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Interactions of polyomavirus structures in the endoplasmic reticulum and on the path to the nucleus
Svobodová, Terezie ; Huerfano Meneses, Sandra (advisor) ; Weber, Jan (referee)
Mouse polyomavirus is a member and model virus of Polyomaviridae family. In order to infect cells and produce viral progeny, the viral chromosome must be transported to the nucleus. Several studies suggest that virions are transporeted to the endoplasmic reticulum, from which they are transferred to the cytosol with assistace of host proteins. Two of these proteins are the chaperon, BiP (binding immunoglobulin protein) and the cochaperone, DNAJ B14. Polyomaviruses probably enter the nucleus through nuclear pores with the assistence of importins. These processes were mainly studied with SV40. In this work, we show that MPyV infection induces a change in distribution of the DNAJ B14 protein, which became clustered into foci, where it co-localizes with the viral capsid protein, VP1. The occurrence of foci varies during infection. With use of proximity ligation assay, we have shown that during an early fase of MPyV infection, DNAJ B14 and BiP get in the close proximity with VP1. It is suggested that negatively charged amino acids at the N-terminus of the minor capsid protein, VP2, are required for targeting virions to translocon and proteins associated with ERAD. We created MPyV with VP2 mutated in these amino acids. The negatively charged amino acid at position 17 is not necessary for successful...
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The image and image-consciousness in relation to the absolute subjectivity and the meontic absolute: the route of the phenomenology according to Eugen Fink
Coli, Anna Luiza ; Sepp, Hans Rainer (advisor) ; Giubilato, Giovanni (referee) ; Weber, José Fernandes (referee)
The present work intends to offer an interpretation key to Eugen Fink's philosophy from the leitmotiv of the negative. The approach taken here considers his philosophical project from a perspective of autonomy and originality regarding Fink's contemporary philosophies - Edmund Husserl and Martin Heidegger. The aim is to provide a counterpoint to the still predominant tendency of considering Fink mainly as an interpret of both Husserl and Heidegger. In the course of the work it becomes clear that the direct dialogue with other philosophies - for instance as Nietzsche's, Kant's and above all Hegel's philosophy - is the methodological procedure through which he raises his own philosophy. The negative accounts for a particular concept that not only traverses the two moments of Fink's philosophy, but in which the metaphysical transformation of his later thinking is most manifest. In order to distinguish these moments in Fink's work, we refer here on the one hand to a meontic matrix of thought, the one that prevailed during the period of assistance to Husserl, and on the other hand to a cosmological matrix of thought, which was developed as such only after Husserl's death. The movement of the negative concept ultimately reveals itself as a development in Fink's own conception of philosophy, which could...
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Music Therapy, Music Education and Adolescence
Weber, Jana
Music plays an important role in our lives. Especially during puberty, music is an important part of an adolescent's influence which supports the development of identity. In today's world, music is disappearing from interpersonal relationships. School is often the last place where young people can learn about themselves and the world around them through music. Looking at music education from this perspective, teaching music could be instrumental in developping soft skills. Unfortunately music education is often given only marginal consideration on the education plan in Czech secondary schools. As a consequence neither school management nor pupils understand its significance and music classes become a gathering of people with no interest. In many schools music education loses its meaning and its potential is wasted. Based on ideas of music therapy and artefiletic, the aim of this work is to find possible ways to encourage the interest of adolescents in this field again. One option is to motivate students to link school life with their life outside of school. They can learn through their own experiences in creative activities and discussions. This work is heuristically oriented, i.e. we do not seek answers, but rather consider the issues and come up with new ideas for future work. Our goal is to offer music...
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65/5000 Analysis and optimization of cycling transport in Bohumin
Weber, Jiří ; Radimský, Michal (referee) ; Matuszková, Radka (advisor)
This diplom thesis deals with the analysis of existing state and prorosal to improve bicycle parking in the part of town Bohumín. Thesis also deals with evalution of critical section of cycling traffic in the Bohumín. It propose the optimalization of the existing state, or a complete change of it. The aim of the proposal is to improve the traffic situation in place e.g. to increase safety and driving comfort.
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Economic Efficiency of Public Construction Project
Weber, Jiří ; Hromádka, Vít (referee) ; Korytárová, Jana (advisor)
This bachelor's thesis deal with economic efficiency of public construction project. Information about efficiency it takes from Cost-benefit analysis. Cost-benefit analysis compares economic cash-flow and it rates project´s usefullness versus total project´s cost. In the economic cash-flow are financial cash-flow, as investment, yield and cost, and socioeconomic cash-flow. In thit part is project´s effect on the community in the formof benefits and savings. This effect we can see in a project as a unit price. For this cash-flow result indexs of ekonomic efficiency, which evaluates efficiency, or demand of grant. The one of part economic efficiency is project life cycle´s financial sustainability.
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The role of Hepatitis B virus capsid protein in the host ubiquitin proteasome pathway
Eliáš, Vratislav ; Weber, Jan (advisor) ; Šmahel, Michal (referee)
Hepatitis B virus (HBV) is a Hepadnaviridae virus infecting mammals. Its infection can result in an acute or chronic infection. Chronic infection can result in hepatocellular carcinoma and liver cirrhosis, potentially leading to death of the patient. HBV is a small 42 nm virus with a genome length of 3.2 kb encoding seven viral proteins. HBV Core protein (HBc) is a capsid forming protein which is pleiotropic in function. We have identified two ubiquitin ligases which could interact with this protein: F-box only protein 3 (FBXO3; E3 ubiquitin ligase) and Ubiquitin conjugating enzyme E2 O (UBE2O; E2/E3 ubiquitin ligase). By employing multiple methods we have confirmed these interactions. Co- immunoprecipitation and further western blot analysis unveiled multiple new insights into the ligases′ impact on HBc: FBXO3-mediated HBc polyubiquitination stimulation and UBE2O-mediated HBc monoubiquitination promotion. FBXO3's and UBE2O's role in HBV life cycle was investigated as well. By silencing the expression of FBXO3 and UBE2O respectively, we have observed changes in HBV replication levels: FBXO3 serves as an inhibitor of HBV replication, while UBE2O stimulates the course of HBV life cycle. Further investigation of these newly-discovered understandings may lead to a whole new HBV - host interplay...
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Rous sarcoma virus replication blocks in mammalian cells
Koslová, Anna ; Hejnar, Jiří (advisor) ; Ruml, Tomáš (referee) ; Weber, Jan (referee)
One of the important tasks of virology and immunology is to explore the species- and cell-barriers preventing virus horizontal transmission and reveal the ways how viruses overcome these barriers and "adapt" to different species. This work is based on a well- established retroviral model - avian Rous sarcoma virus (RSV) and studies virus replication blocks in mammalian cells at both pre- and post-integration level. Interaction of the viral envelope glycoprotein (Env) with a specific cellular receptor mediates virus entry into cells. Although mammalian orthologues of specific chicken receptors do not support RSV entry, it was observed that some RSV strains are able to enter mammalian cells. Several RSV-transformed rodent cells lines were described and analysis of provirus H20- RSV in one these cells lines (hamster H-20 tumor cell line) showed multiple mutations including two crucial amino acid substitutions in different regions of Env. Substitutions D32G and L378S confer virus transmission to hamster, human and also chicken cells lacking the appropriate receptor. Altered conformation of H20-RSV Env is similar to a receptor-primed (activated) state of Env. This observation indicates that virus can circumvent the need of original cell receptor because of spontaneous Env activation caused by single...
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Trends in Medicines Usage in ATC Group L01
Weber, Jakub ; Lešetický, Ondřej (advisor) ; Kostřiba, Jan (referee)
The work analyzes trends in medicines used in specialized medical centers from ATC (Anatomical-Therapeutic-Chemical Classification System) L01 in Czech Republic from perspective of the State Institute of Drug Control (SUKL) and of Czech Health Insurance Companies (Vseobecna zdravotni pojistovna Ceske republiky as the biggest one, and rest joined in Svaz zdravotnich pojišťoven Ceske republiky). SUKL perspective considers medicines delivered to healthcare facilities and the other side providing information regarding payments for L01 medicines between years 2012-2015. Conclusions presented show slightly decreasing trend in terms of number of packages delivered (935 ths. in 2012 and 931 ths. in 2015). On the other hand, value of medicines has increased from 5 067 mil. CZK in 2012 to 6 568 mil. CZK in 2015. The increase in value and decrease in volume in the same time (while prices remain stable or decreasing) indicates trend in increasing size of packaging or entry of new strength (with more active ingredient and therefore more expensive) e.g. entry of Herceptin 600 mg in 2014. This paper further identifies the most expensive medicines in ATC L01 group and starts a discussion about possible reasons of discrepancies between different sources of data which are for the most expensive medicines in hundreds of mil. CZK in value and ths. of packages in terms of volume (e.g. Avastin in 2014: difference 104 mil. CZK and 5 598 packages).
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Interactions of polyomavirus structures in the endoplasmic reticulum and on the path to the nucleus
Svobodová, Terezie ; Huerfano Meneses, Sandra (advisor) ; Weber, Jan (referee)
Mouse polyomavirus is a member and model virus of Polyomaviridae family. In order to infect cells and produce viral progeny, the viral chromosome must be transported to the nucleus. Several studies suggest that virions are transporeted to the endoplasmic reticulum, from which they are transferred to the cytosol with assistace of host proteins. Two of these proteins are the chaperon, BiP (binding immunoglobulin protein) and the cochaperone, DNAJ B14. Polyomaviruses probably enter the nucleus through nuclear pores with the assistence of importins. These processes were mainly studied with SV40. In this work, we show that MPyV infection induces a change in distribution of the DNAJ B14 protein, which became clustered into foci, where it co-localizes with the viral capsid protein, VP1. The occurrence of foci varies during infection. With use of proximity ligation assay, we have shown that during an early fase of MPyV infection, DNAJ B14 and BiP get in the close proximity with VP1. It is suggested that negatively charged amino acids at the N-terminus of the minor capsid protein, VP2, are required for targeting virions to translocon and proteins associated with ERAD. We created MPyV with VP2 mutated in these amino acids. The negatively charged amino acid at position 17 is not necessary for successful...
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Contribution of gag region to overall HIV replicative fitness in patients with different disease progression
Suchý, Tomáš ; Weber, Jan (advisor) ; Šmahel, Michal (referee)
Human immunodeficiency virus (HIV) is globally spread virus without available cure. Since its life-long presence, virus is carefully monitored as well as patient's immunological status. Replicative fitness of the virus is one of important aspects which can be taken into account, when monitoring HIV. Here, we are measuring HIV replicative fitness of gag recombinant viruses and comparing the results with replicative fitness of primary isolates. Further, we are comparing our findings of replicative fitness change over time with disease progression in the patient. We found that gag can be major contributor to overall fitness, although not in all cases. Additionally, we observed a correlation of replicative fitness development and slope of patient's CD4+ T cells. Moreover, this relation was even more noticeable in patients with slow disease progression or in carriers of protective alleles. In summary, our results extend the understanding of replicative fitness and its role in disease progression; and pave the way to use the recombinant HIV for replicative fitness measurement in clinical practice. Keywords: HIV, replicative fitness, recombinant virus, HIV disease progression, gag
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