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This survey provides a general discussion on drug residue analysis in biological material. Quality criteria for the use of thin layer chromatography in residue analysis have been evaluated and compared with those of other methods. The present application of TLC in residue analysis of antibacterials, growth promoters and neuroleptics has been discussed.
A simple and rapid method is described which allows analysis of Carazolol (ß-blocker agent) in tissue of pig by thin layer and liquid chromatography. The method utilizes an octadecyl solid phase column that selectively absorbs this drug and significantly improves sample clean-up procedure. The compound is eluted from the column with acidic acetonitrile. Analysis is performed using TLC for detection and qualitative determination and isocratic reversed-phase LC for confirmation and quantitation. The detection limit was 10 ng g-1 (TLC) and 2 ng g-1 (LC). The recoveries of Carazolol from spiked samples were above 80%. The procedure is suitable for routine residue analysis.
A simple and sensitive method for the determination of ß-agonists in animal tissue and urine using thin layer and liquid chromatography was prepared. The development of the method involves optimization of sample clean-up (solid-phase extraction) and chromatographic separation and detection conditions. The recovery of salbutamol and clenbuterol from tissue and urine samples were above 80% and the limit of detection was 1 ng g-1. The method has been used on a routine scale for residue control in samples from meat-producing animals.
The aim of this study was to demonstrate the bioequivalence of two formulations base on enrofloxacin i.e. between a test formulation Enrofloksacyna 10% per os (Biowet Drwalew) and the reference Baytril 10% (Bayer), according to a framwork required for the registration files. Both formulations were administered in a single oral administration at the recommended dose of 10 mg enrofloxacin per kg of b.w. in chickens. The study was conducted following conventional design, blood samples were collected until 72 h after drug application and analysed by validated method. The plasma disposition kinetics of enrofloxacin in chickens was similar following treatment with both formulations Cmax values (0.92 and 0.98 μg/ml, respectively) and were achieved at the 2nd h after administration of both formulations. None of the estimated pharmacokinetic parameters of both drug formulations were statistically different. Findings show that Enrofloksacyna 10% per os is equivalent to reference Baytril 10%.
Dendritic cells (DC) generated from human umbilical cord blood might replace patients' DC in attempts to elicit tumor-specific immune response in cancer patients. We studied the efficiency of transfection of human cord blood DC with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene, to test if nonviral gene transfer would be a method to load DC with protein antigens for immunotherapy purposes. Cord blood mononuclear cells were cultured in serum-free medium in the presence of granulocyte-monocyte colony stimulating factor (GM-CSF), stem cell factor (SCF) and Flt-3 ligand (FL), to generate DC from their precursors, and thereafter transfected by electroporation. Maturation of DC was induced by stimulation with GM-CSF, SCF, FL and phorbol myristate acetate (PMA). Transfected DC strongly expressed EGFP, but transfection efficiency of DC, defined as HLA-DR+ cells lacking lineage-specific markers, did not exceed 2.5%. Expression of the reporter gene was also demonstrated in the DC generated from transfected, purified CD34+ cord blood cells, by stimulation with GM-CSF, SCF, FL, and tumor necrosis factor α (TNF-α). Transfection of CD34+ cells was very efficient, but proliferation of the transfected cells was much reduced as compared to the untransfected cells. Therefore, the yield of transgene-expressing DC was relatively low. In conclusion, nonviral transfection of cord blood DC proved feasible, but considering the requirements for immunotherapy in cancer patients, transfection of differentiated DC or generation of DC from transfected hematopoietic stem cells provide only a limited number of DC expressing the transgene.
Tests performed in 2013 and 2014 revealed the occurrence of three tick species parasitizing pet cats and dogs in the Wrocław Agglomeration. In total, 1,455 tick specimens were removed from 931 hosts (760 dogs and 171 cats) in 18 veterinary clinics. The dominant tick species was Ixodes ricinus (n=1272; 87.4%), followed by I. hexagonus (n=137; 9.4%) and Dermacentor reticulatus (n=46; 3.2%). Females were the most often collected development stage among I. ricinus and D. reticulatus, and nymphs among I. hexagonus. Additionally, D. reticulatus ticks (n=337) were then collected from vegetation in the Wrocław area to detect Babesia canis; however, none was found positive. Only 9.0% of dog blood samples sent to VETLAB were positive for Babesia spp. Negative results for B. canis from ticks may result from the short period of the occurrence of D. reticulatus in the Wrocław area and therefore the vectorpathogen cycle may not have been fully established at the time of the study. Nevertheless, D. reticulatus is expanding its range, and the size of its population in the Wrocław Agglomeration is increasing. The presence of the pathogenic Babesia spp. combined with the occurrence of its main vector¸ D. reticulatus, suggests that the epizootiological situation in the area can change and may pose a new veterinary problem in the future.
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