oxidase) by the spin-trapping method using electron spin resonance. Bromocriptine had a strong scavenging effect on the 5,5-dimethyl-1-pyrroline- N -oxide hydroxide signal produced from Fenton's reaction. Bromocriptine also attenuated the stable free radical diphenyl- p -picrylhydrazyl signal. This drug inhibited the autooxidation of rat brain homogenates in a dose-dependent manner in vitro. Autooxidation of brain homogenates collected from rats treated with bromocriptine (2.5 mg/kg, i.p., daily for 3 days) was significantly reduced as compared with values in untreated rat homogenates. These observations suggest that bromocriptine is a free radical scavenger and a potent antioxidant.  相似文献   
149.
Abstract: A synthetic peptide corresponding to residues 226–240 (E9 peptide) of human τ, which contains an Lys-Ser-Pro motif, was used to raise a polyclonal antibody. The antibody, E9, was 10-fold less reactive with phospho-E9 peptide than with native E9 peptide. E9 antibody was used to study the extent of phosphorylation in a modified form of τ (PHF-τ) that is found in Alzheimer's disease brain and is incorporated into paired helical filaments (PHFs). E9 immunolabeled Alzheimer's disease neurofibrillary tangles and abnormal neurites in brain sections intensely, with increased immunoreactivity detected after pretreatment of sections with phosphatase. On immunoblots and ELISA, E9 reacted with PHF-τ and recombinant human τ but not with the high and middle molecular weight neurofilament proteins. Phosphatase treatment of PHF-τ improved the E9 immunoreactivity by 30–50%. Dephosphorylated high but not middle molecular weight neurofilament protein became reactive with E9. These results indicate that <50% of the PHF-T is phosphorylated in the subregion corresponding to residues 226–240 of τ and suggest that the phosphorylation of this region may not be essential for PHF formation.  相似文献   
150.
Both naive and vaccinated macaques acquired a virus-specific proliferative helper T-cell reactivity in response to infection with the nonpathogenic human immunodeficiency virus type 2 (HIV-2). In contrast, macaques infected with the pathogenic simian immunodeficiency virus of the macaque strain (SIVmac) did not develop a helper T-cell response. Furthermore, a vaccine-induced preexisting T-cell reactivity was abrogated after SIVmac infection in vaccine failures. These differences may reflect the different pathogenicity of the two closely related viruses.  相似文献   
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