@phdthesis{Varadarajulu2006, author = {Varadarajulu, Jeeva}, title = {Integrin alpha(v) and Focal adhesion kinase - promising targets to limit smooth muscle cell migration}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17484}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2006}, abstract = {Die Pr{\"a}vention einer Restenose nach PTCA ist eines der wichtigsten Ziele f{\"u}r Forscher und Kliniker. Etwa 1,5 Millionen Interventionen werden weltweit j{\"a}hrlich durchgef{\"u}hrt und mit Hilfe von Stentimplantationen k{\"o}nnen die meisten Patienten erfolgreich behandelt werden. Jedoch kommt es in bis zu 60 \% der F{\"a}lle zu einer Restenosierung des behandelten Gef{\"a}sses innerhalb von etwa 6 Monaten. F{\"u}r die Entwicklung der Neointima, Hauptursache der Restenose, sind Wanderung glatter Gef{\"a}ssmuskelzellen (GMZ) und Ablagerungen von Proteinen der extrazellul{\"a}ren Matrix (EZM) verantwortlich. Signalkaskaden, die {\"u}ber Integrin Rezeptoren vermittelt werden, spielen in der Migration von GMZ eine zentrale Rolle. Viele Integrine binden {\"u}ber eine spezifische Aminos{\"a}uresequenz, die sogenannte RGD Sequenz, die in verschiedenen EZM Proteinen und in auf Zelloberfl{\"a}chen gebundenen Immunglobulinen vorkommt. Bisher konnte von verschiedenen Integrinantagonisten, wie Antik{\"o}rpern, zyklischen Peptiden, Peptidomimetika und Nicht-Peptiden, gezeigt werden, dass die die pathologische Reaktion vermindern k{\"o}nnen, was auf die Bedeutung der Integrin vermittelten Signalkaskaden in der GMZ Migration hinweist. Wir konnten zeigen, dass die Wanderung von GMZ sowohl mit einem pharmakologischen Inhibitor, wie auch durch die endogene {\"U}berexpression eines FAK Inhibitors, der {\"u}ber ein AAV Vektorsystem {\"u}bertragen wurde, gehemmt werden konnte. So stellt die Blockade der Integrin vermittelten Signalkaskaden ein vielversprechendes Ziel f{\"u}r die Inhibition der Restenose nach PTCA dar. Im ersten Teil der Arbeit konnten wir nach Stimulation von humanen GMZ mit Vitronektin (VN) eine verst{\"a}rkte Tyrosinphosphorylierung (PTyr) verschiedener zellul{\"a}rer Proteine nachweisen. Dabei zeigte sich eine besonders signifikante Phosphorylierung eines Proteins, das mittels Immunpr{\"a}zipitation als "focal adhesion kinase" (FAK) identifiziert wurde. Die erh{\"o}hte PTyr zeigte sich auch am Tyrosinrest FAK Tyr-397, der Autophosphorylierungsstelle der Kinase. Die erh{\"o}hte PTyr von FAK war abh{\"a}ngig von der Stimulation durch VN und nicht zu beobachten, wenn die GMZ auf Poly-L-Lysin ohne spezifische Rezeptor Ligand Interaktion adh{\"a}rierten. Mit Hilfe eines Integrin \&\#61537;V Inhibitors konnte diese rezeptorvermittelte Aktivierung in einer dosisabh{\"a}ngigen Weise verhindert werden. Die Inhibition der durch VN stimulierten Migration (Haptotaxis) mit Hilfe des \&\#61537;V Inhibitors korrelierte mit der Reduktion der Aktivierung Integrin vermittelter Signalwege, im Besonderen der PTyr von FAK. Interessanterweise konnte die Blockade von Integrin \&\#61537;V nicht nur die durch VN stimulierte Haptotaxis, sondern auch die durch Wachstumsfaktoren induzierte Chemotaxis hemmen. Die Migrationsrate wurde mit Hilfe eines modifizierten Boyden-Migrationskammer Experiments ermittelt, das ein in vitro Modell zur Untersuchung von Zellwanderung darstellt. Der \&\#61537;V Inhibitor hemmte auch die Invasion der GMZ in eine Matrigel Matrix und die Sekretion der Matrixmetalloproteinase 2. Eine Apoptose wurde bei den verwendeten Konzentrationen nicht induziert. FAK stellt ein wichtiges Schl{\"u}sselprotein in vielen zellul{\"a}ren Mechanismen dar. So konnte die Beteiligung von FAK in der Regulation der Zellmigration an verschiedenen Zellarten gezeigt werden. Die {\"U}berexpression von FRNK, der C-terminalen Dom{\"a}ne von FAK, ist in der Lage die in vitro Migration von GMZ wie auch die Neointimabildung in einem Schweinemodell zur Entwicklung der Restenose zu verhindern. FAK stellt somit ein vielversprechendes Ziel f{\"u}r die Inhibition der Restenoseentwicklung nach PTCA dar. Der letzte Teil der Arbeit konzentrierte sich auf die Identifikation von Bindungspartnern der N-terminalen Dom{\"a}ne von FAK mit Hilfe eines bakteriellen „two hybrid" Systems. Es wurde als ein m{\"o}glicher Bindungspartner ein 17,9 kDa grosses Protein gefunden. Das humane Homolog ist als AGS4 bezeichnet und stellt einen GTPase Aktivator dar. Es zeigte sich, dass es in der Lage ist, mit der N-terminalen Dom{\"a}ne von FAK zu interagieren, und dass es stark in h{\"a}matopoetischen Zellen exprimiert wird. Zusammenfassend kann man sagen, dass unsere Ergebnisse FAK als ein vielversprechendes Ziel f{\"u}r die Inhibition der GMZ Migration erscheinen lassen. Das Vorliegen verschiedener induzierter Signalwege kann durch die Rolle der EZM Proteine und der Wachstumsfaktoren in der Zellmigration erkl{\"a}rt werden. Das Ziel dieser Studie war die Signalkaskaden, die zu einer GMZ Migration und somit zu einer Restenose f{\"u}hren, zu unterbrechen. Die Ergebnisse zeigen, dass \&\#61537;V Integrine und Signalkaskaden, die FAK vermittelt sind, wichtig f{\"u}r die Zellmigration sind. Die Unterbrechung dieser FAK vermittelten Signalwege, sei es durch einen pharmakologischen Inhibitor oder durch die {\"U}berexpression von FRNK f{\"u}hrte zu einer Inhibition der Migration.}, subject = {Restenose}, language = {en} } @phdthesis{Wang2006, author = {Wang, Dapeng}, title = {The mechanism of glucocorticoid induced murine thymocyte and peripheral T cell apoptosis}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17317}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2006}, abstract = {Glucocordicoide sind kleine lipophile Verbindungen, die viele biologische Effekte verursachen, wenn sie an den intrazellul{\"a}ren Glukokortikoidrezeptor (GR) binden. Dieser wandert wiederum in den Nucleus, um dort direkt oder indirekt die Transkription der Gene zu regulieren. Glukokortikoide sind der Grundstein in der Behandlung f{\"u}r eine Anzahl von h{\"a}matologischen b{\"o}sartigen Erkrankungen, wie Leuk{\"a}mie, Lymphome und Myelome. In der Literatur wird beschrieben, dass Glukokortikoide {\"u}ber die Vermittlung von Apoptose wirken.die Wirkung. Trotz der enormen Fortschritte im Verst{\"a}ndnis des regulierten Zelltodes, ist der genaue Mechanismus, den Glukokortikoide bei der Apoptose vermitteln, unbekannt. Die Daten, die bis jetzt erzielt wurden, deuten stark darauf hin, dass Gentransaktivierung durch den GR f{\"u}r den Beginn der durch Glukokortikoide verursachten Thymozytenapoptose verantwortlich ist. Außerdem wurde gezeigt, dass das multikatalytische Proteasom, einige Mitglieder der BCL2-Familie, {\"A}nderungen im Kalziumfluss sowie Caspasen eine wichtige Rolle in der Durchf{\"u}hrungsphase des durch Glukokortikoide vermittelten Zelltodes spielen Jedoch ist die genaue Reihenfolge dieses Prozesses bisher nicht bekannt. Ein Hauptschwierigkeit der gegenw{\"a}rtigen Diskussion entsteht aus der Tatsache, dass unterschiedliche Zellarten, wie Thymozyten, reife T-Zellen und Lymphomzellen verglichen werden, ohne ihre unterschiedlichen Eigenschaften und Genexpressionsprofile zu beachten. Obwohl angenommen wird, dass Glukokortikoide Apoptose {\"u}ber einen konservierten Mechanismus, wird dies nicht durch irgendwelche Daten unterst{\"u}tzt. In anderen Worten, es ist m{\"o}glich, dass Apoptose in Thymozyten, reifen T-Zellen und Lymphomzellen {\"u}ber unterschiedliche Signalwege vermittelt wid. Wir fragten uns daher, ob ein einzelner durch Glukokoritkoide eingeleiteter Signaltransduktionsweg daf{\"u}r verantwortlich ist, dass Apoptose in allen T-Lamphozytenarten eingeleitet wird, oder ob noch andere Signalwege existieren. Daher verglichen wir die Rolle des Proteasomes, verschiedener Caspasen, des lysosomalen Kompartements und anderer Faktoren in der durch Glukokortikoide induzierten Apoptose in Mausthymozyten und pepripheren T-Zellen sowie T-ALL Lymphomzellen. Unsere Entdeckungen zeigen, dass die Anfangsphase der durch Glukokortikoide induzierten Apoptose unabh{\"a}ngig von der Differenzierungsstadien der Zelle ist. Apoptose wird sowohl in Thymozyten als auch in reifen T-Zellen durch den GR vermittelt und ist von der Gentranskription abh{\"a}ngig. Im Gegensatz dazu unterscheidet sich die Durchf{\"u}hrungsphase erheblich in ihren Anforderungen f{\"u}r eine Anzahl von Signaltransduktionskomponenten zwischen Thymozyten und peripheren T-Zellen. W{\"a}hrend in Thymozyten das Proteasom, die Caspasen 3, 8 und 9 sowie Cathepsin B eine wichtige Rolle in durch Glukokortikoide induzierten Zelltod spielen, sind diese Faktoren f{\"u}r die Induktion des Zell-Todes in peripheren T- Zellen entbehrlich. Im Gegensatz dazu scheinen {\"A}nderungen in der Expression und intrazellul{\"a}ren Lokalisation von Mitgliedern der Bcl-2 Familie nicht zum durch Glukokortikoide induzierten Zellltod beitzutragen, egal um welchen Zelltyp es sich handelt. Wir haben beobachtet, dass eine Behandlung von Thymozyten mit Glukokortikoiden zu einer Aktivierung der lysosomalen Protease Cathepsin B f{\"u}hrt. Dies ist ein essentieller Schritt zur Einleitung von Apoptose durch Glukortikoide und zeigt zum ersten Mal, dass der lysosomale Amplifikationsloop in diesen Prozess involviert ist. Die Analyse des durch Glukokortikoide induzierten Zelltodes in verschiedenen T-ALL Zelllinien deutet darauf hin, dass die durch Glukokortikoide induzierten Signalwege in Thymozyten und allen Lymphonzelllinien aber nicht in peripheren T Zellen {\"u}bereinstimmen. Da die hoch-dosierte Glukokortikoidbehandlung eine wichtige Rolle in der Behandlung von hematologischen b{\"o}sartigen Erkrankungen spielt, k{\"o}nnen unsere Beobachtungen eine Grundlage f{\"u}r eine neue Anti-Krebs-Stragie bilden, die darauf ausgelegt ist, spezifisch Tumorzellen zu eliminieren aber reife T-Zellen unber{\"u}hrt lassen.}, subject = {T-Lymphozyt}, language = {en} } @phdthesis{Groh2005, author = {Groh, Claudia}, title = {Environmental influences on the development of the female honeybee brain Apis mellifera}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17388}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {F{\"u}r die Honigbiene spielt der Geruchssinn eine entscheidende Rolle bei der Kommunikation innerhalb des Sozialstaates. Kastenspezifische, auf uweltbedingten Einfl{\"u}ssen basierende sowie altersbedingte Unterschiede im olfaktorisch gesteuerten Verhalten liefern ein hervorragendes Modellsystem f{\"u}r diese Studie, um die Entwicklung und Funktion neuronaler Plastizit{\"a}t im olfaktorischen System zu untersuchen. Diese Studie konzentriert sich auf Unterschiede zwischen K{\"o}niginnen und Arbeiterinnen, den beiden weiblichen Kasten innerhalb des Bienestaates, sowie auf umweltbedingte Plastizit{\"a}t. Diploide Eier, aus denen sich K{\"o}niginnen und Arbeiterinnen entwickeln, sind genetisch identisch. Dennoch entwickeln sich K{\"o}niginnen wesentlich schneller zum Adulttier als Arbeiterinnen, sind als Imago gr{\"o}ßer, leben wesentlich l{\"a}nger und zeigen andere Verhaltensweisen. Diese Unterschiede werden durch eine differentielle larvale F{\"u}tterung initiiert. Im Anschluss an das Larvenstadium und somit nach erfolgter Kastendetermination, entwickeln sich die Bienen {\"u}ber eine Puppenphase (verdeckelte Phase) zum Imago. Adulte Bienen klimatisieren das zentrale Brutareal auf einer mittleren Temperatur von 35°C konstant. Bienen, die bei niedrigeren Temperaturen innerhalb des physiologisch relevanten Bereichs aufwachsen, weisen Defizite im olfaktorischen Lernverhalten und in der Tanzkommunikation auf. M{\"o}gliche neuronale Korrelate f{\"u}r altersbedingte, temperatur- und kastenspezifische Unterschiede im olfaktorisch gesteuerten Verhalten sollten in dieser Arbeit betrachtet werden. Die strukturellen Analysen konzentrierten sich dabei auf prim{\"a}re (Antennalloben) und sekund{\"a}re (Pilzk{\"o}rper-Calyces)olfaktorische Verarbeitungszentren im Gehirn von sich entwickelnden und adulten Tieren beider Kasten. Synchron verdeckelte Brutzellen beider Kasten wurden unter kontrollierten Bedingungen im Inkubator herangezogen. Neuroanatomische Untersuchungen wurden an fixierten Gewebeschnitten mittels einer Doppelfluoreszenzf{\"a}rbung mit Fluor-Phalloidin und anti-Synapsin Immuncytochemie durchgef{\"u}hrt. Diese Doppelmarkierung erm{\"o}glichte die Visualisierung und Quantifizierung individueller Synapsenkomplexe (Microglomeruli) im Pilzk{\"o}rper-Calyx. Phalloidin bindet an verschiedene F-Aktin Isoformen und kann zum Nachweis von F-Aktin im Insektennervensystem verwendet werden. F-Aktin wird w{\"a}hrend der Entwicklung in Wachstumskegeln und in adulten Gehirnen in pr{\"a}synaptischen Endigungen und dendritischen Dornen exprimiert. Pr{\"a}synaptische Elemente wurden durch den Einsatz eines spezifischen Antik{\"o}rpers gegen das Drosophila-Vesikeltransportprotein Synapsin I charakterisiert. Mit Hilfe der konfokalen Laser-Scanning Mikroskopie wurde die exakte r{\"a}umliche Zuordnung der Fluoreszenzsignale anhand optischer Schnitte durch die Pr{\"a}parate realisiert. Anhand dieser Methodik konnten erstmals {\"u}ber reine Volumenanalysen hinausgehende Messungen zur synaptischen Strukturplastizit{\"a}t im Pilzk{\"o}rper-Calyx durchgef{\"u}hrt werden. Die Untersuchungen an Gehirnen in den verschiedenen Puppenstadien zeigten Unterschiede im Entwicklungsverlauf der Gehirne mit dem Fokus auf die Bildung antennaler Glomeruli und calycaler Microglomeruli. Unterschiede in der Gehirnentwicklung verdeutlichten die ontogenetische Plastizit{\"a}t des Gehirns der Honigbiene. Entsprechend der k{\"u}rzeren Puppenphase der K{\"o}niginnen bildeten sich sowohl antennale Glomeruli als auch alle Untereinheiten (Lippe, Collar, Basalring) des Calyx etwa drei Tage fr{\"u}her aus. Direkt nach dem Schlupf zeigten quantitative Analysen innerhalb der Pilzk{\"o}rper-Calyces eine signifikant geringere Anzahl an Microglomeruli bei K{\"o}niginnen. Diese neuronale Strukturplastizit{\"a}t auf verschiedenen Ebenen der olfaktorischen Informationsverarbeitung korreliert mit der kastenspezifischen Arbeitsteilung. Die Arbeit liefert Erkenntnisse {\"u}ber den Einfluss eines wichtigen kontrollierten Umweltparameters, der Bruttemperatur, w{\"a}hrend der Puppenphase auf die synaptische Organisation der adulten Pilzk{\"o}rper-Calyces. Bereits geringe Unterschiede in der Aufzuchtstemperatur (1°C) beeinflussten signifikant die Anzahl von Microglomeruli in der Lippenregion des Calyx beider weiblicher Kasten. Die maximale Anzahl an MG entwickelte sich bei Arbeiterinnen bei 34.5°C, bei K{\"o}niginnen aber bei 33.5°C. Neben dieser entwicklungsbedingten neuronalen Plastizit{\"a}t zeigt diese Studie eine starke altersbedingte Strukturplastizit{\"a}t der MG w{\"a}hrend der relativ langen Lebensdauer von Bienenk{\"o}niginnen. Hervorzuheben ist, dass die Anzahl an MG in der olfaktorischen Lippenregion mit dem Alter anstieg (~55\%), in der angrenzenden visuellen Collarregion jedoch abnahm (~33\%). Die in der vorliegenden Arbeite erstmals gezeigte umweltbedingte Entwicklungsplastizit{\"a}t sowie altersbedingte synaptische Strukturplastizit{\"a}t in den sensorischen Eingangsregionen der Pilzk{\"o}rper-Calyces k{\"o}nnte kasten- und altersspezifischen Anpassungen im Verhalten zugrunde liegen.}, subject = {Biene}, language = {en} } @phdthesis{Thakar2006, author = {Thakar, Juilee}, title = {Computational models for the study of responses to infections}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17266}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2006}, abstract = {In diesem Jahrhundert haben neue experimentelle Techniken und Computer-Verfahren enorme Mengen an Information erzeugt, die bereits viele biologische R{\"a}tsel enth{\"u}llt haben. Doch die Komplexit{\"a}t biologischer Systeme wirft immer weitere neue Fragen auf. Um ein System zu verstehen, bestand der Hauptansatz bis jetzt darin, es in Komponenten zu zerlegen, die untersucht werden k{\"o}nnen. Ein neues Paradigma verkn{\"u}pft die einzelnen Informationsteile, um sie auf globaler Ebene verstehen zu k{\"o}nnen. In der vorgelegten Doktorarbeit habe ich deshalb versucht, infekti{\"o}se Krankheiten mit globalen Methoden („Systembiologie") bioinformatisch zu untersuchen. Im ersten Teil wird der Apoptose-Signalweg analysiert. Apoptose (Programmierter Zelltod) wird bei verschiedenen Infektionen, zum Beispiel bei Viruserkrankungen, als Abwehrmaßnahme eingesetzt. Die Interaktionen zwischen Proteinen, die ‚death' Dom{\"a}nen beinhalten, wurden untersucht, um folgende Fragen zu kl{\"a}ren: i) wie wird die Spezifit{\"a}t der Interaktionen erzielt? -sie wird durch Adapter erreicht, ii) wie werden Proliferation/ {\"U}berlebenssignale w{\"a}hrend der Aktivierung der Apoptose eingeleitet? - wir fanden Hinweise f{\"u}r eine entscheidende Rolle des RIP Proteins (Rezeptor-Interagierende Serine/Threonine-Proteinkinase 1). Das Modell erlaubte uns, die Interaktions-Oberfl{\"a}chen von RIP vorherzusagen. Der Signalweg wurde anschließend auf globaler Ebene mit Simulationen f{\"u}r verschiedene Zeitpunkte analysiert, um die Evolution der Aktivatoren und Inhibitoren des Signalwegs und seine Struktur besser zu verstehen. Weiterhin wird die Signalverarbeitung f{\"u}r Apoptosis-Signalwege in der Maus detailliert modelliert, um den Konzentrationsverlauf der Effektor-Kaspasen vorherzusagen. Weitere experimentelle Messungen von Kaspase-3 und die {\"U}berlebenskurven von Zellen best{\"a}tigen das Modell. Der zweite Teil der Resultate konzentriert sich auf das Phagosom, eine Organelle, die eine entscheidende Rolle bei der Eliminierung von Krankheitserregern spielt. Dies wird am Beispiel von M. tuberculosis veranschaulicht. Die Fragestellung wird wiederum in zwei Aspekten behandelt: i) Um die Prozesse, die durch M. tuberculosis inhibiert werden zu verstehen, haben wir uns auf das Phospholipid-Netzwerk konzentriert, das bei der Unterdr{\"u}ckung oder Aktivierung der Aktin-Polymerisation eine große Rolle spielt. Wir haben f{\"u}r diese Netzwerkanalyse eine Simulation f{\"u}r verschiedene Zeitpunkte {\"a}hnlich wie in Teil eins angewandt. ii) Es wird vermutet, dass Aktin-Polymere bei der Fusion des Phagosoms mit dem Lysosom eine Rolle spielen. Um diese Hypothese zu untersuchen, wurde ein in silico Modell von uns entwickelt. Wir fanden heraus, dass in der Anwesenheit von Aktin-Polymeren die Suchzeit f{\"u}r das Lysosom um das F{\"u}nffache reduziert wurde. Weiterhin wurden die Effekte der L{\"a}nge der Aktin-Polymere, die Gr{\"o}ße der Lysosomen sowie der Phagosomen und etliche andere Modellparameter analysiert. Nach der Untersuchung eines Signalwegs und einer Organelle f{\"u}hrte der n{\"a}chste Schritt zur Untersuchung eines komplexen biologischen Systems der Infektabwehr. Dies wurde am Beispiel der Wirt-Pathogen Interaktion bei Bordetella pertussis und Bordetella bronchiseptica dargestellt. Die geringe Menge verf{\"u}gbarer quantitativer Daten war der ausschlaggebende Faktor bei unserer Modellwahl. F{\"u}r die dynamische Simulation wurde ein selbst entwickeltes Bool'sches Modell verwendet. Die Ergebnisse sagen wichtige Faktoren bei der Pathologie von Bordetellen hervor, besonders die Bedeutung der Th1 assoziierten Antworten und dagegen nicht der Th2 assoziierten Antworten f{\"u}r die Eliminierung des Pathogens. Einige der quantitativen Vorhersagen wurden durch Experimente wie die Untersuchung des Verlaufs einer Infektion in verschiedenen Mutanten und Wildtyp-M{\"a}usen {\"u}berpr{\"u}ft. Die begrenzte Verf{\"u}gbarkeit kinetischer Daten war der kritische Faktor bei der Auswahl der computer-gest{\"u}tzten Modelle. Der Erfolg unserer Modelle konnte durch den Vergleich mit experimentellen Beobachtungen belegt werden. Die vergleichenden Modelle in Kapitel 6 und 9 k{\"o}nnen zur Untersuchung neuer Wirt-Pathogen Interaktionen verwendet werden. Beispielsweise f{\"u}hrt in Kapitel 6 die Analyse von Inhibitoren und inhibitorischer Signalwege aus drei Organismen zur Identifikation wichtiger regulatorischer Zentren in komplexen Organismen und in Kapitel 9 erm{\"o}glicht die Identifikation von drei Phasen in B. bronchiseptica und der Inhibition von IFN-\&\#947; durch den Faktor TTSS die Untersuchung {\"a}hnlicher Phasen und die Inhibition von IFN-\&\#947; in B. pertussis. Eine weitere wichtige Bedeutung bekommen diese Modelle durch die m{\"o}gliche Identifikation neuer, essentieller Komponenten in Wirt-Pathogen Interaktionen. In silico Modelle der Effekte von Deletionen zeigen solche Komponenten auf, die anschließend durch experimentelle Mutationen weiter untersucht werden k{\"o}nnen.}, subject = {Bordetella pertussis}, language = {en} } @phdthesis{Duttu2005, author = {Duttu, Vallabhapurapu Subrahmanya}, title = {Regulation of B lymphocyte terminal differentiation and death by the transcription factor Blimp-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17158}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {B lymphocyte induced maturation protein-1 (Blimp-1) und X-box-binding protein-1" (XBP-1) sind als Transkriptionsfaktoren unverzichtbar f{\"u}r die terminale Differenzierung von B-Lymphozyten zu Immunglobulin (Ig)-sezernierenden Plasmazellen. Ebenso stellen die unfolded protein response (UPR) und das Spleißen von XBP-1, beides ausgel{\"o}st durch erh{\"o}hte Ig-Produktion, entscheidende Schritte auf dem Weg zur Plasmazellentstehung dar. Allerdings ist das Molek{\"u}l/ sind die Molek{\"u}le nach wie vor unbekannt, die diesen beiden Ereignissen in der Signalkaskade vorgeschaltet sind. Da die ektope Expression von Blimp-1 in B-Zellen hinreicht, diese zu Plasmazellen zu differenzieren, erscheint es plausibel, dass Blimp-1 das Molek{\"u}l sein k{\"o}nnte, das die Ausl{\"o}sung einer UPR und das Spleißen von XBP-1 steuert. Dieser M{\"o}glichkeit wurde durch ektope Expression von Blimp-1 in der Maus-B-Zell-Lymphomlinie WEHI 231 und in prim{\"a}ren B-Zellen aus der Milz von M{\"a}usen nachgegangen. Die ektope Expression von Blimp-1 f{\"u}hrte in beiden Zelltypen zur Erh{\"o}hung der Ig Produktion, zum Spleißen von XBP-1 und zur Sekretion von Immunglobulinen. Interessanterweise war der N-terminale Anteil von Blimp-1, bestehend aus den Aminos{\"a}uren 1-751, hinreichend, um diese Effekte auszul{\"o}sen, w{\"a}hrend der C-Terminus, der die Aminos{\"a}uren 465-856 umfaßte, keinen Effekt hatte. Dar{\"u}berhinaus, wurde die Expression von BIP, dessen Gen ein UPR-Zielgen ist, durch ektope Expression von Blimp-1 bzw. dessen N-Terminus in prim{\"a}ren B-Zellen erh{\"o}ht. Diese Ergebnisse zeigen deutlich, dass Blimp-1, speziell dessen N-terminale Dom{\"a}ne, hinreichend ist, um eine UPR und die Prozessierung von XBP-1 auszul{\"o}sen, was zur Ig-Sekretion von B-Zellen f{\"u}hrt.}, subject = {B-Lymphozyt}, language = {en} } @phdthesis{Troesken2005, author = {Tr{\"o}sken, Eva-Regina}, title = {Toxicological evaluation of azole fungicides in agriculture and food chemistry}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-17016}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Azole sind wichtige Chemikalien, die als Fungizide in der Landwirtschaft und der Medizin eingesetzt werden. Auch als Zytostatika in der Humanmedizin finden sie Anwendung. Die fungizide Wirkung beruht auf der Hemmung der Lanosterol-14\&\#945;-Demethylase (CYP51), die die Demethylierung von Lanosterol zum „Follicular Fluid Meiosis Activating Steroid (FF-MAS)" katalysiert. F{\"u}r Pilze ist das sp{\"a}ter resultierende Ergosterol ein essentieller Bestandteil der Zellmembran. Exponierten Pilzen fehlt Ergosterol was zu einem Zusammenbruch der Zellmembran f{\"u}hrt. S{\"a}ugetiere k{\"o}nnen Cholesterol, das sp{\"a}tere Produkt der Lanosterol-14\&\#945;-Demethylierung, das zur Synthese von z.B. Gallens{\"a}uren und Sexualhormonen n{\"o}tig ist, mit der Nahrung aufnehmen. FF-MAS und das resultierende T-MAS (Testis Meiosis Activating Steroids), die direkten Produkte der CYP51 katalysierten Reaktion, wirken als Meiose-aktivierende Steroide auf Ovarien und Hoden und werden nicht mit der Nahrung aufgenommen. Eine Hemmung der CYP51 Aktivit{\"a}t k{\"o}nnte das endokrine System beeinflussen und wird daher als unerw{\"u}nschte Nebenwirkung der Azole betrachtet. Aromatase (CYP19) katalysiert die Demethylierung von Testosteron zu {\"O}stradiol und wird durch Azole gehemmt. Die Verringerung der {\"O}strogenspiegel durch CYP19-Inhibition ist das Wirkprinzip der als Zytostatika genutzten Azole, bei den Fungiziden wird es als unerw{\"u}nschte Nebenwirkung angesehen. Ein ideales Azol sollte Pilz-CYP51 stark inhibieren, aber sowohl humanes CYP19 wie auch humanes CYP51 sollten durch ein solches Azol nicht inhibiert werden. Ein ideales Azol-Zytostatikum sollte eine starke inhibitorische Potenz gegen{\"u}ber humanem CYP19 aufweisen, hingegen sollten humanes und Pilz-CYP51 nicht inhibiert werden. Ziel dieser Arbeit war es nun festzustellen: sind Fungizide und Antimykotika starke Inhibitoren von Pilz-CYP51? Zeigen Fungizide und Antimykotika keine Aktivit{\"a}t gegen{\"u}ber humanem CYP19 und humanem CYP51? Sind Zytostatika starke Inhibitoren von humanem CYP19? Zeigen Zytostatika keine Aktivit{\"a}t gegen{\"u}ber humanem CYP51 und Pilz-CYP51? Die inhibitorische Potenz von 22 Azolen, aus den drei Anwendungsgebieten, wurden an vier Systemen getestet: i) an humanem CYP19 und einem fluoreszierenden Pseudosubstrat, ii) an CYP19 und Testosteron als Substrat, iii) an humanem CYP51 und iv) Candida albicans CYP51 und Lanosterol als Substrat. Die Produktbildung wurde mittels Hochdruckfl{\"u}ssigkeitschromatographie gekoppelter Tandem-Massenspektrometrie nach Photosprayionisation gemessen. Das humane CYP51 wurde von „BD Gentest Cooperation" zur Verf{\"u}gung gestellt. Ein katalytisch aktiver Enzymkomplex bestehend aus der Lanosterol-14\&\#945;-Demethylase von Candida albicans und der Oxidoreduktase von Candida tropicalis, wurde im Baculovirussystem exprimiert. Ein Vergleich der inhibitorischen Wirkst{\"a}rke der Substanzen auf menschliches CYP19 und CYP51 und Pilz-CYP51 zeigt, dass einige Azole das erw{\"u}nschte Bild zeigen. Dazu geh{\"o}ren die beiden Zytostatika Fadrozol und Letrozol, sowie Fluconazol und Itraconazol, zwei Antimykotika aus der Humanmedizin, und einige Fungizide z.B. Cyproconazol und Hexaconazol. Ein unerw{\"u}nschtes Bild zeigen z.B. Prochloraz, Bifonazol, Ketoconazol und Miconazol. Sieben Azole weisen ein gemischtes Bild an inhibitorischen Wirkst{\"a}rken auf. Um einen modellartigen Eindruck der R{\"u}ckst{\"a}nde von Azolen in Lebensmitteln zu erhalten, wurde eine auf LC-ESI-MS/MS basierende R{\"u}ckstandsanalytik f{\"u}r Azole im Wein entwickelt. Alle gefunden R{\"u}ckst{\"a}nde lagen unterhalb der beh{\"o}rdlich festgelegten R{\"u}ckstandsh{\"o}chstmengen. Um die inhibitorische Wirkung der Azole auf die verschiedenen Enzymsysteme in einem gr{\"o}ßeren Zusammenhang zu bringen, wurden die IC50 Werte mit Expositionsdaten von Bauern, maximalen Plasmaspiegeln in Patienten nach der Einnahme von Antimykotika und mit Expositionsgrenzwerten f{\"u}r die Langzeitaufnahme von Pflanzenschutzmittelr{\"u}ckst{\"a}nden („Acceptable Daily Intake Levels", ADI) verglichen. Basierend auf den dargestellten Ergebnissen k{\"o}nnen folgende Schlussfolgerungen gezogen werden. Das Risiko f{\"u}r landwirtschaftliche Arbeiter durch Exposition gegen{\"u}ber Azolfungiziden kann im Bezug auf menschliches CYP19 und CYP51 als vernachl{\"a}ssigbar eingestuft werden, wenn die entsprechenden Sicherheitsvorkehrungen getroffen werden. Im medizinischen Bereich muss grunds{\"a}tzlich der Einsatz von Bifonazol, Miconazol und Ketoconazol mit Blick auf die hohe inhibitorische Potenz gegen{\"u}ber menschlichem CYP19 und 51 kritisch betrachtet werden. Unter der Annahme, dass die ADI Werte eingehalten werden, stellen R{\"u}ckst{\"a}nde auf Lebensmitteln in Bezug auf die genannten Enzymsysteme keine Bedrohung f{\"u}r den Verbraucher da. Die Inhibition von CYP19 muss als St{\"o}rung des Hormonsystems angesehen werden. Die Bedeutung von FF-MAS und T-MAS im endokrinen System muss noch abschließend gekl{\"a}rt werden und damit auch die Frage, wie viel Bedeutung der Inhibition von menschlichem CYP51 beigemessen werden muss.}, subject = {Azole}, language = {en} } @phdthesis{Stahl2005, author = {Stahl, Rainer}, title = {Electroactive Conjugated Polymers as Charge-Transport Materials for Optoelectronic Thin-Film Devices}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16980}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {In this work the electrochemical and spectroelectrochemical properties of a series of pi-conjugated organic polymers were studied. The polymers were deposited on platinum electrodes or ITO-coated glass substrates by potentiodynamic electro-polymerisation of the corresponding monomeric precursor molecules. The electro-chemical and photophysical properties of the triarylborane monomers were studied in detail in order to estimate possible influences on the behaviour of the corresponding polymer. The first part of this work aimed at the synthesis and investigation of conjugated donor-acceptor polymers which combine the prerequisites of an OLED within one material: the transport of positive and negative charges and the formation of emissive excited states. With the carbazole-substituted oxadiazoles 1-3 it was shown that on the one hand the carbazole functionality is suitable for enabling the electrochemical polymerisation of the monomers and on the other hand it facilitates reversible p-doping of the resultant polymers. Although n-doping of poly-1-poly-3 is possible due to the electron-deficient oxadiazole rings, it causes the continuous degradation of these electron-acceptor units. Interestingly, this process does not influence the capability of p-doping of the polymers. With respect to its electrochemical and spectroelectrochemical properties the behaviour of the borane polymer poly-4 is absolutely identical with that of the oxadiazole polymers. Moreover, the optical excitation of poly-4 in the solid state leads to the emission of blue-green light which suggests that this polymer might also possess electroluminescent properties. AFM-measurements of poly-4 films on ITO-coated glass substrates revealed, that the film thickness can be controlled to a certain extent by the number of polymerisation redox cycles. It was shown from the electrochemical and photophysical properties of the triarylboranes 4-6 that the pi-pi-interaction between boron and nitrogen atoms is comparably weak in these molecules. This leads to an unexpected ground-state polarisation with a partially positive boron atom and a partially negative nitrogen atom. Moreover, it was found that TAB 4 possesses a lower symmetry than D3 in solution and that excitation energy can be transferred amongst the three subchromophores of 4. By titration experiments it was also demonstrated that TAB 4 can reversibly bind fluoride ions and that the binding event significantly influences the optical absorption characteristics of the chromophore. It can be assumed, that the above mentioned properties, which have a profound influence on the photophysical behaviour of these triarylborane chromophores, also determine the behaviour of the corresponding polymer in a solid state environment. The aim of the second part of this work was the investigation of purely n-conducting materials based on electron-deficient borane and viologen polymers. The corresponding precursor molecules should be polymerised on platinum electrodes by reductive electropolymerisation. However, a reductive polymerisation was not possible for the borane monomer 19 which is thought to be due to a strong localisation of the unpaired electron on the central boron atom of the radical anion. An electropolymerisation of the cyano-substituted bispyridinio-compound 17 failed because of the poor quality of CN- as a leaving group. Thus, a synthesis of the analogous isomer 18 was developed, in which the cyano-substituents were exchanged by the better leaving group Cl-. The viologen polymer poly-18, which can be regarded as an electron-deficient iso-electronic analogue of poly(para-phenylene), was successfully deposited on a platinum electrode by reductive electropolymerisation of 18. Poly-18 can be reversibly n-doped at comparably low potentials; however, at higher potentials the polymer is overcharged and destroyed irreversibly. As the synthetic strategy for 18 allows the variation of both spacer unit and leaving group in the last two steps of the reaction sequence, a series of analogous compounds can be easily synthesised using this route.}, subject = {Polymerhalbleiter}, language = {en} } @phdthesis{Denker2006, author = {Denker, Katrin}, title = {Isolation and characterization of channel-forming proteins in the outer membrane of E. coli and Borrelia species}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16955}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2006}, abstract = {In this study pore forming proteins of the gram-negative bacteria B. burgdorferi, B. duttonii and E.coli were investigated. Therefore the study is subdivided into three parts. In the first part outer membrane preparation of three relapsing fever Borrelia were investigated. In the second part the putative TolC homologue BB0124 of B. burgdorferi, the Lyme borreliosis agent, was studied. In the last part the influence of point mutants within the greasy slide of the maltose specific porin (LamB) of E. coli were shown. In the first part of this study outer membrane preparations of three Borrelia relapsing fever strains have been studied for pore-forming activity in the black lipid bilayer assay. Histograms of conductance fluctuations were obtained from single-channel experiments with outer membrane preparations of B. hermsii, B. recurentis and B. duttonii. All strains had a different conductance fluctuation pattern with a broad range of single-channel conductance values varying from 0.5 nS - 11 nS. Common for all three strains was a high pore-forming activity at around 0.5 nS. Furthermore the proteins of the outer membrane of B. duttonii were separated by chromatographic methods. Some eluate fractions contained a channel-forming protein, which was forming stable channels with a single-channel conductance of 80 pS in 1 M KCl. Characterization of this channel showed that it is slightly anionic selective and voltage independent. The small single-channel conductance suggests that it is a specific pore. However, a substrate specificity could not be determined. In the second part, for the B. burgdorferi HB19 and p66 knock out strain HB19/K02, their outer membrane preparations were characterized in the black lipid bilayer assay. Comparing the histograms of single-channel conductions fluctuations of both strains showed no single-channel activity at 11.5 nS for the p66 knock out strain. This verifies earlier studies that P66 is a pore-forming protein in B. burgdorferi. Furthermore, one fraction obtained by anion exchange chromatography of the p66 knock out outer membrane protein preparation showed a uniform channel-forming activity with a single channel conductance of 300 pS. The electrophysically characterization of the 300 pS channel showed that it is not ionselective or voltage dependent. By mass spectrometry using peptide mass finger prints, BB0142 could be identified as the sole channel forming candidate in the active fraction. A BLAST search and a conserved domain search showed that BB0142 is a putative TolC homologue in B. burgdorferi. Furthermore the location of the bb0142 gene within the chromosome is in an operon encoding a multidrug efflux pump. In this study the expression of an outer membrane component of a putative drug efflux system of B. burgdorferi was shown for the first time. In the third part functional studies of the maltooligosaccharide-specific LamB channel were performed. The 3D-structure of LamB suggests that a number of aromatic residues (Y6, Y41, W74, F229, W358 and W420) within the channel lumen is involved in carbohydrate and ion transport. All aromatic residues were replaced by alanine (A) scanning mutagenesis. Furthermore, LamB mutants were created in which one, two, three, four and five aromatic residues were replaced to study their effects on ion and maltopentaose transport through LamB. The purified mutant proteins were reconstituted into lipid bilayer membranes and the single-channel conductance was studied. The results suggest that all aromatic residues provide some steric hindrance for ion transport through LamB. Highest impact is provided by Y6 and Y41, which are localized opposite to Y118, which forms the central constriction of the LamB channel. Stability constants for binding of maltopentaose to the mutant channels were measured using titration experiments with the carbohydrate. The mutation of one or several aromatic amino acids led to a substantial decrease of the stability constant of binding. The highest effect was observed when all aromatic amino acids were replaced by alanine because no binding of maltopentaose could be detected in this case. However, binding was again possible when Y118 was replaced by tryptophane (W). The carbohydrate-induced block of the channel function could also be used for the study of current noise through the different mutant LamB-channels. The analysis of the power density spectra of some of the mutants allowed the evaluation of the on- and off-rate constants (k1 and k-1) of carbohydrate binding to the binding-site inside the channels. The results suggest that both on- and off-rate constants were affected by the mutations. For most mutants k1 decreased and k-1 increased.}, subject = {Escherichia coli}, language = {en} } @phdthesis{Pils2005, author = {Pils, Birgit}, title = {Insights into the evolution of protein domains give rise to improvements of function prediction}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16805}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The growing number of uncharacterised sequences in public databases has turned the prediction of protein function into a challenging research field. Traditional annotation methods are often error-prone due to the small subset of proteins with experimentally verified function. Goal of this thesis was to analyse the function and evolution of protein domains in order to understand molecular processes in the cell. The focus was on signalling domains of little understood function, as well as on functional sites of protein domains in general. Glucosaminidases (GlcNAcases) represent key enzymes in signal transduction pathways. Together with glucosamine transferases, they serve as molecular switches, similar to kinases and phosphatases. Little was known about the molecular function and structure of the GlcNAcases. In this thesis, the GlcNAcases were identified as remote homologues of N-acetyltransferases. By comparing the homologous sequences, I was able to predict functional sites of the GlcNAcase family and to identify the GlcNAcases as the first family member of the acetyltransferase superfamily with a distinct catalytic mechanism, which is not involved in the transfer of acetyl groups. In a similar approach, the sensor domain of a plant hormone receptor was studied. I was able to predict putative ligand-binding sites by comparing evolutionary constraints in functionally diverged subfamilies. Most of the putative ligand-binding sites have been experimentally confirmed in the meantime. Due to the importance of enzymes involved in cellular signalling, it seems impossible to find substitutions of catalytic amino acids that turn them catalytically inactive. Nevertheless, by scanning catalytic positions of the protein tyrosine phosphatase families, I found many inactive domains among single domain and tandem domain phosphatases in metazoan proteomes. In addition, I found that inactive phosphatases are conserved throughout evolution, which led to the question about the function of these catalytically inactive phosphatase domains. An analysis of evolutionary site rates of amino acid substitutions revealed a cluster of conserved residues in the apparently redundant domain of tandem phosphatases. This putative regulatory center might be responsible for the experimentally verified dimerization of the active and inactive domain in order to control the catalytic activity of the active phosphatase domain. Moreover, I detected a subgroup of inactive phosphatases, which presumably functions in substrate recognition, based on different evolutionary site rates within the phosphatase family. The characterization of these new regulatory modules in the phosphatase family raised the question whether inactivation of enzymes is a more general evolutionary mechanism to enlarge signalling pathways and whether inactive domains are also found in other enzyme families. A large-scale analysis of substitutions at catalytic positions of enzymatic domains was performed in this work. I identified many domains with inactivating substitutions in various enzyme families. Signalling domains harbour a particular high occurrence of catalytically inactive domains indicating that these domains have evolved to modulate existing regulatory pathways. Furthermore, it was shown that inactivation of enzymes by single substitutions happened multiple times independently in evolution. The surprising variability of amino acids at catalytic positions was decisive for a subsequent analysis of the diversity of functional sites in general. Using functional residues extracted from structural complexes I could show that functional sites of protein domains do not only vary in their type of amino acid but also in their structural location within the domain. In the process of evolution, protein domains have arisen from duplication events and subsequently adapted to new binding partners and developed new functions, which is reflected in the high variability of functional sites. However, great differences exist between domain families. The analysis demonstrated that functional sites of nuclear domains are more conserved than functional sites of extracellular domains. Furthermore, the type of ligand influences the degree of conservation, for example ion binding sites are more conserved than peptide binding sites. The work presented in this thesis has led to the detection of functional sites in various protein domains involved in signalling pathways and it has resulted in insights into the molecular function of those domains. In addition, properties of functional sites of protein domains were revealed. This knowledge can be used in the future to improve the prediction of protein function and to identify functional sites of proteins.}, subject = {Dom{\"a}ne }, language = {en} } @phdthesis{Pozgajova2005, author = {Pozgajova, Miroslava}, title = {Studies on formation and stabilization of pathological thrombi in vivo}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16784}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Platelet activation and adhesion resulting in thrombus growth is essential for normal hemostasis, but can lead to irreversible, life-threatening vessel occlusion. In the current study, the contribution of platelet integrins, activation receptors and the contact system of blood coagulation in such pathological conditions was investigated in mice.}, subject = {Thrombose}, language = {en} } @phdthesis{Toben2005, author = {Toben, Catherine Gisela}, title = {Generation and analysis of transgenic mice expressing ovalbumin as a neo-self antigen under control of the myelin basic protein promoter}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16708}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {In this project two novel murine autoimmune models were to be established in an attempt to further investigate the nervous system disorders of Multiple Sclerosis and Guillain Barr{\´e} Syndrome. Previous experimental autoimmune encephalomyelitis (EAE) and experimental autoimmune neuritis (EAN) models have demonstrated that T cells play a major role in these diseases. Which roles CD4 and CD8 T cells specifically have in the initiation, propagation and termination of an autoimmune nervous system disorder remains controversial. To this end two transgenic mice specifically expressing the neo-antigen (Ag) ovalbumin (OVA) in either the central nervous system (CNS) or peripheral nervous system (PNS) were to be generated. The myelin basic protein (MBP) is a major component of the myelin sheath both within the CNS and the PNS. Therefore the MBP promoter was employed for its distinct regulatory elements to facilitate exclusive CNS or PNS OVA expression. The adoptive transfer of OVA specific MHCI restricted (OT-I) and MHCII restricted (OT-II) TCR Tg T cells extended the OVA Tg mouse model by allowing potentially encephalitogenic T cells to be tracked in vivo. Specificity for the target Ag should enable the dynamic role of antigen specific T cells in neuroinflammatory diseases to be revealed in more detail.}, subject = {Multiple Sklerose}, language = {en} } @phdthesis{Kleinsteuber2005, author = {Kleinsteuber, Martin}, title = {Jacobi-type methods on semisimple Lie algebras : a Lie algebraic approach to numerical linear algebra}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16454}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Es wird eine Lie-algebraische Verallgemeinerung sowohl des klassischen als auch des Sortier-Jacobi-Verfahrens f{\"u}r das symmetrische Eigenwertproblem behandelt. Der koordinatenfreie Zugang erm{\"o}glicht durch eine neue Betrachtungsweise die Vereinheitlichung strukturierter Eigen- und Singul{\"a}rwertprobleme, darunter bis dato noch nicht betrachtete F{\"a}lle. F{\"u}r beide Verfahren wird lokal quadratische Konvergenz, sowohl f{\"u}r den regul{\"a}ren als auch f{\"u}r den irregul{\"a}ren Fall, gezeigt. Die Analyse und Verallgemeinerung der sog. speziellen Sweeps f{\"u}r das symmetrische Eigenwertproblem f{\"u}hrt zu neuen Sweep-Methoden f{\"u}r strukturierte Eigen- und Singul{\"a}rwertprobleme, die ein besseres Konvergenzverhalten als die bisher bekannten aufweisen.}, subject = {Eigenwert}, language = {en} } @phdthesis{Bradeanu2005, author = {Bradeanu, Ioana Lavinia}, title = {Photoionization and excitation of free variable size van der Waals clusters in the inner shell regime}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16372}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The studies presented in this thesis deal with resonant and non-resonant excitation of free variable size clusters using synchrotron radiation in the soft X-ray regime. The post collision interaction (PCI) effect is investigated in free variable size krypton and argon clusters near the Kr 3d and Ar 2p ionization energies. The core ionization energies of surface and bulk sites in variable size clusters can be clearly distinguished. This is mostly due to the polarization screening. It is found that the asymmetry, which is a consequence of PCI, is characteristically smaller for clusters than for isolated atoms. Moreover, there is less asymmetry for bulk sites than for surface sites in variable size rare gas clusters. We assign the results in terms of mechanisms that are based on quantum mechanical models of post collision interaction. Complementary experiments on the photoionization of free van der Waals clusters are performed by using zero kinetic energy (ZEKE) photoelectron spectroscopy in the Ar 2p-, Kr 3d-, Ne 1s-, and N2-regimes. The experimental approach is also suitable to detect cluster size dependent changes in electronic structure. This also allows us to study post collision interaction in variable size clusters. The parameters of the PCI profiles deduced for ZEKE experiments indicate that there are no significant changes in core ionization dynamics compared to near-threshold experiments. Results from model calculations in Kr 3d ionization energy indicate that different geometric sites can be clearly distinguished from each other by their substantial shift in Kr 3d ionization energy, though the dimer shows almost the same Kr 3d ionization energy as the free atom. A comparison with the experimental results indicates that there is resemblance with the model calculations, even though close-lying ionization energies are blended and require deconvolutions of the experimental spectra. It is evident from the present work that one can observe distinct shifts in core ionization energies in van der Waals clusters that are formed in wide size distributions of a jet expansion. The emission of ultraviolet fluorescence radiation from variable size argon clusters is investigated with high spectral resolution in the Ar 2p-excitation regime. The fluorescence excitation spectra reveal strong fluorescence intensity in the Ar 2p-continuum, but no evidence for the occurrence of discrete low-lying core-exciton states in the near-edge regime. This finding is different from the absorption and photoionization cross sections of argon clusters and the solid. The dispersed fluorescence shows a broad molecular band centered near 280 nm. The present results are consistent with the formation of singly charged, excited moieties within the clusters, which are assigned as sources of the radiative relaxation in the 280 nm regime. A fast energy transfer process (interatomic Coulombic decay, ICD) is assigned to be primarily the origin of these singly charged, excited cations besides intra-cluster electron impact ionization by Auger electrons. Our findings give possibly the first experimental evidence for ICD in the core level regime. Free, variable size nitrogen clusters are investigated in the N 1s excitation regime in comparison with the free molecule and solid nitrogen. The conversion of Rydberg states into core excitons, surface and bulk, was studied. The experimental results are simulated by ab initio calculations using (N2)13 as a reasonable prototype cluster structure that allows us to simulate both surface and bulk properties in comparison with the isolated molecule. The present results clearly show that there are specific properties, such as molecular orientation, in molecular van der Waals clusters, which do not exist in atomic van der Waals clusters. It is shown that inner and outer surface sites give rise to distinct energy shifts of the low lying surface core excitons.}, subject = {Photoionisation}, language = {en} } @phdthesis{Kackstaetter2003, author = {Kackstaetter, Uwe R.}, title = {Contaminant diffusion and sorption of an artificial leachate in selected geologic barriers of Frankonia, Bavaria, Germany}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16151}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2003}, abstract = {The geologic barrier represents the final contact between a landfill and the environment. Ideally suited are clays and mudstones because of sufficient vertical and lateral extent, low hydraulic conductivities and high sorptive characteristics. Since hydraulic conductivity is no longer the single criteria to determine transport and retardation of contaminants in geologic landfill barrier materials, diffusive and sorptive characteristics of 4 different clay and mudstone lithologies in Northern Bavaria, were investigated. Cored samples from various depths were included in this study and subjected to evaluations of geochemistry, mineralogy, physical parameters, sorption and diffusion. A transient double reservoir with decreasing source concentration was designed and constructed using clear polycarbonate cylinders for undisturbed clay plugs of 2 to 4cm thickness. Samples were also fitted with internal electrical conductivity probes to determine the migration of the diffusive front. A multi chemical species synthetic landfill leachate was contrived to simulate and evaluate natural pollutant conditions. A computational method for determining mineralogy from geochemical data was also developed. It was found that sorptive processes are mostly controlled by the quality and type of fine grained phyllosilicates and the individual chemical species involved exhibited linear, Freundlich, as well as Langmuir sorption properties. Effective diffusion and sorption coefficients were also determined using POLLUTEv6 (GAEA, 1997) software and receptor reservoir concentrations for K, Na, Ca, Cu, NH4, Cl, NO3, SO4, and concentration totals at predetermined time intervals. Anion exclusion proved to be a major factor in the diffusion process and was used to explain many observed anomalies. Furthermore, diffusion coefficients were found not to be static with a multi chemical species leachate, but actually varied during the course of the experiment. Strong indications point toward the major role of pore space quality, shape, and form as control of diffusive properties of a geologic barrier. A correlation of CECNa of the samples with De may point to a possible deduction of diffusive properties for multi species leachates without extensive and time consuming laboratory tests}, language = {en} } @phdthesis{Boehler2005, author = {B{\"o}hler, Elmar}, title = {Algebraic closures in complexity theory}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-16106}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {We use algebraic closures and structures which are derived from these in complexity theory. We classify problems with Boolean circuits and Boolean constraints according to their complexity. We transfer algebraic structures to structural complexity. We use the generation problem to classify important complexity classes.}, subject = {Komplexit{\"a}tstheorie}, language = {en} } @phdthesis{Amthor2005, author = {Amthor, Stephan}, title = {Redox properties of Bis-Triarylamines and ligand properties of Thianthrenophane}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15916}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The one electron oxidation potential of ten TAAs with all permutations of Cl , OMe- and Me-substituents in the three p-positions were determined by CV. The half wave potential of the first oxidation wave correlates linearly with the number of Cl- and OMe-substituents. AM1-CISD derived values of the absorption energies are in good agreement with the experiments but differ strongly for the oscillator strengths as well as for neutral compounds and their corresponding mono radical cations. The small solvent dependence of the experimental UV/Vis spectra in CH2Cl2 and MeCN reflects a minor charge transfer character of the electronic transitions. The UV/Vis/NIR spectra of the series of TAAs and their corresponding radical cations and the AM1 computations reveal that even small substituents may lead to strong symmetry breaking and to a modified electronic structure. The spectroscopic properties of a series of four bis-TAA donor-bridge-donor X-B-X dimers, composed of two asymmetric TAA chromophores (monomers) were investigated. UV/vis-, fluorescence and transient absorption spectra were recorded and compared with those of the corresponding X-B monomers. The excited states of the dimers are described as MV states which show, depending on the chemical nature of the bridge, a varying amount of interactions. It was found that superradiant emission only proceeds in the case of weak and medium coupling. Whether the first excited state potential energy surface of the dimers is a single minimum or a double minimum potential depends on the solvent polarity and the electronic coupling. In the latter case, the dimer relaxes in a symmetry broken CT state. The [2.2]paracyclophane bridged dimer is an example for a weakly coupled system, because the spectroscopic behavior is very similar to the corresponding p xylene monomer. In contrast, anthracene as well as p-xylene bridges mediate a stronger coupling and reveal a significant cooperative influence on the optical properties. A series of [2.2]paracylophane bridged bis-TAA MV radical cations X-B-X+ were analyzed by a GMH three-level model which takes two transitions into account: the IV-CT band and the bridge band. From the GMH analysis, one can conclude that the [2.2]paracyclophane moiety is not the limiting factor which governs the intramolecular charge transfer. The electronic interactions are of course smaller than direct conjugation but from the order of magnitude of the couplings of the [2.2]paracyclophane MV species it can be assumed that this bridge is able to mediate significant through-space and through-bond interactions. From the exponential dependence of the electronic coupling V between the two TAA localized states on the distance r between the two redox centers, it was inferred that the HT proceeds via superexchange mechanism. The analysis reveals that even significantly longer conjugated bridges should still mediate significant electronic interactions, because the decay constant of a series of conjugated MV species is small. The absorption properties of a series of bis-TAA-[2.2]paracyclophane dications X+-B-X+ were presented. The localized and the CT transitions of these dications are explained and analyzed by an exciton coupling model which also considers the photophysical properties of the monomeric TAA radical cations. Together with AM1-CISD calculated transition moments, experimental transition moments and transition energies of the bis-TAA dications were used to calculate electronic couplings by a GMH approach. These couplings are a measure for interactions of the excited MV CT states. The modification of the diabatic states reveals similarities of the GMH three-level model and the exciton coupling model. Comparison of the two models shows that the transition moment between the excited mixed-valence states of the dimer equals the dipole moment difference of the ground and the excited bridge state of the corresponding monomer. Thianthrenophane (1) has a cavity which offers enough room to potentially enable endohedral coordination to small ions or molecules. For the complexation of silver(I) perchlorate, the complex stability constants of thianthrenophane logK1=5.45 and of thianthrene logK2=9.16 were determined by UV/Vis titration. Single competition transport experiments with ten metal salts demonstrate a very high selectivity of thianthrenophane as a carrier for silver(I) and a distinctly higher transport rate compared to carriers such as thianthrene and 14-ane-S4. Although the X-ray crystal structure analysis of the polymeric [Ag(1)]ClO4 shows an exohedral coordination to silver(I), the formation of an endohedral [Ag(1)]+ complex is suggested to be the explanation for the unusual carrier selectivity of silver(I) by 1 in bulk liquid membrane.}, subject = {Triarylamine}, language = {en} } @phdthesis{Kriegisch2005, author = {Kriegisch, Volker}, title = {Electron transfer processes between organic redox centres and electrodes via active bridges in self-assembled monolayers}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15892}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Cyclovoltammetrische Messungen der Ferrocenalkylthiole 1 - 3 belegen, dass homogene, gemischte Monolagen aus redoxaktiven Verbindungen und redoxinaktiven Alkylthiolen gebildet werden. Die von Creager et al. bestimmten ET Raten der Ferrocenalkylthiole 1 - 3 konnten hierbei verifiziert werden. Wie erwartet erfolgt eine Abnahme der ET Geschwindigkeit bei einer Kettenverl{\"a}ngerung des Alkylspacers von 2 nach 3. Eine unterschiedliche Konnektivit{\"a}t zwischen Redoxzentrum und Alkylspacer, z. B. die Einf{\"u}hrung einer Carbonyl-Funktion im Falle von 1, unter Beibehaltung der Kettenl{\"a}nge zeigt keinen bemerkbaren Einfluß auf den ET. Trotzt vergleichbaren Abstands der aromatischen Ferrocenthiole 4 und 5 zu der C8-Alkyl-Verbindung 2 zwischen Redoxzentrum und Elektrode, weisen diese aufgrund ihrer starken Konjugation sehr hohe ET Geschwindigkeiten auf. Die elektronischen Kopplungsfaktoren selbst deuten auf einen nichtadiabtischen ET zwischen Redoxzentrum und Elektrode hin. Wie erwartet kommt es zu einem Anwachsen der Kopplungsfaktoren bei sich verk{\"u}rzender Kettenl{\"a}nge oder bei Einf{\"u}hrung konjugierter Spacersysteme. Zusammenfassend kann gesagt werden, dass Erfahrungen hinsichtlich der Pr{\"a}paration der Monolagen gesammelt, die gemessenen ET Raten f{\"u}r der literaturbekannten Verbindungen 1 - 3 best{\"a}tigt und diese Informationen auf die konjugierten Verbindungen 4 und 5 angewandt werden konnten. Im zweiten Teil wurden die Triarylamin- (29, 32) und Phenothiazinalkylthiole (35) bez{\"u}glich ihres ET Verhaltens in gemischten Monolagen untersucht. Mittels Cyclovoltammetrie konnte gezeigt werden, daß einheitlich geformte, verd{\"u}nnte Monolagen vorliegen. Die ET Raten der Triarylamin- (29, 32) und Phenithiazinalkylthiole (35) sind jedoch um den Faktor 10 bis 100 h{\"o}her als vergleichbare Ferrocenalkylthiole gleicher Kettenl{\"a}nge [1, 2], wohingegen f{\"u}r Monolagen, welche [Ru(bpy)2(pp)]+-Alkythiole enthalten, {\"a}quivalente Werte gefunden wurden [3]. Die ET Geschwindigkeit wird von zwei Parametern beeinflusst: dem elektronischen Kopplungsmatrixelement und der Regorganisationsenergie \&\#61548;\&\#61472; [4]. Die ET Geschwindigkeit in Donor-substituierten Alkylthiolen wird haupts{\"a}chlich durch \&\#61548; beeinflusst und sogar kleine {\"A}nderungen dieser zeigen eine große Auswirkung auf die zu untersuchenden Prozesse. Aus diesem Grund wird eine Zunahme der ET Geschwindigkeit von Ferrocen (hohe Reorganisationsenergie) {\"u}ber die Phenothiazinverbindung 35 und [Ru(bpy)2(pp)]+ zu den Triarylaminchromophoren 29 und 32 (niedrige Reorganisationsenergie) beobachtet. Weiterhin spielt, im Gegensatz zu Beobachtung von Creager et al. an {\"a}quivalenten Ferrocenverbingungen, die Anbindung des Redoxzentrums an den Alkylspacer eine bedeutende Rolle. Im Falle der elektronenreichen Ether-verbr{\"u}ckten Verbindung 29 wird der ET nicht alleine durch \&\#61548;, sondern ebenso durch mesomere Effekte bestimmt. Bei 29 kommt es durch Lokalisation der positiven Ladung nahe der Ether Funktion formal zu einer Kettenverk{\"u}rzung um eine „Methyleneinheit", welche schließlich in h{\"o}heren ET Geschwindigkeiten resultiert. Im dritten Teil dieser Dissertation wurde ein Serie „molekularer Dr{\"a}hte" bestehend aus Methoxy- oder Chlorid-substituierten Triarylamin- und Phenothiazinverbindungen mit unterschiedlichen Br{\"u}ckeneinheiten und Br{\"u}ckenl{\"a}ngen zwischen Redoxzentrum und Ankerfunktion dargestellt und im Hinblick auf ihr ET Verhalten untersucht. Durch cyclovoltammetrische und UV/Vis-spektroskopische Untersuchungen konnte gezeigt werden, dass sowohl die Oxidationspotentiale als auch die energetischen Zust{\"a}nde der Chromophore recht gut durch Einf{\"u}hrung unterschiedlicher Redoxzentren und Br{\"u}ckeneinheiten beeinflusst werden k{\"o}nnen. Trotz erfolgreicher Kontrolle der Dichte der Chromophoreinheiten in den gemischten Monolagen konnte nur f{\"u}r die Verbindungen 49, 52 und 87 mit Nitril-substituierten Br{\"u}ckeneinheiten verl{\"a}ssliche ET Geschwindigkeiten erhalten werden. Bei diesen Chromphoren ist ein Absinken der ET Geschwindigkeit bei zunehmender Dichte der redoxaktiven Molek{\"u}le in den gemischten Monolagen zu beobachten, welche auf eine {\"A}nderung der Adsorptionsgeometrie hindeutet. Bei zunehmender Packungsdichte der Chromophore f{\"u}hrt dies zu einer aufrechteren Stellung der redoxaktiven Spezies. F{\"u}r alle anderen Verbindungen konnten keine Werte aufgrund der zu schnellen ET Geschwindigkeiten ermittelt werden. Konformelle, wie auch die sehr geringe Abstandsabh{\"a}ngigkeit des ET, resultieren in hohen ET Geschwindigkeiten oder auch ung{\"u}nstige HOMO-LUMO Energien bez{\"u}glich des Donors, der Br{\"u}cke und der Elektrode sind Gr{\"u}nde f{\"u}r dieses Verhalten. Die Tatsache, dass Verbindung 49 und 52 beinahe die gleichen Geschwindigkeitskonstanten des ETs unabh{\"a}ngig von der Anzahl der Br{\"u}ckeneinheiten (n = 2, n = 3) besitzen, deutet darauf hin, dass ein Hopping-Prozess stattfindet, bei welchem eine geringere L{\"a}ngenabh{\"a}ngigkeit des ETs als bei eine Superexchange-Mechanismus zu erwarten ist.}, subject = {Monoschicht}, language = {en} } @phdthesis{Ruester2005, author = {R{\"u}ster, Christian}, title = {Magnetotransport effects in lateral and vertical ferromagnetic semiconductor junctions}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15554}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {This work is an investigation of giant magnetoresistance (GMR), tunneling magnetoresistance (TMR) and tunneling anisotropic magnetoresistance (TAMR)effects in (Ga,Mn) based ferromagnetic semiconductor junctions. Detailed results are published in the following articles: [1] C. R{\"u}ster, T. Borzenko, C. Gould, G. Schmidt, L.W. Molenkamp, X. Liu, T.J.Wojtowicz, J.K. Furdyna, Z.G. Yu and M. Flatt´e, Very Large Magnetoresistance in Lateral Ferromagnetic (Ga,Mn)As Wires with Nanoconstrictions, Physical Review Letters 91, 216602 (2003). [2] C. Gould, C. R{\"u}ster, T. Jungwirth, E. Girgis, G.M. Schott, R. Giraud, K. Brunner, G. Schmidt and L.W. Molenkamp, Tunneling Anisotropic Magnetoresistance: A Spin-Valve-Like Tunnel Magnetoresistance Using a Single Magnetic Layer, Physical Review Letters 93, 117203 (2004). [3] C. R{\"u}ster, C. Gould, T. Jungwirth, J. Sinova, G.M. Schott, R. Giraud, K. Brunner, G. Schmidt and L.W. Molenkamp, Very Large Tunneling Anisotropic Magnetoresistance of a (Ga,Mn)As/GaAs/(Ga,Mn)As Stack, Physical Review Letters 94, 027203 (2005). [4] C. R{\"u}ster and C. Gould, T. Jungwirth, E. Girgis, G.M. Schott, R. Giraud, K. Brunner, G. Schmidt and L.W. Molenkamp, Tunneling anisotropic magnetoresistance: Creating a spin-valve-like signal using a single ferromagnetic semiconductor layer, Journal of Applied Physics 97, 10C506 (2005).}, subject = {Galliumarsenid}, language = {en} } @phdthesis{Sohns2005, author = {Sohns, Andreas}, title = {Halfsandwich iron complexes with Silanol-functionalized Cyclopentadienyl ligands}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15809}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {...}, subject = {Halbsandwich-Verbindungen}, language = {en} } @phdthesis{Schmitt2005, author = {Schmitt, Rainer}, title = {P-H-functionalized Phosphenium Tungsten Complexes : exchange reactions at the phosphorus and cycloadditions with heteroallenes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15818}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {...}, subject = {Phospheniumkomplexe}, language = {en} } @phdthesis{Schaefer2005, author = {Sch{\"a}fer, Matthias}, title = {Molecular mechanisms of floor plate formation and neural patterning in zebrafish}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15789}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The vertebrate spinal cord is composed of billions of neurons and glia cells, which are formed in a highly coordinated manner during early neurogenesis. Specification of these cells at distinct positions along the dorsoventral (DV) axis of the developing spinal cord is controlled by a ventrally located signaling center, the medial floor plate (MFP). Currently, the origin and time frame of specification of this important organizer are not clear. During my PhD thesis, I have analyzed the function of the novel secreted growth factor Midkine-a (Mdka) in zebrafish. In higher vertebrates, mdk and the related factor pleiotrophin (ptn) are widely expressed during embryogenesis and are implicated in a variety of processes. The in-vivo function of both factors, however, is unclear, as knock-out mice show no embryonic phenotype. We have isolated two mdk co-orthologs, mdka and mdkb, and one single ptn gene in zebrafish. Molecular phylogenetic analyses have shown that these genes evolved after two large gene block duplications. In contrast to higher vertebrates, zebrafish mdk and ptn genes have undergone functional divergence, resulting in mostly non-redundant expression patterns and functions. I have shown by overexpression and knock-down analyses that Mdka is required for MFP formation during zebrafish neurulation. Unlike the previously known MFP inducing factors, mdka is not expressed within the embryonic shield or tailbud but is dynamically expressed in the paraxial mesoderm. I used epistatic and mutant analyses to show that Mdka acts independently from these factors. This indicates a novel mechanism of Mdka dependent MFP formation during zebrafish neurulation. To get insight into the signaling properties of zebrafish Mdka, the function of both Mdk proteins and the candidate receptor Anaplastic lymphoma kinase (Alk) have been compared. Knock-down of mdka and mdkb resulted in the same reduction of iridophores as in mutants deficient for Alk. This indicates that Alk could be a putative receptor of Mdks during zebrafish embryogenesis. In most vertebrate species a lateral floor plate (LFP) domain adjacent to the MFP has been defined. In higher vertebrates it has been shown that the LFP is located within the p3 domain, which forms V3 interneurons. It is unclear, how different cell types in this domain are organized during early embryogenesis. I have analyzed a novel homeobox gene in zebrafish, nkx2.2b, which is exclusively expressed in the LFP. Overexpression, mutant and inhibitor analyses showed that nkx2.2b is activated by Sonic hedgehog (Shh), but repressed by retinoids and the motoneuron-inducing factor Islet-1 (Isl1). I could show that in zebrafish LFP and p3 neuronal cells are located at the same level along the DV axis, but alternate along the anteroposterior (AP) axis. Moreover, these two different cell populations require different levels of HH signaling and nkx2.2 activities. This provides new insights into the structure of the vertebrate spinal cord and suggests a novel mechanism of neural patterning.}, subject = {Zebrab{\"a}rbling}, language = {en} } @phdthesis{Bayer2005, author = {Bayer, Tanja}, title = {Toxicity and biotransformation of 1,1,1,3,3-Pentafluoropropane, 3,3,3-Trifluoropropionic acid and 1,1,1,3-Tetrachloropropane}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15731}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The biotransformation of 1,1,1,3,3-pentafluoropropane was investigated in rats and in in vitro systems. First, the metabolites were identified in vivo using GC/MS and 19F NMR analysis. The main metabolite was identified as trifluoroacetic acid, the minor metabolite as 3,3,3-trifluoropropionic acid and as a cleavage product, inorganic fluoride was found. As the in vitro system, liver microsomes from rat and human samples and rat liver homogenates were used. Trifluoroacetic acid and 3,3,3 trifluoropropionic acid were confirmed in vitro as metabolic intermediates, following biotransformation of 1,1,1,3,3-pentafluoropropane by the cytochrome P-450-system. Studies, designed for clarifying the cardiotoxicity of 1,1,1,3,3-pentafluoropropane were driven by the hypothesis that 3,3,3-trifluoropropionic acid is the toxic agent. This was based on the lethal toxicity, which was observed in previous in vivo experiments. In addition, the point of its structural similarity to toxic agents as for example monofluoroacetic acid or of possible metabolic intermediates like difluoroacrylic acid with known toxicity were considered to support this assumption. However, trifluoroacetic acid was neglected as the sought-after toxic agent because of its different toxic effects, known from literature. Investigations on the biotransformation of 3,3,3-trifluoropropionic acid were performed and resulted in no metabolic activity and in poor elimination of 3,3,3-trifluoropropionic acid in vivo. The histopathological effects on the heart, which were observed in the 90-day oral toxicity study of 1,1,1,3,3-pentafluoropropane in rats, namely mononuclear inflammatory cell infiltrations and degenerated myocardial fibers, were not observed after a 28 day repeated exposure of up to 10 mg/kg b.w. of 3,3,3-trifluoropropionic acid. However, a single high dose of 3,3,3-trifluoropropionic acid lead to severe toxicological effects. The difference in the observed toxic effects after a single and repeated administration may be due to adaptive mechanisms in rats. The toxicological effects included clinical signs like ataxia, coma and cramps. The conditions of the rats suggested possible inhibition of the energy supply to the organism. Furthermore, the interference of 3,3,3-trifluoropropionic acid in the functionality of the organism was investigated. Experiments were performed in vitro in rat liver and heart mitochondria to investigate effects on the mitochondrial ß-oxidation. However, the transformation of the substrate [U14C] palmitic acid in the ß oxidation pathway was not inhibited by 3,3,3-trifluoropropionic acid. In addition, no cytotoxicity of 3,3,3 trifluoropropionic acid was observed in the cell culture systems. The main effect after a single dose of 3,3,3-trifluoropropionic acid was seen in clinical pathology and metabonomic analysis. The decrease in blood glucose is considered to have the most far-reaching consequences for the toxicity of 3,3,3-trifluoropropionic acid. If considering this change as the primary effect after a single dose, secondary effects, for example, the above-mentioned clinical signs could be explained. In addition, the observed high level of ketone bodies might have been responsible for life-threatening possible ketoacidosis. In general, ketoacidosis occurs after an imbalance between glycolysis, lipolysis, TCA cycle activity and respiratory function. Based on the results, ß-oxidation of fatty acids was not affected, and due to the decrease in glucose levels and the high levels of acetyl CoA, glycolysis was considered not to be impaired. Increased amounts of acetyl CoA might be a result of insufficient activity of the TCA cycle. However, the inhibition of the TCA cycle can be based on the impairment of specific enzymes and/or on the involvement of messenger substrates like insulin. Supporting the first mentioned aspect are decreased levels of TCA cycle intermediates, like \&\#945;-ketoglutarate or citrate, as seen in 1H-NMR spectra of urine. However, the second aspect would explain the drop in blood glucose with the impairment of glucose transporters or the impairment of the insulin balance. If a single dose of 3,3,3-trifluoropropionic acid had stimulated the insulin release, glycolysis would be activated, and high amounts of acetyl CoA would be produced. In case of impaired use by the TCA cycle, levels of ketone bodies would be increased. Experiments were designed to characterize the direct effect of 3,3,3-trifluoropropionic acid on rat insulinoma-derived INS-1 cells as possible increase in insulin release. Further investigations are necessary to answer in which step of the metabolic pathway 3,3,3-trifluoropropionic acid interferes or finally which specific enzyme is inhibited or activated by 3,3,3-trifluoropropionic acid, leading to the drop in blood glucose and finally in lethal toxicity.}, subject = {Fluorkohlenwasserstoffe}, language = {en} } @phdthesis{JimenezPearson2005, author = {Jim{\´e}nez-Pearson, Mar{\´i}a-Antonieta}, title = {Characterization of the mechanisms of two-component signal transduction involved in motility and chemotaxis of Helicobacter pylori}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15698}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Flagellen-basierte Motilit{\"a}t und Chemotaxis stellen essentielle Pathogenit{\"a}tsfaktoren dar, die f{\"u}r die erfolgreiche Kolonisierung der Magenschleimhaut durch H. pylori notwendig sind. Die Mechanismen der Regulation der Flagellensynthese und das Chemotaxis-System von H. pylori weisen trotz einiger {\"A}hnlichkeiten fundamentale Unterschiede zu den Systemen anderer Bakterien auf. In H. pylori ist die Flagellensynthese durch eine komplex regulierte Kaskade kontrolliert, die Regulatorkomponenten wie das Zweikomponentensystem HP244/FlgR, die Sigma Faktoren 54 und 28 und den Sigma Faktor28-Antagonisten FlgM enth{\"a}lt. Das Signal, welches {\"u}ber die Histidinkinase des Zweikomponentensystems HP244/FlgR die Expression der Sigma Faktor54-abh{\"a}ngigen Klasse 2 Flagellengene reguliert, ist bisher noch nicht bekannt. Allerdings konnte mit HP137 ein Protein identifiziert werden, das im „yeast two-hybrid" System sowohl mit der korrespondierenden Kinase HP244 des Flagellenregulators FlgR, als auch mit der Flagellenkomponente FlgE´ interagiert (Rain et al., 2001). In dieser Arbeit wurde eine m{\"o}gliche Rolle von HP137 in einem R{\"u}ckkopplungsmechanismus untersucht, welcher die Aktivit{\"a}t der Histidinkinase in der Flagellenregulation kontrollieren k{\"o}nnte. Obwohl die Deletion des ORF hp137 zu einer unbeweglichen Mutante f{\"u}hrte, legen die erfolglosen Komplementations Experimente, sowie die Beobachtung, dass HP137 in vitro keinen bedeutenden Effekt auf die Aktivit{\"a}t der Histidinkinase HP244 hat nahe, dass HP137 weder in H. pylori noch im nahe verwandten C. jejuni direkt an der Flagellenregulation beteiligt ist. Das Chemotaxis-System von H. pylori unterscheidet sich vom gutuntersuchten Chemotaxis-System der Enterobakterien in einigen Aspekten. Zus{\"a}tzlich zu dem CheY Response Regulator Protein (CheY1) besitzt H. pylori eine weitere CheY-artige Receiver-Dom{\"a}ne (CheY2) welche C-terminal an die Histidinkinase CheA fusioniert ist. Zus{\"a}tzlich finden sich im Genom von H. pylori Gene, die f{\"u}r drei CheV Proteine kodieren die aus einer N-terminalen Dom{\"a}ne {\"a}hnlich CheW und einer C-terminalen Receiver Dom{\"a}ne bestehen, w{\"a}hrend man keine Orthologen zu den Genen cheB, cheR, and cheZ findet. Um einen Einblick in den Mechanismus zu erhalten, welcher die chemotaktische Reaktion von H. pylori kontrolliert, wurden Phosphotransferreaktionen zwischen den gereinigten Signalmodulen des Zweikomponentensystems in vitro untersucht. Durch in vitro-Phosphorylierungsexperimente wurde eine ATP-abh{\"a}ngige Autophosphorylierung der bifunktionellen Histidinkinase CheAY2 und von CheA´, welches ein verk{\"u}rztes Derivat von ChAY2 ohne Receiver-Dom{\"a}ne darstellt, nachgewiesen. CheA´ zeigt eine f{\"u}r an der Chemotaxis beteiligte Histidinkinasen typische Phosphorylierungskinetik mit einer ausgepr{\"a}gten exponentiellen Phase, w{\"a}hrend die Phosphorylierungskinetik von CheAY2 nur eine kurze exponentielle Phase aufweist, gefolgt von einer Phase in der die Hydrolyse von CheAY2~P {\"u}berwiegt. Es wurde gezeigt, dass die Anwesenheit einer der CheY2 Dom{\"a}ne die Stabilit{\"a}t der phosphorylierten P1 Dom{\"a}ne im CheA Teil des bifunktionellen Proteins beeinflusst. Außerdem wurde gezeigt, dass sowohl CheY1 als auch CheY2 durch CheAY2 phosphoryliert werden und dass die drei CheV Proteine die Histidinkinase CheA´~P dephosphorylieren, wenn auch mit einer im Vergleich zu CheY1 und CheY2 geringeren Affinit{\"a}t. Außerdem ist CheA´ in der Lage seine Phosphatgruppen auf CheY1 aus C. jejuni und CheY aus E. coli zu {\"u}bertragen. Retrophosphorylierungsexperimente weisen darauf hin, dass CheY1~P die Phosphatgruppe zur{\"u}ck auf die Histidinkinase CheAY2 {\"u}bertragen kann und dass die CheY2-Dom{\"a}ne in dem bifunktionellen Protein CheAY2 als „Phosphat Sink" agiert der den Phosphorylierungszustand und damit die Aktivit{\"a}t des frei diffundierbaren Proteins CheY1 reguliert, das vermutlich es mit dem Flagellenmotor interagiert. Es konnte weiterhin gezeigt werden, dass die unabh{\"a}ngige Funktion der beiden Dom{\"a}nen CheA´ und CheY2 f{\"u}r eine normale chemotaktische Signalgebung in vivo nicht ausreicht. In dieser Arbeit wurden also Hinweise auf eine komplexe Kaskade Phosphat{\"u}bertragungsreaktionen im chemotaktischen System von H. pylori gefunden, welches {\"A}hnlichkeiten zu dem Syteme-Chemotaxis von S. meliloti aufweist an denen multiple CheY Proteine beteiligt sind. Die Rolle der CheV Proteine bleibt im Moment unklar, jedoch k{\"o}nnte es sein, dass sie an einer weiteren Feinregulierung der Phosphatgruppen{\"u}bertragungsreaktionen in diesem komplexen chemotaktischen System beteiligt sind}, subject = {Helicobacter pylori}, language = {en} } @phdthesis{Dietzek2005, author = {Dietzek, Benjamin}, title = {Ultrafast linear and non-linear spectroscopy : from biological light-receptors to artificial light-harvesting systems}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15684}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {In the experiments presented in this work, linear and non-linear femtosecond time-resolved spectrsocopy were applied to investigate the structure-function and functiondynamics relationship in biological and artificially designed systems. The experiments presented in this work utilize femtosecond time-resolved transient absorption and transient grating as well as picosecond time-resolved fluorescence spectroscopy to investigate the photophysics and photochemistry of biological photoreceptors and address the light-induced excited-state processes in a particular molecular device that serves as a - structurally - very simple light-harvesting antenna and potentially as a catalysis-switch for the production of hydrogen in solution. The combination of white-light probe transient absorption and coherent transient grating spectroscopies yields spectral information about the excited state absorption in concert with high quality, high signal-to-noise kinetic transients, which allow for precise fitting and therefore very accurate time-constants to be extracted from the data. The use of femtosecond time-resolved transient grating spectroscopy is relatively uncommon in addressing questions concerning the excited-state reaction pathways of complex (biological) systems, and therefore the experiments presented in this work constitute according to the literature the first studies applying this technique to a a metalloporphyrin and an artificial light-harvesting antenna.}, subject = {Femtosekundenspektroskopie}, language = {en} } @phdthesis{Daumer2005, author = {Daumer, Volker}, title = {Phase coherent transport phenomena in HgTe quantum well structures}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15538}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Although spintronics has aroused increasing interest, much fundamental research has to be done. One important issue is the control over the electronic spin. Therefore, spin and phase coherent transport are very important phenomena. This thesis describes experiments with mercury based quantum well structures. This narrow gap material provides a very good template to study spin related effects. It exhibits large Zeeman spin splitting and Rashba spin-orbit splitting. The latter is at least four to five times larger than in III-V semiconductors. Initially a short review on the transport theory was presented. The main focus as on quantisation effects that are important to understand the related experiments. Thus, Shubnikov-de Haas and the quantum Hall effect have been analysed. Due to the first fabrication of nanostructures on Hg-based quantum well samples, the observation of ballistic transport effects could be expected. Hence, the Landauer-B¨uttiker theory has been introduced which gives the theoretical background to understand such effects. With respect to the main topic of this thesis, phase coherence has been introduced in detail. Experiments, where coherence effects could be observed, have been explained theoretically. Here, possible measurement setups have been discussed, e.g., a ring shaped structure to investigate the Aharonov-Bohm and related effects. Due to the fact, that all experiments, described in this thesis, were performed on Hg-based samples, the exceptional position of such samples among the \&\#147;classical\&\#148; semiconductors has been clarified. Hg1-xMnx Te quantum wells are type-III QWs in contrast to the type-I QWs formed by e.g., GaAs/AlGaAs heterostructures. With a well width of more than 6 nm and a manganese content of less than 7\% they exhibit an inverted band alignment. Band structure calculations based on self consistent Hartree calculations have been presented. The common description of a diluted magnetic semiconductor with the Brillouin function has been introduced and the experiments to obtain the empiric parameters T0 and S0 have been presented. Rashba spin-orbit splitting and giant Zeeman splitting have been explained theoretically and the magnetic ordering of a spin glass as well as the relevant interactions therein have been discussed. The next chapter describes the first realisation of nanostructures on Hg-based heterostructures. Several material specific problems have been solved, but the unique features of this material system mentioned above justify the effort. Interesting new insight could be found and will be found with these structures. Onto a series of QW samples, cross-shaped structures with several lead widths have been patterned. With the non-local resistance measurement setup, evidence for quasiballistic transport was demonstrated in cross-shaped structures with lead widths down to 0.45 mm. The non-local bend resistance and a regime of rebound trajectories as well as the anomalous Hall effect could be identified. Monte-Carlo simulations of the classical electron trajectories have been performed. A good agreement with the experimental data has been achieved by taking a random scattering process into account. Encouraged by this success the technology has been improved and ring-shaped structures with radii down to 1 mm have been fabricated. Low temperature (below 100 mK), four terminal resistance measurements exhibit clear Aharonov-Bohm oscillations. The period of the oscillations agrees very well with a calculation that takes only the sample geometry into account. One goal using such a structure is the experimental prove of the spin-orbit Berry phase. Therefore an additional Shottky gate on top of the ring was needed. With this structure evidence for the Aharonov-Casher effect was observed. Here, a perpendicular applied electric field causes analogous oscillations as does the magnetic field in the AB effect. A subsequent change in the Rashba SO splitting due to several applied gate voltages while measuring the AB effect should reveal the SO Berry phase. Although initially evidence of a phase change was detected, a clear proof for the direct measurement of the SO Berry phase could not be found. In the future, with an advanced sample structure, e.g., with an additional Hall bar next to the ring, which permits a synchronous measurement of the Rashba splitting, it might be possible to measure the SO Berry phase directly. In manganese doped HgTe QWs two different effects simultaneously cause spin splitting: the giant Zeeman and the Rashba effect. By analysing the Shubnikovde Haas oscillations and the node positions of their beating pattern, it has been possible to separate these two effects. Whereas the Rashba effect can be identified by its dependence on the structure inversion asymmetry, varied by the applied gate voltage, the giant Zeeman splitting is extracted from its strong temperature dependence, because Rashba splitting is temperature independent. The analysis revealed, that the Rashba splitting is larger than or comparable to the giant Zeeman splitting even at moderately high magnetic fields. In an extraordinary HgMnTe QW sample, that exhibits the n= 1 quantum Hall plateau from less than 1 T up to 28 T, the anomalous Hall effect could be excluded. Intense studies on the temperature dependence of the QHE as well as band structure calculations have revealed this extraordinary behaviour to be an ordinary band structure effect of this system. In a series of mesoscopic structures on nonmagnetic and magnetic QWs, an investigation of the universal conductance uctuations have been carried out. In the}, subject = {Quecksilbertellurid}, language = {en} } @phdthesis{Masek2005, author = {Masek, Pavel}, title = {Odor intensity learning in Drosophila}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15546}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {It has been known for a long time that Drosophila can learn to discriminate not only between different odorants but also between different concentrations of the same odor. Olfactory associative learning has been described as a pairing between odorant and electric shock and since then, most of the experiments conducted in this respect have largely neglected the dual properties of odors: quality and intensity. For odorant-coupled short-term memory, a biochemical model has been proposed that mainly relies on the known cAMP signaling pathway. Mushroom bodies (MB) have been shown to be necessary and sufficient for this type of memory, and the MB-model of odor learning and short-term memory was established. Yet, theoretically, based on the MB-model, flies should not be able to learn concentrations if trained to the lower of the two concentrations in the test. In this thesis, I investigate the role of concentration-dependent learning, establishment of a concentration-dependent memory and their correlation to the standard two-odor learning as described by the MB-model. In order to highlight the difference between learning of quality and learning of intensity of the same odor I have tried to characterize the nature of the stimulus that is actually learned by the flies, leading to the conclusion that during the training flies learn all possible cues that are presented at the time. The type of the following test seems to govern the usage of the information available. This revealed a distinction between what flies learned and what is actually measured. Furthermore, I have shown that learning of concentration is associative and that it is symmetrical between high and low concentrations. I have also shown how the subjective quality perception of an odor changes with changing intensity, suggesting that one odor can have more than one scent. There is no proof that flies perceive a range of concentrations of one odorant as one (odor) quality. Flies display a certain level of concentration invariance that is limited and related to the particular concentration. Learning of concentration is relevant only to a limited range of concentrations within the boundaries of concentration invariance. Moreover, under certain conditions, two chemically distinct odorants could smell sufficiently similarly such, that they can be generalized between each other like if they would be of the same quality. Therefore, the abilities of the fly to identify the difference in quality or in intensity of the stimuli need to be distinguished. The way how the stimulus is analyzed and processed speaks in favor of a concept postulating the existence of two separated memories. To follow this concept, I have proposed a new form of memory called odor intensity memory (OIM), characterized it and compared it to other olfactory memories. OIM is independent of some members of the known cAMP signaling pathway and very likely forms the rutabaga-independent component of the standard two-odor memory. The rutabaga-dependent odor memory requires qualitatively different olfactory stimuli. OIM is revealed within the limits of concentration invariance where the memory test gives only sub-optimal performance for the concentration differences but discrimination of odor quality is not possible at all. Based on the available experimental tools, OIM seems to require the mushroom bodies the same as odor-quality memory but its properties are different. Flies can memorize the quality of several odorants at a given time but a newly formed memory of one odor interferes with the OIM stored before. In addition, the OIM lasts only 1 to 3 hours - much shorter than the odor-quality memory.}, subject = {Taufliege}, language = {en} } @phdthesis{Motsch2005, author = {Motsch, Isabell}, title = {Lamin A and lamin C are differentially dysfunctional in autosomal dominant Emery-Dreifuss muscular dystrophy}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-15360}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Emery-Dreifuss muscular dystrophy (EDMD) is a rare genetic disorder characterised by early contractures of the elbows, Achilles tendons and spine, slowly progressive muscle wasting and cardiomyopathy associated with cardiac conduction defect. The autosomal dominant form is caused by mutations in the LMNA gene which gives rise to lamin A and lamin C proteins by alternative splicing. These A-type lamins, together with B-type lamins, form the nuclear lamina, a network of intermediate filament proteins underlining the nuclear envelope. In order to ascertain the role lamin A and C separately contribute to the molecular phenotype, we analysed ten LMNA mutations and one single nucleotide polymorphism (SNP) in transfection studies in COS7 fibroblasts and, partially, in C2C12 myoblasts. The EGFP or DsRed2 tagged lamins were exogenously expressed either individually or both A-types together and examined by light and electron microscopy. The protein mobility of lamin A mutants was determined by FRAP analysis. Additionally, a co-immunoprecipitation binding assay of in vitro synthesised A-type lamins and emerin was performed.Eight of the LMNA mutations (R50S, R133P, E358K, E358K+C}, language = {en} } @phdthesis{Pietsch2005, author = {Pietsch, Christof}, title = {The genetics of species differences within the genus Nasonia ASHMEAD 1904 (Hymenoptera: Pteromalidae)}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-14348}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {The genetics of species differences is an outstanding question in evolutionary biology. How do species evolve to become phenotypically distinct and how is the genetic architecture organized that underlie species differences? Phenotypic diverged traits are supposed to be frequently involved in prezygotic isolation, i.e. they prevent the formation of hybrids, whereas postzygotic isolation occurs when hybrids experience a fitness reduction. The parasitic wasp genus Nasonia represents an appropriate model system to investigate the genetics of species differences as well as the genetics of postzygotic isolation. The genus consists of three species N. vitripennis, N. longicornis and N. giraulti that differ particularly in male traits that are assumed to posses an adaptive significance: courtship behaviour and wing size differences. The courtship behaviour consists of cyclically repeated series of head nods that are separated by pauses. The stereotypic performance allowed to split up the display into distinct courtship components. Males of N. vitripennis bear vestigial forewings and are incapable of flight, whereas N. longicornis wear intermediate sized wings and N. giraulti is fully capable of flying. Nasonia species can produce interspecific hybrids after removing Wolbachia bacteria induced hybrid incompatibilities with antibiotics. Postzygotic isolation occurs to different extent and is asymmetric among reciprocal crosses, e.g. inviability is stronger in the N. vitripennis (\&\#9792;) x N. longicornis (\&\#9794;) cross than in the N. longicornis (\&\#9792;) x N. vitripennis (\&\#9794;) cross. The formation of hybrids allow to study the genetic of species differences in QTL (quantitative trait locus) analyses as well as the genetics of postzygotic isolation causing hybrid inviability. The aim of the study was to investigate the genetic architecture of differences in courtship behaviour and wing size between N. vitripennis and N. longicornis and to assess the genetics of postzygotic isolation to gain clues about the evolutionary processes underlying trait divergence and establishment of reproductive isolation between taxa. In a QTL analysis based on 94 F2-hybrid individuals of an LV cross only few QTL for wing size differences have been found with relatively large effects, although a large proportion of the phenotypic variance remained unexplained. The QTL on courtship behaviour analysis based on 94-F2 hybrid males revealed a complex genetic architecture of courtship behaviour with QTL of large phenotypic effects that explained more than 40 \% of the phenotypic variance in one case. Additionally, an epistatic analysis (non-additive interlocus interaction) of courtship QTL revealed frequent genetic interchromsomal relations leading in some instances to hybrid specific effects, e.g. reversion of phenotypic effects or the transgression of phenotypes. A QTL analysis based on a threefold sample size revealed, however, an overestimation of QTL effects in the analysis based on smaller sample size pointing towards a genetic architecture of many loci with small effects governing the phenotypic differences in courtship behaviour. Furthermore, the the study comprised the analysis of postzygotic isolation in the reciprocal crosses N. vitripennis (\&\#9792;) x N. longicornis (\&\#9794;) versus N. longicornis (\&\#9792;) x N. vitripennis (\&\#9794;) located several loci distributed over different chromosomes that are involved in hybrid incompatibility. The mapping of hybrid incompatibility regions reproduced for the first time the observed asymmetries in the strength of postzygotic isolation in reciprocal crosses of between the more distant related taxa within the genus Nasonia. Stronger postzygotic incompatibilities in the VL cross are supposed to result from the superposition of nuclear-nuclear incompatibilities with nuclear-cytoplasmic incompatibilities, whereas the coincidences of these to types of incompatibilities were found to be much weaker in the reciprocal LV cross.}, subject = {Pteromalidae}, language = {en} } @phdthesis{Rabie2005, author = {Rabie, Tamer}, title = {Cellular regulation of platelet glycoprotein VI : in vivo and in vitro studies in mice}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-14267}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Platelet interaction with the subendothelium is essential to limit blood loss after tissue injury. However, upon rupture of atherosclerotic plaques, this interaction may result in blood vessel occlusion leading to life threatening diseases such as myocardial infarction or stroke. Among the subendothelial matrix proteins, collagen is considered to be the most thrombogenic component as it directly activates platelets. Platelets interact with collagen, either indirectly through glycoprotein (GP) Ib-V-IX receptor complex, or directly through the major collagen receptor on the platelet surface, GPVI. The work presented here focused on studying the cellular regulation of GPVI. In addition, a possible role for GPVI in thrombus formation induced by atherosclerotic plaque material was investigated and it was found that GPVI plays an important role in this process. Using a recently published mitochondrial injury model, it was found that GPVI contains a cleavage site for a platelet-expressed metalloproteinase. Further studies showed that platelet activation by CRP, or thrombin induced down-regulation of GPIb\&\#61537;, but not GPVI. In parallel, cellular regulation of GPV was studied and it was found that GPV is cleaved in vitro by the metalloproteinase ADAM17. In previous studies it was shown that injection of mice with the anti-GPVI mAb, JAQ1, induces GPVI down-regulation, which is associated with a strong, but transient, thrombocytopenia. Using new anti-GPVI mAbs, which bind different epitopes on the receptor, it is shown in this study that GPVI down-regulation occurs in an epitope-independent manner. Further experiments showed that antibody treatment induces a transient, but significant increase in bleeding time. Using different genetically modified mice, it is shown that, upon antibody injection, GPVI is both, shed from the platelet surface and internalized into the platelet. Signaling through the immunoreceptor tyrosine-based activation motif (ITAM) of the FcR\&\#61543; chain is essential for both processes, while LAT and PLC\&\#61543;2 are essential for the shedding process only. Antibody-induced increase in bleeding time and thrombocytopenia were absent in LAT deficient mice, showing that it is possible to uncouple the associated side effects from the down-regulation process. As antibody-induced GPVI internalization still occurs in LAT and PLC\&\#61543;2 deficient mice, this suggests a novel signaling pathway downstream of GPVI that has not been described so far.}, subject = {Maus}, language = {en} } @phdthesis{Hipondoka2005, author = {Hipondoka, Martin H.T.}, title = {The development and evolution of Etosha Pan, Namibia}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-14351}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {This study explores and examines the geomorphology of a large endorheic basin, approximately twice the size of Luxemburg, situated in the Etosha National Park, Namibia. The main focus is directed on how and when this depression, known as Etosha Pan, came into being. Geomorphological investigation was complemented and guided primarily by the application and interpretation of satellite-derived information. Etosha Pan has attracted scientific investigations for nearly a century. Unfortunately, their efforts resulted into two diverging and mutually exclusive views with respect to its development. The first and oldest view dates back to the 1920s. It hypothesized Etosha Pan as a desiccated palaeolake which was abandoned following the river capture of its major fluvial system, the Kunene River. The river capture was assumed to have taken place in the Pliocene/Early Pleistocene. In spite of the absence of fluvial input that the Kunene contributed, the original lake was thought to have persisted until some 35 ka ago, long after the Kunene severed its ties with the basin. The current size of the basin and its playa status was interpreted to have resulted from deteriorating climatic conditions. The opposing view emerged in the 1980s and gained prominence in the 1990s. This view assumed that there were an innumerable number of small pans on the then surface of what later to become Etosha Pan. Since the turn of the Pliocene to Early Pleistocene, these individual pans started to experience a combined effect of fluvial erosion during the rainy season and wind deflation during the dry period. The climatic regime during that entire period was postulated to be semi-arid as today. This climatic status was used to rule out any existence of a perennial lake within the boundary of Etosha since the Quaternary. Ultimately, these denudational processes, taking place in a seasonal rhythm, caused the individual pans to deepen and widen laterally into each other and formed a super-pan that we call Etosha today. Thus the Kunene River had no role to play in the development of the Etosha Pan according to this model. However, proponents of this model acknowledged that the Kunene once fed into the Owambo Basin and assigned the end of the Tertiary to the terminal phase of that inflow. Findings of this study included field evidence endorsing the postulation that the Kunene River had once flowed into the Owambo Basin. Its infilled valley, bounding with the contemporary valley of the Kunene near Calueque, was identified and points towards the Etosha Pan. It is deliberated that a large lake, called Lake Kunene, existed in the basin during the time. Following the deflection of the Kunene River to the coast under the influence of river incision and neo-tectonic during the Late Pliocene, new dynamics were introduced over the Owambo Basin surface. After the basin was deprived of its major water and sediment budget that the Kunene River contributed, it was left with only smaller rivers, most notably the Cuvelai System, as the only remaining supplier. This resulted in the Cuvelai System concentrating and limiting its collective load deposition to a lobe of Lake Kunene basin floor. The accident of that lobe is unclear, but it is likely that it constituted the deepest part of the basin at the time or it was influenced by neo-tectonic that helped divert the Kunene River or both. Against the backdrop of fluvial action that was initiating the new lake, most parts of the rest of the basin, then denied of lacustrine activity, were intermittently riddled with a veneer of sediment, especially during phases of intensified aeolian activity. In the mean time, the area that was regularly receiving fluvial input started to shape up as a distinct lake with the depositions of sediments around the water-body, primarily via littoral action, serving as embankment. Gradually, a shoreline is formed and assisted in fixing and delineating the spatial extent of the new and much smaller lake, called Lake Etosha. That Lake Etosha is the predecessor of the modern day Etosha Pan. Indicators for a perennial lake found in this study at Etosha include fossil fragments of Clariidae species comparable to modern species measuring some 90 cm, and those of sitatunga dated to approximately 5 ka. None of these creatures exist today at Etosha because of their ecological requirements, which among others, include permanent water. The sitatunga, in addition, is known as the only truly amphibious antelope in the world. Since its inception, the new lake underwent a number of geomorphological modifications. A prominent character amongst these modifications is the orientation of the lake, which has its long-axis oriented in the ENE-WSW direction. It resulted from wave action affected by the prevailing dominant northeasterly wind, which is believed to have been in force since the Middle Pleistocene. Lake Etosha has also witnessed phases of waning and waxing under the influence of the prevailing climatic regime. Over the last 150 ka, the available data intercepted about seven phases of high lake levels. These data are generally in agreement with regional palaeoclimatic data, particularly when compared with those obtained from neighbouring Makgadikgadi Pans in Botswana. The last recorded episode of the wet phase at Etosha was some 2,400 years before the present.}, subject = {Etoschapfanne}, language = {en} } @phdthesis{Strehl2005, author = {Strehl, Christoph-Peter}, title = {Evolution of colony characteristics in the harvester ant genus Pogonomyrmex}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-14324}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Die Gattung Pogonomyrmex ist besonders gut geeignet, um die Evolution der Charakteristika von Ameisenkolonien allgemein zu untersuchen, und insbesondere deren soziogenetische Struktur, da die Biologie f{\"u}r einige ihrer Arten sehr gut bekannt ist, und eine Diversit{\"a}t an Paarungsh{\"a}ufigkeiten und K{\"o}nniginnenzahlen vorkommt. Diese Variation in der soziogenetischen Struktur der Kolonien erzeugt eine hohe Varianz an Verwandschaftsgraden innerhalb von Kolonien, und kann eine Hauptkomponente darstellen, welche die Evolution verschiedenster Koloniecharakteristika vorantreibt. Um die Variabilit{\"a}t intrakolonialer Verwandschaftsgrade innerhalb der Gattung Pogonomyrmex genau zu bestimmen, wurde f{\"u}r ausgew{\"a}hlte Mitglieder der Gattung, n{\"a}mlich f{\"u}r P. (sensu stricto) rugosus, P. (sensu stricto) badius and P. (Ephebomyrmex) pima, mit Hilfe der Technik des DNA-Fingerabdruckes die Anzahl an Matrilinien und Patrilinien bestimmt. Es wurde versucht die Evolution dieser Koloniecharakteristika vor dem Hintergrund einer Phylogenie zu erkl{\"a}ren. Zu diesem Zweck wurde ein Gen-Stammbaum f{\"u}r 39 Arten der Gattung Pogonomyrmex erstellt. Die Artabdeckung betrug 83\% bei den Nord-Amerikanischen, und 43\% bei den S{\"u}d-Amerikanischen Arten. Effektive Mehrfachpaarung von K{\"o}niginnen wurde f{\"u}r P. rugosus (me=4.1) und P. badius (me=6.7) best{\"a}tigt. Zus{\"a}tzlich wurde gezeigt, dass beide Arten monogyn sind. Diese Ergebnisse best{\"a}tigen Verhaltensbeobachtungen von Mehrfachpaarungen in diesen Arten. Mittlerweile ist Mehrfachpaarung in 9 Pogonomyrmex Arten bekannt (bei 3 Arten durch Verhaltensbeobachtungen - bei 6 Arten durch genetischen Nachweis). In P. (E.) pima hingegen waren alle der untersuchten K{\"o}niginnen einfach gepaart (me=1.0). Daher k{\"o}nnte es sein, dass Mehrfachpaarung entweder fr{\"u}h in der Evolution der Gattung Pogonomyrmex entstand und nachtr{\"a}glich in der Untergattung Ephebomyrmex verloren wurde (Plesiomorphie-Hypothese), oder sie entstand zum ersten mal in der Untergattung Pogonomyrmex sensu stricto (Apomorphie-Hypothese). In P. huachucanus, einer Art, die basal zu dem Nord-Amerikanischen sensu stricto Komplex ist, k{\"o}nnten die im Vergleich zu ihren sensu stricto Verwandten geringeren effektiven Paarungsh{\"a}ufigkeiten der K{\"o}niginnen (J. Gadau and C.-P. Strehl, unver{\"o}ffentlicht) einen Wechsel von Monandrie zu Polyandrie im Verlauf der Entstehung der fortschrittlicheren sensu stricto Arten widerspiegeln, was die Apomorphie-Hypothese unterst{\"u}tzen w{\"u}rde. Die intrakolonialen Verwandtschaftsgrade sind dennoch in P. (E.) pima niedrig. Dies ist m{\"o}glicherweise auf mehrere reproduktive K{\"o}niginnen (Polygynie) zur{\"u}ckzuf{\"u}hren. Polygynie ist ebenfalls dokumentiert f{\"u}r mindestens vier weitere Arten der Untergattung Ephebomyrmex, mit genetischer Evidenz allerdings bisher nur f{\"u}r P. (E.) pima. Es k{\"o}nnte sein, dass es einen evolution{\"a}ren Ausgleich (trade-off) zwischen Polyandrie und Polygynie innerhalb der Untergattung Ephebomyrmex gab, und daher beide Untergattungen eine hohe genetische Vielfalt innerhalb der Kolonien behielten. Diese hohe genetische Vielfalt k{\"o}nnte einer der Gr{\"u}nde sein f{\"u}r den Erfolg und die Radiation der Gattung Pogonomyrmex in Trockengebieten. Evolution k{\"o}nnte eine hohe genetische Vielfalt von Pogonomyrmex Kolonien beg{\"u}nstigt haben, da sie den Kolonien hilft die Organisation der Kolonie und die Effizienz mit der externe Aufgaben ausgef{\"u}hrt werden zu verbessern. Wenigstens in P. badius konnte eine Verkn{\"u}pfung zwischen Patrilinien und physischem Polyethismus gefunden werden, was auf eine Verbesserung der Kolonieorganisation mit Hilfe von Polyandrie hindeutet. Dar{\"u}ber hinaus k{\"o}nnten die dargelegten extremen Polyandrie- Werte den P. badius-Weibchen helfen die M{\"o}glichkeit der Inzucht aufgrund eingeschr{\"a}nkter Ausbreitung zu bew{\"a}ltigen. Eine eingeschr{\"a}nkte Ausbreitung wird auch in P. (E.) pima durch fl{\"u}gellose, intermorphe K{\"o}niginnen beobachtet. Jedoch wird bei P. (E.) pima die Inzucht durch Auskreuzen mittels M{\"a}nnchen m{\"o}glicherweise verhindert, da keine signifikante Inzucht gefunden wurde. In den vorliegenden Gen-Stammb{\"a}umen war die Untergattung Pogonomyrmex Ephebomyrmex von der Untergattung Pogonomyrmex sensu stricto getrennt. Daher k{\"o}nnte es sein, dass P. Ephebomyrmex in den Status einer Gattung erhoben wird, auch aufgrund distinkter morphologischer und lebensgeschichtlicher Charaktere. F{\"u}r eine pr{\"a}zise taxonomische Revision m{\"u}sste allerdings eine breite Erg{\"a}nzung an Arten vorgenommen werden. Es wurde in P. rugosus Kolonien normalerweise eine geringe Anzahl von unverwandten Arbeiterinnen vorgefunden, die m{\"o}glicherweise aus Brutraub ausgewachsener Kolonien auf G{\"u}ndungs-Kolonien stammen. Es ist allseits bekannt, dass die meisten Gr{\"u}ndungskolonien von benachbarten, ausgewachsenen Kolonien der eigenen Art zerst{\"o}rt werden, aber es wurde bisher angenommen, dass die Brut dieser Kolonien ebenfalls zerst{\"o}rt wurde. Dieser oft vernachl{\"a}ssigte Aspekt k{\"o}nnte einen wichtigen St{\"a}rke-Bonus f{\"u}r ausgewachsene Kolonien darstellen.}, subject = {Pogonomyrmex}, language = {en} } @phdthesis{AlOuran2005, author = {Al-Ouran, Nedal}, title = {Environmental assessment, documentation and spatial modeling of heavy metal pollution along the Jordan Gulf of Aqaba using coral reefs as environmental indicator}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-14090}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Two phases of reef sampling were carried out. The first included regular samples taken along the coastline of Aqaba (27km long) at depths of 4-15m, and used to determine spatial distribution of pollution. The second phase included three 20cm-deep cores obtained from within the industrial zone. These cores were drilled from pre-dated communities, where the growth rate was determined earlier to be 10mm y-1, therefore the core obtained represented a period of 20 years (i.e. 1980-2000). The cores were used to reconstruct the metal pollution history at the most heavily used site along the coast (industrial zone).All samples were examined with respect to their metal content of Cd, Pb, Cu, Zn, Ni, and Cr. Almost all of them have shown records above the calculated background values. Mean values of Cd, Pb, Cu, Zn, Ni and Cr recorded along the coast were 1,25; 4,26; 9,76; 11,40; 2,29 and 10,522, µg g-1 respectively, and for core samples 1.4; 4.2; 5.7; 6.4; 2.3 and 8.21 µg g-1 respectively. Spatial distribution of metal enrichment in reef samples have shown a general and clear increasing trend towards the south. Same increasing trend was also in core samples where the six metals have shown a prominent increasing trend towards the core surface indicating an increase of coastal activities during the last twenty years. High and relatively high values were recorded at the oil port, the industrial area and main port, and thus categorized as highly impacted areas. Intermediate metal content were recorded in samples of the north beach, and thus classified as being relatively impacted, where the lowest metal concentrations were observed at the marine reserve, the least impacted site along the coast. The high enrichment of metal is attributed mainly to anthropogenic impacts. The natural inputs of the six metals studied in the Gulf of Aqaba are generally very low, due to the geographic positions and the absence of wadi discharge and as a result of low rainfall. Several potential sources of heavy metals were investigated. The industrial-related activities, port operations and phosphate dust were among the main sources currently threatening the marine ecosystem in Aqaba. Applying the Principle Components Analysis method (PCA) to all samples taken along the coastline has resulted in categorizing three different groups according to their metal enrichment, the first is composed of samples taken from the north beach and the main port with intermediate to high enrichment, the second joined the samples of the marine park and the marine reserve with low and relatively low enrichment, and the last group joined samples of the industrial zone and the oil port with high enrichment. The Principle Component Scores were also utilized to confirm the spatial distribution and relationships of the examined heavy metals along the coast. Two models (interpolated by SURFER \&\#61666; 7.0 and ArcView\&\#61666; 3.2a) were developed, the first was based on the PC scores of the first component, and shows clearly the positive anomalies in metal concentrations along the coast. The second model was developed by plotting the second factor scores on a landuse map of Aqaba. According to these models, it has shown that the positive anomalies are associated with three different zones; industrial area, the main port and the oil port. The results have shown that coral reefs can be used as good environmental indicator for assessments and monitoring processes, and they can provide data and information on both the spatial distribution of pollution and their history. The present work is the first to document the environmental status along the whole coast of Aqaba and the first to use coral reef as a tool/ indicator.}, subject = {Golf von Akaba}, language = {en} } @phdthesis{Huenten2005, author = {H{\"u}nten, Peter}, title = {Channel-forming proteins in the cell wall of amino acid-producing Corynebacteria}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-13890}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2005}, abstract = {Corynebacterium glutamicum is together with C. callunae and C. efficiens a member of the diverse group of mycolic-acid containing actinomycetes, the mycolata. These bacteria are potent producer of glutamate, lysine and other amino acids on industrial scale. The cell walls of most actinomycetes contain besides an arabinogalactan-peptidoglycan complex large amounts of mycolic acids. This three-layer envelope is called MAP (mycolyl-arabinogalactan-peptidoglycan) complex and it represents a second permeability barrier beside the cytoplasmic membrane similar to the outer membrane of Gram-negative bacteria. In analogy to the situation in the outer membrane of Gram-negative bacteria, channels are present in the mycolic acid layer of the mycobacterial cell wall for the passage of hydrophilic solutes. Molecular studies have provided far-reaching findings on the amino acid flux and its balance in C. glutamicum in general, but the L-glutamate export still remains unknown. The properties of the outer layers, typical of mycolata, seem to be of major importance in this process, and diffusion seems to play a key role for this part of the cell wall. The major aim of this thesis was to identify and study novel channel-forming proteins of the amino acid producers C. glutamicum, C. callunae and C. efficiens. Cell wall extracts of the organisms were investigated and a novel pore-forming protein, named PorH, that is homologue in all three organisms, was detected and characterized. PorHC.glut was isolated from C. glutamicum cells cultivated in minimal medium. The protein was identified in lipid bilayer experiments and purified to homogeneity by fast-protein liquid chromatography across a HiTrap-Q column. The purified protein forms cation-selective channels with a diameter of about 2.2 nm and an average single-channel conductance of about 2.5 nS in 1 M KCl in the lipid bilayer assay. Organic solvent extracts were used to study the permeability properties of the cell wall of C. callunae and C.efficiens. The cell extracts contained channel-forming activity, the corresponding proteins were purified to homogeneity by fast-protein liquid chromatography across a HiTrap-Q column and named PorHC.call and PorHC.eff. Channels formed by PorHC.call are cation-selective with a diameter of about 2.2 nm and an average single-channel conductance of 3 nS, whereas PorHC.eff forms slightly anion selective channels with an average single-channel conductance of 2.3 nS in 1 M KCl in the lipid bilayer assay. The PorH proteins were partially sequenced and the corresponding genes, which were designated as porH, were identified in the published genome sequence of C. glutamicum and C. efficiens. The chromosome of C. callunae is not sequenced, but PorHC.call shows a high homology to PorHC.eff and PorHC.glut. The proteins have no N-terminal extension, only the inducer methionine, which suggests that secretion of the proteins could be very similar to that of PorAC.glut of C. glutamicum. PorHC.glut is coded in the bacterial chromosome by a gene that is localized in the vincinity of the porAC.glut gene, within a putative operon formed by 13 genes that are encoded by the minus strand. Both porins are cotranscribed and coexist in the cell wall, which was demonstrated in RT-PCR and immunological detection experiments. The arrangement of porHC.glut and porAC.glut on the chromosome is similar to that of porBC.glut and porCC.glut and it was found that PorAC.glut, PorHC.glut, PorBC.glut and PorCC.glut coexist in the cell wall of C. glutamicum. The molecular mass of about 6 kDa of the PorH channel forming proteins is rather small and suggests that the cell wall channels are formed by oligomers. A possibly hexameric form was demonstrated for PorHC.glut in Western blot analysis with anti- PorHC.glut antibodies. Secondary structure predictions for PorHC.glut, PorHC.call and PorHC.eff predict that a stretch of about 42 amino acids of PorHC.glut and 28 amino acids of PorHC.call and PorHC.eff forms amphipathic \&\#61537;-helices with a total length of 6.3 nm and 4.2 nm respectively. This should be sufficient to cross the mycolic acid layer. Another objective of this work was to establish an heterologous expression system for corynebacterial channel-forming proteins, to investigate the channel-forming properties of the up to now only hypothetical porins PorA, PorB, PorC from C. efficiens and PorC from C. glutamicum. We could demonstrate with recombinant expression experiments in E. coli that porBC.eff and porCC.eff encode for channel-forming proteins. They are, like PorBC.glut, anion-selective with a similar single-channel conductance of 1 nS in 1 M KCl.}, subject = {Corynebacterium callunae}, language = {en} }