The development of the central nervous system is dysregulated in neurodevelopmental disorders such as intellectual disability, autism spectrum disorder, and epilepsy. These three disorders have different clinical features, yet there is high comorbidity between them. They can be difficult to study due to their highly complex aetiologies, however there are various monogenic diseases that can cause all of them, including SYNGAP1 haploinsufficiency where the synaptic guanosine triphosphatase (GTPase)-activating protein (SYNGAP) protein levels are highly reduced; Fragile X syndrome where the fragile X mental retardation protein (FMRP) is no longer translated; and DNM1 epileptic encephalopathy where mutations in the Dynamin1 gene alter the protein function. These monogenic conditions are synaptopathies as the proteins affected play important roles in synapse stability and neurotransmission. Because of the high comorbidity between these disorders, it is hypothesised that there may be a common mechanism underlying them. We hypothesise that a deficit in presynaptic vesicle recycling may be part of a common mechanism underlying intellectual disability, autism spectrum disorder, and epilepsy especially in SYNGAP1 haploinsufficiency, Fragile X syndrome, and DNM1 epileptic encephalopathy. Using various fluorescent presynaptic activity reporters including synaptic pHluorins, tetramethylrhodamine dextran and calcium dyes to compare presynaptic activity in in vitro models of these monogenic conditions, we found differences in synaptic vesicle (SV) endocytosis in the genetically altered conditions compared to wildtype controls. We observed various SV endocytosis defects in clathrin-mediated endocytosis (CME) or activity-dependent bulk endocytosis (ADBE) in our models. We observed enhanced CME in SynGAP1 KO mouse hippocampal neurons. This enhanced SV endocytosis was accompanied by decreased SV cargo on the plasma membrane. Rat SynGAP1 KO hippocampal neurons did not display enhanced SV endocytosis, nor did neurons with the GTPase-activating (GAP) domain of SynGAP deleted. This was perhaps due to the altered time course of development between these rodent species. In mouse and rat models of Fragile X syndrome, CME was not altered compared to wildtype controls. However, in a rat model, we observed fewer nerve terminals undergoing ADBE which is the dominant SV endocytosis mode during elevated neuronal activity. De novo epileptic encephalopathy-associated mutations in DNM1 had differential effects on SV recycling through both CME and ADBE. Mouse hippocampal neurons overexpressing Dyn1R237W, Dyn1I289F and Dyn1H396D all showed less CME compared to overexpression of Dyn1WT. Moreover, fewer nerve terminals overexpressing Dyn1H396D were found to undergo ADBE. We also found that a large-conductance potassium (BK) channel opener can accelerate clathrin-mediated endocytosis and thus may be able to rescue the impaired SV endocytosis caused by these mutants. Although there is not yet a common underlying pathway at the presynaptic level between these conditions, SV recycling dysfunction is present across all of these models. Furthermore, we propose an axis of pathophysiology model where optimal SV endocytosis is required for optimised neural performance. We propose that either decreased or increased SV endocytosis can lead to the synaptic dysfunction observed in these models.

At the neuronal synapse, neurotransmitter-filled synaptic vesicles (SVs) fuse with the presynaptic plasma membrane during activity. Following exocytosis, SVs must be retrieved for neurotransmission to be maintained. Several modes of SV recycling have been identified. During mild neuronal activity, clathrin-mediated endocytosis has been regarded as the dominant SV retrieval mode, however the recently identified ultrafast endocytosis mode may also be important in this condition. During elevated activity, activity-dependent bulk endocytosis (ADBE) is the dominant SV retrieval pathway. In ADBE, large invaginations are formed from the plasma membrane, which then undergo scission to create bulk endosomes. In a second distinct step, SVs bud from these endosomes and specifically repopulate the reserve SV pool. However, since its first identification, only few molecules have been shown to participate in ADBE. The aim of this PhD was to identify novel molecules and elucidate the molecular mechanism of ADBE. To achieve this, two independent biochemical approaches were designed to purify and enrich bulk endosomes from primary neuronal cultures. In the first approach, bulk endosomes and SVs were labelled with a dye, FM1-43, using a strong stimulus. Cells were broken mechanically and the post nuclear supernatant, that contains all intracellular organelles, was collected. The supernatant was then subjected to subcellular fractionation using discontinuous Nycodenz gradients. This stimulated sample was always processed in parallel with a basal sample, where no neuronal stimulus was applied, in order to visualise activity dependent FM loading. After different fractionation protocols were applied, bulk endosomes were efficiently separated from SVs, as revealed by tracking fluorescence in different fractions. The fractionation results were further validated by electron microscopy, where bulk endosomes and SVs were labelled with horseradish peroxidase and purified using the established protocol. Immunoblotting against selected SV cargo proteins from stimulated bulk endosome and SV samples, indicated the specific and preferential localisation of VAMP4 on bulk endosomes, in contrast to other SV cargo. The molecular identity of bulk endosomes was also approached by submitting the bulk endosome fractions to semi-quantitative mass spectrometry. This analysis revealed many different proteins that were identified in bulk endosome samples and quantification approaches further indicated proteins that can be localised on bulk endosomes and have a potential role in ADBE. A second magnetic isolation approach was designed, to purify bulk endosomes using a completely different methodology. In this case, bulk endosomes were specifically labelled with iron nanoparticles, which are preferentially taken up by bulk endosomes since they are larger than SVs. The cells were broken as before and post nuclear supernatant was acquired. In this case, the supernatant was submitted to magnetic isolation that separated iron beads labelled structures from all other intracellular organelles. An extensive immunoblotting analysis of magnetic bulk endosomes validated that VAMP4 and syndapin I, two essential ADBE proteins, were enriched in these purified samples. These magnetic bulk endosomes were also analysed using semi-quantitative MS and revealed many proteins with a potential role in ADBE. Significant overlap between the two independent methods was observed, further validating these approaches. Combining these two methods with bioinformatics tools allowed the identification of the molecular signature of ADBE as well as novel key candidates for this process. Specific molecules were investigated for their role in ADBE and SV recycling using a variety of different real-time fluorescent imaging assays. A major focus was on rab small GTPases. High molecular weight dextran uptake was used to specifically study the role of these proteins in ADBE, as it preferentially reports uptake via larger bulk endosomes. A pH sensitive chimeric protein, synaptophysin-pHluorin, was used to investigate the role of these proteins in CME. Additional imaging assays were used to answer emerging questions regarding the function and localisation of these targets in the presynapse. Using these approaches, rab11A and rab35 were found to promote ADBE and accelerate clathrin-mediated endocytosis. This effect was specific to high intensity stimulation, while SV exocytosis was not affected. Further research on the role of both novel and established ADBE molecules will provide key future insights into the mechanism of both bulk endosome generation/scission and subsequent SV reformation. A very promising group is rab proteins and now evidence for their implication in SV recycling is presented here. Identification and characterisation of new targets will allow to investigate the role of ADBE in neurotransmission in both physiology and pathophysiology.

<p>У нашој дисертацији Интертекстуално раслојавање романа &bdquo;Опсада цркве Св. Спаса&ldquo; (контекст &bdquo;Библије&ldquo; и средњовјековне књижевности у настави) указујемо на функционисање интертекстуалне методологије у настави на примјеру српског постмодернистичког романа с краја двадесетог вијека.<br />На почетку рада, у поглављу Преглед интертекстуалних теорија, дајемо кратак преглед најзначајнијих теорија интертекстуалности и потом указујемо како смо их, зависно од Петровићеве поетике, користили приликом анализе романа Опсада цркве Св. Спаса.<br />Поглавље Опсада и &bdquo;Библија&ldquo; осмишљено је преко типологије Нортропа Фраја и стога се дијели на седам потпоглавља: стварање, излазак, закон, мудрост, пророчанство, јеванђеље, апокалипса, унутар којих се указује на низ цитата, алузија, реминисценција и мотива потеклих из библијског текста, па стога и на формирање постмодернистичких антитипова наведених библијских књига у роману.<br />Опсада и средњовјековна књижевност&nbsp; најсложеније је и најразуђеније поглавље у дисертацији. У првом потпоглављу указујемо како је философија Дионисија Псеудо-Ареопагита дубински уграђена у савремени текст те се препознаје како на семантичком, тако и на стилском нивоу &ndash; отуд и наше компаративно истраживање стила плетенија словес средњовјековне књижевности према стилу Петровићевог романа. Посебну пажњу посветили смо композицији књиге и, самјеривши је са средњовјековним предлошцима, указали на постмодернистички &bdquo;живот&ldquo; житија, слова, повијести, хронике, плача, записа. У посљедњем дијелу проучавамо ликове Данила Другог, Драгутина и Милутина и то највише преко Даниловог зборника који је писац обилно имао за подтекст свом роману. Лик Данила Другог уобличава се поетичким поступком метанарације, Драгутин преко житија из Даниловог зборника, док је Милутинов лик остварен преко интермедијалних релација постмодернистичког романа са средњовјековним фрескама.<br />У посљедњем поглављу, Опсада и ХХ вијек, указујемо на који начин,&nbsp; преко библијског и средњовјековног подтекста, Опсада успоставља интертекстуалне везе са књижевношћу двадесетог вијека. Преко два неисторијска лика, Андрије Скадранина и Блашка, односно два типа &ndash; ђавола и јуродивог &ndash; успостављају се релације са романима На Дрини ћуприја Иве Андрића, Хазарски речник Милорада Павића и Страх и његов слуга Мирјане Новаковић, као и са збирком прича Божји људи Борисава Станковића и приповијетком Запис о даровима моје рођаке Марије Момчила Настасијевића.</p> / <p>U našoj disertaciji Intertekstualno raslojavanje romana &bdquo;Opsada crkve Sv. Spasa&ldquo; (kontekst &bdquo;Biblije&ldquo; i srednjovjekovne književnosti u nastavi) ukazujemo na funkcionisanje intertekstualne metodologije u nastavi na primjeru srpskog postmodernističkog romana s kraja dvadesetog vijeka.<br />Na početku rada, u poglavlju Pregled intertekstualnih teorija, dajemo kratak pregled najznačajnijih teorija intertekstualnosti i potom ukazujemo kako smo ih, zavisno od Petrovićeve poetike, koristili prilikom analize romana Opsada crkve Sv. Spasa.<br />Poglavlje Opsada i &bdquo;Biblija&ldquo; osmišljeno je preko tipologije Nortropa Fraja i stoga se dijeli na sedam potpoglavlja: stvaranje, izlazak, zakon, mudrost, proročanstvo, jevanđelje, apokalipsa, unutar kojih se ukazuje na niz citata, aluzija, reminiscencija i motiva poteklih iz biblijskog teksta, pa stoga i na formiranje postmodernističkih antitipova navedenih biblijskih knjiga u romanu.<br />Opsada i srednjovjekovna književnost&nbsp; najsloženije je i najrazuđenije poglavlje u disertaciji. U prvom potpoglavlju ukazujemo kako je filosofija Dionisija Pseudo-Areopagita dubinski ugrađena u savremeni tekst te se prepoznaje kako na semantičkom, tako i na stilskom nivou &ndash; otud i naše komparativno istraživanje stila pletenija sloves srednjovjekovne književnosti prema stilu Petrovićevog romana. Posebnu pažnju posvetili smo kompoziciji knjige i, samjerivši je sa srednjovjekovnim predlošcima, ukazali na postmodernistički &bdquo;život&ldquo; žitija, slova, povijesti, hronike, plača, zapisa. U posljednjem dijelu proučavamo likove Danila Drugog, Dragutina i Milutina i to najviše preko Danilovog zbornika koji je pisac obilno imao za podtekst svom romanu. Lik Danila Drugog uobličava se poetičkim postupkom metanaracije, Dragutin preko žitija iz Danilovog zbornika, dok je Milutinov lik ostvaren preko intermedijalnih relacija postmodernističkog romana sa srednjovjekovnim freskama.<br />U posljednjem poglavlju, Opsada i HH vijek, ukazujemo na koji način,&nbsp; preko biblijskog i srednjovjekovnog podteksta, Opsada uspostavlja intertekstualne veze sa književnošću dvadesetog vijeka. Preko dva neistorijska lika, Andrije Skadranina i Blaška, odnosno dva tipa &ndash; đavola i jurodivog &ndash; uspostavljaju se relacije sa romanima Na Drini ćuprija Ive Andrića, Hazarski rečnik Milorada Pavića i Strah i njegov sluga Mirjane Novaković, kao i sa zbirkom priča Božji ljudi Borisava Stankovića i pripovijetkom Zapis o darovima moje rođake Marije Momčila Nastasijevića.</p>

This thesis investigates the recently developed technique of spin-torque-driven ferromagentic resonance (ST-FMR). Contrary to conventional FMR techniques where the magnetodynamics are excited by torques on the magentic moments produced by microwave fields, ST-FMR uses the spin-transfer torque acting on a nanomagnet. Here we present two experiments which exploit ST-FMR in the standard AFM/FM/NM/FM exchange-biased spin valve (EBSV) structures, where the ferromagnetic (FM) layers are separated by a nonmagnetic (NM) spacer, and one ferromagnet is pinned with an antiferromagnetic (AFM) layer. In our experiments microwave currents are applied to a mechanical point contact between a sharpened Cu tip and a SV (IrMn/Py/Cu/Py) multilayer film. While most ST-FMR experiments require noncollinear orientation of FM-layer magnetizations, we studied ST-FMR in SVs above saturation, where the two FM layers have parallel magnetizations. The resulting magnetodynamics are detected electrically by a small rectified dc voltage, which appears across the structure during resonance. Studies of the resonance frequencies, amplitudes, line widths, and line shapes as a function of microwave power, microwave frequency, dc current and magentic field are presented. The results are analyzed in terms of ST-FMR and rectification based on GMR. However the origin of the observed voltage cannot be fully explained by the resistance changes which come from the giant magnetoresistance (GMR) effect of the spin valve. To investigate other sources of rectified dc voltages at resonance we have performed the second set of measurements with lithographically patterned pairs of (Py/Cu/Co/IrMn)-SV microstripes. These measurements also revealed a resonance in the rectified voltage at FMR frequencies, and showed additional structures which might be related to spin wave excitations. The observations can be tentatively attributed to additional rectification effects due to anisotropic magnetoresistance (AMR). The line pair structure allows us to use different measurement geometries to investigate the magnetodynamics in the SV. In this experiment FMR can either be excited by spin transfer or by a rf magnetic field created by the microwave current, depending on the used geometry. Qualitative studies of the FMR dependencies and characteristics are presented for different measurement geometries. / text

Pieter C. van den Toorn has shown that one can label sections of Igor Stravinsky's Russian-period music based on two types of melodic-line (block) interactions: Type I and Type II. This paper analyzes Stravinsky's Rite of Spring and proposes the further classification of Type II sections of music into three categories. Organizing the music into these three categories depends on two aspects:1) the type of texture of the interacting blocks: melody and accompaniment or contrapuntal, and 2) the overall alignment of the interacting blocks' group-level rhythmic accents, which create either a unified or disunified metric accent. Furthermore, these three categories of Type II block structure also occur in other Stravinsky works, as well as works of minimalist composers. / School of Music

The diploma thesis focuses on the analysis of the woman's ideal during the High Middle Ages, how it was represented by St. Elizabeth of Hungary as the main character of the old-Bohemian Život svaté Alžběty that was credited to Tomáš Štítný ze Štítného. Elizabeth belongs to a new type of saintesses which during the contemporary times embodied important ideals of Christianity (above all poverty and love toward a fellowman realized by charitable and social work) and they offered a model for some overshadowed aspects of feminity (mainly a model of a perfect wife and a widow). We concentrate especially on the formation of the main character in all three women's statuses typical for the Middle Ages, that is virginity, married life and viduity, and also on the revision of their traditional hierarchy in the Elizabethean hagiography.

The topic of the diploma thesis is A Linguistic Analysis of a Baroque Sermon. The text, written by Václav Ignác Blovský, dates back to 1724. The diploma thesis continues in my bachelor work. It´s aim was to give the analysis of phonetic and morphological features of the same text. The diploma thesis is divided into two parts - the theory and the research. The first one deals with the author´s life and his work, the cult of St. John of Nepomuk and sermons dedicated to this Czech patron saint in general. The analytical part summaries the results of the previous research and includes the syntactic analysis. The aim of the diploma thesis is to find out the level of the Czech language in the early 18th century on the basis of the findings.

The topic of the diploma thesis is the impact of the St. Wenceslas'cult to the tourism in Brandýs nad Labem -- Stará Boleslav. First part of the thesis consists of basic information about St. Wenceslas and the development of his cult in periods of the Czech history. Second part represents the survey of the Czech nation about St. Wenceslas. Last part of the thesis forms the analysis of the tourism industry in Brandýs nad Labem - Stará Boleslav and the impact of St. Wenceslas cult to the tourism of the town.

My aim is to make a financial analysis of the private health care establishment: to assess whether the organisation finance its projects effectively and to identify its potential weaknesses. I would like to point out some measures of correction and submit them to the management of the assessed establishment.

The aim of this thesis is the documentation of the church of James the Greater in the South Moravian village Čučice. It is complete drawings of the structure, consisting of the topographic situation around the church, floor plans, vertical sections and views on the facade. Text part contents writing about working activities. It also discusses the historical and cultural characteristics of the church as immovable monument, the description of the preparatory work, a way of building surveying networks and methods of measurement. Finally, it highlights the processing of measured values in the computing and graphics phases. Use of this thesis and its evaluation is given in the final chapter. Graphic documentation, along with sketches, notebooks, calculation protocols, lists of coordinates, and testing the accuracy are in the annex section.