Univerzita Karlova v Praze Filozofická fakulta katedra kulturologie DIPLOMOVÁ PRÁCE Jana Velichová Volný čas jako fenomén dnešní doby Praha 2012 Vedoucí práce: PhDr. Ondřej Hubáček, Ph.D. 2 Abstrakt Leisure time is topical problem of the modern society, as well as a multidimensional phenomenon. It is associated with a number of other current issues that affect various areas of the life of the society. The thesis focuses on their identification, their historical development, and their reflection from the point of view of social sciences. It attempts to interpret them as socio-cultural phenomena. These problems manifest themselves in quite divergent areas, such as gender differences, influence of technology and the media on the society, economic behaviour of people, self- presentation, social activities, impact of new technology, dissemination of socially pathological phenomena, escapist functions of the media, creation and reproduction of social networks and social groups. The thesis applies an axiological and non- axiological approach to the examined phenomena. The solution of the topic is complemented with findings relating to the specified problems, which have been obtained through an empirical survery.

Human Caprin-1 and Caprin-2 are prototypic members of the caprin (cytoplasmic activation/proliferation-associated protein) protein family. Vertebrate caprin proteins contain two highly conserved homologous regions (HR1 and HR2) and C-terminal RGG motifs. Drosophila caprin (dCaprin) shares HR1 and RGG motifs but lacks HR2. Caprin-1 and Caprin-2 have important and non-redundant functions. The detailed molecular mechanisms of their actions remain largely unknown.

One of the key events in a virus-infected host cell is the activation and repression of a large number of host genes. In recent years, such differentially expressed host genes have been identified for several viruses, bacteria and parasites. Such studies indicate that reprogramming of host transcriptome during infection by a pathogen is a major component of host response and many of the reprogrammed genes may promote or prevent pathogen infection or may contribute to pathogen-induced pathological changes. Host gene expression changes have been studied for a number of viruses in cell lines. However, in case of neurotropic viruses which infect nonrenewable populations of central nervous system (CNS), changes in the host gene expression need to be studied in the intact host rather than cells grown in culture. Since such studies are reported only for a few neurotropic viruses, an attempt has been made in this thesis to identify and characterize genes that are differentially expressed in the mouse brain during infection by Japanese encephalitis virus (JEV) and rabies virus. Using subtraction hybridization technique, subtraction cDNA libraries were generated representing mRNAs that are induced or repressed in the mouse brain during JEV infection. Sequence analysis of the 350 isolated clones resulted in the identification of 73 unique genes. Out of these, 66 were of forward library clones (upregulated genes) and 7 of reverse library clones (downregulated genes). The forward library clones was clustered in different functional groups such as, proteins involved in immune response and interferon-inducible proteins, GTPase and GTP binding proteins, transcriptional regulators, enzymes, ribosomal proteins, neuronal proteins, carrier proteins, DNA-binding proteins, miscellaneous and proteins of unknown function. The differential expression of all these genes was further validated by northern blot analysis of brain RNA isolated from normal and JEV-infected mice, which indicate that out of 66 forward library clones 33 were genuinely upregulated in JEV-infected mouse brain, whereas all 7 reverse library clones were repressed in JEV infection. Since vaccination is known to prevent virus replication in the brain, host gene expression changes in mice immunized with BIKEN JE vaccine was also examined. There was a good correlation between inhibition of JEV replication and reduced expression of JEV-inducible CNS genes in the vaccinated mice. To check whether JEV-induced CNS genes identified in this study are specific to JEV or can be induced by any other neurotropic virus, expression patterns of 15 randomly chosen genes were checked in RV infected mouse brain. Results indicated that all the chosen genes are modulated in the same way during RV infection as well. Comparison of JEV-induced gene expression changes with those induced by other neurotropic viruses indicated that 83% of the JEV-inducible mouse CNS genes are also induced by Sindbis virus, a neurotropic virus of the family alphaviridae, indicating that despite diverse life cycles, these two viruses may activate common host signaling pathways. This study also led to the identification of 9 unique JEV-inducible genes (LRG-21, VHSV induced gene1, Tpt1, SLC25A3, Olfm1, Ina/NF-66, Dst/Bpag1, Mdm2 and Gbp5) which are not reported to be activated by any other neurotropic virus. Since it is beyond the scope of this study to characterize all the JEV-induced and repressed genes, two genes were chosen for a detailed analysis. These are: JEV-inducible gene encoding GARG-39 protein which is a member of the glucocorticoid attenuated response gene family and an unannotated, JEV-repressible gene designated in this study as clone # 137. The gene encoding GARG-39 identified as a JEV-inducible gene in this study was originally discovered as lipopolysaccharide- and interferon-inducible gene in macrophages. This protein contains tetratricopeptide repeat (TPR) motifs that are known to be involved in protein-protein interactions. However, the function of this protein remains unknown till date. Therefore the gene was cloned and over-expressed in E. coli and antibodies were raised against the recombinant protein. Western blot analysis revealed that GARG-39 protein is detectable only in JEV-infected but not in the normal mouse brain. Surprisingly, immunoflourescence studies carried out in NIH3T3 cells revealed that GARG-39 is localized in the cytosol of normal cells and it colocalizes with α-tubulin in the mitotic spindle in a small fraction of cells which are in the mitotic stage. Further, in an in vitro assay, GARG-39 was found to interact with taxol-stabilized tubulin polymers. Since microtubules are known to play an important role in virus assembly, it is possible that GARG-39 may have a role in virus assembly and maturation. Alternatively, microtubule-associated proteins are implicated in several neurodegenerative disorders including Parkinson’s, Alzheimer’s and mental retardation and therefore, a role for GARG-39 in virus-induced neuropathogenesis cannot be ruled out. In addition, the expression of GARG-39 in normal dividing cells in the culture indicates a role for this protein in mitosis. In a normal mouse brain, mitotically active cells are very low in number and hence GARG-39 expression (both at the RNA and protein levels) is below the detection limits. JEV infection may trigger mitotic activity in brain leading to increased expression of GARG-39. One of the cDNA clones identified in this study, designated as clone # 137, hybridized to a ~2.6 kb transcript which was found to be down regulated in the mouse brain by JEV as well as rabies virus. A series of investigations led to the conclusion that clone #137 corresponds to the 3′ end of a ~2.6 kb transcript encoding mouse calcium calmodulin kinase inhibitor II α (mCaMKIINα). Interestingly, only the α isoform but not the β isoform of mCaMKIINα mRNA is down regulated in the mouse brain during JEV infection. Since the physiological function of mCaMKIINα is not known, the gene encoding 8 kDa mouse mCaMKIINα open reading frame was cloned into an E. coli expression vector and antibodies were raised against the purified recombinant protein. Surprisingly, antibodies raised against the ~8 kDa recombinant mouse CaMKIINα reacted with a ∼37 kDa mouse brain protein. This protein designated as CaMKIINα-immunoreactive protein (CaMKIINα-IRP) is also down regulated during JEV infection and is localized in the post synaptic density (PSD) of normal mouse brain. In addition, distinct changes are also observed in the subcellular localization and phosphorylation of CaMKIIα leading to an increase in cytosolic CaMKII activity in JEV-infected mouse brain. The differential regulation of CaMKIIα and CaMKIINα during JEV infection suggests a possible role for CaMKII signaling pathway in JEV infection and/or JEV-induced neuropathogenesis in the CNS. Conclusions: • A number of host genes whose expression is modulated in the mouse brain during JEV and/or rabies virus infection have been identified. • One of the JEV-inducible genes encoding the GARG-39 protein was shown to be a microtubule-associated protein with a possible role in mitosis. • One of the JEV-repressible genes was found to encode the mouse CaMKIINα mRNA. • A novel JEV-repressible ∼37 kDa protein immunoreactive to antibodies raised against the recombinant CaMKIINα was identified in the post synaptic density of the mouse brain.

This thesis has both a theoretical and a research section. The theoretical section investigates how literature defines the basic concepts relating to unemployment?. An analysis of unemployment in various social groups is also included. The research section comprises of personal experiences both from being a candidate applying for a job position, as well as working as an employment officer. The quantitative research focuses on candidates of all social groups applying for a job. Qualitative research of the labour office's employees is undertaken with figures and entries from the department of the labour office (KoP ÚP ČR) completing this empirical study. The acquired data is then evaluated and an conclusion formed as to how the current situation can be improved.

In this thesis, the single-level correlated quantum dot attached to two BCS superconducting leads is analyzed. A difference in the superconducting phases of the leads induces the DC Josephson supercurrent in the junction. In this setup, the influence of asymmetrical dot-lead couplings on transport properties is clarified analytically. The coupling asymmetry and the phase difference can be combined into one function, which allows us to calculate physical properties of a system with coupling asymmetry from the properties of its effective symmetric counterpart. The coupling asymmetry turns out to be an important parameter which influences the position of the 0 − �෰ quantum phase transition even in the strongly correlated Kondo regime. Further, this thesis contributes to the interpretation of an AC Josephson current measurement, in which a surprising drop in the amplitude was observed in the Kondo regime. The experimental setup is characterized using numerical renormalization group calculations of the equilibrium many-body spectra. Possible quantum-point-contact- based interpretations are discussed. Although a drop in the AC Josephson current at the experimental bias voltage is also expected in a quantum point contact, we conclude that the physical mechanisms causing it in the quantum dot system are likely not...

This Master thesis deals with usage of radar signal for vehicle classification. The thesis uses radar modules with continuous wave based on Doppler effect. Radar signal is processed by a series of signal processing method finished by Fourier transform. Data produced by FFT is used to create SVM and AdaBoost classifier which can be used to classify vehicles into groups.

The purpose of this paper is to assemble measurement chain dedicated to contactless measuring of acoustic fields emitted by test sources and to validate its results. At first author discusses possible optic methods which can be used to measure acoustic fields, then he talks about theoretical foundations which are needed to measure data and to reconstruct final data. The last two chapters are dedicated to cover basics about created program and to analyse various aspects which influence measured and reconstructed data. Created measurement chain is fully functional, at the end of this thesis are proposed some ways of improving

This thesis focuses on simulation modelling of a piezoelectric MEMS filter, which may be used as a part of cochlear implant. A model of this filter is created in the programme ANSYS. A research on experimental cochlear implants and materials suitable for this application is also included.

This thesis is focused on a study of CdTe and CdZnTe semiconductor detectors working under high flux of radiation. We studied experimentally an influence of high flux of X-rays and optical radiation on polarization of the detector. The polarization phenomenon decreases the efficiency of the detector due to a screening of an applied electric field by a space charge accumulated at deep levels due to a trapping of photogenerated carriers. In order to measure the electric field profiles in the detectors we employed a method based on cross polarizers technique and Pockels effect. The main objective of this work was to study the possibilities of an optical de-polarization of CdTe and CdZnTe detectors for different photon energies of additional light, its dynamics and physical origin. We have found that detectors can be de-polarized by above bandgap light. Moreover, CdZnTe detector can be depolarized by near infrared light and in a pulse mode. The de- polarization is associated with a compensation of the space charge at deep traps.

This work presents study of magnetocaloric properties of compounds exhibiting unusual magnetic characteristics. Several systems were studied: TbNiAl, (Tb,Y)NiAl, TbNi(Al,In), TbFeAl, GdFeAl, Ni2MnGa based compounds and amorphous metallic alloy Gd-Co-Al-Y. Influence of magneto-crystalline anisotropy on magnetocaloric effect was studied on singlecrystalline sample of TbNiAl compound. The maximum of entropy change was measured with orientation of magnetic field along c axis and compared to polycrystalline sample a 100% increase was observed. Substitutions of Y and In in TbNiAl led to a change of magnetic ordering. Both substitution led to increase of RCP values of studied material. TbFeAl and GdFeAl compounds, characterized by partial disorder of Fe and Al atoms, showed magnetocaloric effect in wide temperature region which led to interesting values of RCP~350 J kg-1 . Very broad region of magnetocaloric effect was also observed on amorphous metallic material of Gd-Co-Al-Y. The effect of Er substitution in Ni2MnGa based compound on its magneto-structural transition, connected with inverse magnetocaloric effect, was studied. The direct measurement method of adiabatic temperature change is described and instrument for such measurement, developed in collaboration with FZU AVČR, v.v.i., is presented.