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Characterization of the effect of human mutated huntingtin on the neuronal stem cell differentiation.
Budková, Kateřina ; Vodičková, Kateřina (advisor) ; Romanyuk, Natalyia (referee)
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expansion of the CAG codon repeat in the huntingtin gene (HTT). This expansion causes a change in the biochemical properties of the huntingtin protein (HTT), its aggregation and cellular toxicity, which leads to the degeneration of brain neurons, especially in the striatum. Induced pluripotent cells (iPSC) derived directly from HD patient cells can serve as a model system for in vitro modeling of this disease. Because neuronal dysfunctions occur in HD patients years before the first clinical symptoms manifest, this model system may help elucidate the mechanisms that precede the onset of the disease. The aim of this thesis was to differentiate iPSCs (derived from fibroblasts of HD patients and healthy controls) into neural stem cells (NSCs) and subsequently into neuronal cell populations and to monitor molecular changes in their differentiation associated with the effect of mutated HTT. The differentiation process was monitored based on selected markers using immunofluorescence, western blot and qRT-PCR. We were able to generate stable NSC lines derived from 3 control and 3 HD iPSC lines. All 6 NSC lines were able to further differentiate into neural populations. At the transcriptional level, we found a higher...
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Monasticism of Martin Luther and some of its echoes in the history of Protestantism
Vodičková, Kateřina ; Ventura, Václav (advisor) ; Hrdlička, Jaroslav (referee)
The writer in his work deals with the question, if the ideals of Catholic monasticism were preserved also in the Protestant movements and Churches. Theoretical backgrounds for this research were the thoughts of Martin Luther, who wrote several papers on the topic of monasticism. Work is introduced with historical overview of the beginnings of Christian monasticism and a brief biography of Martin Luther to reach better understanding of his thought background. Consecutively begins the analysis of the Luthers works and other theological and historical writings, whose purpose is to bring information about the reflection of Luthers standpoints in later Protestantism. As a specific case is shortly described the community of Taizé. The result is the corroboration of the hypothesis and description of some distinctive residua of monastic ideals, particularly the chastity, obedience and poverty.
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Interaction of proteins with bromphenol blue - possible analytical uses
Vodičková, Kateřina ; Hudeček, Jiří (advisor) ; Moserová, Michaela (referee)
The so called protein error of pH indicators is often used for protein determination in body fluids. A popular dye for this purpose is the bromophenol blue. A most common calibration standard, bovine serum albumin (or human serum albumin) is used. In this Thesis, I examined the influence of urea on the interaction between bovine serum albumin and bromophenol blue at pH 3,23. Addition of urea in amount corresponding to physiological values in urine (0,2-0,4 mol/l) does not cause changes in absorbance exceeding the experimental error (1 %); the same is true up to 4 mol/l of urea. Higher concentration of urea (6 mol/l) causes a considerable decrease of absorbance (up to 29 %), probably as a result of bovine serum albumin denaturation.
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Protein determination - Effect of protein composition, application of small-volume spectrophotometer
Vodičková, Kateřina ; Hudeček, Jiří (advisor) ; Moserová, Michaela (referee)
Recently, several spectrometers for small volume measurements in order of microliter have been introduced. They are primarily intended for protein determination (or determination of proteins and nucleic acids in one measurement) by direct spectrophotometry or other spectral methods. One of such instruments is the NanoVueTM Plus (GE Healthcare). In this work, we first tried to characterize the instrument in general terms (stability) and to optimize measurement condititions (sample volume). Proteins have been determined by direct spectrophotometry using internal programs of the instrument, data were controlled by an independent computation. We studied also influence of differences in composition of various proteins on the results. According to the results of this Thesis, the most accurate values could be obtained using the internal program E 1%, using the E 1% value from an experiment. On the other hand, the program Protein UV is producing often inaccurate values, strongly infleunced by the protein amino acid composition. Keywords: protein determination, spectrophotometer NanoVueTM Plus, influence of amino acid composition
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Interdiurnal variability of air temperature
Vodičková, Kateřina ; Holtanová, Eva (advisor) ; Mikšovský, Jiří (referee)
Title: Interdiurnal variability of air temperature Author: Kateřina Vodičková Department: Department of Atmospheric Physics Supervisor: RNDr. Eva Holtanová, Ph.D., Department of Atmospheric Physics Abstract: One of important climate characteristics is interdiurnal variability of air temperature. Significant attention of the bachelor thesis was paid to anomalies from a smoothed annual course of daily mean air temperature and their interdiurnal and multidiurnal variability. Changes of anomalies and their variability during period 1900-2015 on various climatologic stations in central Europe were studied. The changes of anomalies and their interdiurnal variability fits the changes of the air temperature in the 20th century. An increasing trend of interdiurnal variability of anomalies of air temperature in last decades was found. Keywords: air temperature, variability, interdiurnal changes, mean annual cycle, anomaly from mean annual cycle
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Protein determination - Effect of protein composition, application of small-volume spectrophotometer
Vodičková, Kateřina ; Hudeček, Jiří (advisor) ; Moserová, Michaela (referee)
Recently, several spectrometers for small volume measurements in order of microliter have been introduced. They are primarily intended for protein determination (or determination of proteins and nucleic acids in one measurement) by direct spectrophotometry or other spectral methods. One of such instruments is the NanoVueTM Plus (GE Healthcare). In this work, we first tried to characterize the instrument in general terms (stability) and to optimize measurement condititions (sample volume). Proteins have been determined by direct spectrophotometry using internal programs of the instrument, data were controlled by an independent computation. We studied also influence of differences in composition of various proteins on the results. According to the results of this Thesis, the most accurate values could be obtained using the internal program E 1%, using the E 1% value from an experiment. On the other hand, the program Protein UV is producing often inaccurate values, strongly infleunced by the protein amino acid composition. Keywords: protein determination, spectrophotometer NanoVueTM Plus, influence of amino acid composition
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Interaction of proteins with bromphenol blue - possible analytical uses
Vodičková, Kateřina ; Hudeček, Jiří (advisor) ; Moserová, Michaela (referee)
The so called protein error of pH indicators is often used for protein determination in body fluids. A popular dye for this purpose is the bromophenol blue. A most common calibration standard, bovine serum albumin (or human serum albumin) is used. In this Thesis, I examined the influence of urea on the interaction between bovine serum albumin and bromophenol blue at pH 3,23. Addition of urea in amount corresponding to physiological values in urine (0,2-0,4 mol/l) does not cause changes in absorbance exceeding the experimental error (1 %); the same is true up to 4 mol/l of urea. Higher concentration of urea (6 mol/l) causes a considerable decrease of absorbance (up to 29 %), probably as a result of bovine serum albumin denaturation.
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Transcriptomics of bovine preimplantation embryo genome activation in vivo and in in vitro culture conditions
Vodičková Kepková, Kateřina ; Kaňka, Jiří (advisor) ; Petr, Jaroslav (referee) ; Krylov, Vladimír (referee)
The goal of the thesis was to characterize transcriptional profiles of in vivo and in vitro derived embryos during bovine minor and major embryonic genome activation and to identify mRNA transcripts newly synthesized during these stages. In our first work we have concentrated on the study of minor genome activation at the 4-cell stage of embryo. Using SSH, we have identified 31 amplicons homologous with already identified genes. We have selected 5 of these for detailed study of their expression during the whole period of preimplantation development: centromere protein, 350/400 kDa (CENPF, mitosin), splicing factor arginine/serine-rich 3 (SRFS3), high mobility group nucleosomal binding domain 2 (HMGN2) protein and eukaryotic translation initiation factors EIF4A2 a EIF4E. All these genes play an important role in the early embryo development. SRFS3 is the first described gene with an important function in preimplantation development, which is expressed already during bovine minor genome activation, and its transcription is α-amanitin sensitive during this period. We have selected CENPF gene for a more thorough study. By silencing its expression by the injection of CENPF dsRNA into the zygote, we have studied its function throughout the whole preimplantation development of bovine embryo....
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Monasticism of Martin Luther and some of its echoes in the history of Protestantism
Vodičková, Kateřina ; Ventura, Václav (advisor) ; Hrdlička, Jaroslav (referee)
The writer in his work deals with the question, if the ideals of Catholic monasticism were preserved also in the Protestant movements and Churches. Theoretical backgrounds for this research were the thoughts of Martin Luther, who wrote several papers on the topic of monasticism. Work is introduced with historical overview of the beginnings of Christian monasticism and a brief biography of Martin Luther to reach better understanding of his thought background. Consecutively begins the analysis of the Luthers works and other theological and historical writings, whose purpose is to bring information about the reflection of Luthers standpoints in later Protestantism. As a specific case is shortly described the community of Taizé. The result is the corroboration of the hypothesis and description of some distinctive residua of monastic ideals, particularly the chastity, obedience and poverty.
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