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Establishment of method for determination of selected triazine pesticides in water using GC-MS/MS
OPEKAR, Jan
This bachelor thesis deals with the determination of triazine pesticides in water using GC-MS/MS. Firstly, the development and optimisation of an analytical method was performed. The following parameters of the analytical method were chosen for optimisation: carrier gas flow rate, temperature gradient, selection of SRM transitions and collision energy of SRM transitions. Then, several extraction parametres such as SPE column drying time, selection of elution solvent, sample pH and sample volume were optimised. Finally, the method was successfully validated and applied to the analysis of a real tap water sample.
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Vliv růstových regulátorů na proteom rostlin v průběhu klíčení
Habánová, Hana
Seed germination is a complex process during which an embryonic plant contained within a seed transforms into a seedling. The majority of species, including model plant Arabidopsis thaliana produce dry seeds that will fully re-establish metabolism after imbibition. This early phase of a plant's life is highly important for its survival and its progress depends on both internal and external stimuli. This thesis entitled "Effects of plant growth regulators on proteome dynamics during seed germination" is focused on effects of light and plant growth regulators on germination. To get an insight into molecular mechanism of germination, proteome dynamics in germinating Arabidopsis were investigated. Seeds were imbibed in presence of plant growth regulators in continuous light or dark and material for LC-MS proteome profiling was harvested after 24 and 48 hours. Altogether, abundances of more than 1100 proteins were followed. PCA analysis and a detailed pair-wise comparison of mock-treated seeds provided evidence that the modern proteome-wide analysis is a promising tool for plant germination research.
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Metody frakcionace a obohacování rostlinného proteomu
Breineková, Alžběta
The proteome is a complex and dynamic mixture of proteins produced by an organism. It represents tens of thousands different protein species and its characterization is a difficult task even with state-of-the-art techniques. This thesis entitled "Methods for plant proteome fractionation and enrichment" is focused on plant proteomics and approaches that can improve detection limits and increase proteome coverage. In the experimental part, model plant Arabidopsis thaliana proteome was analysed. The tested techniques include crude fractionations on the tissue level, a subcellular fractionation, and separations on the protein level. Finally, sets of samples were analysed via LC-MS and the method for peptide analysis was optimized. Combining the LC-MS datasets revealed more than 4900 identified Arabidopsis proteins (FDR<1%). Of these, 248 were detected only in equalized proteome. The results of this work helped to establish a robust proteome analysis workflow that will be implemented in plant proteome research at MENDELU.
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Efekt inhibice proteasomu na proteom Arabidopsis thaliana
Dufek, Martin
Proteasome is highly conserved in its structure among all eukaryotes. The main function of this multi-protein complex is to facilitate protein turnover and degrade misfolded, altered, aged or unneeded proteins. Proteasome-dependant degradation predominantly operates in cooperation with a complex system that conjugates proteins destined for degradation to ubiquitin. Emerging evidence suggests that the proteasome-dependant degradation is crucially involved in plant hormone signalling and as such could play a central role in plant growth and development. This bachelor thesis entitled "Effects of proteasome inhibition on Arabidopsis thaliana proteome" reviews proteasome-ubiquitin pathway with the focus on plant hormone signalling. In the experimental part, the effects of proteasome inhibition were investigated via LC-MS profiling. Altogether, abundances of more than 1400 proteins were followed. PCA analysis and a detailed pair-wise comparison of MG-132 and mock treated seedlings provided an insight into the molecular mechanism behind processes in the response to proteasome inhibition.
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Applicability of Mass spectrometry ( MALDI-TOF ) to classification and differentiation of other bacteria from genus Enterobacteriacae. Identification and incidence of Serovars Salmonela Enteritidis in matter from Synlab Č.B.
WIMMEROVÁ, Zuzana
The aim of my bachelor thesis, are diagnostics and presence of genus Salmonella. Salmonella is a relatively common cause of diarrheal disease, which occurs in our country very often. Reason to choose this topic? Diagnostics of salmonella is very difficult because of the possibility of confusion with other bacteria of Enterobacteriaceae family and large number of Salmonella serotypes. I wanted to compare the methods that are used to diagnose Salmonella and detect the best method for laboratories in term of time, financial, but also which method is the most accurate. The variety of diagnostic possibilities were also on of the key elements in decision making. This bachelor thesis is divided into two major sections. The first section deals with the literary searchers related to the genus Salmonella. In the theoretical part I described prokaryotic cell. That is difficult because according to cell membrane we differentiate gram positive and gram negative bacterium. It will facilitate the identification of bacterium. Then I dealt with Kauffmann - White's scheme, by which are distinguish the deferent serotypes of the genus Salmonella. This scheme is very important in terms of diagnosis and classification of serotype. After that follows dividing of the genus Salmonella according the infection types.I dealt with genus Salmonella, which can infect humans and animals. I mentioned about salmonellas disease. Thereafter was solved incidence of genus Salmonella, way of spreading, etc. In the last chapter was mentioned the treatment of salmonellosis. I introduced a simple listing of known antibiotics and drugs used to treat salmonella. I also noted known medicament Endiaron, which is convenient and has no side effects. The second part is dedicated to methodology, summarizing the procedure to be used by determination of salmonella. Here are present all parts of the laboratory research, beginning pre-analytical parts, such as the collection and transport. Then follow the laboratory and post laboratory part. The largest part is the laboratory part, in which are cultivation, biochemical method. There is something about MALDI TOFF and biochemical characterization. The aim of this work was to compare the method MALDI TOFF with other biochemics methods at determination the genus Salmonella. The best solution in diagnosis of salmonella is in my opinion a device MALDI TOFF, which is the fastest method in comparison with the slide agglutination and number of biochemics API 20E and biochemical wedge. The next aim of my work was to find out occurrence of genus Salmonella in a all seasons. I have found out, that's the most by found serotype is Salmonella Enteritidis, next is Salmonella Montevideo, and then close behind Salmonella Typhimurium. Salmonella Enteridis appeared in material synlab czech,s.r.o. 601 times total. Salmonella Typhimurium appeared in material synlab czech,s.r.o. 31 times total and Salmonella Montevideo 13 times. Overall I computed 701 tribes for year 2013. Also I also looked at the "exotic" serotypes that were imported here in my opinion from a tourist's destinations with lower levels of hygiene. The highest incidence was in the fall (September, October, November) - 268 tribes. The second most frequent occurrence was in sommer months (June - August). In this cases was diagnosed 224 salmonella in total. In the winter there were 124 genus and in spring there were 105. The most frequent occurrence was surpisingly in September - in total 128. The less result was measured in March with count of 24 genuses.
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