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Mammalian proteins carrying zinc finger arrays
Vobruba, Adam ; Svoboda, Petr (advisor) ; Mašek, Jan (referee)
A zinc finger is a small peptide motif stabilised by a single zinc ion, best known for its capability to specifically bind a 3-nucleotide sequence of DNA, depending on the exact amino acids present in the DNA-binding positions. Zinc fingers are unique in their ability to freely link together and form longer tandem arrays, which can bind DNA targets of any length and sequence determined by the combination of individual fingers. These arrays can easily mutate and be rebuilt to change binding specificity, allowing great flexibility and helping zinc fingers to their widespread presence in numerous endogenous proteins of various functions. This property of zinc finger arrays also made them a suitable tool for the creation of custom DNA- binding domains for genetic engineering. This thesis provides an overview of the discovery, structure and function of these domains and then reviews and discusses selected naturally occurring mammalian zinc finger proteins and their properties, showcasing diverse uses zinc finger arrays have been adapted for throughout evolution. The history and future of zinc fingers in artificial proteins created for gene therapy and research are discussed as well. Keywords: zinc finger, ZnF, KRAB, KZFP, CTCF, PRDM9, ZFN, mammals
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Political trials of the 1950s: a case study
Svoboda, David ; Kuklík, Jan (advisor) ; Blažek, Lukáš (referee)
81 Political trials of the 1950s - a concrete case study (The Babice Case) Abstract The subject of the thesis "Political Trials of the 1950s - a Specific Case Study", subtitled The Babice Case, was a description of the application of the general tools of political persecution used in the Czechoslovak Republic in the 1950s to the specific case of the shooting at the Babice school and the subsequent political trials. Also significant is the aim of placing the political trials in Czechoslovakia in the context of the times and the international political situation, which played an important role in the Babice case. The first chapter of the thesis formulates the aims of the thesis and the background on which it is based. The second chapter of the thesis describes the significance of the political processes and the general background of their organisation. An important part of Chapter 2 of the thesis is a description of the legal framework for the conduct of political trials, focusing both on substantive aspects, such as the newly defined offences, and on procedural issues, especially the limitations on the rights of the accused. The legal biennium, an initiative aimed at changing the legal system to reflect the needs and ideology of the Communist Party, is also described as a significant phenomenon. The third...
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Function of hippocampal formation in avoidance of a moving object in rat
Zitta, Pavel ; Svoboda, Jan (advisor) ; Blahna, Karel (referee)
In the last decade, there has been a significant shift in hippocampal research from static tasks to dynamic environments that better model real-life animal situations in nature. At the same time, special attention has recently been paid to inhibitory parvalbumin interneurons (PVIs) that modulate hippocampal oscillations. Therefore, the aim of this work was to investigate the role of PVIs in orientation in dynamic environments, or in behavior relative to a potential threat posed by a moving object. In this context, chemogenetic methods were used to allow experimental manipulation of PVIs. In this work, we trained transgenic PV-cre rats in a robot avoidance task. Despite intensive behavioral training, the rats appeared to learn the task only very slowly. Chemogenetic activation of PVI with substance C21 resulted only in increased locomotor activity with a stable robot. The results show that C21 does not otherwise have a major effect on rat behavior, at least not with the relatively low number of hM3Dq expressed in the PVI. Based on local potential recording (LFP) results, we further showed that PVI activation had an effect on theta frequency band power. Keywords: Behavioral task, Hippocampus, Chemogenetics, LFP, Parvalbumin, Robot
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Cortical microcircuits and synaptic transmission in epilepsy
Doležal, Filip ; Králíková, Michaela (advisor) ; Svoboda, Jan (referee)
Epilepsy is a permanent predisposition of the brain to generate epileptic seizures. This widespread disease significantly impairs patients' life. Since epilepsy is defined only by its common symptoms, the research is complicated due to the variety of possible causes that can result in this disorder. Although many forms of therapy targeting different etiologies of this disorder exist, it is often challenging to select the right treatment and about one third of the patients still suffer from refractory seizures. Nevertheless, several proteins that participate in synaptic transmission and may undergo specific changes in their function have been identified. These changes in function affect both the activity of synapse and the involvement of synaptic connections in neuronal microcircuits. Microcircuits are common motifs of connections amidst excitatory and inhibitory neurons that are necessary for the transmission and processing of information in the brain. Microcircuits can be temporarily or permanently impaired by pathogenic processes on the molecular level, which may be responsible for the origination and propagation of seizures. Keywords epilepsy, ictogenesis, microcircuits, synaptic transmission
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Pharmacokinetics, systemic toxicity, and acute behavioural effects of hexahydrocannabinol (HHC)- an animal study.
Mallarino, Barbara ; Šíchová, Klára (advisor) ; Svoboda, Jan (referee)
The emergence of semi-synthetic cannabinoids, exemplified by hexahydrocannabinol (HHC), poses a notable public health concern. Despite its growing popularity among cannabis users, HHC remains inadequately studied in both animal models and human subjects. To this day, only a few studies examining the drug's pharmacology, toxicology, and behavioral effects have been published. The present study therefore aims to delineate the pharmacokinetic profile, acute toxicity, and behavioral effects across a spectrum of dosages (1, 5, and 10 mg/kg, p.o.) in Wistar rats. Two hours after oral administration, the concentration of HHC peak in both blood serum and brain tissue. Toxicity was established according to the OECD toxicity assay, where HHC was categorized as a compound of class 4 toxicity, with an estimated lethal dose of 1 g/kg in a rat model. Further, HHC exhibited dose-dependent effects on locomotor activity and anxiety levels, since high doses induced a significant reduction in locomotor activity in the Open field test, concurrent with elevated anxiety. Moreover, HHC administration impaired sensorimotor gating, as evidenced by deficits in Prepulse Inhibition, with the severity of these effects correlating positively with dosage escalation. This thesis is one of the first studies focused on the acute...
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The role of the HPA axis in the development of affective disorders
Fiedlerová, Alžběta ; Telenský, Petr (advisor) ; Svoboda, Jan (referee)
This bachelor thesis focuses on the influence of the hypothalamic-pituitary-adrenal axis (HPA axis) on the development of affective disorders. Affective disorders such as Major Depressive Disorder or Posttraumatic Stress Syndrom are serious mental illnesses that have a serious impact on the life of the individual suffering from the disorder. The HPA axis plays a key role in the adaptation of individual to stressful situations. Dysfunction of the HPA axis can significantly affect the homeostasis of the organism, due to increased or decreased levels of hormones and neurotransmitters or deregulation of the stress system. This can lead to the development of affective disorders. This thesis focuses on the analysis of the connections between the activity of the HPA axis and the emergence of affective disorders and examines the possibilities of therapeutic treatment of this axis as a potential target for the treatment of these disorders. The results of this review of individual scientific articles support the understanding of the mechanism that links the HPA axis to affective disorders and offer a perspective for future researh. Key words: HPA axis, cortisol, major depressive disorder, anxiety disorder, posttraumatic stress disorder, bipolar disorder
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Behavioral phenotype of APPswe/PSEN1dE9 mice model of Alzheimer's disease and a role of parvalbumin interneurons
Klausová, Tereza ; Svoboda, Jan (advisor) ; Telenský, Petr (referee)
Alzheimer's disease is a notable burden to the contemporary society concerning not only healthcare but also economy. Its aetiology remains unknown, but we know the mechanisms of pathogenesis, which allow us to create animal models (especially mice). In this thesis, we investigate the behavioral phenotype of the frequently used APPswe/PSEN1 murine model. We find that neither six-month-old nor ten-month-old animals show impairment in anxiety (open field test and elevated plus maze) or social behavior tests (three chambre test and social interaction test) compared to controls. However, the prepulse inhibition test revealed that PPI completely disappear in Alzheimer mice, indicating a significant aberration in sensorimotoric gating. The second part of this thesis focuses on a chemogenetic study of the role of parvalbumin interneurons in Alzheimer's disease and the effect of their excitation on sociability, social memory and sensorimotor gating. Here we use the aforementioned model crossed with Pv-Cre mice, which allowed us to introduce Cre-dependent DREADDs and in this group of animals we investigated social behaviour and the presence of disturbances in sensorimotor gating. However, the results obtained so far do not provide any evidence for an effect of parvalbumin interneuron excitation on animal...
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Data Analysis on the Position of Public Transport Vehicles: A Case Study of Most and Litvínov
Zátka, Josef ; Lysák, Jakub (advisor) ; Svoboda, Jan (referee)
The thesis deals with ex post (i.e. not real time) analysis of data on the position of public transport vehicles. The aim of the work is to design and implement spatio-temporal analyses of this data that will be practically applicable to real public transport networks. Their functionality is verified on data from public transport in the towns of Most and Litvínov, recorded during October 2023. The data is a point record with information about the location of the vehicle, the time and date of recording, the line number, the final stop, the last passed stop, the azimuth and speed of the vehicle movement and its delay. A total of 5 analyses are proposed. The first two analyses deal with the possibilities of effective control of the input data, within them algorithms for positional and attribute control of data are presented. The third analysis monitors and evaluates the spatiotemporal differentiation of delays, focusing on trends in the distribution of delays along the routes of individual lines over time of day and over longer time scales. The fourth analysis looks at the occurrence of delays at individual stops. The final analysis studies the reliability of transfers between public transport links at interchanges. The proposed methods were implemented through scripts written in the Python...
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Design of a single-purpose clamping device for mass production
Okénka, Jan ; Michalec, Michal (referee) ; Svoboda, Petr (advisor)
Bachelor thesis deals with the problem of clamping of a complex cast part on a four-axis CNC machine in serial production. The aim was to design a clamping jig to reduce the total number of clamping operations required. A total of 4 concepts were presented and the best design was selected according to the criteria. The proposed solution is a hydraulic centering vise, equipped with a center piece to locate two workpieces, and two rotating jaws that adjust by the workpiece. A series of 250,000 pieces per year can be fulfilled with this fixture. The jig can be easily converted to another type of production by changing the jaws and the middle part. The necessary clamping force for secure clamping was also calculated and it was verified by finite element analysis that no permanent deformation of the workpiece would occur during clamping. The work also includes a review of commercially available jigs, aluminium alloys for die casting and selected CNC milling machines.
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