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Optimization of biologic therapy in children with inflammatory bowel disease (IBD) using modern biomarkers
Ohem, Jan ; Bronský, Jiří (advisor) ; Kotaška, Karel (referee) ; Bajerová, Kateřina (referee)
Optimization of biologic therapy in children with inflammatory bowel disease (IBD) using modern biomarkers Abstract to thesis Study programme: Biochemistry and Pathobiochemistry Introduction: In adults, infliximab (IFX) levels correlate with disease activity and antibodies to IFX (ATIs) predict treatment failure. We aimed to determine the association of IFX levels and ATIs with disease activity in paediatric population. Methods: This study was performed as a prospective observational study. We prospectively collected blood, stool, and clinical data from 65 patients (age 10.5-15.1 years) with Crohn's disease (CD) before IFX administration, and measured IFX trough levels, ATIs, and faecal calprotectin levels (CPT). We used multivariate analysis to identify the predictors of IFX levels. IFX and ATIs levels were meassured using ELISA. Results: Lower levels of IFX were associated with ATIs positivity (OR [odds ratio] 0.027, CI [confidence interval] 0.009-0.077). Higher C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR) and CPT levels were found in patients with lower IFX levels. The optimal combination of specificity (50%) and sensitivity (74%) for disease activity was calculated for IFX levels ≥ 1.1 µg/ml using CRP level < 5 mg/l as a marker of laboratory remission. In a model that used CPT ≤...
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Determination of vancomycin and its crystalline degradation products by HPLC
Neščáková, Monika ; Klapková, Eva (advisor) ; Kotaška, Karel (referee)
The aim of this bachelor thesis was to introduce and validate the method for the determination of vancomycin and its crystalline degradation products by high performance liquid chromatography with detection by diode-array detector. This method is able to reliably detect and distinguish the crystalline degradation products of vancomycin, which lack the necessary therapeutic effect. The long-term using of vancomycin or renal insufficiency lead to their accumulation in the patient's body. Another target of this bachelor thesis was to monitor the release profile of vancomycin concentration and its crystalline degradation products from local carrier of antibiotics that are used in some complicated orthopedic surgeries such as prevention of inflammatory infections caused by Staphylococcus aureus. We compared in vitro release of vancomycin from two bone grafts, two different bone cements and from one synthetic bone graft. The aim was to determine if the examine material releases the quantity of vancomycin, which ensure the long-term local concentrations of vancomycin higher than is the MIC of vancomycin-resistant Staphylococcus aureus (VRSA). Powered by TCPDF (www.tcpdf.org)
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Determination of selenium and manganese in cerebrospinal fluid by atomic absorption spectrometry
Jiráková, Lucie ; Kotaška, Karel (advisor) ; Klapková, Eva (referee)
The aim of this study was to confirm the use of electrothermal atomic absorption spectrometry for the determination of selenium and manganese in the cerebrospinal fluid and to investigate concentrations of selenium and manganese in selected group of patients. For selenium was determined the detection limit 2,9 μg/l, for manganese 0,26 μg/l. 73 patients were examined (31 women, 42 men) whose average age was 14,1 years. The patients were divided into two groups according to age (56 children, 17 adults) and into two subgroups according to diagnoses (oncological, neurological). The control group consists of 18 subjects (5 women, 13 men, average age 21,7 years) with non-oncological and non-neurological diagnoses. We found significantly increased selenium concentrations in cerebrospinal fluid in the control group, compared with the group with neurological diagnoses (median = 14,4 μg/l vs. 12,4 μg/l, p < 0.05). Elevated levels of manganese in cerebrospinal fluid was observed in a group of children with oncological diseases compared with control group (1,2 μg/l vs. 0,5 μg/l, p < 0.05). Determination of selenium and manganese in the cerebrospinal fluid may have diagnostic importance in selected groups of patients. Powered by TCPDF (www.tcpdf.org)
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Determination of adrenaline and noradrenaline in plasma by HPLC
Kučerová, Tereza ; Klapková, Eva (advisor) ; Kotaška, Karel (referee)
The target of this bacalary work was to validate the metod for determination of adrenaline and noradrenaline in plasma by high performance liquid chromatography with electrochemical (coulometric) detection. The metod was first necessary to optimize for the coulometric detection because it was used the Bio-Rad kit, which was designed to determine with amperometric detection. The principle of this metod is the chromatographic separation of substances with subsequent oxidation of the determined analyte passes through the analytic cell with coulometric detection. The part of the analytical procedure is the solid phase extraction. The part of this work was the optimization of the metod for the coulometric detector. For the analysis were selected following potentials: channel 1 - 350 mV with range 100 nA, channel 2 - 0 mV with range 50 nA, guard channel 0 mV with range 50 nA. Furthermore, the chromatographic metod was fully validated (accuracy, precision, limit of detection, robustness). In the conclusion was made interlaboratory comparison of results of 42 patients samples with those obtained in the hospital Šternberk.
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Comparison of gel and capillary electrophoresis of proteins
Krbec, Michal ; Franěk, Tomáš (advisor) ; Kotaška, Karel (referee)
Electrophoresis of serum proteins is one of the methods used in laboratory diagnosis of pathological conditions, but also for research purposes. This is a one-step method, which involves isolation of individual proteins from a mixture (mostly blood proteins) in the electric field in the presence of a suitable buffer and a suitable carrier based on their different mobility, dependent on their surface charge and molecular weight. Originally the carrieer used to be a paper, later cellulose acetate membrane and now it is almost exclusively agarose (purified agar with low endoosmosis) or polyacrylamide (PAA). My task in this work was to analyze a serie of samples by gel electrophoresis and subsequently analyze the same serie by capillary electrophoresis. Results obtained by both methods I worked numerically, compared with each other and statistically evaluated. The output of my work is a comparison of gel and capillary electrophoresis, evaluation of the advantages, disadvantages and especially the differences between these two methods.
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