Silicon carbide (SiC), is a wide band gap (2.4 eV < Eg < 3.3 eV) semiconducting material well known for its potential applications in high-temperature, high-power, high-frequency or hard radiation resistant devices. In this thesis, we are broadening elementary knowledge about this material. We identify energy levels in the material, using Photo-Hall effect spectroscopy supported by the temperature dependency of classic Hall effect measurement and temperature dependent photoluminescence. This knowledge is essential to allow SiC application as a radiation detector.

Nela Dvořáková 19.7.2019 1 Abstract We study a representative of a diverse group of stars exhibiting the B[e] phe- nomenon, designated MWC 939. This object might be an important link be- tween the stage of the asymptotic giant branch and planetary nebula stage as it is one of a few such objects observed. Our focus is aimed at spectral analysis of the circumstellar matter around the central star. We assemble a line list for MWC 939 and observe variations of its spectrum. Electron density and temper- ature are estimated using nebular diagnostics of ratios of [S II] λλ 6716, 6731 ˚A and [O I] λλ 6300, 6364 ˚A and 5577 ˚A. These estimates are then used for detailed calculations of the structure of circumstellar envelope. 1

This thesis is an attempt to reflect on the theme of phenomena and sensuous world in the context of environmentalism. It deals with the condemnation of phenomena in the modern era science and especially in the contemporary neo-Darwinian biology, and with various attempts at rehabilitation of sensory perception as a relevant way of relating to the world in biology (Adolf Portmann, Hans Driesch) and philosophy (phenomenology, particularly Maurice Merleau-Ponty). The main subject of the thesis is the eco- phenomenology of David Abram and a comparison of his approach with the one held by contemporary biology. Based on this comparison it is argued in what manner may the thoughts of David Abram, and the effort to rehabilitate phenomena in general, be of benefit to the environmental discourse, particularly in terms of forming an original epistemology and ontology and concerning the ethical motivations. KEYWORDS: Abram, phenomenon, environmentalism, neo-Darwinism, phenomenology, eco-phenomenology

Japanese encephalitis virus (JEV) is one of the major causes of encephalitis in Asia. JEV causes serious inflammation of the brain, which may lead to permanent brain damage and has a high mortality rate. Almost 3 billion people live in JE endemic areas and JEV causes an estimated 20,000 cases of disease and 6000 deaths per year. JEV is a positive stranded RNA virus belonging to the Flavivirus genus of the family Flaviviridae. The genome of JEV is about 11 kb long and codes for a polyprotein which is cleaved by both host and viral encoded proteases to form 3 structural and 7 non-structural proteins. JEV transmission occurs through a zoonotic cycle involving mosquitoes and vertebrate amplifying hosts, chiefly pigs and ardeid birds. Humans are infected when bitten by an infected mosquito and are dead end hosts. The role of humoral and cell mediated immune responses during JEV infection have been studied by several groups. While the humoral responses play a central role in protection against JEV, the cell mediated immune responses contributing to this end are not fully understood. The MHC molecules have been known to play predominant roles in host responses to viral infections and the consequences of virus infection on the expression of MHC molecules are varied. The expression of MHC-I molecules is known to decrease upon infection with many viruses such as HIV, MCMV, HCMV, Adv, and EBV. In contrast, infection with flavivirus such as West Nile Virus (WNV) has been shown to increase the cell surface expression of both MHC-I and MHC-II molecules. It has been reported previously that WNV infection increases the cell surface expression of adhesion molecules such as ICAM-1, VCAM-1 as well as E-Selectin and these changes were mediated directly by WNV and not by soluble cytokines. In contrast to classical MHC-I molecules, the nonclassical MHC-I molecules do not belong to a single group of structurally and functionally homologous proteins and normally have lower cell surface expression. Earlier studies have shown that the expression of nonclassical MHC-I molecules were induced during infection with JHM strain of mouse hepatitis virus (MHV). However, the functional significance of this induction is unclear. Expression of nonclassical MHC-I molecules upon flaviviral infection is not very well understood. In this thesis, evidence is presented that JEV infection induces the expression of both classical and nonclassical MHC-I molecules on primary mouse brain astrocytes, mouse embryonic fibroblasts (MEFs) and H6 (hepatoma cell). The levels of adhesion molecules as well as molecules involved in antigen processing and presentation were also analyzed and our results clearly demonstrate that JEV infection induces their expression on astrocytes, MEFs and H6. The role of NF-κB and type-I IFNs in the induction of classical and nonclassical MHC-I molecules as well as molecules involved in antigen processing and presentation were also analyzed and our results demonstrated that type-I IFN mediated signaling is responsible for the induction of these molecules during JEV infection. Chapter 1 discusses the innate and adaptive immune system, the role of classical and nonclassical MHC molecules in the initiation of immune response and diverse strategies adapted by different viruses to evade the immune response. It also includes a detailed discussion about the IFN and NF-κB signaling pathways and their modulation by viral infection. Finally, the genome organization, epidemiology, transmission cycle, pathogenesis and pathology, clinical features, humoral as well as cell mediated immune response to JEV infection and the current vaccine status to JEV infection are briefly discussed. Chapter 2 describes the general materials and methods used in this study. It includes the details of the reagents and cell lines used in the experiments. It also discusses the various techniques such as RT-PCR, FACS analysis, EMSA and ELISA. Chapter 3 focusses on the validation of different knockout MEFs used in the study as well as confirming the purity of primary astrocyte cultures established from pub brains. The susceptibility of various cells to JEV infection has also been investigated. Our results confirmed the authenticity of all the cells and the purity of primary astrocyte cultures used in the study. Our results also indicated that all the cells used in the study are susceptible to JEV infection. Chapter 4 discusses the expression of MHC and related genes involved in immune response upon JEV infection of primary mouse brain astrocytes, MEFs and H6. Chapter 4 demonstrates for the first time that JEV infection induces the expression of nonclassical MHC-I or class Ib molecules namely Qa-1, Qb1 and T10 in addition to the induction of classical MHC-I molecules. In contrast to WNV, there was no increase in the cell surface expression of MHC-II molecules upon JEV infection of primary mouse brain astrocytes. JEV infection also induces the expression of adhesion molecules as well as molecules involved in antigen processing and presentation namely Tap1, Tap2, Tapasin, Lmp2, Lmp7 and Lmp10. Chapter 5 demonstrates that JEV infection induces NF-κB activation in astrocytes and MEFs. Studies using MEFs deficient in classical and alternate pathways of NF-κB activation indicate that JEV activates the classical pathway of NF-κB activation and is dependent on canonical lKKβ/IKK2 activity. JEV infection of astrocytes, MEFs and H6 induces the production of type-I IFNs. To determine the mechanism of type-I IFN induction during JEV infection, MEFs deficient in NF-κB signaling and IFN signaling were used. Results indicate that type-I IFN production in MEFs occurs by both NF-κB dependent and independent mechanisms. In contrast, the production of IFN-α was completely abrogated in IFNAR-\- MEFs whereas IFN-β production was greatly reduced. Production of type-I IFNs in IFNGR-\- MEFs is also reduced upon JEV infection but the reason for this is unclear. Chapter 6 demonstrates that JEV induced expression of classical MHC-I molecules occurs by type-I IFN mediated signaling. This result is in contrast to WNV infection, in which both NF-κB and type-I IFNs are involved in the induction of classical MHC-I molecules. Type-I IFNs were also shown to be involved in the induction of nonclassical MHC molecules namely, Qa-1 and Qb1 during JEV infection. In contrast, the expression of T10, another nonclassical MHC molecule occurs independent of type-I IFN signaling. The expression of molecules involved in antigen processing and presentation namely, Tap1, Tap2, Lmp2 and Lmp7 was type-I IFN-mediated, whereas the expression of Tapasin and Lmp10 was mediated by both type-I IFN dependent and independent mechanisms. The expression of VCAM-1 was dependent on NF-κB mediated signaling. Chapter 7 precisely describes the underlying mechanism of induction of MHC and various other related molecules and their significance during JEV infection. In addition, it also includes a working model for the induction of these molecules during JEV infection. In summary, this is the first study in which the mechanism of JEV mediated induction of classical as well as nonclassical MHC molecules has been studied in detail. This study clearly demonstrated that type-I IFNs are involved in the induction of classical and nonclassical MHC-I molecules during JEV infection. The functional significance of this JEV mediated induction of classical MHC-I molecules is unclear, but it has been proposed that this is to escape from the action of NK cells. The absence of MHC-II induction during JEV infection could be important because it may lead to the initiation of an immune response which is different from that induced during other viral infections which induce the expression of MHC-II molecules. In contrast to classical MHC-I molecules, the functional and biological significance of nonclassical MHC-I molecules are poorly studied. Nonclassical MHC-I molecules play an important role in bridging adaptive and innate immune response. So the nonclassical MHC molecules induced during JEV infection may play an important role in the initiation of immune response during JEV infection. The role these nonclassical MHC-I molecules in antigen presentation during JEV infection is not known. These nonclassical antigens are also recognized by NK and γδT cells, thus the expression of nonclassical MHC-I molecules during JEV infection might also confer a protective role.

This thesis deals with the aspects of methods for pulsed high-level EM quantities measurement. Methods for current and voltage measurement in pulsed power generator and power measurement in pulse microwave generator are discussed. New approaches to single-shot measurement methods application are proposed. The theoretical analysis of suitable sensor designs is performed. The magneto-optic measurement method has been experimentally realized. On the basis of experimental results a fiber-optic current sensor has been designed and theoretically analyzed. For identification and measurement of the free-space electromagnetic pulse a combined calorimetric sensor has been designed and built.

David Wagenknecht: Abstract of a diploma thesis Optical response of magnetic materials, 2014 Magnetooptical properties of anisotropic semiconductors are studied to describe asymmetry of Ga1−xMnxAs, because theoretical calculations predict extraordinary behaviour of reflectivity. Analytical formulae to describe materials with non-diagonal permittivity are derived and they are used for the numerical calculations to describe the optical response of the samples available for the measurement. The transversal Kerr effect is calculated and it exhibits asymmetry in both rotation of the plane of polarization and ellipticity of circularly polarized light due to asymmetry in reflectivity. Moreover, longitudinal and polar magnetization are studied because of the influence on the observability of the phenomena. Results are not only used to discuss conditions, which must be satisfied to prove the asymmetry, but also the actual experimental setup is designed to prepare the measurement. 1

Cytotoxic T Lymphocytes (CTL) are known to play an important role in clearing infectious virus from infected hosts in a variety of viral infections. Depending on the type of virus and mode of virus entry both class I and class II restricted CTL can contribute to protection from virus-induced disease. Although CD8 positive CTL are associated with virus elimination and control in many viral infections, elimination of neurotropic viruses from the Central Nervous system (CNS) is more complex due to the lowered expression of MHC antigens on neuronal cells. This failure to constitutively express high levels of MHC antigens by neurons could serve as an advantage to avoid damage to this differentiated and non-renewable tissue. However, abnormal induction of MHC antigens in the CNS mediated by CD4 positive lymphocytes or by astrocytes have also been shown to cause destructive inflammation in the CNS. The present study deals with CTL responses against one such neurotropic virus called Japanese Encephalitis Virus (JEV). JEV is a positive-stranded RNA virus that belongs to the flavivirus group, a group that is among the most important agents causing human encephalitis worldwide. Although passive transfer of monoclonal antibodies against this virus has been shown to confer protection of mice from lethal challenge with virus, neither the presence of CTL against this virus nor its role in conferring protection has been reported so far. Understanding the CTL responses against these viruses acquired importance in light of recent reports that neurovirulence of JEV and yellow fever viruses can be enhanced by the administration of virus specific antibodies. Hence this study was undertaken to examine the possibility of raising CTL specific to JEV. The specificity of the CTL raised, their therapeutic value and the ability of different lymphocyte subsets to mediate protection in vivo are dealt with in this study. Generation of CTL against JEV The generation of CTL against JEV in BALB/c mice, requires MHC defined cell lines that not only support virus infection but are also histocompatible. Several cell lines were initially examined for their ability to support JEV infection as a prc-rcquisitc before their utilization in in vivo and in vitro stimulation protocols aimed at generating JEV-specific CTL. Virus infection was monitored by immunofluorescence using JEV envelope-specific monoclonal antibodies as well as by titration of virus produced from infected cells by plaque assays. These different cell lines that were characterised for their ability to support JEV infection were then utilised to generate and monitor antiviral CTL. Several in vivo immunisation protocols were examined initially find out which of these infected cells prime BALB/c mice efficiently for generation of virus-specific CTL upon secondary stimulation in vitro with infected syngeneic cells. Immunisation of mice with infected cells per se was preferred over free virus since this was thought to facilitate priming against some viral non-structural proteins preferentially found on infected cells in addition to other viral structural proteins. It was observed that not only infected syngeneic and allogeneic cells but also infected xenogeneic cells prime BALB/c mice for the generation of JEV- specific CTL upon secondary restimulation in vitro. An optimal protocol was standardised for the generation of CTL against JEV. This included primary in vivo immunisation of mice followed by secondary in vitro restimulation of splenocytes with infected syngeneic cells. Either immunisation alone or in vitro stimulation of naive splenocytes alone was unsuccessful. The effector cells generated specifically lysed JEV-infccted P388D1 targets but not uninfected P388D1 or YAC-1 targets suggesting that the lysis on infected targets is not mediated by Natural Killer activity. Specificity and MHC restriction of anti JEV Effectors Cell depletion studies using complement mediated lysis were performed to examine the phenotype of the cells mediating virus specific lysis of infected targets. Depletion of Lyt 2.2+ or Thy 1+ but not L3T4+ sub-populations of effector cells inhibited lysis of infected targets showing that the effectors mediating virus-specific lysis were Lyt-2+ T cells. Examination of target specificities and MHC restriction of the antiviral CTL generated showed that although infected xenogeneic cells were used for immunisation, the effector cells recognised only infected syngeneic (P388D1, Sp2/0) and semisyngeneic (Neuro 2a, YAC-1) cells. Virus-specific recognition was found to be class I Kd and class I Dd restricted. These effector cells were also found to recognise cells infected with a closely related flavivirus, West Nile Virus (WNV) suggesting that they were crossreactive to some degree. Based on the consensus motif that has been established for H-2Kd associated peptides, several nonamers were predicted as possible CTL epitopes by scanning the deduced amino acid sequences of three strains of JEV and WNV. Among several predicted nonamers, three peptides were examined for their ability to reconstitute lysis of uninfected targets by polyclonal anti JEV CTL populations. Results demonstrate that peptides derived from NS1 and NS3 but not NS5 protein of JEV were able to partially reconstitute lysis of uninfected targets by effectors when pulsed with the appropriate peptide. Protective ability of the CTL raised against JEV To examine whether anti-JEV effectors raised in vitro could confer protection from intracerebral challenge with JEV, these effectors were adoptively transferred into adult BALB/c mice intracerebrally along with 10 x LDJ0 dose of JEV. More than 55% of these animals were protected from death and survived beyond 100 days after JEV challenge demonstrating that adoptively transferred anti-JEV effectors could indeed confer protection from lethal challenge with JEV. However, adoptive transfer of effectors by either intravenous or intraperitoneal routes did not protect adult mice from the lethal effects of intracerebral challenge with JEV. In contrast to adult mice, newborn mice were not protected from death by the adoptively transferred effector cells. This was also supported by experiments where a correlation was observed with the increasing age of mice and the success of protection conferred by the adoptively transferred effector cells. To establish the identity of cell subsets responsible for protection, Lyt 2, L3T4 or Thy 1 positive cells were specifically depleted from the polyclonal CTL by multiple cycles of complement mediated lysis and the remaining cells were adoptively transferred intracerebrally along with 10 x LD of JEV. These results demonstrate that both Lyt 2 and L3T4 positive T cells present in the effector population were necessary to confer protection of adult mice. Examination of virus-specific neutralising antibodies in the sera of protected and unprotected mice revealed that presence of L3T4 positive cells in the adoptively transferred population increases virus-specific neutralising antibodies. However presence of neutralising antibodies alone was not sufficient to confer protection. The protection required both Lyt-2 and L3T4 positive cells together. These studies could in the long term throw some light on similar observations about age dependant susceptibility to JEV in humans.

Popis magnetodynamických vlastností nanomagnetů a nanostrukturovaných magnetických materiálů vyžaduje metody vhodné pro zkoumání typické časové odezvy těchto systémů, tj. v řádu nanosekund a méně. Nedostatek technik, vhodných právě pro charakterizaci v časové doméně, je spojen s možnostmi současné elektroniky. Další možný přístup, jak popsat vlastnosti nanomagnetů, je charakterizace ve frekvenční doméně v pásmu GHz. Nejrozšířenější technikou charakterizace ve frekvenční doméně je měření feromagnetické rezonance (FMR). Ze spekter FMR lze získat cenné informace o systému: parametr tlumení, saturační magnetizace atd. Metoda, kterou využíváme k detekci excitací spinových vln, má za cíl zjednodušení charakterizace. Využíváme termoelektrickou detekci spinových vln v magnetických drátech prostřednictvím anomálního Nernstova jevu. Metoda je založena na disipaci tepla uvnitř magnetické vrstvy v důsledku útlumu spinových vln, a proto dochází k vytvoření teplotního gradientu směrem k substrátu (kolmo k povrchu). To vede k vytvoření elektrického pole kolmého jak na teplotní gradient, tak na směr magnetizace. Napětí je obvykle v řádu V, proto může být měřeno obvyklým laboratorním vybavením. Navzdory své jednoduchosti poskytuje tato metoda velmi zajímavé výsledky a může být použita pro charakterizaci magnonických vlnovodů, magnonických metamateriálů, emitorů spinových vln a dalších zařízení, pracujících se spinovými vlnami.

This diploma thesis describes the piezoelectric phenomenon and its use for positioning with nanometric precision in laboratory use. In the thesis is description of direct and indirect piezoelectric phenomenon, various types of piezoelectric actuators and ways of their control with the aim of eliminating their hysteresis. The goal is to design a controller for piezo actuator built in the three-axis nanometric manipulator Thorlabs MAX341/M.

Magnonika je novým odvětvím výzkumu, který se zabývá fyzikou spinových vln. Magnonika jako vědní obor nabízí nové možnosti například v nediskrétních výpočtech na základě vlnového charakteru spinových vln. Při výrobě magnonických prvků klasickými metodami není možné příliš měnit charakter materiálů, ze kterých jsou jednotlivé prvky vyrobeny. Tento fakt silně omezuje univerzálnost vyrobených struktur. Cílem této práce je aplikovat nový typ materiálu do oboru magnoniky. Specifikum daného materiálu je možnost zápisu magnetických struktur pomocí iontového svazku. Ukazuje se, že tyto struktury mají velice zajímavé magnetické vlastnosti, které lze velice přesně řídit právě strategií ozařování iontovým svazkem. Na základě fázově rozlišené Brillouinovy spektroskopie jsme získali disperzní relaci spinových vln v tomto systému a tím i důležité parametry systému. Pozorování podkládáme mikromagnetickými simulacemi a analytickými modely. Vlastnosti systému pro magnonické aplikace prezentujeme na třech prototypických sadách struktur, které nelze vyrobit pomocí klasických materiálů.