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71.
Effects of medium perfusion on matrix production by bovine chondrocytes in three-dimensional collagen sponges 总被引:6,自引:0,他引:6
Various culture systems have been used for examining the anabolic and catabolic functions of isolated chondrocytes as well as for tissue engineering purposes. Perfusion or frequent medium change is beneficial for three-dimensional (3D) cultures of many cell types. In this study, bovine articular chondrocytes (bACs) were grown in 3D collagen sponges with or without medium perfusion (0.33 mL/min) for up to 15 days. The influence of medium perfusion was evaluated using markers of cartilage matrix accumulation, synthesis, and gene expression. Metachromatic matrix, collagen type II, and hyaluronan accumulated around the cells within the collagen sponges. Sulfated glycosaminoglycans (S-GAGs) that accumulated in the sponge exposed to nonperfused control were 130% of that in the perfused sponge at day 7. S-GAG accumulation after 15 days in the nonperfused control was 230% more than at day 7 (p < 0.01). (35)S-sulfate incorporation during the final 18 h of culture in the sponge exposed to nonperfusion was 180% greater than that in the perfused sponge (p < 0.01). Quantitative analyses show that at day 7, aggrecan and collagen type II gene expression were 350% and 240% greater, respectively, in the nonperfused culture than in the perfused one. These results indicate that perfused conditions that are beneficial for other cell types inhibit chondrogenesis by articular chondrocytes in 3D culture. 相似文献
72.
73.
Moore DJ Zhang L Troncoso J Lee MK Hattori N Mizuno Y Dawson TM Dawson VL 《Human molecular genetics》2005,14(1):71-84
The identification of rare monogenic forms of Parkinson's disease (PD) has provided tremendous insight into the molecular pathogenesis of this disorder. Heritable mutations in alpha-synuclein, parkin, DJ-1 and PINK1 cause familial forms of PD. In the more common sporadic form of PD, oxidative stress and derangements in mitochondrial complex-I function are considered to play a prominent role in disease pathogenesis. However, the relationship of DJ-1 with other PD-linked genes and oxidative stress has not been explored. Here, we show that pathogenic mutant forms of DJ-1 specifically but differentially associate with parkin, an E3 ubiquitin ligase. Chemical cross-linking shows that pathogenic DJ-1 mutants exhibit impairments in homo-dimer formation, suggesting that parkin may bind to monomeric DJ-1. Parkin fails to specifically ubiquitinate and enhance the degradation of L166P and M26I mutant DJ-1, but instead promotes their stability in cultured cells. The interaction of parkin with L166P DJ-1 may involve a larger protein complex that contains CHIP and Hsp70, perhaps accounting for the lack of parkin-mediated ubiquitination. Oxidative stress also promotes an interaction between DJ-1 and parkin, but this does not result in the ubiquitination or degradation of DJ-1. Parkin-mediated alterations in DJ-1 protein stability may be pathogenically relevant as DJ-1 levels are dramatically increased in the detergent-insoluble fraction from sporadic PD/DLB brains, but are reduced in the insoluble fraction from parkin-linked autosomal recessive juvenile-onset PD brains. These data potentially link DJ-1 and parkin in a common molecular pathway at multiple levels that may have important implications for understanding the pathogenesis of inherited and sporadic PD. 相似文献
74.
75.
Watanabe Y Hashimoto S Kakita A Takahashi H Ko J Mizuno M Someya T Patterson PH Nawa H 《Neuroscience research》2004,48(3):345-353
Cytokines have been implicated in the etiology or pathology of various psychiatric diseases of developmental origin such as autism and schizophrenia. Leukemia inhibitory factor (LIF) is induced by a variety of brain insults and known to have many influences on mature and immature nervous system. Here, we assessed the neurobehavioral and pathological consequences of peripheral administration of LIF in newborn rats. Subcutaneous LIF injection induced STAT3 phosphorylation in many brain regions and increased glial fibrillary acidic protein (GFAP) immunoreactivity in the neocortex, suggesting that LIF had direct effects in the central nervous system. The LIF-treated rats displayed decreased motor activity during juvenile stages, and developed abnormal prepulse inhibition in the acoustic startle test during and after adolescence. They displayed normal learning ability in active avoidance test, however. Brain neuronal structures and startle responses were grossly normal, except for the cortical astrogliosis during neonatal LIF administration. These results indicate that LIF induction in the periphery of the infant has a significant, but discrete impact on neurobehavioral development. 相似文献
76.
Shimamura K Zhou M Toba M Kimura S Higuchi T Kawaguchi H Sekiguchi F Sunano S 《Pflügers Archiv : European journal of physiology》2003,446(1):30-35
The effects of L-arginine on spontaneous contraction of endothelium-denuded longitudinal preparations of the rat portal vein were studied. L-arginine increased the frequency of spontaneous contraction concentration-dependently between 10 microM and 1 mM. Changes in contraction amplitude and duration were not remarkable. D-arginine had a negligible effect on spontaneous contraction. N(omega)-nitro-L-arginine (1 mM) did not affect spontaneous contraction or the response to L-arginine. Addition of N(G)-monomethyl-L-arginine (1 mM), l-lysine (1 mM) or N-ethymaleimide (0.1 mM) increased the frequency of spontaneous contractions and inhibited the effect of L-arginine. Glibenclamide (10 microM) did not affect spontaneous contraction or the response to L-arginine. Spontaneous increase in concentration of intracellular Ca2+, estimated as the ratio of Fura-PE3 fluorescence occurred synchronously with spontaneous contraction. Spontaneous increase in concentration of intracellular Ca2+ occurred more frequently in the presence of L-arginine (1 mM). L-arginine (1 mM) also increased the number of action potential bursts/min in the longitudinal smooth muscle layer. L-arginine (1 mM) also depolarized cell membranes. This study indicates that L-arginine increases the frequency of spontaneous contraction of longitudinal muscle in the rat portal vein by membrane depolarization through mechanisms that do not involve nitric oxide or the inhibition of ATP-sensitive K+ channels. 相似文献
77.
Tanaka R Komine-Kobayashi M Mochizuki H Yamada M Furuya T Migita M Shimada T Mizuno Y Urabe T 《Neuroscience》2003,117(3):531-539
Brain ischemia induces a marked response of resident microglia and hematopoietic cells including monocytes/macrophages. The present study was designed to assess the distribution of microglia/macrophages in cerebral ischemia using bone marrow chimera mice known to express enhanced green fluorescent protein (EGFP). At 24 h after middle cerebral artery occlusion (MCAO), many round-shaped EGFP-positive cells migrated to the ischemic core and peri-infarct area. At 48-72 h after MCAO, irregular round- or oval-shaped EGFP/ionized calcium-binding adapter molecule 1 (Iba 1)-positive cells increased in the transition zone, while many amoeboid-shaped or large-cell-body EGFP/Iba 1-positive cells were increased in number in the innermost area of ischemia. At 7 days after MCAO, many process-bearing ramified shaped EGFP/Iba 1-positive cells were detected in the transition to the peri-infarct area, while phagocytic cells were distributed in the transition to the core area of the infarction. The distribution of these morphologically variable EGFP/Iba 1-positive cells was similar up to 14 days from MCAO. The present study directly showed the migration and distribution of bone marrow-derived monocytes/macrophages and the relationship between resident microglia and infiltrated hematogenous element in ischemic mouse brain. It is important to study the distribution of intrinsic and extrinsic microglia/macrophage in ischemic brain, since such findings may allow the design of appropriate gene-delivery system using exogenous microglia/macrophages to the ischemic brain area. 相似文献
78.
Effects of mandibular advancement on supine airway size in normal subjects during sleep 总被引:2,自引:0,他引:2
STUDY OBJECTIVES: To examine changes in the upper-airway dimension and its surrounding structures induced by mandibular advancement during sleep. DESIGN: Eleven nonapneic adult males participated in the study. A set of supine lateral cephalograms was taken for each subject at the end of expiration during stage 1 and 2 non-rapid-eye-movement sleep with and without a Klearway appliance (Great Lakes Orthodontics, NY, USA), which was adjusted to 67% of the maximum protrusion position. The Wilcoxon signed-rank test was used to compare changes in the anteroposterior width of the upper airway and the positions of the hyoid bone and third cervical vertebra with and without the appliance. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The amount of jaw opening was significantly increased by wearing the titratable oral appliance, and the mandibular symphysis moved backward. The sagittal dimension of the superior pharyngeal airway was significantly increased; however, no significant changes were found in the middle and inferior pharyngeal airway. Significant posterior displacement of the hyoid bone and third cervical vertebra was seen. Moreover, significant inferior displacement of the hyoid bone was also seen. The relationship among the mandibular symphysis, the hyoid bone, and the third cervical vertebra remained constant. CONCLUSIONS: Mandibular advancement significantly increases the size of the upper airway in the velopharynx and results in posteroinferior displacement of the hyoid bone and posterior displacement of the third cervical vertebra during sleep. 相似文献
79.
80.
Yamamoto T Matsushita T Marui T Mizuno K Minami R Hayashi Y Hanioka K 《Pathology international》2000,50(7):558-561
We describe a rare case of dedifferentiated liposarcoma with features resembling chondroblastic osteosarcoma in the dedifferentiated component. The tumor was removed from the left thigh in a 78-year-old male. It consisted of a well-differentiated liposarcoma and an anaplastic component that contained numerous osteoid and cartilaginous tissues surrounded by high-grade spindle cell sarcoma. To our knowledge, only two cases similar to the divergent chondroblastic osteosarcomatous dedifferentiation of this disease have been reported in the literature. 相似文献