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Determination of genetic markers for kidney complications of diabetes mellitus.
Vedralová, Marcela ; Kotrbová - Kozak, Anna Katarzyna (advisor) ; Kalousová, Marta (referee)
In this work we chose to study genes whose protein products significantly affect calcium- phosphate metabolism in kidneys and are hypothesized to be involved in the pathogenesis of diabetes as well as kidney damage. Vitamin D receptor (VDR) belongs to a large family of nuclear receptors and transcription factors. VDR is expressed in many cells in different tissues and activation of this receptor by its ligand 1,25 - dihydroxyvitamin D3 affects expression of many genes. Using restriction endonucleases Bsm I, Fok I, Taq I and Apa I, there were identified polymorphic sites in VDR gene that were found to be associated with certain chronic diseases, like type 1 and type 2 diabetes and lung carcinoma. Parathyreoid hormone is one of the most important regulators of calcium and phosphate metabolism. Allelic polymorphism of PTH gene involves B/b and D/d alleles that can participate in pathogenesis of kidney complications as well. Aims: We aimed to determine whether there is a difference in frequency of certain combinations of VDR and PTH genotypes between group of diabetic patients with kidney complications and diabetic patients without kidney complications. We also assessed influence of single-nucleotide polymorphisms at VDR gene mRNA expression. Methods: DNA was isolated by salt-extraction method and...
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Preparation of monoclonal antibodies against Borrelia
ŠVEJDOVÁ, Veronika
In my bachelor thesis I deal with preparation of monoclonal antibodies against antigens of Borrelia. In theoretical section I describe genus Borrelia and antibodies. The introduction deals with the history of discovery of the spirochete that causes Lyme disease, vectors of this disease and classification of Borreliae according to geographical occurrence. Taxonomic categorization and description of the bacterium followers. It is focused on flagellum, arrangement of genetic material and on external surface antigens important for this bacterium. In the next section I describe Borrelia vector - host interactions and the development of immune response during the disease. Dermal symptoms of the Lyme disease are described in detail. A few words about relaxing fever are added. This chapter ends with description of laboratory diagnostics of Borrelia burgdorferi. Next, I focused on the characterization of antibodies and on preparation of monoclonal antibodies by hybridoma technology. I briefly describe ways of their utilization. Practical part of Bachelor thesis was carried out in the laboratories of Faculty of Science. Here, with the help of hybridoma technology, I fused myeloma cells with splenocytes of immunized mice producing hybridomas. These hybridomas I cultivated in medium, which I subsequently tested for occurrence of antibodies by ELISA. Hybridomas having high productivity were cloned, frozen and stored in liquid nitrogen. Finally I purified monoclonal antibodies, concentrated them and tested their titre.
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Characterization of Mitochondrial Respiratory Chain Defects Using DNA Microarrays
Čížková, Alena ; Kmoch, Stanislav (advisor) ; Kováč, Ladislav (referee) ; Červinková, Zuzana (referee)
A mitochondrion is a organelle under nuclear and mitochondrial genetic control and its maintence requires a nucleo-mitochnodrial cross-talk. Better understanding of mitochondrial pahologies using various gene expression techniques. This thesis presents the implementation and application of microarray technology for identification of disease causing genes in respiratory chain disorders, expecially in patients with ATP synthase deficiency of nuclear origin. The introduction to this thesis provides an overview of mitochondrial structure and biogenesis, respiratory chain complexes and their disorders as well as different types of microarrays. A study of mitochondrial ATP synthase deficiency represents the major experimental part of this thesis and is accordingly that were used in further studies centered on characterization of inherited metabolic disorders. This is documented in positional cloning of patientes with lysosomal storage disorders - mucopolysaccharidosis IIIC.
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Phenotypic and molecular approaches in the systematics of the Palearctic and Neotropic tree frogs, Hyla and Osteocephalus (Amphibia: Hylidae)
Gvoždík, Václav ; Moravec, Jiří (advisor) ; Mikulíček, Peter (referee) ; Hulva, Pavel (referee)
The systematics and taxonomy of tree frogs of the family Hylidae has undergone turbulent changes both at higher (subfamilies, genera) and lower (species) level during the last decades. The original approach used morphological characters to distinguish tree frogs. It has been completed by bioacoustics, since the advertisement calls of the males were in some cases found to represent an important behavioural reproductive barrier and therefore useful for systematic evaluations. However, a completely new impulse was given to systematics by the introduction of modern genetic methods. These methods have allowed to distinguish not only morphologically but even acoustically cryptic taxa. The most reliable method to distinguish tree frogs at present is a combination of data from the different disciplines. The present dissertation thesis aims to evaluate the systematics of tree frogs of the genus Hyla from Eastern Europe and the Middle East as well as the systematics of the Amazonian genus Osteocephalus using a combination of morphological, bioacoustic and molecular approaches. The thesis is composed of a general introduction, three published papers, two manuscripts under reviews and a conclusion chapter. The first three papers deal with the phenotypic (morphological plus bioacustic) approach in systematic...
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Population biology and genetics of rare plant species
Dostálek, Tomáš ; Münzbergová, Zuzana (advisor) ; Oostermeijer, Gerhard (referee) ; Jersáková, Jana (referee)
5 Summary In this thesis I synthesized various approaches to rare species research which is necessary for their effective conservation. First three papers (paper 1, 2, 3) are dealing with Dracocephalum austriacum, which is rare and endangered species throughout the whole Europe. In the first paper I studied complete life cycle of this species and focused on differences between two distant regions. The results suggest that the knowledge of the most important life cycle transitions can be transferred between regions. However, the long term predictions of population growth and their sensitivity to external disturbance are largely region specific. In the second paper I estimated genetic diversity and its distribution in populations and regions using allozyme analyses. The results showed large variation in genetic diversity between populations and indicated which populations should get a priority in attempts to conserve all the genetic diversity within the region. The second paper also revealed significant positive relationship between seed production and genetic diversity. The third paper combines the two previous studies. It is the first study explicitly linking changes in population dynamics and changes in genetic diversity. I found that decrease of expected heterozygosity caused strong decrease in long term...
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Regulation of calcium influx via voltage-dependent calcium channels (L-VDCC) into the vascular smooth muscle of genetically hypertensive rats
Surovcová, Mária ; Kuneš, Jaroslav (advisor) ; Hampl, Václav (referee) ; Pácha, Jiří (referee)
The present in vivo study was focused on better understanding of the pathophysiological mechanisms contributing to high blood pressure maintenance in spontaneously hypertensive rats (SHR). Genetic hypertension is characterised by abnormally elevated activity of sympathetic nervous system (SNS) and increased sensitivity to catecholamine-induced vasoconstriction comprising both enhanced calcium influx to vascular smooth muscle cells via L-type of voltage-dependent calcium channels (L-VDCC) and altered vascular sensitivity to intracellular calcium mediated by RhoA/Rho-kinase pathway. Thus, our aims were to study the regulation of L-VDCC channels, to determine the role of "calcium sensitization" in hypertension and finally to evaluate which of these pathways is important in the maintenance of high blood pressure. Using conscious SHR rats and their normotensive controls, WKY, we have confirmed that high sympathetic tone is responsible for increased calcium influx via L-VDCC in hypertension. The experiments based on the pertussis toxin-induced inactivation of inhibitory G proteins (Gi) have revealed that the control of L-VDCC by SNS is mediated by Gi protein- coupled pathway, the elimination of which leads to the attenuation of sympathetic vasoconstriction and to the decrease of blood pressure response to...
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The comparison of biolistic and Agrobacterium mediated transformation of tomato (Solanum lycopersicum L.)
DĚDOUCHOVÁ, Martina
Currently we can do genetic manipulation by means of direct and indirect methods. Indirect methods use strategy of gram-negative soil bacteria Agrobacterium tumefaciens, which introduces part of its own genes carried by the section of plasmid Ti called T-DNA into the genome of infected plants. It is difficult to reach the transformation of monocotyledonous plants using this method. Application of direct methods of transformation offers the possibility for transformation of monocotyledonous plants. One of the most effective methods of direct transformation is biolistic transformation. This diploma thesis dealt with the comparison of efficacy of biolistic and A. tumefaciens mediated transformation of tomato (Solanum lycopersicum L.).
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Quasi-parallel optimizing of simulated systems by means of genetic algorithms
Konopa, Michal ; Neruda, Roman (referee) ; Kindler, Evžen (advisor)
This work goals are SIMULA classes to model experts sessions so that each expert has his own start ideas the optimal system's parameters. All the experts are simulating the system (each expert with his own parameters), but during the simulation they are mutually exchanging information about behavior of theirs models and - in accordance with this information - they are learning, changing their own system parameters. The learning process is performed by means of genetic algorithms. The resulting optimizer is tested on concrete examples, both from the mathematical theory and real practice (e.g. optimizing of a given project). This work reassumes the dissertation of RNDR. Jiří Weinberger. CSc., who made similar type of optimizer in the 80es, when nothing concrete was known on genetic algorithms and so the experts learning scheme was modeled by techniques, which had great success in solution of real problems afterwards; but nowadays it is worth to replace them by genetic algorithms, or at least to compare both methods. Genetic algorithms are successfully used in systems optimization, where the model run is algorithmic-managed, but never in the way of directly changing the models during their running times. Parallel run of multiple simulating models on the computer equipped with only one processor requires...
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Simulation of Entities Collective Behavior in Virtual World
Vymazal, Tomáš ; Žák, Pavel (referee) ; Láník, Aleš (advisor)
Theme of this work is to evaluate and compare aviable paradigms for entity control in virtual worlds, and to implement one of these paradigms in application. Dynamic finite state machine upgraded using genetic algorithms has been chosen. This paradigm should make agent's behavior better and adapt agent to required state: i.e. make agent harvest resources in virtual world. Output of this work is application for running evolution and application for 3D view of agent's behavior.
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Learning Restricted Restarting Automata using Genetic Algorithm
Basovník, Stanislav ; Plátek, Martin (referee) ; Mráz, František (advisor)
Restarting automata are linguistically motivated models for language representation. The main goal of this work is to propose a suitable version of restarting automaton for learning from positive and negative samples using genetic algorithms. We also characterize the class of languages accepted by limited context restarting automata with respect to the Chomsky hierarchy. The proposed learning algorithm is compared to two well-known methods for learning languages from positive and negative samples - RPNI and LARS. A tool for learning the restricted version of restarting automaton is developed as a part of this work. Examples of usage and user guide are included in this work.
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