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The evaluation of microbiological quality of game meat
TŮMOVÁ, Lucie
Game meat has been representing an important part of human nutrition for a long time. However, hygienic requirements for its acquisition, storing and preparation have developer more slowly in comparison with the requirements for the livestock meat. Nevertheless, a proper manipulation with this kind of meat is absolutely essential. The objective of this diploma thesis was to evaluate the microbiological quality of game meat samples (n=76) depending on the selected internal (species, age, sex), and external (the location of bullet penetration, the place of sampling, the length of storage) factors. It has been found out that the total number of microorganisms in samples ranged from 0.00 to 8.11 log CFU/g with the mean value of 6.08 log CFU/g. Numbers of coliform bacteria in the samples were most significantly affected by the external factors and ranged from 0.00 to 7.28 log CFU/g with the mean value of 3.30 log CFU/g. Statistically significant effect (p<0,05) on the microbiological quality of game meat had been found in all external factors and in sex of game animals.
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Plant alkaloid sanguinarine and its derivatives
Tůmová, Lucie ; Stiborová, Marie (advisor) ; Hýsková, Veronika (referee)
This work is summarizing actual knowledge about sanguinarine and quaternary benzo[c]fenanthridine alkaloids. The quaternary benzo[c]fenanthridine alkaloids were found in roots plants Sanguinaria canadensis and Macleaya cordata. This plants are used in tradicional Chinese medicine for its antimycotic, antibacterial and anti-inflammatory activities since antiquity. Regarding to possibility quaternary benzo[c]fenanthridine alkaloids to induce apoptosis these investigated such as possible agents for cancer treatment. The quaternary benzo[c]fenanthridine alkaloids interact with DNA and proteins. They are able to intercalate to the DNA. The alkaloids can be used like fluorescence DNA probe. Metabolism by sanguinarine and next quaternary benzo[c]fenanthridine alkaloids has not yet completely determinated. The first step sanguinarine detoxication is its conversion to less toxic dihydrosanguinarine. Sanguinarine oxidation is mediated by cytochrome P450 1A1. Key words: Quaternary benzo[c]fenanthridine alkaloids, sanguinarine, apoptosis, intercalate, heterogenous substances, enzymes, cytochrom P450
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Physical mapping of centromeric and terminal markers of linkage groups in Xenopus tropicalis
Tůmová, Lucie ; Krylov, Vladimír (advisor) ; Nedvídek, Josef (referee)
Xenopus tropicalis belongs together with its related genus Xenopus laevis to important model organisms on the field of study of developmental processes. In comparison to well introduced models (i.e. Melanogaster, Mus etc.) for Xenopus tropicalis there is an absence of linkage and physical maps that have invaluable importance for the identification of developmentally relevant genes, including their syntheny with human. Currently, there exists provisional linkage map constructed by scientific group of Department of Biology and Biochemistry, University of Huston, Texas, USA. It is constituted of ten linkage groups (LG1-LG10) and six linkage clusters (A-F) composed by total of 1713 Simple Sequence Repeat (SSR) polymorphic markers. The laboratory of developmental biology, Charles University in Prague, participates in completing the linkage map by physical mapping of chosen polymorphic markers on chromosomes Xenopus tropicalis, which led to allocation of each linkage group and cluster to the right chromosome and to assessment of the orientation in most of them. The physical mapping of terminal markers will show us, how far chromosomes are covered with linkage groups and which areas still remain unmapped. The cooperative laboratory from National Institute for Medical Research, London, UK, originated a list of...
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Structural characterization of Nkr-p1f/Clrg interaction
Tůmová, Lucie ; Kavan, Daniel (advisor) ; Ptáčková, Renata (referee)
NK cells are ranked among lymphocytes, which are involved in primary immune responses to virus infected and tumour cells. They produce cytokines at the same time and are involved in the formation of secondary immune responses together with T and B lymphocytes. This diploma thesis is engaged in the study of the structure and interaction of the transmembrane receptor Nkr-p1f with its ligand Clr-g. This receptor Nkr-p1f has an activation function and is able to initiate the cytotoxic function of the NK cells. The three-dimensional structure has been solved as a Clr-g homodimer, but its interaction with the Nkr-p1f receptor remains mysterious. The aim was to prepare an expression vector coding the entire extracellular region of the receptor NKR-P1F and carry out the production of the receptor Nkr-p1f and Clr-g in prokaryotic expression system, subsequently renaturation and purification of the proteins in vitro. This way prepared proteins were analyzed by electrophoresis and mass spectrometry. Their interaction was studied with biophysical method, surface plasmon resonance (SPR). However, the interaction between them was not revealed by SPR technology. (In Czech)
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Detection viability of bacteria population using flow cytometry
TŮMOVÁ, Lucie
This work deals with the detection of viability of Xanthomonas euvesicatoria using a flow cytometer. The center of this work was tp optimize the method and the treatment of bacterial cultures with high temperature at given time intervals. The heat-treated cultures were dyed with an intercalating dye called propidium iodide which marks the permeabilized membranes of the cells and thus provides information on the number of dead (apoptotic) cells. The evaluation of the results confirmed the assumption that the number of surviving cells would decrease with the increasing treatment time. A temperature of 90 ° C was used, which according to other works, seems to be the most evident for the Xanthomonas genus. Flow cytometry has proven to be a simple, fast and efficient method for the viability detection of Xanthomonas euvesicatoria.
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Structural characterization of Nkr-p1f/Clrg interaction
Tůmová, Lucie ; Kavan, Daniel (advisor) ; Ptáčková, Renata (referee)
NK cells are ranked among lymphocytes, which are involved in primary immune responses to virus infected and tumour cells. They produce cytokines at the same time and are involved in the formation of secondary immune responses together with T and B lymphocytes. This diploma thesis is engaged in the study of the structure and interaction of the transmembrane receptor Nkr-p1f with its ligand Clr-g. This receptor Nkr-p1f has an activation function and is able to initiate the cytotoxic function of the NK cells. The three-dimensional structure has been solved as a Clr-g homodimer, but its interaction with the Nkr-p1f receptor remains mysterious. The aim was to prepare an expression vector coding the entire extracellular region of the receptor NKR-P1F and carry out the production of the receptor Nkr-p1f and Clr-g in prokaryotic expression system, subsequently renaturation and purification of the proteins in vitro. This way prepared proteins were analyzed by electrophoresis and mass spectrometry. Their interaction was studied with biophysical method, surface plasmon resonance (SPR). However, the interaction between them was not revealed by SPR technology. (In Czech)
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Recombinant expression of immunoglobulin superfamily receptor NKp30
Tůmová, Lucie ; Kavan, Daniel (advisor) ; Šulc, Miroslav (referee)
1 NK cells belong to lymphocytes, which are able to kill some tumor cells and virus-infected cells, produce cytokines and participate in the formation of the adaptive immune response. This bachelor thesis deals with human receptor of NK cells, specifically NKp30. This protein belongs to the group of activating receptors which can activate the cytotoxic function of NK cells. Its three-dimensional structure and the interaction between NKp30 and its physiological ligand B7-H6 has been recently resolved. Howewer the interaction with the other ligands, BAT3 and pp65, remains unknown. The aim of the work was optimalization of expression of the NKp30 receptor extracellular domain in prokaryotic expression system. The structural studies with ligands of this protein may provide further details on the interaction between the receptor NKp30 and ligands BAT3 and pp65. (In Czech)
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Plant alkaloid sanguinarine and its derivatives
Tůmová, Lucie ; Hýsková, Veronika (referee) ; Stiborová, Marie (advisor)
This work is summarizing actual knowledge about sanguinarine and quaternary benzo[c]fenanthridine alkaloids. The quaternary benzo[c]fenanthridine alkaloids were found in roots plants Sanguinaria canadensis and Macleaya cordata. This plants are used in tradicional Chinese medicine for its antimycotic, antibacterial and anti-inflammatory activities since antiquity. Regarding to possibility quaternary benzo[c]fenanthridine alkaloids to induce apoptosis these investigated such as possible agents for cancer treatment. The quaternary benzo[c]fenanthridine alkaloids interact with DNA and proteins. They are able to intercalate to the DNA. The alkaloids can be used like fluorescence DNA probe. Metabolism by sanguinarine and next quaternary benzo[c]fenanthridine alkaloids has not yet completely determinated. The first step sanguinarine detoxication is its conversion to less toxic dihydrosanguinarine. Sanguinarine oxidation is mediated by cytochrome P450 1A1. Key words: Quaternary benzo[c]fenanthridine alkaloids, sanguinarine, apoptosis, intercalate, heterogenous substances, enzymes, cytochrom P450
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Physical mapping of centromeric and terminal markers of linkage groups in Xenopus tropicalis
Tůmová, Lucie ; Krylov, Vladimír (advisor) ; Nedvídek, Josef (referee)
Xenopus tropicalis belongs together with its related genus Xenopus laevis to important model organisms on the field of study of developmental processes. In comparison to well introduced models (i.e. Melanogaster, Mus etc.) for Xenopus tropicalis there is an absence of linkage and physical maps that have invaluable importance for the identification of developmentally relevant genes, including their syntheny with human. Currently, there exists provisional linkage map constructed by scientific group of Department of Biology and Biochemistry, University of Huston, Texas, USA. It is constituted of ten linkage groups (LG1-LG10) and six linkage clusters (A-F) composed by total of 1713 Simple Sequence Repeat (SSR) polymorphic markers. The laboratory of developmental biology, Charles University in Prague, participates in completing the linkage map by physical mapping of chosen polymorphic markers on chromosomes Xenopus tropicalis, which led to allocation of each linkage group and cluster to the right chromosome and to assessment of the orientation in most of them. The physical mapping of terminal markers will show us, how far chromosomes are covered with linkage groups and which areas still remain unmapped. The cooperative laboratory from National Institute for Medical Research, London, UK, originated a list of...
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