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An enhanced microdialysis method for neuropeptides is described and some prelimi­nary results of this novel approach are presented. The enhancement is achieved by adding a vehicle (solid support) to the perfusion fluid in order to increase the diffusion coefficient across the membrane and efficiently transport the analytes towards the de­tector. The microdialysis samples are desalted and then analyzed on an electrospray ionization orthogonal time-of-flight mass spectrometer. The preliminary results show major increase in signal when comparing this new approach of microdialysis with or­dinary microdialysis.
Drugs of abuse may cause acute as well as chronic damage to the nervous system, and a common mechanism of neurotoxicity is to induce disturbances in mitochondrial function. The mitochondrion is also an important source for cytotoxic reactive oxygen species (ROS). If the mitochondrial membrane potential (MMP) becomes depolarized, it can increase the production of ROS. This project has evaluated whether the fluorophore JC-1, which measures the depolarization of MMP, and the fluorophore H2DCFDA that oxidizes and produce fluorescence in the presence of oxygen radicals, are useful tools to screen for drug-induced neurotoxicity. The studies have been performed in embryonal carcinoma (EC) P19 cells that are pluripotent and upon retinoic acid (RA)-treatment will differentiate in culture into neurons, astrocytes and oligodendrocytes. In order to determine the predictive validity of the model/methods, a number of compounds known to cause oxidative stress and mitochondrial dysfunction have been examined (hydrogen peroxide, ionomycin, sodium azide). Main techniques employed culturing, induction and differentiation of neuronal cells, pharmacological dose-response experiments, detection and quantification of fluorescence using microplate reader and fluorescence microscopy, microplate-based colorimetric methods for assessment of cell viability, pharmacological/toxicological data and statistical analyses using the GraphPad prism software.
The presence of catecholamines in the nervous system of a plagiorchiid cercaria of Opisthioglyphe ranae (Frölich, 1791) is demonstrated using fluorescence methods (SPG method after De la Torre and Surgeon 1976, exciting filter of 400-410 nm max. wave length). Comparison of the distribution of catecholamines and acetylcholinesterases in the cercaria of the same species showed closely similar patterns. However, some differences in distribution were identified and these are discussed.
Galanin is a peptide present in the nervous system and peripheral tissues which exerts a broad range of physiological functions. The influence of centrally administered galanin (Gal; 100 pM i.c.v.) on arginine vasopressin (AVP) and oxytocin (OT) content in the hypothalamus and neurohypophysis as well as on their blood plasma concentration was estimated in male Wistar rats drinking ad libitum 2% solution of natrium chloride per 48 hours. In euhydrated rats and subsequently applied i.c.v. with Gal a significant fall in the hypothalamic and neurohypophysial content of OT but not AVP was observed, however, without simultaneous changes in these neurohormones blood plasma concentration. On the contrary, i.c.v. injection of Gal to salt-loaded rats caused a marked raise in AVP and OT level in the hypothalamus and neurohypophysis with subsequent diminution of both neurohormones concentration in blood plasma. These results suggest that in euhydrated rats Gal has an inhibitory influence on the biosynthesis as well as axonal transport of OT, but not AVP. On the contrary, in salt-loaded rats galanin restricts secretion of both neurohormones into the systemic circulation.
The nervous system of daughter sporocyst of D. pseudospathaceum is described on a basis of acetylcholinesterase activity, histofluorescence of biogenic amines and serotonin immunoreactivity. Description of the posterior part of the nervous system is added to the hitherto described brain ganglia and irregular network of nerve fibres surrounding anterior end of the sporocyst. The immunoreactivity of 5-HT was observed in laterally situated main nerve cords, which run along the whole length of the body and unite at its posterior end. Some ramifications of the main cords were also visible. A possible role of messenger molecules observed in daughter sporocyst nervous system is discussed.
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