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81.
82.
Uchida Y Ohshima T Sasaki Y Suzuki H Yanai S Yamashita N Nakamura F Takei K Ihara Y Mikoshiba K Kolattukudy P Honnorat J Goshima Y 《Genes to cells : devoted to molecular & cellular mechanisms》2005,10(2):165-179
Collapsin response mediating protein-2 (CRMP2) has been identified as an intracellular protein mediating Semaphorin3A (Sema3A), a repulsive guidance molecule. In this study, we demonstrate that cyclin-dependent kinase 5 (Cdk5) and glycogen synthase kinase 3beta (GSK3beta) plays a critical role in Sema3A signalling. In In vitro kinase assay, Cdk5 phosphorylated CRMP2 at Ser522, while GSK3beta did not induce any phosphorylation of CRMP2. Phosphorylation by GSK3beta was exclusively observed in Cdk5-phosphorylated CRMP2, but barely in CRMP2T509A. These results indicate that Cdk5 primarily phosphorylates CRMP2 at Ser522 and GSK3beta secondarily phosphorylates at Thr509. The dual-phosphorylated CRMP2, but not non-phosphorylated or single-phosphorylated CRMP2, is recognized with the antibody 3F4, which is highly reactive with the neurofibrillary tangles of Alzheimer's disease. 3F4 recognized the CRMP2 in the wild-type but not cdk5-/- mouse embryonic brain lysates. The phosphorylation of CRMP2 at Ser522 caused reduction of its affinity to tubulin. In dorsal root ganglion neurones, Sema3A stimulation enhanced the levels of the phosphorylated form of CRMP2 detected by 3F4. Over-expression of CRMP2 mutant substituting either Ser522 or Thr509 to Ala attenuates Sema3A-induced growth cone collapse response. These results suggest that the sequential phosphorylation of CRMP is an important process of Sema3A signalling and the same mechanism may have some relevance to the pathological aggregation of the microtubule-associated proteins. 相似文献
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85.
Takasaki T Nawa M Yamada KI Harada M Takeda A Kurane I 《Journal of virological methods》2002,102(1-2):61-66
Three commercial dengue IgM test kits and 'in-house' IgM-capture enzyme-linked immunosorbent assay (ELISA) were examined for false positive reactions, using 49 serum samples from patients with autoimmune diseases. All the samples were found to be negative by the 'in-house' IgM-capture ELISA. Five samples were determined to be positive by the immunochromatographic test and three of the five samples were also found positive by one commercial IgM-capture ELISA kit. These results suggest that a possibility of false positive reaction should be considered when serum samples from autoimmune disease patients are tested for dengue IgM by some commercial dengue IgM test kits. 相似文献
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87.
Tsukasa Tsunoda Toshifumi Eto Takahito Tsurifune Shigeki Tokunaga Tatsuhiro Ishii Koichi Motojima Teiji Matsumoto Tohru Segawa Kazuhide Ura Hiroshi Fukui 《Pathology international》1991,41(10):763-770
A solid and cystic tumor (SCT) was located at the head of the pancreas in a 43-year-old Japanese male, and pancreatoduodenectomy was performed on the suspicion of papillary carcinoma or cystadenocarcinoma of the pancreas. The lesion, which measured 4.5 X 4.5 X 4.0 cm, was clearly demarcated by connective tissue. The cut surface showed solid grayish-white areas with central cystic degenerative changes. The solid areas consisted of small round cells proliferating in a small solid or a pseudopapillary pattern. The tumor cells partially invaded the surrounding normal pancreatic parenchyma. Immunohistochemical studies revealed positive staining for alpha-1-antitrypsin and neuron-specific enolase, but no staining for known pancreatic hormones. Moreover, ultrastructural studies showed the absence of zymogen granules and the presence of anullate lamellae and neurosecretory granules. On the basis of these findings, a diagnosis of SCT of the pancreas was established. In order to clarify the histogenesis and biological behavior of the tumor, it is necessary to accumulate and analyze similar cases, an endeavor which in turn will contribute to the successful management of this disease. Acta Pathol Jpn 41: 763-770, 1991. 相似文献
88.
T Kawamata S Nakamura I Akiguchi J Kimura M Kameyama H Kimura T Takeda 《Neurobiology of aging》1990,11(3):185-192
Age-related changes of reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d)-containing neurons were examined quantitatively in the laterodorsal tegmental nucleus (TLD) and the caudate-putamen of mice. Six 2-month-old and six 25- to 30-month-old DDD mice were studied using computer-assisted image analysis. Although no age-related changes in neuronal counts were found in the TLD, the cell size in this nucleus showed a statistically significant reduction with aging. In addition, the degree of the age-related neuronal shrinkage differed within the TLD; the most significant occurring in the rostral, less in the caudal third and no significant alteration being found in the middle third portion of TLD. In contrast, NADPH-d-positive neurons in the striatum did not show distinct age-related changes. NADPH-d-containing neurons in the TLD correspond to cholinergic cells, which project to the forebrain. Thus, the age-related shrinkage of NADPH-d neurons in the TLD may be related to the cholinergic dysfunctions seen in the forebrain of senescent mice. 相似文献
89.
Purification and partial characterization of heat-stable enterotoxin of enterotoxigenic Escherichia coli. 总被引:6,自引:10,他引:6 下载免费PDF全文
Heat-stable enterotoxin was purified from a strain of enterotoxigenic Escherichia coli 53402 A-1 from human intestine. The cells were cultured in Casamino Acids-yeast extract-salts medium, and the purification procedure consisted of protamine sulfate treatment of the culture supernatant, ultrafiltration with an Amicon PM-10 membrane, diethylaminoethyl-cellulose column chromatography, hydroxyapatite column chromatography, Bio-Gel P-10 gel filtration, 90% ethanol extraction, and preparative polyacrylamide gel disc electrophoresis. About 300-fold purification was achieved, with a yield of about 12%. However, the homogeneity of the purified heat-stable enterotoxin was not rigorously demonstrated. The purified heat-stable enterotoxin had an absorption maximum at about 275 nm, and its isoelectric point was about 3.90. The molecular weight of the purified heat-stable enterotoxin was ca. 4,000 by Sephadex G-100 gel filtration. The minimum effective dose of purified heat-stable enterotoxin was about 2.5 ng in the suckling mouse assay. The purified heat-stable enterotoxin gave a positive reaction in not only the suckling mouse assay but also the mouse intestinal loop test and the guinea pig skin permeability test. 相似文献
90.
Chakraborty S Garg P Ramamurthy T Thungapathra M Gautam JK Kumar C Maiti S Yamasaki S Shimada T Takeda Y Ghosh A Nair GB 《Journal of medical microbiology》2001,50(10):879-888
This study identified 17 matching serogroups of Vibrio cholerae belonging to serogroups other than O1 and O139 isolated from human cases and from the environment during a concurrent clinical and environmental study conducted in Calcutta, a cholera endemic area. Isolates within these matching serogroups were compared by various phenotypic and genotypic traits to determine if the environment was the source of the organisms associated with the disease. Clinical strains of V. cholerae were resistant to a greater number of drugs and exhibited multi-drug resistance compared with their environmental counterparts. Except for the presence of the genes for the El Tor haemolysin and the regulatory element ToxR in most of the strains of V. cholerae examined, non-O1, non-O139 V. cholerae strains lacked most of the other known virulence traits associated with toxigenic V. cholerae O1 or O139. Restriction fragment-length polymorphism of virulence-associated genes, ribotypes and DNA fingerprints of strains of matched serogroups showed considerable diversity, although some gene polymorphisms and ribotypes of a few strains of different serogroups were similar. It is concluded that despite sharing the same serogroup, environmental and clinical isolates were genetically heterogeneous and were of different lineages. 相似文献