The aim of this dissertation thesis is to emphasize the sense of extended cultivation of embryos to the stadium of blastocyst and its influence on success of assisted reproduction and facilitation of pre implantation diagnosis, analysis of cultivation media and derivation of human embryonic stem cells. Author summarizes current literary findings in assisted reproduction and examines the currently used methods. Author also submits his own published experimental works, in which he compares his own results of infertility treatment with usage of extended cultivation to blastocyst with results of other techniques. Furthermore author submits his own published experimental works which are using extended cultivation for pre implantation diagnosis and its improvement. Another experimental works includes possibility of stem cells derivation. Usage of extended cultivation to blastocyst convincingly leads, according to author's own experiments and simultaneously to available literary findings, to higher success of infertility treatment. This is especially significant by middle-aged mothers. Sufficient term of cultivation enables not just selection, but also biopsy and its generic treatment. Long-term cultivation also enables analysis of cultivation media - but these didn't met the expectations for increase of...

The data from public herbal collections were used to determine the frequency of the occurence of fertile bracken populations. Samples of bracken were collected to assess the degree of fertility and also to reveal the frequency of both diploid and triploid cytotypes using flow cytometry. Collected samples were also used for SSR analysis to characterize the population structure of both cytotypes and the propagation of triploid plants of bracken.

This diploma thesis deals with a serious honey bee (Apis mellifera) disease, the American foulbrood (AFB), and with possibilities of its control. The thesis is divided into two parts, the theoretical and the experimental. The theoretical part is written as a research and contains two big chapters. The first one describes American foulbrood disease and its causative agent, bacteria Paenibacillus larvae. The second part deals with the wax moth (Galleria mellonella). The experimental part then combines above mentioned themes in an effort to find the relationship between the wax moth caterpillars and bacteria P. larvae that could possibly help to control the AFB. The aim of this work was to determine whether the wax moth can disrupt resistant layers of P. larvae spores thanks to its well adapted digestive tract and whether it could change their germination.

Cytogenetic analysis of cells of chronic lymphocytic leukemia (CLL) is difficult because of their low proliferative activity. To obtain sufficient number of mitoses for performing chromosomal analysis a suitable stimulation of cell division is needed. Using DSP30/IL2 stimulated cultivation 391 CLL samples were investigated in 5 years' period. The cultivation was showed to have high success rate (96%; 375/391) with also high rate of detection of pathological clones by both karyotype and metaphase FISH analyses (in 84% of samples; 329/391). Almost in half of samples (44%; 171/391) other aberrations than recurrent FISH (i.e. 13q14 deletion, trisomy 12, TP53, ATM genes deletions) were found. Also high frequency of translocations (37%; 144/391), complex karyotypes (28%; 111/391) and clonal evolution, which was detected in one third of all samples (34% of samples with presence of more than two clones; 133/391) and like a new event in disease duration even more frequently (in 39% of samples repeatedly investigated after stimulated cultivation; 21/54), was revealed. The presence of translocations, complex karyotypes and clonal evolution was associated with progressive form of disease (P 0,000003, resp. P 0,0002 and P 0,05/P 0,04). In cases of the recurrent deletions the detailed analysis of metaphase...

The class Trematoda includes many pathogenic representatives. Main subject of this thesis, avian schistosome Trichobilharzia regenti, is a close relative to the important human pathogen Schistosoma mansoni (family Schistosomatidae). In vitro cultivation of trematodes enables closer understanding of their biology and parasite- host interactions; however, no trematode species has been successfully kept in vitro from the egg stage to the adults producing eggs. Many studies are focused on the problematic of S. mansoni cultivation, but data concerning T. regenti cultivation remain scarce. Only the ability of T. regenti cercariae to transform into schistosomula in vitro was documented, with following survival in a culture medium for a few days. Comparison of eight transformation methods was performed with T. regenti cercariae. Based on the number of tailless cercarial bodies obtained, five transformation methods were selected for further evaluation of the early schistosomula characteristics (glycocalyx shedding, penetration glands emptying and survival in vitro). It was observed that the largest quantity of cercarial bodies can be obtained by using a syringe needle or the BeadBeater cell disrupter. The largest quantity of schistosomula meeting the criteria of early schistosomulum was recorded after...

The master thesis addresses the issue of cultivation of selected strains of carotenogenic yeasts on waste materials of the food industry using a laboratory bioreactor. Carotenogenic yeasts are able to produce highly valuable metabolites during cultivation, which are located predominantly in the lipid part of the cells. Particularly, they are carotenoids, ergosterol, coenzyme Q and fatty acids. The thesis is divided into two main parts, the theoretical part and the practical part. The theoretical part describes individual yeast strains, types of waste materials, produced metabolites and methods of their analysis. The experimental part deals with the processing of waste materials of the food industry, specifically animal fat, whey and spent coffee grounds into the form of substrates usable as nutrition sources for yeast cultivation. Furthermore, cultivations focused on the recovery of the monitored metabolites and their analysis by using HPLC/PDA and GC/FID assemblies were studied as well. The yeast strains Rhodotorula mucilaginosa (CCY 19-4-6), Rhodotorula kratochvilae (CCY 20-2-26), Rhodosporidium toruloides (CCY 062-002-001), Sporidiobolus pararoseus (CCY 19-9-6) a Cystofilobasidium macerans (CCY 10-1-2) were used in this work. As one of the best producing strains Sporidiobolus pararoseus (CCY 19-9-6) was found, which achieved very high productions of carotenoids, coenzyme Q and ergosterol.

The aim of this dissertation study was to optimize the isolation and long term cultivation protocols for human dental pulp stem cells. The protocols which showed best results were used for cultivation of dental pulp stem cell isolated from exfoliated teeth (SHED). Additional aims were to characterize DPSC and SHED and prove their ability to proliferate over Hayflick's limit and differentiate into mature cell lines (osteoblasts, chondroblasts and adipocytes). In order to find optimal protocols for isolation of dental pulp from tooth, we tried three different approaches. During optimization of cultivation protocol we focused on decreasing amount of fetal calf serum (FCS) from 10 % FCS in cultivation media (most often used in literature) into 2 % and thus get closer to cultivation conditions suitable for clinical usage. We compared DPSC cultivated in three different media (medium with 10 % FCS, 2 % FCS supplemented with growth factors and media with 2 % FCS supplemented with ITS and growth factors). For characterization of DPSC and SHED we used basic biological properties (proliferation activity, viability, morphology), their phenotype and karyotype. The study demonstrated that the best protocol for isolation of dental pulp from tooth was to break the roots and extract the pulp throw this aperture. We...

Cytogenetic analysis of cells of chronic lymphocytic leukemia (CLL) is difficult because of their low proliferative activity. To obtain sufficient number of mitoses for performing chromosomal analysis a suitable stimulation of cell division is needed. Using DSP30/IL2 stimulated cultivation 391 CLL samples were investigated in 5 years' period. The cultivation was showed to have high success rate (96%; 375/391) with also high rate of detection of pathological clones by both karyotype and metaphase FISH analyses (in 84% of samples; 329/391). Almost in half of samples (44%; 171/391) other aberrations than recurrent FISH (i.e. 13q14 deletion, trisomy 12, TP53, ATM genes deletions) were found. Also high frequency of translocations (37%; 144/391), complex karyotypes (28%; 111/391) and clonal evolution, which was detected in one third of all samples (34% of samples with presence of more than two clones; 133/391) and like a new event in disease duration even more frequently (in 39% of samples repeatedly investigated after stimulated cultivation; 21/54), was revealed. The presence of translocations, complex karyotypes and clonal evolution was associated with progressive form of disease (P 0,000003, resp. P 0,0002 and P 0,05/P 0,04). In cases of the recurrent deletions the detailed analysis of metaphase...

7 Annotation Graduation thesis deal with problems of behavioral disturbances about children and adolescents. First part describes professional notions about behavioral disturbances in next chapters describes particular behavioral disturbances it's etiology and diagnosis. Final parts focus on kinds of primary prevention predominantly in elementary school and in non-for- profit organization. Thesis deal with specific and nonspecific prevention of behavioral disturbances.

Snow algae are psychrophilic microorganisms, that inhabit snow fields in mountains and polar regions, which creates colored snow in good conditions. Most species belong to order Chlamydomonadales (Chlorophyta) with complicated life cycles, containing flagellates and immobile stages (cysts). Extreme environment of snow is characteristed by low temperature, problems with availability of water and lack of nutrition. Depending on location and phase of life cycle, snow algae have to deal with excess or lack of solar radiation, the important component of which is also dangerous UV radiation. The light conditions differ substantially from open location above the forest level or polar regions to forest habitats. In the first part of this work I compared reactions of photosynthetic apparatus of snow algae strains from forest and forestless habitats to different intensity of radiation, then I dealt with assessing any changes related to long-term cultivation in laboratory. Based on measurement of rapid light curves on PAM fluorometer I have determined some characteristics of photosynthetic apparatus of individual strains (parameters α and Ik), which indicate adaptation to low or high light intensity. For some strains, it was possible to compare the results obtained at a three-year interval. In the second part...