Forkhead transcription factor FOXO4 is involved in a wide range of processes including metabolism control, cell cycle regulation, apoptosis, DNA repair or oxidative stress resistance. The crystal structure of FOXO4 DNA-binding domain bound to the target DNA revealed that FOXO4 interacts with DNA in the same manner as other forkhead proteins. The DNA-binding interface consists of helix H3, which binds to the major groove of DNA, the N- terminal segment and flexible loops located at the C-terminus of forkhead domain. However, many questions concerning the regulation of DNA-binding specificity and the role of different parts of forkhead domain in this process remain elusive. In this diploma thesis, the interactions between the DNA-binding domain of FOXO4 and DNA with various sequences were studied. The DNA-binding domain of FOXO4 and its mutants were expressed and purified and their DNA-binding properties were studied using fluorescence spectroscopy techniques and surface plasmon resonance. Results of these experiments revealed that the nucleotide sequence at the 5′ end of the consensus binding motif affects the stability of the complex between the DNA-binding domain of FOXO4 and DNA. In addition, the importance of individual amino acid residues of FOXO4 that are involved in DNA binding for the...

In the present work we study gravitational waves in the approximation of high frequency, and their interaction with matter waves with comparable frequency. We generalize the formalism of R. A. Isaacson to the case of non-vacuum spacetimes. We formulate and study a technique of so called averaging, which helps us to separate slow changes and high-frequency fluctuations of various tensor fields. The described methods are used in examples with a given particular metric - so called background metric. We thus study the interaction of high-frequency gravitational waves on the Robinson- Trautman background and the high-frequency electromagnetic waves. Further, we seek a perfect fluid with high-frequency energy-momentum tensor that could coexist with high-frequency gravitational waves on the flat Friedmann-Lemaˆıtre-Robertson-Walker background; again, we examine the relation between those potential gravitational and matter waves.

Title: Coaching of the football Objectives: The main goal of this thesis was to explore how players perceive their coach during the length of a season, and his/her strengths and weaknesses. Another objective was to compare the results of male and female squads in the selected areas. Because the players' individual preferences and expectations can vary, the study aims to compare differences between players who evaluated their season as successful, and players who did not have a successful season. At the end the results were fed back to the players to evaluate whether the CBAS system can be used as a form of feedback, and whether the coach can use this tool to work collectively with the team, as well with individuals according to their specific needs. Methods: A structured questionnaire was used as the research method as well as individual interviews with players during their season. The main part of this research, identifying 12 assessing categories of a coach, was obtained by using a structured questionnaire. Interviews with individual players were conducted to clarify opinions of these players towards the evaluated coach. Results: After the interpretation of the results it was found that the differences between male and female players are apparent in area General encouragement. More evident...

This dissertation is focused on 30 kDa protein phosducin (Pdc) and on the regulation of its function through the interaction with 28 kDa adaptor protein 14-3-3. These two proteins participate in G-protein signal transduction pathways, especially in the process of light signal transduction. It is assumed that Pdc binds to the Gtβγ complex of G-protein called transducin and through this interaction it inhibits the reassociation of Gtβγ with Gtα thus reducing the visual signal transfer. This process is thought to participate in a long- term light adaptation. The regulation of Pdc function is further regulated by its phosphorylation and subsequent binding to the 14-3-3 protein. It has been speculated that the 14-3-3 binding plays a key role in the inhibition of the interaction between phosphorylated Pdc (Pdc-PP) and Gtβγ. The formation of the 14-3-3/Pdc-PP complex leads to the reassociation of Gtβγ with Gtα and consequently to the amplification of visual signal transfer. Nevertheless, the mechanism by which the 14-3-3 protein binding inhibits the interaction between Pdc and Gtβγ remains elusive. The main aims of this dissertation were: (i) to investigate the structure of Pdc in its apo-state (in the absence of the binding partner) and in the complex with 14-3-3, and (ii) to suggest the mechanism of the...

Fish muscle contains omega-3 fatty acids and high quality protein and other valuable nutrients. On the other hand, due to the high content of water, neutral pH, presence of PUFA and sensitive amino acids, fish muscle is a very perishable food. The progress of lipid and protein oxidation as well as interaction between the oxidation products could have a negative role during storage time. Negative effect of oxidation on the fish fillet quality has been reported by several authors, but still the pathways and products of protein oxidation owning to the diversity of the protein oxidation products is not clear. Pathways of lipid and protein interaction, target amino acids, storage conditions and also, final impact on the fish muscle quality have been investigated in this study. We examined the quality of fish muscle in common carp and perch during different storage conditions (+4°C and -20°C). Also, we compared some quality parameters in perch muscle from two different rearing systems to identify the pathways and products of oxidation. The present study investigated the oxidation progress with respect to TBARS and carbonyls as markers of lipid and protein oxidation respectively as well as contribution of PUFA on the lipid oxidation development. Further, the effect of storage conditions and oxidation on the textural and sensorial parameters were examined. Possible changes of pH, rigor index and biogenic amines, during post mortem were studied. Additionally, we performed proteomic analysis in order to investigate more deeply the protein alteration and find a correlation between protein structures and fish muscle quality. Our results showed a good stability in common carp (Cyprinus carpio) fillet during 6 months storage at -20oC. According to our results, we observed a significant decrease in the firmness of common carp muscle after 1 week and then it was constant while liquid loss increased significantly with storage time. Regarding the sensory parameters, we could not find any differences in the examined parameters in both stored raw and cooked fillets compared to the fresh one. Although, both lipid oxidation parameters (TBARS and PV) were increased significantly but it did not reach the critical level for human consumption. On the other hand, protein oxidation which is involved in WHC deterioration was increased significantly. In addition, we investigated the muscle quality of Eurasian perch (Perca fluviatilis L.) from two rearing systems (RAS and pond). During frozen and refrigerated storage, we observed a significant increase in the amount of TBARS and carbonyls in both rearing systems. Comparison between both rearing systems did not show any differences in the amount of TBARS while carbonyl was significantly higher in the RAS fillets rather than pond fillets particularly, until 4 month storage at -20oC. Also, we found a significant increase in liquid loss and considerable decrease in firmness in the fillets from both rearing systems. Our results showed higher firmness and liquid loss in the RAS fillets compared to the pond one. Proteomic analysis on the fillets from both rearing systems during frozen storage showed more stability in the RAS fillet however, western blot revealed more oxidized carbonyls in the RAS fillets rather than pond fillet specially, at time 0 and 4. Probably, formation of ice crystals and accumulation of ROS have key roles on the development of oxidation during frozen storage while enzymatic activity follow by protein degradation are important during refrigerated storage.

The Thesis deals with interactions of the trachet and artist. The theoretical section contains selected educational-psychological perspective oninteraction, complemented by a sociological perspective in the kontext of the changing times of the communication world by John Thompson. The practical part narrows to specifically interact using touch, and tactile perception of visual artifacts. The real impulse comes from the work of Jan Svankmajer and interactive exhibitions of Peter Nikl. The result of this work is a set of interactive, visual haptic objects for presentation in the form of a separate exhibition.

The focus of this master thesis is on the phenomenon of neighborhood and social relations between residents of the housing estate Cerny Most. The main objective of this thesis is to determinate, what social relations and interactions are between residents of Cerny Most and how different are these relations and interactions between established residents and newcomers. The research is a combination of quantitative and qualitative approaches. The quantitative part is based on a survey, which is focused on determinates of neighborhood, for example the residential stability, the resident´s connection to Cerny Most and spending of free time in the locality depending on location of residence and length of staying in the location. Research is also focused on neighborly relations and social interactions between residents of Cerny Most, but also between established residents and newcomers. Research deals with the satisfaction in the place of residence, quality of neighborly relations, purposeful meeting of neighbors and involvement in the community life. The qualitative part is consist of field research, own photographs of the area and semistructured interviews which help to better understanding to detected areas. From the survey results, statistic calculations and semistructured interviews is compile the plastic image of neighborly relations and interactions between the residents of Cerny Most.

The thesis deals with the mutational analysis of conserved amino acids of Potato virus X coat protein (PVX CP). The importance of selected amino acids for the spread of the virus in the plant should be determined. Nicotiana benthamiana was selected as an experimental plant. Mutations of the PVX CP were based on the comparison of PVX CP amino acid sequence with the sequence of Papaya mosaic virus coat protein (PapMV CP), the only representative of potexvirus, which includes PVX, with the described crystal structure of the CP. The importance of certain amino acids for interaction of coat protein subunits PapMV CP,CP and PapMV CP,RNA and thus for virus particles formation was experimentally determined. The available amino acid sequences of isolates and strains of PVX CP were obtained from the National Center for Biotechnology Information (NCBI) database, compared with each other and alsowith the sequence of PVX CP used in the Laboratory of Virology, Institute of Experimental Botany of the CAS. Codons encoding conserved phenylalanine and lysine at positions 33 and 118 in the PVX CP amino acid sequence were mutated using methods of molecular biology. Five constructs of PVX CP mutants were prepared, Two deletion mutants of the PVX CP N-terminus which were created in the vector derived from PVX (pGR106) in which the cDNA sequence of PVX is integrated, the remaining 3 point mutants were prepared only as a product of SOE PCR. The reporter gene GFP for monitoring of infection and spread of mutants in plant tissue was cloned into pGR106 carrying the deletion mutants of the N-terminus of PVX CP (deletion of 2.-32. or 2.-33. amino acid). Both mutants are able to move only in a short distance from infected cells to adjacent cells within the inoculated N. benthamiana leaf. These two deletion mutants showed difference in the speed and in the extent of the GFP signal spread. Deletion mutant still possessing the codon for F33 showed faster onset of the GFP signal and was able to spread more rapidly to surrounding cells in comparison with deletion mutant, where the codon for F33 was removed. Other mutants carrying the point mutations were also prepared: the deletion mutant of the codon for F33 (deletion F33), the substitution mutant, in which the codon for phenylalanine at position 33 was replaced by the codon for alanine (F33A) and substitution mutant, where the codon for lysine at position 118 was replaced by the codon for alanine (K118A). Unfortunately, all three point mutants could not be cloned into the vector pGR106, therefore, the evaluation of their spread in N. benthamiana plants was not possible. Based on obtained results it is possible to conclude that the amino acid F33 is important for the intercellular movement within the neighboring cells. To assess the importance of the amino acids F33 and K118 in the systemic infection, it would be necessary to evaluate also the point mutants deletion F33, F33A, and K118.

Vít Neznal’s PhD thesis begins by defining theatre as a specific sphere of media. According to the author, such notion is made possible by following up to the tradition of Czech theatre theory (immediacy as a specific feature of the medium, a synthetic basis, a non-substantial conception of a work, acting as a decisive art, a total three-dimensional feedback-based communication with cardinal impact on the specific nature of stage acting, and an active position of the spectator). On this basis, Neznal proceeds to explain why there is no justifiable reason for theatre theory to abolish the dichotomy between art and reality. His premise is that while theatre is rooted in life, it is at the same time automatically and irreversibly taken out of it as a medium. As a result, there is, from the point of view of theatre theory, a difference between an intensified “life” entering the stage and becoming the basis of expression (intensification of “life” is the basis of means of expression and is therefore made intentional) and between its integration with other pillars forming the constitutive basis of a theatre act as such on the actual level of media (the artificial pervaded by “life”).

Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Supervisor: doc. PharmDr. Lenka Tůmová, CSc. Student: Tereza Sigmundová Title of Thesis: Flavonoids and natural coumarins interactions with conventional medicine Keywords: medicinal plant, flavonoids, coumarins, interactions The aim of this diploma thesis is to give an overview of the interactions between conventional drugs and simultaneous use of flavonoids, coumarins or medicinal plants containing flavonoids and coumarins. Cited literature includes articles published since 2000 till 2018, that were collected mostly using online databases (Web of Science, Science Direct, PubMed, Google Scholar). Flavonoids can affect the metabolism of drugs through interaction with CYP450 enzymes, P-glycoprotein, ABC or SLC transporters. This type of interaction is particularly important for drugs with narrow therapeutic index such as cyclosporin, digoxin or warfarin. Furanocoumarins contained for example in grapefruit juice cause irreversible inhibition of CYP3A4 enzyme and affect the pharmacokinetics of many drugs. The major drug classes that have been reported to present pharmacodynamic interactions with flavonoids or natural coumarins are anticoagulants, cytostatics and central nervous system agents. The thesis also describes...