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A liquid chromatographic method coupled with UV-Vis and fluorescence detectors for the determination of azaperone, carazolol, and chlorpromazine residues in urine has been described. Analytes were isolated from biological samples with acetonitrvle. The obtained extracts were applied to solid phase extraction Strata X-C 33 µn Polimeric Strong Cation columns. The analytes were eluted with alkalised acetonitrvle. The whole procedure was validated according to European Decision 657/2002/EC. The detection limits were established at the level 2 µg/L for Carazolol and 5 µg/L for azaperone and chlorpromazine. Detection capabilities for carazolol, azaperone, and chlorpromazine were 8.62, 18.9, and 6.53 µg/L, respectively. Recoveries were above 90%.
The involvement of the clathrin-mediated endocytic internalization route in the uptake of cholera toxin (CT) was investigated using different cell lines, including the human intestinal Caco-2 and T84 cell lines, green monkey Vero cells, SH-SY5Y neuroblastoma cells and Madin-Darby canine kidney cells. Suppression of the clathrin-mediated endocytic pathway by classical biochemical procedures, like intracellular acidification and potassium depletion, inhibited cholera toxin up take by up to about 50% as well as its ability to raise intracellular levels of cAMP. Also prior exposure of these cell types to the cationic amphiphilic drug chlorpromazine reduced the functional uptake of cholera toxin, even to a greater extent. These effects were dose- and cell type-dependent, suggesting an involvement of clathrin-mediated endocytosis in the functional uptake of cholera toxin. For a more straightforward approach to study the role of the clathrin-mediated uptake in the internalization of cholera toxin, a Caco-2eps-cell line was exploited. These Caco-2eps-cells constitutively suppress the expression of epsin, an essential accessory protein of clathrin-mediated endocytosis, thereby selectively blocking this internalization route. CT uptake was found to be reduced by over 60% in Caco-2eps-paralleled by a diminished ability of CT to raise the level of cAMP. The data presented suggest that the clathrin-mediated uptake route fulfils an important role in the functional internalization of cholera toxin in several cell types.
The use of veterinary medicines and medicated feed has a potential for the exposure of agricultural workers to pharmaceuticals with phototoxic and photoallergic sideeffects. We present a 67-year-old self-employed farmer and pig breeder with a 22-year history of severe persistent photosensitivity following photoallergic contact dermatitis due to direct occupational dermal and airborne contact to chlorpromazine (sedative) and olaquindox (antibiotic and animal growth promoter, AGP). His fi rst dermatitis symptoms appeared at the age of 45 when the pig breeding was intensifi ed. He showed erythematous, scaly, and pruritic plaques localized symmetrically on the sun-exposed backs of his hands, fi ngers, and forearms, spreading to his face and other sun-exposed body sites. Without protective measures, he injected the animals with chlorpromazine. Besides, for several years he mixed by hand a powder containing olaquindox into the pigs’ dry food. Epicutaneous and photo-patch tests showed positive reactions to promethazine, chlorpromazine, and olaquindox. In spite of the complete avoidance of the identifi ed photoallergens for several years, his life is still extremely disabled due to the persistent photosensitivity. Our case report stresses the observation that olaquindox and chlorpromazine as phototoxic agents and photoallergens are capable of inducing a persistent and severe photosensitivity for many years, even after termination of exposure. Although the use of phenothiazine derivates and APGs for animals has meanwhile been banned in the European Union (EU), AGPs are still widely used in Asia. Physicians, especially occupational physicians, should be still aware of these phototoxic and photoallergic agents to reduce the burden of skin disease at work.
The effect of chlorpromazine (CPZ) on the neurosecretory action of the hypothalamo-hypophyseal system was investigated in 72 male rats. The experimental animals received CPZ in a dose of 0.4 mg, 4.0 mg and 20.0 mg/kg b.w. for 30 days. The rats were sacrificed by decapitation at 24 h and 7 days after the last dose of the drug. The neurosecretory material was stained with paraldehyde fuchsin in the supraoptic nucleus, paraventricular nucleus, eminentia mediana and neurohypophysis, the tigroid was stained with toluidine blue and the acid phosphatase activity was evaluated histoenzymatically. It was found that CPZ reduced the content of the neurosecretory material after 24 h, while an increase was observed 7 days after the last drug administration.
Photopheresis (ECP) is an immunomodulatory therapy that involves extracorporeal exposure of isolated peripheral blood leukocytes to UVA irradiation in the presence of 8-methoxypsoralen (8-MOP) followed by their reinfusion to the patient. However, the underlying mechanism of ECP is not well understood yet. We selected 8-methoxypsoralen (8-MOP), chlorpromazine (CPZ) and 4,6,4'-trimethylangelicine (TMA) because of differences in their ability to induce immune suppression in rats in vivo. In this study, we investigated the role of UVA irradiation of lymphocytes in the presence of TMA, CPZ or 8-MOP on cell apoptosis, and their impact on adhesion of lymphocytes to monocytes in vitro. Apoptosis of lymphocytes and their sub-populations (lymphocytes T and B, NK cells) were determined by a flow cytometry, using AnnexinV-FITC, TUNEL assay and DNA content analysis and antibodies CD3, CD56, CD19. Mitochondrial potential was measured using CMXRos staining and the interaction of monocytes with lymphocytes was monitored by PKH26 Red Sigma staining of lymphocytes and subsequent use of flow cytometry. Our results show a significant increase of apoptosis of the photochemically treated lymphocytes and a decrease of their mitochondrial potential that depended on the dose and time after the treatment. Our data also reveal an increased recognition of apoptotic lymphocytes by freshly isolated monocytes.
Multidrug resistance (MDR) of cancer cells poses a serious obstacle to successful chemotherapy. The overexpression of multispecific ATP-binding cassette transporters appears to be the main mechanism of MDR. A search for MDR-reversing agents able to sensitize resistant cells to chemotherapy is ongoing in the hope of their possible clinical use. Studies of MDR modulators, although they have not produced clinically beneficial effects yet, may greatly enrich our knowledge about MDR transporters, their specificity and mechanism of action, especially substrate and/or inhibitor recognition. In the present review, interactions of three groups of modulators: phenothiazines, flavonoids and stilbenes with both P-glycoprotein and MRP1 are discussed. Each group of compounds is likely to interact with the MDR transporters by a different mechanism. Phenothiazines probably interact with drug binding sites, but they also could indirectly affect the transporter's activity by perturbing lipid bilayers. Flavonoids mainly interact with ABC proteins within their nucleotide-binding domains, though the more hydrophobic flavonoids may bind to regions within transmembrane domains. The possible mechanism of MDR reversal by stilbenes may result from their direct interaction with the transporter (possibly within substrate recognition sites) but some indirect effects such as stilbene-induced changes in gene expression pattern and in apoptotic pathways should also be considered. Literature data as well as some of our recent results are discussed. Special emphasis is put on cases when the interactions of a given compound with both P-glycoprotein and MRP1 have been studied simultaneously.
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