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81.
Główka FK Karaźniewicz M Lipnicka E 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2006,839(1-2):54-61
A new indirect RP-HPLC method was developed for determination of small, ng/ml, concentrations of triamcinolone (TMC) in human plasma, in presence of endogenous corticosteroids: cortisol (hydrocortisone, F), cortisone (E) and their metabolites, after administration of TMC in a free alcohol form. After solid phase extraction (SPE) in cartridges with octadecyl phase, TMC and prednisolone (I.S.) were derivatized by treatment with 9-anthroyl nitrile (9-AN) in a basic mixture, consisting of triethanolamine and quinuclidine, to receive fluorescent esters at 21-hydroxyl group of the steroid chain. Optimal conditions were also established to purify fluorescent TMC and I.S. derivatives before injection into HPLC column. The fluorescent esters were determined using an isocratic RP-HPLC technique in a C18 column. The mobile phase consisted of acetonitrile and 0.3 mM ortho-phoshoric acid. The method was validated before using to pharmacokinetic studies. Calibration curve of TMC was linear in the range of 2.5-100.0 ng/ml. Intra- and inter-day measurement precision and accuracy were equal to or lower than 15%. Percent recovery, and limits of detection (LOD) and quantification (LOQ) of TMC were also determined. The method was applied for in vivo conditions after administration of tablets containing TMC to healthy volunteers. Moreover, the method provided potential to determine TMC and, simultaneously, other glucocorticoids: E, F and their metabolites in one analytical run. Column interactions were observed between endogenous metabolites of E. Usefulness of the elaborated method was confirmed in pharmacokinetic studies following administration of a small (4 mg) dose of TMC to human volunteers. The method can provide an alternative to HPLC coupled with RIA in determination of small quantities of TMC. 相似文献
82.
F Burdan Z Siezieniewska R Maciejewski K Burski Z Wójtowicz 《Journal of physiology and pharmacology》2000,51(3):463-470
Omeprazole is one of the substituted benzimidazoles, which is not free of side effects. The aim of the study was to evaluate the influence of omeprazole therapy on pancreas. Omeprazole was administered intraperitoneally, twice a day, for 3 days to the male rats in 0.571 mg/kg b.w. and 5.71 mg/kg b.w. doses. Half of animals were sacrificed in the 4th day of the experiment. The remaining rats were raised for another 6 weeks, without any xenobiotics, and sacrificed on the 47th day. The activity of acid phosphatase, beta-galactosidase, cathepsin B, and L, lipase, N-acetyl-glucosaminidase, and sulphatase was evaluated. The slides of the pancreas were examined in light microcopy in hematoxylin-eosin, asan, periodic acid-Schiff (paS) stains. Statistical increase in total activities of acid phosphatase, beta-galactosidase, lipase, N-acetyl-glucosaminidase, sulphatase, and acute inflammatory infiltration in peripancreatic fat tissue without histological pancreas impairment, were observed after the higher dose on the 4th day of experiment. Histological picture and enzymatic profiles were normalized during the next 6 weeks. We concluded that intraperitoneal administration of omeprazole causes tissue inflammation in the peripancreatic lipid tissue and reactive elevation of some pancreatic lysosomal enzymes. 相似文献
83.
R Maciejewski F Burdan B Madej G Ki? P Szkodziak K Burski 《Acta physiologica Hungarica》1997,85(4):335-345
The aim of this study was to establish and quantify the changes of the level of cathepsin B, D and L in the spleen during experimental pancreatitis. The experiment was carried out in 115 male Wistar rats, randomly divided into three groups: intact (n = 15), injected with 0.9% NaCl solution into the common bile pancreatic duct (n = 50) and injected with 5% sodium taurocholate into this duct to induce acute pancreatitis (n = 50). After 2, 6, 12, 24 and 48 hours rats were anaesthetised, and blood was taken for amylase determination from the heart, and the spleen was removed. Alpha-amylase level in the blood serum samples was measured by enzymatic method. Cathepsin activity was established by spectrophotometric methods using substrates which form coloured complexes when they react with these proteases. The specific free fraction activity of cathepsin B, D and L in the spleen changed during the course of experiment, but there was no correlation between their activity and the intensity of pancreatitis established by serum amylase level. 相似文献
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Justyna Bobrowska Joanna Pabijan Joanna Wiltowska-Zuber Benedykt R. Jany Franciszek Krok Kamil Awsiuk Jakub Rysz Andrzej Budkowski Malgorzata Lekka 《Analytical biochemistry》2016
There are several techniques like time of flight secondary ion mass spectrometry (ToF SIMS) that require a special protocol for preparation of biological samples, in particular, those containing single cells due to high vacuum conditions that must be kept during the experiment. Frequently, preparation methodology involves liquid nitrogen freezing what is not always convenient. In our studies, we propose and validate a protocol for preparation of single cells. It consists of four steps: (i) paraformaldehyde fixation, (ii) salt removal, (iii) dehydrating, and (iv) sample drying under ambient conditions. The protocol was applied to samples with single melanoma cells i.e. WM115 and WM266-4 characterized by similar morphology. The surface and internal structures of cells were monitored using atomic force, scanning electron and fluorescent microscopes, used to follow any potential protocol-induced alterations. To validate the proposed methodology for sample preparation, ToF SIMS experiments were carried out using C60+ cluster ion beam. The applied principal component analysis (PCA) revealed that chemical changes on cell surface of melanoma cells were large enough to differentiate between primary and secondary tumor sites. 相似文献
87.
Iwona Żur Ewa Dubas Elżbieta Golemiec Magdalena Szechyńska-Hebda Franciszek Janowiak Maria Wędzony 《Plant Cell, Tissue and Organ Culture》2008,94(3):319-328
The accumulation of abscisic acid (ABA) and the activities of antioxidative enzymes along with cell metabolic activity were
monitored during androgenesis induction in triticale (×Triticosecale Wittm.). Tested cultivars ‘Mieszko’ and ‘Wanad’ were selected due to their significantly different responses to androgenic
induction. Significant variation was observed in respect of superoxide dismutase activity and endogenous ABA content in anthers
isolated from freshly cut tillers. For both cultivars, tillers pretreatment with low temperature decreased peroxidase activity
by 36%, highly accelerated respiration rate and reduced heat production. At the same time, the level of ABA in ‘Mieszko’ was
increased to the level measured in ‘Wanad’. This effect was associated with higher microspore culture viability and increased
stress tolerance in ‘Mieszko’. Low temperature and metabolic starvation during 4-day anther preculture did not influence activities
of antioxidative enzymes, while it resulted in slight decrease in respiration rate and heat emission. The importance of these
changes for effective androgenesis induction is discussed. 相似文献