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Construction of mouse polyomavirus chimeric VLP bearing melanoma epitopes
Kojzarová, Martina ; Drda Morávková, Alena (advisor) ; Tachezy, Ruth (referee)
Major capside protein of Polyomaviridae family viruses is able to selfassemble into virus-like particle (VLP) even without the presence of minor proteins, bind exogenous DNA non-specifically and recognise the receptor on the cellular surface. These characteristics determine its use as vector in gene therapy or immunotherapy. It was discovered before that MPyV VLPs significantly stimulate immune system and have strong adjuvant effect. Chimeric VLP derived from mouse polyomavirus carrying exogenous antigene or epitop is supposed to elicit specifically targeted immune response after immunisation. The main obstacle is choice of immunogene that is strong enough to cause adequate immune response. The goal of this thesis was to construct chimeric particles carrying epitop of malignant melanoma, one of the most immunogenic tumours, on their surface, using methods of genetic engineering. For future research of particle's immunogenic properties three types of particles were developed - particles with human and mouse melanoma epitopes, respectively and control particles with ovalbumine epitop. For the purpose of production of chimeric protein was used baculovirus expression system. It was verified then, with the use of electron microscopy, that introduction of tumour antigen into one of surface loops of VP1...
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Location of Balbani cytoplasm in sturgeon eggs as primordial gonocytes precursor during fertilization
DVOŘÁK, Matěj
The oocytes of many organisms, including fish, contain germ plasm. Germ plasm is maternally inherited determinants of germ cells. New-found germ plasm has been also described in previtellogenic sturgeon oocytes as Balbani cytoplasm. It contains precursors of germinal granules so called nuage. Germinal granules have been detected as a part of primordial gonocytes (PGCs) and according to some studies it play the role in the formation PGCs. PGCs are embryonic cells from which the gametes are differentiated. These cells has potential to transmit genetic information to the next generation. PGCs or subsequently spermatogonia therefore have the potential to be of value for gene banking and cryopreservation, particularly via the production of donor gametes with germ?line chimeras. Nevertheless the first step for successfully transplantation is visualization of PGCs. It is usually done using injection of GFP (green fluorescent protein) mRNA conjugated with mRNA, such as vasa and nanos, to the area of egg with PGC precursor localization (nuage) early after fertilization. Electron microscopic observation at this study revealed three main principal layers (a topmost alveolar layer, a zona radiata externa and a downmost layer as an zona radiata interna), a layer of cortical granules in unfertilized eggs and a perivitelline space in fertilized eggs; then germ plasm, containing nuage, which was dispersed only in the vegetal pole of ovulated sturgeon eggs. In addition, we observed formation interspace between the zona radiata externa and zona radiata interna. Sturgeon eggs have too strong envelopes for injection by glass needle. The glass capillary needle penetration could be facilitated by separation of parts of the egg envelopes as the alveolar layer and the zona radiata externa from interspace, which created by separation of the zona radiata externa and zona radiate interna by tweezers.
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Phylogeography of Rousettus aegyptiacus in the Mediterranean region
Dundarova, Cheliana ; Hulva, Pavel (advisor) ; Janko, Karel (referee)
The genus Rousettus has distributional pattern unique among fruitbats comprising both Asia and Africa and reaching northern distributional limits of the family in Persia, Arabia and Mediterranean basin. This could be ascribed to the ability of echolocation, consequent cave dwelling, and presumably other site-specific adaptations, which enabled dispersal independent of forest block and surviving in Mediterranean type of climate. Using fastly evolving mitochondrial marker, we aimed to assess genetic variability, its geographic distribution and demography of northern populations of the Egyptian fruitbat (Rousettus aegyptiacus). Mitochondrial network indicates deep genetic divergence between disjunct Mediterranean and eastern African parts of the range. Basal position of Sinaic and Jordanian haplotypes within northern clade indicate important role of these regions in colonization of eastern Mediterranean. Generally, the northern haplogroup is moderately diversified with partial geographic localization of particular haplotypes. Significant isolation by distance pattern suggests relatively pronounced site fidelity of particular colonies, at least in terms of maternal gene flow. Landscape genetics analyses indicate discontinuities in distribution of mitochondrial genetic variability, in some cases correlating with...
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Design of PMSG for micro hydro power plant
Skurka, Tomáš ; Ondrůšek, Čestmír (referee) ; Huzlík, Rostislav (advisor)
This bachelor’s thesis deals with a design and simulation of various implementation of the synchronous generator for a low-speed waterwheel. The construction of the generator was implemented as the radial synchronous generator with the permanent magnets on the rotor’s surface. In the introduction the reader gets familiar with the theory describing the construction of the synchronous generator with the permanent magnets intended for the small hydroelectric power plants. Within the thesis the author shows his own design of the generator and the optimization using the genetic algorithm. Next, three proposals with the predefined sheets are presented. Every single proposition of the generator was simulated in the software ANSYS Maxwell. Finally, the heat calculation was made for the certain designed variations of the generator.
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Multiple sequence alignment using genetic algorithms
Pátek, Zdeněk ; Mráz, František (advisor) ; Pešková, Klára (referee)
Title: Multiple sequence alignment using genetic algorithms Author: Zdeněk Pátek Department: Department of Software and Computer Science Education Supervisor: RNDr. František Mráz, CSc. Abstract: The thesis adresses the problem of multiple sequence alignment (MSA). It contains the specication of the proposed method MSAMS that allows to find motifs in biological sequences, to split sequences to blocks using the motifs, to solve MSA on the blocks and nally to assemble the global alignment from the aligned blocks and motifs. Motif search and MSA are both solved using genetic algorithms. The thesis describes the implementation of the method, conguration of its settings, benchmarking on the BAliBASE database and comparison to the ClustalW program. Experimental results showed that MSAMS can discover better alignments than ClustalW. Keywords: multiple sequence alignment, motif nding, genetic algorithms, ClustalW
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Using artificial neural networks to control genetic algorithms
Dörfler, Martin ; Holeňa, Martin (advisor) ; Gemrot, Jakub (referee)
Genetic algorithms are some of the most flexible among optimization methods. Because of their low requirements on input data, they are able to solve a wide array of problems. The flexibility is balanced by their lower effectiveness. When compared to more specialized methods, their results are inferior. This thesis examines the possibility of increasing their effectiveness by means of controlling their run by an artificial neural network. Presented inside are means of controlling a run of a genetic algorithm by a self-organizing map. The thesis contains an algorithm proposal, a prototype implementation of such algorithm and a series of tests to assess its efficiency. While the results on benchmark functions show some positive properties, the problems of greater complexity yield less optimistic results.
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Type 1 diabetes mellitus: pathogenesis and advances in therapy.
Andrýsová, Jana ; Hochmann, Jiří (referee) ; Semecký, Vladimír (advisor)
Type 1 diabetes mellitus is an autoimmune disease resulting from the destruction of insulin-producing beta cells in the pancreas leading to insulin deficience. It is multifactorial disease, in its genesis are participating interactions of genetic background and environmental factors. Type 1 diabetes is a life-long autoimmune disease that usually occurs in childhood but can be diagnosed at any age. Diabetes mellitus type 1 is primarily characterized by hyperglycemia. The others classical symptoms include: polyuria, polydipsia and weight loss. Long-term complications can result from type 1 diabetes, including nephropathy, retinopathy, neuropathy and vascular disease. Treatment goals for diabetic patients are related to control of blood glukose, blood pressure and lipids to minimize the risk of long-term consequences. Type 1 diabetes is treated with insulin replacement therapy - usually by insulin injection or insulin pump. An integral part of therapy is the patient's involvement in the therapeutical process, education, frequent glucose monitoring, adjustments in diet and exercise. The review discuss the up-to date view of treatment and continuous subcutaneous insulin infusion (insulin pump) therapy. The rewiew compare effects of continuous subcutaneous insulin infusion with multiple insulin injection....
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Regulation of pre-mRNA splicing in S. cerevisiae: where RNA cooperates with proteins.
Gahura, Ondřej ; Půta, František (advisor) ; Pospíšek, Martin (referee) ; Staněk, David (referee)
Ondřej Gahura, PhD Thesis 2011 Regulation of pre-mRNA splicing in S. cerevisiae: where RNA cooperates with proteins Abstract Removal of introns from protein coding transcripts occurs in two splicing reactions catalyzed by a large nuclear complex, spliceosome. The spliceosome is an extremely intricate and dynamic machine, wherein contributions of small RNA molecules and multiple proteins are coordinated to meet the requirements of absolute precision and high flexibility. For an intimate understanding of pre-mRNA splicing, it is necessary to unravel roles of individual components and to dissect the partial mechanisms. In the first part of this work, we describe the role of the Prp45 splicing factor in Saccharomyces cerevisiae. Mapping of genetic interactions of a conditionally lethal allele prp45(1-169) suggests a relationship of Prp45 to the NTC complex and to the second transesterification. Two-hybrid assay and purification of spliceosomal complexes reveal a contribution of the Prp45 C-terminus in the Prp22 helicase recruitment and/or regulation. Numerous experiments with reporter substrates document the need of Prp45 for the efficient splicing of a specific subset of introns. Our observations suggest that the function of Prp45 in splicing is conserved in evolution. The second part is devoted to the role of...
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Molecular Genetic Applications in Archeological and Anthropological Context
Urbanová, Markéta ; Černý, Viktor (advisor) ; Brdička, Radim (referee) ; Brůžek, Jaroslav (referee)
6 Coxcr-usroN The aim of our studywas to establisha laboratoryprotocolfor isolationand analysisof aDNA. By studying three different skeletal collections answersabout sex, kinship and presenceof pathogenicbacteriawere searched. During the course of this thesisnew protocols for aDNA study were developedand continuously optimized'We wereableto evaluateinformationaboutsexof 37 subadultindividuals(.l8'7%)from Mikulčice settlementcemetery(9thcentury)andto confirmpÍesenceof Mycobacteriumlepraeon 2 skeletonsfromŽatec cemetery(l3th century).Subadult skeletonsexaminationbrought interestingresults of extremelyhigher presenceof maleindividuals. Unfortunately,notall aimswerefulťrlled.By establishedprotocolfor aDNA kinshipanalysisit wasnot possibleto retrievesufficientinťormationfor explanationof familial relationshipof individualsfrom thesame doubleburialsite. All acquiredexperienceand establishedprotocols are very valuable and in fururewe supposetheir full utilizationandimorovementin thefield of kinshioanalvsisof aDNA molecule. 28
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Psychological and geographical profiling of serious crime
Veverková, Lucie ; Gillernová, Ilona (referee) ; Boukalová, Hedvika (referee)
The topic of this thesis is the psychological and geographical profiling of serious crimes such as rape, homicide and arson. The thesis is divided into four parts. The first of them introduces the basic concepts and terminology with respect to the offender's personality. Attention is focused on personality disorders and on some sexual paraphilias that are the most important for forensics. The topics that relate to offender's behavior are discussed as well - motivation, modus operandi, offender's signature, and the crime scene. The second part is focused on rape, homicide and arson and the characteristics of offender's personality. The third part deals with psychological and geographical profiling. Various attitudes and theories are discussed as well the base of a geographical information system. Fourth and at the same time the closing part of the thesis focuses on practical applications of knowledge derived from psychological and geographical profiling in a geographical information system. Real cases of serious crimes are analyzed. The evaluation of the findings, and of the discussed methods is presented in the discussion. The appendices are placed at the back of the thesis and involve map outputs of the geographical profiling and a vocabulary of important words.
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