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Study of selected mushroom toxins of Amanita muscaria.
Adámek, Michal ; Bosáková, Zuzana (advisor) ; Hložek, Tomáš (referee)
The presented bachelor's thesis was focused on the development of an HPLC-MS/MS method for the separation and eventual quantification of the main toxins of Amanita muscaria (fly agaric). After optimization of tandem mass detection conditions, MRM transitions were found for muscarine (174 → 57), ibotenic acid (159 → 113), muscimol (115 → 98) and cycloserine (103 → 75), which was used as an internal standard. Measurements were performed on Luna Omega 1.6 µm polar C18 column in RP-HPLC mode with an optimized mobile phase composition of 2:98 (v/v) acetonitrile/deionized water with the addition of 0.1% formic acid in both components and Ascentis Express RP- Amide in HILIC mode with an optimized mobile phase composed of 70:30 (v/v) methanol/deionized water with the addition of 0.5% formic acid in both components. Calibration curves with a range of 0.01 to 10 µg/ml for RP-HPLC and 0.01 to 25 µg/ml for HILIC were measured under optimized conditions. Urine and blood serum solutions of a patient with Amanita muscaria poisoning, including diluted versions of these solutions, were measured. Ibotenic acid and muscimol were found in the patient's urine, muscarine, ibotenic acid and muscimol in the blood serum. The RP-HPLC method gave narrower peaks with lower retentions and lower noise levels. The studied toxins...
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Raman spectroscopy as the tool for Alzheimer's disease diagnostics
Tesař, Adam ; Kopecký, Vladimír (advisor) ; Jelínek, Otakar (referee)
Alzheimer disease (AD) is the most frequent dementia. The prevalence is approximately 10% in 65 years old people. The current treatment is only progression protective, therefore it is crucial to find a new diagnostic approach for diagnosing AD in early stage. We analysed a set of 55 patients by the drop coating deposition Raman spectroscopy with the goal to verify previously published high sensitivity of the AD spectroscopic diagnosis in cerebral spinal fluid (CSF) and to find a new diagnostic method for blood serum (BS). We optimized measurement conditions for BS. The results were evaluated by the cluster analysis and the principal component analysis. The small set of samples exhibited high sensitivity in both CSF and BS but that distinctly decreased in the whole set. The results for CSF were affected by the choice of the analysed spectral interval. The best for AD diagnose was the interval containing peaks at 980, 1080 and 1249 cm-1.The results for BS have been the most sensitive in the whole spectral range. They have low sensitivity but high specificity for AD (92%). The usage of neural networks has conversely high sensitivity and low specificity in both sets of samples of BS and CSF. Powered by TCPDF (www.tcpdf.org)
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Microbiological Aspects of Infectious Diseases Therapy
Paterová, Pavla ; Buchta, Vladimír (advisor) ; Kolář, Milan (referee) ; Jílek, Petr (referee)
Background: The method of serum bactericidal assay represents an alternative possibility of optimization of anti-infectious therapy and administration of antibiotics. It mirrors the real activity of one or more administered antibiotics in the complex system of the antibacterial effect of patient's serum. The paper aimed to confirm non-inferiority of bactericidal testing using the broth dilution method according to CLSI M21-A Guidelines (time to results 48, 72 hours) in comparison with modified methods of testing on the basis of turbidimetry (time to result 6, 8, 24 hours) and resazurin color (time to results 8, 24 hours). Methods: Four antibiotics were tested: gentamicin, amikacin, piperacillin/tazobactam and meropenem with 30 Escherichia coli strains isolated from blood cultures of 29 pacients hospitalised in different wards, University Hospital in Hradec Kralove. Human blood sera (n = 76) from ten hematological patients (4th Department of Clinical Medicine, University Hospital, Hradec Kralove) were tested to establish bactericidal titer. Patients' blood was withdrawn prior to and in the course of the first and third day of antibiotic therapy of febrile neutropenia. Testing employed the reference strain Escherichia coli ATCC 25922. Results: A comparison with the standard CSLI showed that the...
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Studium změn chemického složení a kvality kravského mleziva v závislosti na čase, věku plemenice a způsobu výživy v době stání na sucho
Fröhdeová, Martina
The dissertation is focused on assessing the influence of yeast culture and selenium addition on yeast culture in feed diet, on colostrum quality and blood serum parameters. The content of immunoglobulins G and the density of colostrum, as the main parameters of the quality of the colostrum, were evaluated in the first born cows and other cows on the second and higher lactation and compared with each other. At the same time, blood serum parameters were also monitored as supplementary indications of feeding intervention in the feed diet of breeds. The main aim of the work was to evaluate the influence of additions of feed additives on the quality and composition of colostrum. Immunoglobulins G were analyzed in all colostrum and blood serum samples. IgG were determined by the ELISA method. The results obtained indicate that the addition of yeast culture and selenium to yeast culture did not have the expected positive effect on improving the quality and composition of cows' colostrum or on blood serum indicators.
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Raman spectroscopy as the tool for Alzheimer's disease diagnostics
Tesař, Adam ; Kopecký, Vladimír (advisor) ; Jelínek, Otakar (referee)
Alzheimer disease (AD) is the most frequent dementia. The prevalence is approximately 10% in 65 years old people. The current treatment is only progression protective, therefore it is crucial to find a new diagnostic approach for diagnosing AD in early stage. We analysed a set of 55 patients by the drop coating deposition Raman spectroscopy with the goal to verify previously published high sensitivity of the AD spectroscopic diagnosis in cerebral spinal fluid (CSF) and to find a new diagnostic method for blood serum (BS). We optimized measurement conditions for BS. The results were evaluated by the cluster analysis and the principal component analysis. The small set of samples exhibited high sensitivity in both CSF and BS but that distinctly decreased in the whole set. The results for CSF were affected by the choice of the analysed spectral interval. The best for AD diagnose was the interval containing peaks at 980, 1080 and 1249 cm-1.The results for BS have been the most sensitive in the whole spectral range. They have low sensitivity but high specificity for AD (92%). The usage of neural networks has conversely high sensitivity and low specificity in both sets of samples of BS and CSF. Powered by TCPDF (www.tcpdf.org)
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Se-Metabolism inside the mammalian organism fed Se-supplemented Brassica napus forage
Žíla, Ondřej ; Čadková, Zuzana (advisor) ; Václav, Václav (referee)
The aim of the thesis was to determine whether the individual Se-speciation in the mammalian organism are affected by the form of received selenium. Selenium is an essential micronutrient important for humans and animals. It plays an important role in the antioxidant protection of the organism and in the conversion of thyroid hormones. In our experiment the laboratory Wistar rats were divided into three groups. Each group had a different diet. The rats were fed with selenium in the form of soy, sodium selenite and extracted rapeseed meal. Urine samples were regularly collected during the four-week experiment and in the end of the feeding study, the blood serum was also collected. The total selenium content was measured by ICP-MS, while the individual Se-speciation in urine and serum by HPLC coupled with ICP-MS. In the urine the identified speciation were methylselenocystein (MeSeCys), trimethylselenium (TMSe) and selenosugar 1 and 3. In the blood serum the measured speciation were TMSe, selenite, selenate and selenosugar 1. For the group fed with sodium selenite the measured values in the urine were generally higher, this might be due to a higher overall intake and also an inorganic form of selenium with a lower absorbency. Groups that received selenium from plant sources took in several Se-compounds and the total measured content of Se-speciation and secretion dynamics were not significantly different. Additionally speciation of selenosugar 2 was measured for the group fed with rapeseed meals, which in the other groups did not appear. When receiving selenium from plant sources the biotransformation in the mammalian organism differs in comparison to receiving selenium from mineral salts. The initial hypothesis that Se-speciation is influenced by the form of selenium administered in the diet was confirm by our results. Since the group fed rapeseed showed similar results as the group fed a standard feed with soy, the extracted rapeseed meal could serve as a good source in livestock nutrition.
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