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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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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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