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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
171

Mikroparaziti a plodnost perlooček rodu Daphnia na gradientech v korytovitých přehradních nádržích / Microparasites and fecundity of Daphnia at environmental gradients of canyon-shaped reservoirs

Hubová, Jana January 2015 (has links)
The original aim of my diploma thesis was examination of fixed samples of cladocerans from the Daphnia longispina species complex from reservoirs Vír and Vranov for the presence of 4 groups of microparasites: microsporidia, oomycetes, the protozoan Caullerya mesnili, and the yeast Metschnikowia bicuspidata. The next aim was to determine the effect of parasites on Daphnia fecundity, and determination of their spatial and temporal distribution within reservoirs. From the total number 4452 of examined Daphnia females, however, only 56 individuals were infected. This number was not sufficient for the planned analyses. As I recorded for all examined individuals the clutch size, I thus dealt to a large extent with an alternative issue: the temporal and spatial variation of, and the influence of environmental factors on Daphnia fecundity. Both studied reservoirs are characteristic by canyon-shaped profile that allows formation of environmental gradients on the horizontal as well as vertical axis. The results confirm that reservoir identity, season, and location within the reservoir (or gradient of food supply) have all significant effects on fecundity. During my work I have encountered difficulties associated with determining microparazites from fixed zooplankton samples. The appendix section of my thesis...
172

Ekofyziologická diferenciace kryptických druhů komplexu Synura petersenii (Synurophyceae) / Ecophysiological differentiation of Synura petersenii cryptic species (Synurophyceae)

Jelínková, Lucie January 2014 (has links)
Synura petersenii represents a complex of pseudo-cryptic species. These are planktonic colonial organisms belonging to the class Chrysophyceae. The species are abundant in spring and autumn, when they produce golden-brown blooms in oligo-mesotrophic waters. In this thesis, I focused on investigating the effect of temperature to growth parameters of four selected species: S. petersenii, S. americana, S. glabra and S. conopea. Several temperature experiments were performed and statistically analyzed. According to growth curves the temperature 25 žC was mostly stressful for these organisms. Only S. petersenii sensu stricto was sometimes able to growth at this temperature. In general, the significant differences in growth rates were detected between S. petersenii and S. americana. In all experiments, S. petersenii had the lowest growth rate. This ubiquitous species probably acts as a K-strategist. S. americana grew generally very quickly, especially at temperature 13 žC that is obviously optimal for it. Further, I observed a significant strain specificity, which is largely a well-known phenomenon in the ecophysiology. Significant differences in growth rates were noted between temperatures 13 žC and 16 žC, where all tested species grew best, as well. This is in accordance with a general recognition of...
173

Syntéza pi-elektronových oligomerů a studium jejich vlastností / The pi-electron oligomers: Their synthesis and properties

Warzecha, Tomáš January 2012 (has links)
The pi-electron oligomers: Their synthesis and properties This diploma thesis is focused on the synthesis of p-oligophenyleneethynylene rods (dimer and trimer) containing laterally attached naphtalenediimide units as electron acceptor groups. These functionalised short oligomers are intended to serve as model oligodentate acceptors in the study on multiple interactions with electron rich molecules. The oligomers were assembled from building blocks such as aryl iodides and aryl alkynes bearing a naphthalenediimide unit. The functionalised monomers were combined in a stepwise way by using Sonogashira reaction to form the target p-oligophenyleneethynylene oligomers. The introductory UV-VIS spectroscopic studies on charge transfer complexes between electron donor (pyrene) and electron acceptor (a naphthalenediimide derivative) were performed. In addition, multiple noncovalent interaction between the electron acceptor dimer containing two naphthalenedimide moieties and the electron donor dimer containing two pyrene moieties were investigated by using 1 H NMR titration. The resulting oligomers and their precursors were characterized by using 1 H NMR, 13 C NMR, MS and IR spectroscopy.
174

Úloha N-terminální domény a/TIF32 podjednotky iniciačního faktoru eIF3 ve vazbě mRNA na 43S pre-iniciační komplexy. / The role of the N-terminal domain of the a/TIF32 subunit of eIF3 in mRNA recruitment to the 43S pre-initiation complexes.

Vlčková, Vladislava January 2013 (has links)
Translation initiation is a complex process which results in the assembly of the elongation competent 80S ribosome from the 40S and 60S ribosomal subunits, the initiator tRNA and mRNA, and is orchestrated by numerous eukaryotic initiation factors (eIFs). Although it represents one of the most regulated processes of gene expression, the exact mechanism of one of the key steps of translation initiation - mRNA recruitment to the 43S pre-initiation complex (PIC) - is still only poorly understood. Recent studies indicated that besides eIF4F and poly(A)-binding protein, also eIF3 might play an important, if not crucial, role in this step. In our laboratory, we recently identified a 10 Ala substitution (Box37) in the a/TIF32 subunit of Saccharomyces cerevisiae eIF3, which interfered with translation initiation rates. Detailed analysis showed that this mutation significantly reduces the amounts of model mRNA in the gradient fractions containing 48S PICs as the only detectable effect in vivo. Moreover, a recently solved crystal structure of the N-terminal part of a/TIF32 pointed to two Box37 residues, Arg363 and Lys364, both proposed to contribute to one of the positive, potentially RNA-binding areas on the a/TIF32 surface. The fact that also their substitutions with alanines severely impaired the mRNA recruitment...
175

Strukturní vývoj, magnetická stavba a mechanismus exhumace podolského komplexu / Structural evolution, magnetic fabric and mechanism of exhumation of the Podolsko complex

Burjak, Miroslav January 2013 (has links)
This thesis concentrates on a detailed field and structural analysis of the Podolsko complex, Moldanubian unit, Bohemian Massif, complemented by a microstructural study and analysis of magnetic susceptibility (AMS). The Podolsko complex occupies the footwall of a major Variscan normal shear zone and is juxtaposed against the southern to southeastern margin of the Central Bohemian Plutonic complex. During the field work, more than 160 outcrops were examined, the AMS samples were taken at 25 stations, and samples for microstructural studies were taken from 12 localities. On the micro-scale, leucocratic migmatites contain abundant garnet grains which may represent relics of an earlier (ultra-)high preassure metamorphic phase. Retrogression is obvious in other samples of biotite migmatites of the Moldanubian Variegated unit. The retrogression is marked by the presence of sillimanite and chlorite. The main tectonometamorphic event in the Podolsko complex is extensive migmatization coeval with formation of pervasive flat-laying fabric. This is corroborated by the AMS study which indicates concordant steep to flat-laying magnetic and mesoscopic foliations striking NNW-SSE. The AMS also shows that the subhorizontal N-S to NNW-SSE trending magnetic lineations in the Podolsko complex correspond to those in the...
176

Studium teplotně citlivých porfyrinů a jejich supramolekulárních komplexů / Studium teplotně citlivých porfyrinů a jejich supramolekulárních komplexů

Hrubovský, Martin January 2016 (has links)
Study of thermoresponsive porphyrins and their supramolecular complexes Abstract: We studied the water-soluble artificial compound meso-tetrakis{3,4,5-tris[2-(2-(2- methoxyethoxy)ethoxy)ethoxy]phenyl}porphyrin prepared at NIMS, Japan, using the high-resolution NMR spectroscopy experimental method. We observed its LCST-type phase separation and applied the Flory-Huggins theory of polymer solutions in order to find its phase diagram (binodal and spinodal curves of the phase separation) and we also obtained molar enthalpies, entropies and critical temperatures of its phase separation; from the Flory-Huggins theory we discovered that its molecules form dimers in aqueous solutions. We also studied its host-guest interactions with the S-camphorsulfonic acid; we learned that the porphyrin binds cations and the porphyrin dimers break down when dissolved cations are available for complexation. We observed no phase separation in chloroform. We obtained no proof of the existence of molecular stacks larger than dimers. 1
177

The IkB kinase complex is a regulator of mRNA stability

Mikuda, Nadine 26 April 2018 (has links)
Bisher wurde davon ausgegangen, dass der IKK-komplex durch Regulation des Transkriptionsfaktors NF-kappaB die stressinduzierte Expression von Zielgenen steuert. Im Rahmen der hier vorgelegten Dissertation konnte jedoch gezeigt werden, dass der IKK-Komplex unabhängig von seiner Rolle in der NF-kappaB-Aktivierung die Stabilität einer Vielzahl von mRNAs kontrolliert. Mittels der Kombination von Ko-Immunopräzipitationsstudien und SILAC-MS konnte die induzierte Interaktion der regulatorischen Untereinheit des IKK-Komplexes IKKgamma mit dem Gerüstprotein EDC4 (Enhancer of Decapping 4) nachgewiesen werden. EDC4 ist eine essentielle Komponente sogenannter zytoplasmatischer „Processing Bodies“ (P-Bodies). Diese fungieren als Depots für die Speicherung von mRNAs, aber auch als Orte der mRNA-Degradation und der miRNA-vermittelten Repression spezifischer Zielgene. Die Interaktion von IKKgamma mit EDC4 konnte durch verschiedene Stimuli induziert werden. Dazu zählen DNA-Schäden durch Doppelstrangbrüche, aber auch die Aktivierung von Oberflächenrezeptoren durch TNFalpha und IL-1beta. EDC4 dient darüber hinaus als Substrat der Kinase IKKbeta. Mittels Massenspektrometrie und Kinaseassays konnten vier IKK-abhängige Phosphorylierungsstellen identifiziert werden. Die IKK-vermittelte Phosphorylierung von EDC4 ist essentiell für die Regulation von mRNAs und die damit verbundene Bildung der zytoplasmatischen P-Bodies. Diese Befunde konnten sowohl in stabilen induzierbaren Zelllinien, mittels transienter Transfektion und durch den Gebrauch von Kinaseinhibitoren in primären als auch in Krebszelllinien bestätigt werden. mRNA-Stabilitätsassays und eine RNA-Seq Analyse bestätigten die stressinduzierten Änderungen in den Halbwertszeiten spezifischer Transkripte und offenbarten einen gemeinsamen Regulationsmechanismus des IKK-Komplexes mit EDC4. / The IKK complex is deemed to regulate gene expression through the activation of the transcription factor NF-kappaB. Here I describe an NF-kappaB-independent function of the IKK complex in regulating mRNA stability across different cell types and stimuli. A SILAC-MS screen for interaction partners of the regulatory subunit IKKgamma revealed an inducible interaction with Enhancer of mRNA Decapping 4 (EDC4). EDC4 is an essential component of cytoplasmic processing bodies (P-bodies). P-bodies function as sites of mRNA storage, degradation and miRNA-mediated silencing. Interaction between IKKgamma and EDC4 can be induced by various stimuli, including DNA damage, TNFalpha and IL-1beta. EDC4 was identified as a novel IKK substrate and four IKKbeta phosphorylation sites were determined by mass spectrometry and in kinase assays. Stable inducible cell lines, transient transfection and kinase inhibitors were used in different human cancer and in primary cell lines and demonstrated that phosphorylation of EDC4 by IKK is essential for formation of P-Bodies in response to numerous stimuli. mRNA stability assays confirmed stress-induced changes in the half-life of target mRNAs and revealed common regulation of mRNA stability by IKK and EDC4. The transcriptome-wide reach of this joint regulation was assessed via RNA-Seq analysis.
178

IFRS 9 FAS 2 : EN UNDERSÖKNING AV INTRESSENTERNAS ATTITYDER / IFRS 9 FAS 2

JINGMARK, JOHANNA, LOJEK, CHRISTIAN January 2011 (has links)
Bakgrund och problem: IASB beslutade 2008 att ersätta IAS 39 med IFRS 9 då IAS 39 ses som alltför omfattande och komplex. Processen har delats upp i tre faser. Fas 1 behandlar klassificering och värdering, fas 2 behandlar nedskrivningar och fas 3 behandlar säkringsredovisning. Denna uppsats behandlar endast fas 2. Både redovisare, revisorer och användare av finansiella rapporter anser standarden för finansiella instrument vara komplex. En orsak till komplexiteten är att finansiella instrument i sig är komplexa och begreppet finansiella instrument innefattar en vid variation av instrument. Det konstaterades även under den globala finansiella krisen att IAS 39 innehöll betydande brister.Syfte: Syftet med denna uppsats är att studera delar av arbetet IASB har gjort för att ersätta IAS 39 med IFRS 9, vilket ger oss möjlighet att studera fas 2 i detta examensarbete. Syftet är att se vad intressenter förväntar sig i redovisning av finansiella instrument och vad det finns för attityder angående förslaget till IFRS 9 fas 2.Avgränsningar: Eftersom det krävs att EU antar standarden för att den ska börja gälla i Sverige har vi även avgränsat oss till att titta på remissvar från enbart EU-länder.Metod: Studien har utförts med en kvalitativ metod där den empiriska undersökningen bestått av 22 remissvar som finns att tillgå på IASB:s hemsida.Resultat och slutsatser: Den genomförda undersökningen visar att det finns ett flertal olika åsikter angående IASB:s arbete med Exposure Draft. Många organisationer är överens om att man vill ha en mer principbaserad standard men vill samtidigt ha mer exempel att tillgå vilket återigen drar mot en mer regelbaserad standard. Slutsatsen är att Exposure Draft fortfarande ses som komplex dock inte i samma bemärkelse som IAS 39. Hur IASB löser denna problematik blir intressant att följa. / Program: Civilekonomprogrammet
179

Supramolecular structures of dendronized polymers and DNA on solid substrates

Gössl, Illdiko Maria 30 June 2003 (has links)
Komplexe aus entgegengesetzt geladenen Polyelektrolyten haben sowohl in der Biologie als auch in den Materialwissenschaften eine große Bedeutung. Im Mittelpunkt des Interesses stehen besonders die Kondensation der DNA in vitro, die Struktur des Nukleosoms im Zellkern, nicht-virale Systeme zur Transfektion von DNA in Zellen oder der Vorgang der layer-by-layer Adsorption. Verschiedene Theorien befassen sich mit den treibenden Kräften solcher Komplexbildungen. Allerdings standen experimentelle Untersuchungen auf diesem Gebiet bisher noch aus. Dieser Arbeit liegt die Fragestellung zu Grunde, ob es mit Hilfe der Rasterkraftmikroskopie möglich ist, die Struktur einzelner Polyelektrolytkomplexe, bestehend aus den beiden Polyelektrolyten DNA und dendronisierten Polymer, aufzuklären und ihre Komplexbildung zu untersuchen. Die Komplexe bildeten sich in Lösung und wurden anschließend auf einer unbeschichteten oder mit positiven Polymeren beschichteten Glimmeroberfläche adsorbiert. Auf der positiv beschichteten Glimmeroberfläche hafteten DNA-dendronisierte Polymer Komplexe mit einem Ladungsverhältnis von 1:1 bis 1:0.7 (DNA:dendronisiertes Polymer). Anhand der hochaufgelösten rasterkraftmikroskopischen Aufnahmen wurde ein Modell entwickelt, das die Umwicklung der DNA um das dendronisierte Polymer beschreibt. Der DNA-DNA Abstand ergab sich zu (2.30 ± 0.27) nm für den Komplex mit DNA und zweiter Generation dendronisierter Polymere und zu (2.16 ± 0.27) nm mit vierter Generation. Die theoretische Vorhersage der Überladung der Komplexe konnte experimentell bestätigt werden. Mit Hilfe der Rasterkraftmikroskopie konnte überdies der Einfluss des Salzgehaltes der Lösung auf die Bildung der Komplexe mit DNA und zweiter Generation dendronisierter Polymere untersucht werden. Wie man anhand des Zusammenwirkens von elektrostatischen Kräften und entropischen Wechselwirkungen bei der Adsorption von Polyelektrolyten vorhersagen kann, durchlief der DNA-DNA Abstand ein Minimum bei ansteigendem Salzgehalt. Bei sehr hohem Salzgehalt (2.4 M NaCl) konnte das Ablösen der DNA von dem Komplex beobachtet werden. Die untersuchten DNA/dendroniserten Polymer Komplexe bilden ein neues Modellsystem, mit dem einzelne Polyelektrolyt-Wechselwirkungen direkt untersucht werden können. Ein Vergleich der experimentellen Daten mit den vorhandenen Theorien zeigte, dass der Prozess des Überladens weitgehend durch elektrostatische Wechselwirkung zwischen den beiden Polyelektrolyten beschrieben werden kann. Sowohl entropische Beiträge als auch die Biegeenergie der umwickelnden DNA sind vernachlässigbar. Basierend auf diesen Ergebnissen können neue Trägerstrukturen für effizientere nicht-virale DNA-Transfektionssysteme entwickelt werden. / Complexes of oppositely charged polyelectrolytes play an important role in both biology and material science, for instance DNA condensation in vitro, nucleosomal structure, non-viral gene transfection systems as well as layer-by-layer adsorption. Although there are theories predicting overcharging of polyelectrolyte complexes, the driving forces are still under debate and systematic experimental studies on single polyelectrolytes remain challenging. Therefore the question arose if it is possible to analyze single polyelectrolyte complexes, using DNA and dendronized polymers, with the scanning force microscope in order to investigate the complexation in detail. For the complex analysis, the polyelectrolytes were allowed to interact in solution and then to adsorb on negatively charged mica or on mica coated with a positively charged polymer. Scanning force microscopy was used to investigate the adsorbed species. DNA/dendronized polymer complexes of charge ratio of 1/1 through 1/0.7 adsorbed on mica coated with a positively charged polymer. The analysis of high resolution molecular images indicated that DNA wraps around the dendronized polymer with an estimated pitch of (2.30 ± 0.27) nm and (2.16 ± 0.27) nm for dendronized polymers of generation two and four, respectively. In the proposed model the polyelectrolyte with the smaller linear charge density is wrapped around the more highly charged dendronized polymer, resulting in a negatively overcharged complex. This overcharging is consistent within recent theories of spontaneous overcharging of complexes of one polyelectrolyte wrapping around the other. Using the complex of DNA and dendronized polymers of second generation, the influence of monovalent salt concentration on the molecular structure was studied. By increasing the salt concentration the pitch showed a minimum as predicted by the interplay of electrostatic forces and entropic interactions of polyelectrolyte adsorption. At high salt concentration (2.4 M NaCl) the release of DNA from the complex can be observed. The results showed that the DNA/dendronized polymer system can be used as a new, high potential model system to investigate single polyelectrolyte interactions. With regard to recent theories, the experimental results indicate that the overcharging of the complex is mainly driven by electrostatic forces whereas contributions of counterion entropy and bending energy seem to be negligible. This understanding may be useful for the design of single polyelectrolyte complexes for non-viral gene delivery systems and might help to optimize the transfection efficiency based on the structure of the vector system.
180

Early steps in cotranslational translocation of proteins across the ER membrane

Neuhof, Andrea 10 July 2000 (has links)
Sekretorische Proteine und Proteine der Kompartimente des sekretorischen Transportweges müssen die Membran des Endoplasmatischen Retikulums überqueren, um an ihren Wirkungsort zu gelangen. In der vorliegenden Arbeit wurden frühe Schritte des kotranslationalen Transports von Proteinen durch die ER-Membran untersucht. Signalsequenzen leiten diese Proteine als ribosomengebundene Intermediate an die ER-Membran. Die Ribosomen binden dort an den Sec61p-Komplex, der als Ribosomenrezeptor wirkt und gleichzeitig den proteinleitenden Kanal in der Membran bildet. Die Assoziation von Ribosomen mit dem Sec61p-Komplex verläuft in zwei Phasen. Die initiale Bindung ist sensitiv gegenüber hohen Salzkonzentrationen. Die Ribosomenbindung wird salzresistent, wenn die naszierende Kette in den Kanal inseriert und der Sec61p-Komplex die Signalsequenz erkennt. Sowohl Ribosomen ohne naszierende Kette als auch Ribosomen, die Proteine ohne Signalsequenzen synthetisieren, sind nur zur initialen salz-sensitiven Bindung an den Sec61p-Komplex fähig. Signalsequenzen interagieren im Cytosol mit SRP (engl.: Signal Recognition Particle). In dieser Arbeit wurde gezeigt, daß Signalsequenzen außerdem von Calmodulin gebunden werden. SRP und Calmodulin scheinen für die Interaktion mit Signalsequenzen einen ähnlichen Mechanismus zu benutzen, der wiederum mit der Signalsequenzerkennung durch den Sec61p-Komplex verwandt ist. Alle Ribosomen, unabhängig davon ob und welches Protein sie translatieren, können mit dem Sec61p-Komplex interagieren und daher um Bindungsplätze an der ER-Membran kompetitieren. Wenn SRP an die Signalsequenz einer naszierenden Kette gebunden ist, erhalten diese Ribosomen jedoch einen Vorteil in der Kompetition. Nur sie können Ribosomen ohne naszierende Kette oder Ribosomen, die ein cytosolisches Protein translatieren, vom Sec61p-Komplex verdrängen und sich selbst dann einen Translokationsort sichern, wenn alle Bindingsplätze an der Membran besetzt sind. In der vorliegenden Arbeit wurden dreidimensionale Strukturen von Komplexen aus Ribosom und proteinleitendem Translokationskanal vorgestellt, die der ersten und zweiten Phase der Ribosomenbindung entsprechen. Überraschenderweise unterscheiden sich diese beiden Stadien strukturell nicht. In beiden Fällen existieren definierte Verbindungen zwischen Ribosom und Kanal, die eine Lücke von etwa 20 Angström zwischen dem Ribosom und der Membranoberfläche überbrücken. Die Lücke stellt eine Verbindung zum Cytosol her, die eventuell dazu dient, naszierende Ketten ins Cytosol zu entlassen, wenn diese nicht ins Lumen des ER transportiert werden sollen. Weiterhin zeigen wir, daß der Kanal in nativen Membranen größer ist als der Kanal, der nur aus gereinigtem Sec61p-Komplex besteht. Dieser größere Kanal besitzt eine zusätzliche lumenale Domäne, die von der Oligosaccharyltransferase oder vom TRAP-Komplex gebildet wird. / The first step in the secretory pathway is the translocation of proteins across the membrane of the endoplasmic reticulum (ER). In this thesis project, early stages of cotranslational protein translocation in mammalian cells were studied. Proteins following the secretory pathway are targeted to the ER as ribosome-nascent chain complexes by their N-terminal hydrophobic signal sequences. The nascent chain is translocated across the ER membrane through a hydrophilic channel formed by the Sec61p complex, which also functions as the ribosome receptor. The initial binding of ribosomes to the ER membrane is salt-sensitive. After insertion of the nascent chain into the translocation channel and signal sequence recognition by the Sec61p complex, the ribosome is bound in a salt-resistant manner. The membrane binding of ribosomes lacking nascent chains and of ribosomes carrying nascent chains without signal sequences is always salt-sensitive. It is known that in the cytosol, the signal sequence binds to the signal recognition particle (SRP). Here we show that another cytosolic factor, the small regulatory protein calmodulin, can interact with signal sequences. Our data suggest that both SRP and calmodulin use a similar mechanism for substrate binding and recognition. In fact, this mechanism may be related to signal sequence recognition by the Sec61p complex. Previously the question has been raised of how efficient targeting of ribosome-nascent chain complexes (RNCs) carrying a signal sequence is possible when all ribosomes, regardless of the presence or nature of a nascent chain, can bind to the Sec61p complex. We demonstrate that all ribosomes compete for common binding sites at the ER membrane and that SRP functions as a positive effector to give RNCs carrying a signal sequence an advantage over other ribosomes. RNCs with a signal sequence and bound SRP can displace ribosomes without a nascent chain and ribosomes synthesizing cytosolic proteins from the membrane and can therefore secure a translocation site even when all ribosome binding sites at the ER membrane are occupied. A structural analysis by single particle cryo electron microscopy revealed that ribosome-translocation channel complexes do not differ in the salt-sensitive or the salt-resistant stage of ribosome binding to the ER membrane. Furthermore our data show that the ribosome is linked to the translocation channel by a discrete number of connections. Even in the presence of a translocating nascent chain the ribosome-membrane junction is not completely sealed towards the cytosol. Instead, a sizable gap exists between the ribosome and the surface of the membrane that may allow nascent polypeptide chains to enter the cytosol when their translocation across the ER membrane is prevented. We also show that translocation channels derived from native microsomes are larger than channels derived from purified Sec61p complex. These larger channels contain a wider central pore and an additional lumenal domain, which is formed by the oligosaccharyl transferase or by the TRAP complex.

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