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261.
262.
Modulation of Ganglioside Biosynthesis in Primary Cultured Neurons 总被引:11,自引:4,他引:7
Murine cerebellar cells were pulse labeled with [14C]galactose, and the incorporation of radioactivity into gangliosides and neutral glycosphingolipids was examined under different experimental conditions. In the presence of drugs affecting intracellular membrane flow, as well as at 15 degrees C, labeled GlcCer was found to accumulate in the cells, whereas the labeling of higher glycosphingolipids and gangliosides was reduced. Monensin and modulators of the cytoskeleton effectively blocked biosynthesis of the complex gangliosides GM1, GD1a, GD1b, GT1b, and GQ1b, whereas incorporation of radioactivity into neutral glycosphingolipids, such as glucosylceramide and lactosylceramide, as well as GM3, GM2, and GD3 was either increased or unaltered. As monensin has been reported to interfere with the flow of molecules from the cis to the trans stacks of the Golgi apparatus, this result highlights at least one subcompartmentalization of ganglioside biosynthesis within the Golgi system. Inhibitors of energy metabolism affected, predominantly, the biosynthesis of the b-series gangliosides, whereas a reduced temperature (15 degrees C) more effectively blocked incorporation of radiolabel into the a-series gangliosides, a result suggesting the importance of GM3, as the principal branching point, for the regulation of ganglioside biosynthesis. 相似文献
263.
We studied the hexose transporter protein of the frontal and temporal neocortex, hippocampus, putamen, cerebellum, and cerebral microvessels (which constitute the blood-brain barrier) in Alzheimer disease and control subjects by reversible and covalent binding with [3H]cytochalasin B and by immunological reactivity. In Alzheimer disease subjects, we found a marked decrease in the hexose transporter in brain microvessels and in the cerebral neocortex and hippocampus, regions that are most affected in Alzheimer disease, but there were no abnormalities in the putamen or cerebellum. Hexose transporter reduction in cerebral microvessels of Alzheimer subjects is relatively specific because other enzyme markers of brain endothelium were not significantly altered. The low density of the hexose transporter at the blood-brain barrier and in the cerebral cortex in Alzheimer disease may be related to decreased in vivo measurements of cerebral oxidative metabolism. 相似文献
264.
The possibility that an increased intracellular concentration of cyclic AMP (cAMP) can regulate the extent of muscarinic receptor-stimulated phosphoinositide (PPI) turnover in the human neuroblastoma cell line SK-N-SH was examined. Addition of either forskolin (or its water-soluble analog, L-85,8051), theophylline, isobutylmethylxanthine, or cholera toxin, agents that interact with either the catalytic unit of adenylate cyclase, cAMP phosphodiesterase, or the guanine nucleotide binding protein linked to adenylate cyclase activation, resulted in a 45-181% increase in cAMP concentration and a 27-70% inhibition of carbachol-stimulated inositol phosphate release. Through the use of digitonin-permeabilized cells, the site of inhibition was localized to a step at, or distal to, the guanine nucleotide binding protein that regulates phospholipase C activity. In contrast, when intact SK-N-SH cells were exposed to prostaglandin E1, the ensuing increases in cAMP were not accompanied by an inhibition of stimulated PPI turnover. These differential effects of increased cAMP concentrations on stimulated PPI turnover may reflect the compartmentation of cAMP within SK-N-SH cells. 相似文献
265.
The biomechanical events which accompany functional loading of the human mandible are not fully understood. The techniques normally used to record them are highly invasive. Computer modelling offers a promising alternative approach in this regard, with the additional ability to predict regional stresses and strains in inaccessible locations. In this study, we built two three-dimensional finite element (FE) models of a human mandible reconstructed from tomographs of a dry dentate jaw. The first model was used for a complete mechanical characterization of physical events. It also provided comparative data for the second model, which had an increased vertical corpus depth. In both cases, boundary conditions included rigid restraints at the first right molar and endosteal cortical surfaces of the articular eminences of temporal bones. Groups of parallel multiple vectors simulated individual masticatory muscle loads. The models were solved for displacements, stresses, strains, and forces. The simulated muscle loads in the first model deformed the mandible helically upward and toward its right (working) side. The highest principal stresses occurred at the bite point, anterior aspects of the coronoid processes, symphyseal region, and right and left sides of the mandibular corpus. In general, the observed principal stresses and strains were highest on the periosteal cortical surface and alveolar bone. At the symphyseal region, maximum principal stresses and strains were highest on the lower lingual mandibular aspect, whereas minimum principal stresses and strains were highest on its upper labial side. Subcondylar principal strains and condylar forces were higher on the left (balancing or nonbiting) side than on the right mandibular side, with condylar forces more concentrated on the anteromedial aspect of the working-side condyle and on the central and lateral aspects of the left. When compared with in vivo strain data from macaques during comparable biting events, the predictive strain values from the first model were qualitatively similar. In the second model, the reduced tensile stress on the working-side, and decreased shear stress bilaterally, confirmed that lower stresses occurred on the lower mandibular border with increased jaw depth. Our results suggested that although the mandible behaved in a beam-like manner, its corpus acted more like a combination of open and closed cross sections due to the presence of tooth sockets, at least for the task modelled.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
266.
The "squeeze" form of power grip is investigated for the purposes of clarifying the hand posture and activities associated with the grip, assessing the potential in chimpanzees for using the grip, and identifying morphological correlates of an effective power grip that may be recognized in fossil hominid species. Our approaches include: (1) the analysis of the human grip, focusing on both the hand posture involved and hand movements associated with use of the grip in hammering; (2) the analysis of similar chimpanzee grips and associated movements; (3) comparative functional analysis of regions in the hand exploited and stressed by the grip and its associated movements in humans; and (4) a review of the literature on the power grip and its morphological correlates. Results of the study indicate that humans use a squeeze form of power grip effectively to wield cylindrical tools forcefully as extensions of the forearm. Several morphological features occur in high frequency among humans which facilitate the grip and are consistent with the large internal and external forces associated with it in hammering and in other tool-using activities. Chimpanzee hand postures resembling this form of human power grip are not fully comparable and lack some of these morphological correlates that facilitate its use. The hand of Australopithecus afarensis does not appear to have been stressed by use of the grip, but there is some evidence for this type of stress in the metacarpals from Sterkfontein Member 4. Hands from Olduvai and Swartkrans do not provide sufficient evidence for assessment of power grip capabilities. 相似文献
267.
Occlusal morphology of permanent dentitions in 29 men with a 47,XXY chromosome complement (Klinefelter syndrome) was determined from dental casts. The results showed that a relatively frequent occlusal anomaly was mesial molar occlusion. Incisal open bite was also more common than in controls. Based on the present and previous observations of occlusal anomalies in various sex chromosome anomaly groups and normal controls, it is suggested that the presence of the Y chromosome in the genome is at least as important as the X chromosome for the development of harmonious occlusal morphology. The tendency towards sexual dimorphism in occlusal phenotype might result from a differential effect of the X and Y chromosomes on cellular activity which leads to different growth patterns. 相似文献
268.
The effect of loading renal tubule cells with cystine was studied by incubating them with cystine dimethylester. Proline uptake into brushborder membrane vesicles isolated from the cystine loaded cells was not different from that observed into brushborder vesicles isolated from tubules incubated in buffer alone. Incubating brushborder membranes with 2 mM cystine dimethylester for 10 minutes reduced the uptake of proline by 27% after 15 seconds of incubation and by 21% after 60 seconds of incubation. There was no effect after 20 minutes of incubation. Pre-incubating brushborder membrane vesicles with cystine dimethylester had no statistically significant effect on the affinity of priline for the carrier, but did reduce the maximal rate of proline uptake by 49%. 相似文献
269.
The membrane-bound photooxidizable cytochrome c-554 from Chloroflexus aurantiacus has been purified. The purified protein runs as a single heme staining band on SDS-PAGE with an apparent molecular mass of 43 000 daltons. An extinction coefficient of 28 ± 1 mM–1 cm–1 per heme at 554 nm was found for the dithionite-reduced protein. The potentiometric titration of the hemes takes place over an extended range, showing clearly that the protein does not contain a single heme in a well-defined site. The titration can be fit to a Nernst curve with midpoint potentials at 0, +120, +220 and +300 mV vs the standard hydrogen electrode. Pyridine hemochrome analysis combined with a Lowry protein assay and the SDS-PAGE molecular weight indicates that there are a minimum of three, and probably four hemes per peptide. Amino acid analysis shows 5 histidine residues and 29% hydrophobic residues in the protein. This cytochrome appears to be functionally similar to the bound cytochrome from Rhodopseudomonas viridis. Both cytochrome c-554 from C. aurantiacus and the four-heme cytochrome c-558-553 from R. viridis appear to act as direct electron donors to the special bacteriochlorophyll pair of the photosynthetic reaction center. They have a similar content of hydrophobic amino acids, but differ in isoelectric point, thermodynamic characteristics, spectral properties, and in their ability to be photooxidized at low temperature.Abbreviations LDAO
lauryl dimethyl amine-N-oxide
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- mV
millivolt
- Em.8
midpoint potential at pH 8.0
- ODV
optical density x volume in ml 相似文献
270.
In order to gain further support for the concept that a homo-oligomeric protein-complex may be sufficient to form a functional ligand-activated ion channel and to explore additional possibilities for the reconstitution of channel activity, a single polypeptide band of the purified neuronal AChR from insects has been electroeluted from SDS-polyacrylamide gels, the SDS removed and the polypeptides incorporated into liposomes. Liposomes were fused into planar lipid bilayers which were subsequently analysed for channel activity. Fluctuations of cation-channels were detected after addition of agonists (carbamylcholine); channel activity was blocked by antagonists (d-tubocurarine). The channels formed by electroeluted polypeptides gave conductance values, as well as kinetic data, quite similar to channels formed by the native receptor protein. Sedimentation experiments using sucrose density gradient centrifugation revealed that a considerable portion of the electroeluted polypeptides assembled during the reconstitution process to form oligomeric complexes with a sedimentation coefficient of about 10 S; thus resembling the native receptor complex.
Offprint requests to: W. Hanke 相似文献