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141.
Junkins EP Knight S Lightfoot AC Cazier CF Dean JM Corneli HM 《The Journal of school health》1999,69(10):409-412
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A comparative proteome analysis of hippocampal tissue from schizophrenic and Alzheimer's disease individuals 总被引:4,自引:0,他引:4
The proteins expressed by a genome have been termed the proteome. Comparative proteome analysis of brain tissue offers a novel means to identify biologically significant gene products that underlie psychopathology. In this study we collected post mortem hippocampal tissue from the brains of seven schizophrenic, seven Alzheimer's disease (AD) and seven control individuals. Hippocampal proteomes were visualised by two-dimensional gel electrophoresis of homogenised tissue. A mean of 549 (s.d. 35) proteins were successfully matched between each disease group and the control group. In comparison with the control hippocampal proteome, eight proteins in the schizophrenic hippocampal proteome were found to be decreased and eight increased in concentration, whereas, in the AD hippocampal proteome, 35 proteins were decreased and 73 were increased in concentration (P<0.05). One protein, which was decreased in concentration in both diseases, was characterised as diazepam binding inhibitor (DBI) by N-terminal sequence analysis. DBI can regulate the action of the GABA(A) receptor. Protein changes involved 6% of the assessed AD hippocampal proteome, whereas, in schizophrenia protein changes involved less than 1% of the assessed hippocampal proteome. We conclude that schizophrenia has a subtle neuropathological presentation and comparative proteome analysis is a viable means by which to investigate diseases of the brain at the molecular level. 相似文献
145.
The effect of ultra-high molecular weight polyethylene wear debris on MG63 osteosarcoma cells in vitro 总被引:7,自引:0,他引:7
Dean DD Schwartz Z Liu Y Blanchard CR Agrawal CM Mabrey JD Sylvia VL Lohmann CH Boyan BD 《The Journal of bone and joint surgery. American volume》1999,81(4):452-461
BACKGROUND: Focal osteolysis due to ultra-high molecular weight polyethylene wear debris involves effects on both bone resorption and bone formation. METHODS: The response of MG63 osteoblast-like osteosarcoma cells to ultra-high molecular weight polyethylene wear debris isolated by enzymatic digestion of granulomatous tissue obtained from the sites of failed total hip arthroplasties was examined. Scanning electron microscopy, particle-size analysis, and Fourier transform infrared spectroscopy were used to characterize the number, morphology, size distribution, and chemical composition of the particles. Cell response was assessed by adding particles at varying dilutions to confluent cultures and measuring changes in cell proliferation (number of cells and [3H]-thymidine incorporation), osteoblast function (alkaline-phosphatase-specific activity and osteocalcin production), matrix production (collagen production and proteoglycan sulfation), and local cytokine production (prostaglandin-E2 production). RESULTS: The mean size of the particles was 0.60 micrometer, and 95 percent of the particles had a size of less than 1.5 micrometers. The number of particles per gram of tissue ranged from 1.39 to 3.38x10(9). Three of the four batches of particles were endotoxin-free. Exposure of the cells to particles of wear debris significantly increased the number of cells (p<0.05) and the [3H]-thymidine incorporation (p<0.05) in a dose-dependent manner. In contrast, the addition of particles decreased alkaline-phosphatase-specific activity and osteocalcin production. Collagen production and proteoglycan sulfation were also decreased, while prostaglandin-E2 synthesis was increased by the addition of particles. CONCLUSIONS: Ultra-high molecular weight polyethylene particles isolated from human tissue stimulated osteoblast proliferation and prostaglandin-E2 production and inhibited cell differentiation and matrix production. These results indicate that particles of wear debris inhibit cell functions associated with bone formation and that osteoblasts may produce factors in response to wear debris that influence neighboring cells, such as osteoclasts and macrophages. CLINICAL RELEVANCE: Particles of wear debris, especially ultra-high molecular weight polyethylene, have been implicated in the loosening of implants and the development of osteolysis. The present study shows that particles of ultra-high molecular weight polyethylene isolated from human tissue inhibit osteoblast functions associated with bone formation. In addition, particles of wear debris induced osteoblasts to secrete factors capable of influencing neighboring cells, such as osteoclasts and macrophages. These results suggest that osteoblasts may play a role in the cascade of events leading to granuloma formation, osteolysis, and failure of orthopaedic implants. 相似文献
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147.
Gao CL Dean RC Pinto A Mooneyhan R Connelly RR McLeod DG Srivastava S Moul JW 《The Journal of urology》1999,161(4):1070-1076
148.
Y Nakai W L Dean Y Hou N A Delamere 《Investigative ophthalmology & visual science》1999,40(7):1460-1466
PURPOSE: To determine whether dopamine receptor stimulation regulates Na,K-ATPase-mediated ion transport in cultured nonpigmented ciliary epithelium (NPE). METHODS: Using a rabbit NPE cell line, active Na-K transport activity was determined by measuring ouabain-sensitive potassium (86Rb) uptake in cell monolayers. Western blot analysis of membrane material obtained from cell homogenates was conducted to examine tyrosine phosphorylation of membrane proteins. RESULTS: Ouabain-sensitive potassium (86Rb) uptake was inhibited in the presence of either dopamine or the D1-selective agonist SKF82958. The response was suppressed by SCH23390, a D1 antagonist, but not by sulpiride, a D2-selective antagonist. Quinpirole, a D2-selective agonist, did not cause inhibition of ouabain-sensitive potassium (86Rb) uptake. Cyclic adenosine monophosphate (cAMP) was detectably increased in SKF82958-treated cells, although the concentration of SKF required to elevate cell cAMP was higher than the concentration needed to inhibit ouabain-sensitive potassium (86Rb) uptake. The protein kinase A inhibitor H89 prevented the 86Rb uptake response to SKF82958. Genistein, an inhibitor of tyrosine kinases, also prevented the 86Rb uptake response to SKF82958. Membrane material isolated from cells exposed to SKF82958 showed an increase in the density of several phosphotyrosine bands. These changes in phosphotyrosine immunoblot density were not observed in material isolated from cells that received either genistein or SCH23390 before SKF82958 treatment. CONCLUSIONS: The results of this study suggest D1 agonists cause a reduction of Na,K-ATPase-mediated ion transport by a mechanism that could involve a tyrosine kinase step. 相似文献
149.
PURPOSE: To determine whether sequence analysis of 16S ribosomal DNA (rDNA) can be used to detect bacterial pathogens in patients with postoperative endophthalmitis. METHODS: In 10 eyes of 10 patients, vitreous specimens were collected for culture and rDNA typing. Variable segments of each ribosomal DNA specimen were amplified by polymerase chain reaction (PCR), sequenced, and aligned by BLAST, a computer alignment program, against sequences in GenBank at the National Institutes of Health. RESULTS: Specimens were available from five eyes with bacterial endophthalmitis diagnosed by Gram stain or culture. Amplified 16s rDNA sequences from the eyes of three patients were identical to microbiologic results. Polymerase chain reaction results were negative in two cases in which unusual organisms were detected. All five control specimens from patients with nonbacterial endophthalmitis or uveitis were PCR negative. Approximately 48 to 72 hours are required under ideal conditions for final species identification with this ribosomal typing technique. CONCLUSIONS: 16S rDNA typing shows potential as a relatively rapid technique for identifying bacteria in vitreous samples. 相似文献
150.
Smith DE Kaye AD Feng CJ Kang B Kadowitz PJ Nossaman BD 《American journal of therapeutics》1996,3(4):314-320
Comparative effects of the nitric oxide (NO) synthase inhibitors, N(omega)-L-nitro-L-arginine methyl ester, and N(omega)-L-nitro-L-arginine benzyl ester on baseline arterial tone and on vasodilator responses to acetylcholine, isoproterenol, prostaglandin E(1), histamine, and nitroglycerin were investigated in the isolated mesenteric vascular bed of the rat. Under constant-flow conditions, intra-arterial (IA) injections of acetylcholine (100 ng--1 &mgr;g), isoproterenol (100 ng--1 &mgr;g), prostaglandin E(1) (0.3--3 &mgr;g), histamine (300 ng--3 &mgr;g), and nitroglycerin (100 ng--1 &mgr;g) caused dose-related decreases in mesenteric arterial perfusion pressure and decreases in systemic arterial pressure. Following administration of the NO synthase inhibitors, N(omega)-L-nitro-L-arginine methyl ester or N(omega)-L-nitro-L-arginine benzyl ester, mesenteric vascular resistance, and systemic arterial pressure were increased and mesenteric vasodilator responses to acetylcholine were significantly decreased, whereas N(omega)-L-nitro-L-arginine methyl ester and N(omega)-nitro-L-arginine benzyl ester did not significantly decrease vasodilator responses to nitroglycerin, histamine, isoproterenol, or prostaglandin E(1). The inhibitory effects of N(omega)-L-nitro-L-arginine methyl ester and N(omega)-L-nitro-L-arginine benzyl ester on vasodilator responses to acetylcholine suggest that acetylcholine-induced vasodilation in the mesenteric circulation of the rat is dependent on the release of NO from the endothelium. The increase in mesenteric vascular resistance following administration of N(omega)-L-nitro-L-arginine methyl ester and N(omega)-L-nitro-L-arginine benzyl ester suggest that tonic production of NO by the endothelium serves to maintain the mesenteric vascular bed of the rat in a dilated state. 相似文献