<p>Für das Verständnis der Strukturbildung bei Proteinen ist es wichtig, allgemein geltende Prinzipien der Stabilität und Faltung zu verstehen. Bisher wurde viel Arbeit in die Erörterung von Gesetzmäßigkeiten zu den Faltungseigenschaften von globulären Proteinen investiert. Die große Proteinklasse der solenoiden Proteine, zu denen z. B. Leucine-Rich Repeat- (LRR-) oder Ankyrin-Proteine gehören, wurde dahingegen noch wenig untersucht. Die Proteine dieser Klasse sind durch einen stapelförmigen Aufbau von sich wiederholenden typischen Sequenzeinheiten gekennzeichnet, was in der Ausbildung einer elongierten Tertiärstruktur resultiert. In der vorliegenden Arbeit sollte versucht werden, die Stabilität und Faltung eines LRR-Proteins mittels verschiedener biophysikalischer Methoden zu charakterisieren. Als Untersuchungsobjekt diente die für die Infektion ausreichende zentrale LRR-Domäne des Invasionsproteins Internalin B (InlB₂₄₁) des Bakteriums <i>Listeria monocytogenes</i>. Des weiteren sollten die Integrität und die Stabilitäts- und Faltungseigenschaften der sogenannten Internalin-Domäne (InlB₃₂₁) untersucht werden. Hierbei handelt es sich um die bei allen Mitgliedern der Internalinfamilie vorkommende Domäne, welche aus einer direkten Fusion des C-terminalen Endes der LRR-Domäne mit einer Immunglobulin (Ig)-ähnlichen Domäne besteht.</p> <p>Von beiden Konstrukten konnte eine vollständige thermodynamische Charakterisierung, mit Hilfe von chemisch- bzw. thermisch-induzierten Faltungs- und Entfaltungsübergängen durchgeführt werden. Sowohl InlB₂₄₁ als auch InlB₃₂₁ zeigen einen reversiblen und kooperativen Verlauf der chemisch-induzierten Gleichgewichtsübergänge, was die Anwendung eines Zweizustandsmodells zur Beschreibung der Daten erlaubte. Die zusätzliche Ig-ähnliche Domäne im InlB₃₂₁ resultierte im Vergleich zum InlB₂₄₁ in einer Erhöhung der freien Enthalpie der Entfaltung (8.8 kcal/mol im Vergleich zu 4.7 kcal/mol). Diese Stabilitätszunahme äußerte sich sowohl in einer Verschiebung des Übergangsmittelpunktes zu höheren Guanidiniumchlorid-Konzentrationen als auch in einer Erhöhung der Kooperativität des Gleichgewichtsübergangs (9.7 kcal/mol/M im Vergleich zu 7.1 kcal/mol/M). Diese Beobachtungen zeigen dass die einzelnen Sequenzeinheiten der LRR-Domäne nicht unabhängig voneinander falten und dass die Ig-ähnliche Domäne, obwohl sie nicht direkt mit dem Wirtszellrezeptor während der Invasion interagiert, eine kritische Rolle für die <i style='mso-bidi-font-style:normal'>in&nbsp;vivo</i> Stabilität des Internalin B spielt. Des weiteren spiegelt die Kooperativität des Übergangs die Integrität der Internalin-Domäne wieder und deutet darauf hin, dass bei beiden Proteinen keine Intermediate vorliegen.</p> <p>Kinetische Messungen über Tryptophanfluoreszenz und Fern-UV<span style='color:red'> </span>Circulardichroismus deuteten auf die Existenz eines relativ stabilen Intermediates auf dem Faltungsweg der LRR-Domäne hin. Faltungskinetiken aus einem in pH&nbsp;2 denaturierten Zustand zeigten ein reversibles Verhalten und verliefen über ein Intermediat. Eine Erhöhung der Salzkonzentration des sauer-denaturierten Proteins führte zu einer Kompaktierung der entfalteten Struktur und resultierte im Übergang zu einem alternativ gefalteten Zustand. Bei der Internalin-Domäne deuteten kinetische Messungen des Fluoreszenz- und Fern-UV Circulardichroismus-Signals während der Entfaltung möglicherweise auf die Präsenz von zwei Prozessen hin. Der erste langsame Entfaltungsprozess kurz nach dem Übergangsmittelpunkt zeigte eine starke Abhängigkeit von der Temperatur, während der zweite schnellere Prozess der Entfaltung stärker von der Guanidiniumchlorid-Konzentration abhing. Renaturierungskinetiken zeigten das Auftreten von mindestens einem Faltungsintermediat. Kinetische Daten aus Doppelsprungexperimenten lieferten für die Erklärung der langsamen Faltungsphase zunächst keinen Hinweis auf dass Vorliegen einer Prolinisomerisierungsreaktion. Die vollständige Amplitude während der Renaturierung konnte nicht detektiert werden, weswegen von einer zweiten schnellen Phase im Submillisekundenbereich ausgegangen werden kann.</p> <p>Die Ergebnisse der Faltungskinetiken zeigen, dass die InlB-Konstrukte als Modelle für die Untersuchung der Faltung von Solenoidproteinen verwendet werden können.<span lang=EN-GB style='mso-ansi-language: EN-GB'><o:p></o:p></span></p> / <p class=MsoBodyText><span lang=EN-GB style='mso-ansi-language:EN-GB'>To understand the processes of protein structure formation, it is necessary to investigate protein stability and protein folding kinetics. The focus of many folding studies has been directed at small, globular proteins. The larger class of solenoid proteins, including leucine-rich repeat (LRR) and ankyrin proteins, has not been extensively investigated. These proteins contain tandem repeat motifs, and their tertiary structure consists of a regular linear array of modules that stack to form non-globular elongated or supercoiled structures. In the present work, the folding and stability of the central LRR domain of the invasion protein internalin B (InlB₂₄₁) from the bacterium <i>Listeria monocytogenes</i> was characterized using different biophysical techniques. In addition, the integrity, stability and folding behavior of the so-called internalin-domain (InlB₃₂₁) was investigated. In this single domain, which is found in all members of the internalin-family, an immunoglobulin (Ig)-like domain is directly fused to the C-terminal end of the LRR domain.<span style='color:red'><o:p></o:p></span></span></p> <p class=MsoBodyText><span lang=EN-GB style='mso-ansi-language:EN-GB'>A complete thermodynamic characterization of the stability of both constructs was performed, using chemical- and temperature-induced folding and unfolding transitions. The reversible and cooperative equilibrium transition of InlB₂₄₁ and InlB₃₂₁ allowed the use of a two-state model for the description of the data points. The additional Ig-like domain present in InlB₃₂₁ resulted in an increase of the unfolding free energy (8.8 kcal/mol compared to 4.7 kcal/mol). This resulted both, from a shift of the transition midpoint to higher denaturant concentration, and from an increase in the <i>m</i>-value, the denaturant dependence of the unfolding free energy (9.7 kcal/mol/M compared to 7.1 kcal/mol/M). These observations suggest that the unravelling of the individual structural repeats in the LRR region is a cooperative process and that the tight fusion with the Ig-like domain leads to a dramatically increased stability <i>in vivo</i> without interfering with the functionality of the protein. In addition, the cooperativity of the equilibrium transition reflects the integrity of the internalin-domain, and suggests that both InlB fragments unfold without significantly populated equilibrium intermediates.<o:p></o:p></span></p> <p class=MsoBodyText><span lang=EN-GB style='mso-ansi-language:EN-GB'>Kinetic measurements with tryptophan fluorescence and far-UV circular dichroism are indicative for the existence of a relative stable intermediate on the folding pathway of the LRR domain. Refolding kinetics from an acid-denatured state showed a reversible behavior and passes off an intermediate. An increase in the salt concentration of the acid-denatured protein results in a transition of the unfolded structure to a compact and alternatively folded state. Unfolding kinetics of the internalin-domain measured by fluorescence and far-UV circular dichroism are indicative for the possible presence of two processes. The first slow unfolding process after the transition midpoint showed a strong dependence on temperature, whereas the second and faster unfolding process showed a stronger dependence on the denaturant concentration. Renaturation kinetics indicated the existence of at least one folding intermediate. Preliminary double-mixing experiments revealed no evidence for a rate-limiting proline isomerization reaction. It was not possible to detect the complete amplitude of the renaturation reaction, suggesting existence of a second faster phase occuring in the submillisecond range.<o:p></o:p></span></p> <p class=MsoBodyText><span lang=EN-GB style='mso-ansi-language:EN-GB'>The results on folding kinetics prove the InlB constructs to be suitable models for the investigation of solenoid protein folding by techniques of high structural resolution.<o:p></o:p></span></p>

Internalin J (InlJ) gehört zu der Klasse der bakteriellen, cysteinhaltigen (leucine-rich repeat) LRR Proteine. Bei den Internalinen handelt es sich um meist invasions-assoziierte Proteine der Listerien. Die LRR-Domäne von InlJ ist aus 15 regelmäßig wiederkehrenden, stark konservierten Sequenzeinheiten (repeats, 21 Aminosäuren) aufgebaut. Ein interessantes Detail dieses Internalins ist das stark konservierte Cystein innerhalb der repeats. Daraus ergibt sich eine ungewöhnliche Anordnung von 12 Cysteinen in einem Stapel. Die Häufigkeit von Cysteinen in InlJ ist für ein extrazelluläres Protein von L. monocytogenes außergewöhnlich, und die Frage nach ihrer Funktion daher umso brennender. Im Vergleich zum ubiquitären Vorkommen der sogenannten repeat-Proteine in der Natur sind Studien zu ihrer Stabilität und Faltung nicht äquivalent vertreten. Die zentrale Eigenschaft der repeat-Proteine ist ihr modularer Aufbau, der durch einfache Topologie gekennzeichnet ist und auf kurzreichenden Wechselwirkungen basiert. Diese Topologie macht repeat-Proteine zu idealen Modellproteinen, um die stabilitätsrelevanten Wechselwirkungen zu separieren und zuzuordnen. In der vorliegenden Arbeit wurde die Faltung und Entfaltung von InlJ umfassend charakterisiert und die Relevanz der Cysteine näher beleuchtet. Die spektroskopische Charakterisierung von InlJ zeigte, dass dessen Faltungszustand durch zwei Tryptophane im N- und C-Terminus fluoreszenzspektroskopisch gut zugänglich ist. Die thermodynamische Stabilität wurde mittels fluoreszenz-detektierten, Guanidiniumchlorid-induzierten Gleichgewichtsexperimenten bestimmt. Um die kinetischen Eigenschaften von InlJ zu erfassen, wurden die Faltungs- sowie die Entfaltungsreaktion spektroskopisch untersucht. Die Identifizierung der produktiven Faltungsreaktion war lediglich durch die Anwendung des reversen Doppelsprungexperiments möglich. Die Auswertung erfolgte nach dem Zweizustandsmodell, wonach die Faltung dem „Alles-oder-Nichts“ Prinzip folgt. Die Gültigkeit dieser Annahme wurde durch die kinetische Charakterisierung bestätigt. Es wurde sowohl in den Gleichgewichtsexperimenten als auch in den kinetisch erhaltenen Daten eine hohe freie Stabilisierungsenthalpie festgestellt. Die hohe Stabilität von InlJ geht mit hoher Kooperativität einher. Die kinetischen Daten zeigen zudem, dass die hohe Kooperativität hauptsächlich der Faltungsreaktion entstammt. Der Tanford-Wert von 0.93 impliziert, dass die Oberflächenänderung während der Faltung bereits zum größten Teil erfolgt ist, bevor der Übergangszustand ausgebildet wurde. Direkte strukturelle Informationen über den Übergangszustand wurden mit Hilfe von Mutationsstudien erhalten. Zu diesem Zweck wurden 12 der 14 Cysteine gegen ein Alanin ausgetauscht. Die repeats 1 bis 11 von InlJ beinhalten jeweils ein Cystein, deren Anordnung eine Leiter ergibt. Deren Substitutionen haben einen vergleichbar destabilisierenden Effekt auf InlJ von durchschnittlich 4.8 kJ/mol. Die Verlangsamung der Faltung deutet daraufhin, dass die Interaktionen der repeats 5 bis 11 im Übergangszustand bereits voll ausgebildet sind. Demnach liegt bei InlJ ein zentraler Faltungsnukleus vor. Im Rahmen dieser Promotionsarbeit wurde eine hohe Stabilität und ein stark-kooperatives Verhalten für das extrazelluläre Protein InlJ beobachtet. Diese Erkenntnisse könnten wichtige Beiträge zur Entwicklung artifizieller repeat-Proteine leisten, deren Verwendung sich stetig ausweitet. / Internalin J (InlJ) is a member of the family of bacterial cysteine-containing leucine-rich repeat (LRR) proteins. Internalins are invasion-associated surface proteins of Listeria monocytogenes. The LRR domain of InlJ consists of 15 repeating units, which are arranged in tandem. The consensus sequence consists of 21 residues. Interestingly, a leucine residue which is highly conserved among the Internalins is replaced by cysteine. This results in a continuous cysteine ladder of 12 repeats. This frequency of cysteines is remarkable for an extracellular protein of L. monocytogenes. Stability and folding of repeat proteins are not equivalently studied considering their ubiquitous distribution in nature. Their modular structure results in simple topology and is dominated by short-range interactions. These characteristic features of repeat proteins facilitate the separation and identification of stabilizing interactions, making repeat proteins to ideal model systems for folding studies. In this work the folding and unfolding of InlJ has been extensively characterized, shedding light on the relevance of the cysteines. Two tryptophans located in the N- and C-terminus allowed monitoring the folding state of the entire protein via fluorescence. Thermodynamic stability was therefore derived by guanidinium chloride induced equilibrium experiments. Furthermore, the chemically induced unfolding and folding reactions were characterized with respect to their kinetics. Interrupted refolding experiments were essential for tracking the productive folding reaction of InlJ. Analysis of the kinetic and equilibrium data leads to the conclusion that the results are compatible with a two-state model. The study presented here reveals high stability of the protein InlJ in conjunction with high cooperativity. Kinetic data disclosed the origin of high cooperativity in the folding reaction; with a Tanford value of about 0.93. This high value implicates that the major change of the accessible surface area occurs before the transition state is formed. Mutational studies provided more detailed structural information about the transition state. 12 of 14 cysteine residues were mutated to alanine for this purpose. The cysteines in repeats 1 to 11 stack over each other and form a ladder of reduced cysteines. The substitution of one of these cysteines has an average destabilizing effect of 4.8 kJ/mol. The deceleration of the folding reaction by the substitution shows that repeats 5 to 11 are already fully structured in the transition state, pointing to a central nucleus in the folding of the LRR-protein InlJ. The extracellular protein InlJ reveals extreme stability and high cooperativity. The insights into the folding of this LRR motif could facilitate the design of further artificial repeat proteins.

Nesta tese são apresentados um novo modelo estatístico para cálculo de probabilidade de paternidade e sua implementação em software. O modelo proposto utiliza o genótipo como informação básica, em contraste com outros modelos que usam alelos. Por esta diferença, o modelo proposto resulta mais abrangente, mas que, sob certas restrições, reproduz os resultados dos modelos que usam alelos. Este modelo foi implementado em um software que recebe descrições da genealogia e dos marcadores em uma linguagem dedicada a isso e constrói uma rede bayesiana para cada marcador. O usuário pode definir livremente a genealogia e os marcadores. O cálculo da probabilidade de paternidade é feito, sobre as redes construídas, por um software para inferência em redes bayesianas e a probabilidade de paternidade combinada considerando todos os marcadores é calculada, resultando em um \"índice de paternidade. / This thesis presents a novel statistical model for calculation of the probability of paternity and its implementation as a software. The proposed model uses genotype as basic information. Other models use alleles as basic information. As a result the proposed model is broader, in the sense that, under certain constraints the results from the other models are reproduced. The software implementation receives pedigree and markers data, in a specifically designed language, as input and builds one bayesian network for each marker. The user can freely define any pedigree and any marker. Paternity probabilities for each locus are calculated, from the built networks, by a software for inference on Bayesian Networks and these probabilities are combined into a single \"paternity index\".

Chlorophyll spielt eine unabdingbare Rolle für die lichtabhängige Reaktion der Photosynthese. Die adäquate Versorgung mit Chlorophyll wird dabei durch die Tetrapyrrolbiosynthese (TBS) gewährleistet. In den letzten Jahrzehnten wurde eine Vielzahl von Proteinen identifiziert, welche an der Anpassung der TBS an wechselnde (a)biotische Wachstumsbedingungen der Pflanze beteiligt sind. Allerdings konnte bislang nicht zweifelsfrei geklärt werden, wie die TBS mit der Integration von Chlorophyllen in die Photosysteme koordiniert wird. Vor einigen Jahren wurde ein Interaktionspartner der Protochlorophyllid-Oxidoreduktase (POR) in Synechocystis identifiziert, welcher als potenzieller Faktor dieser Koordination in Frage kommt. Das POR-INTERACTING TPR-Protein (Pitt) stabilisiert POR an der Thylakoidmembran und interagiert auch mit dem Vorstufenprotein des D1. Pitt gehört zur Familie der tetratricopeptide repeat (TPR) Proteine, deren Vertreter vorrangig für die Vermittlung von Protein-Protein-Interaktionen zuständig sind. Aus diesem Grund war, neben der Identifikation des potenziellen Pitt-Homologs im Modelorganismus Arabidopsis thaliana, die Analyse von anderen Vertretern dieser Proteinklasse ein vielversprechender Ansatz bei der Identifikation von weiteren Regulatoren der TBS oder Photosynthese. Von den fünf ausgewählten TPR-Proteinen aus Arabidopsis thaliana mit einer hohen Sequenzähnlichkeit zu Pitt waren vier in der Lage, physisch mit POR zu interagieren. Von diesen vier Kandidaten ist das durch das Gen At1g78915 kodierte, membranintegrale TPR-Protein (TPR1) der beste Kandidat des putativen Pitt-Homologs in Arabidopsis. Vergleichbar zu Pitt interagiert TPR1 mit POR und stabilisiert das Enzym an den plastidären Membranen. Die Stabilisierung von POR durch TPR1 spielt eine entscheidende Rolle während der Etiolierung und Ergrünung von Keimlingen. Darüber hinaus steht TPR1 im Zusammenhang mit der schnellen Inaktivierung der 5-Aminolävulinsäuresynthese. / Chlorophyll plays an indispensable role in the light reaction of the photosynthesis. The adequate supply of chlorophyll is ensured by tetrapyrrole biosynthesis (TBS). Within the last decades, multiple proteins were identified, which are involved in adjusting the TBS-pathway to changing (a)biotic plant growth conditions. Nevertheless, it is not fully understood how the TBS-pathway is coordinated parallel to the assembly of the photosystems and the integration of chlorophylls into the pigment-binding subunits of the photosystems. Several years ago, an interaction partner of the protochlorophyllide-oxidoreductase (POR) was identified in Synechocystis which was proposed to be involved in the coordination of these mechanisms. The POR-INTERACTING TPR-Protein (Pitt) binds and stabilizes POR at the thylakoid membranes and interacts with the precursor protein of D1. Therefore, Pitt could facilitate the incorporation of chlorophylls into the plastid-encoded nascent photosynthetic subunits. Pitt belongs to the tetratricopeptide repeat (TPR) protein family, whose members mediate protein-protein-interactions. Besides the identification of the potential Pitt-homolog in the model organism Arabidopsis thaliana, analysis of additional members of the TPR-protein superfamily was a promising approach for the identification of further posttranslational regulators of TBS and photosynthesis. Five Arabidopsis thaliana TPR-proteins with a high sequence similarity to Pitt were selected. Four of those proteins are able to interact physically with POR. Among them, the TPR-protein encoded by the gene At1g78915 (TPR1) was the best candidate to represent a putative Pitt homolog in Arabidopsis. Similar to Pitt, TPR1 is a plastid-localized integral membrane protein, which interacts with POR at the thylakoid membranes. The stabilizing effect of TPR1 on POR is especially needed during etioliation and greening. Additionally, TPR1 is required for a inactivation of the 5'-aminolevulinic acid synthesis.

Over the past several years destination loyalty has begun to take up an increasing amount of focus within the tourism research field. While there are still a number of question marks of its different aspects and true value to the destination, the subject has been described as prioritized. On that account, the purpose of this study has been to seek understanding of how Stockholm relate to this, and how their expressed strategies towards the international market can be understood within a destination loyalty perspective. Furthermore, the DMO Visit Stockholm’s marketing and communication strategies towards foreign followers on digital platforms have been examined. To reach knowledge of this, a document study of Visit Stockholm and Svensk Turism was carried out, whereafter three interviews took place with representatives from these two organizations. Lastly, a netnographic examination was carried out of Visit Stockholm’s official webpage and their accounts on Facebook and Instagram. The results showed that strategies have yet to be expressed formally regarding destination loyalty, but that there is an awareness of the issue and that it most likely will play a bigger role in the future. In addition to that, there have been empirical findings which show that marketing and communication in fact is conducted, which can relate to destination loyalty. A challenge for Stockholm in the future will be to further take advantage of the possibilities that the digital channels offer. An active approach will be required in imposing new relations to followers and visitors, and in the retaining of existing relations. / Under de senaste åren har destinationslojalitet börjat ta upp alltmer fokus inom den turistiska forskningen. Medan det ännu finns kvar flera frågetecken över dess olika aspekter och egentliga värde för destinationen, så har ämnet beskrivits som speciellt prioriterat. Med anledning av det har syftet med denna undersökning varit att söka förståelse för hur Stockholm förhåller sig till detta, och hur deras uttalade strategier mot den internationella marknaden kan förstås utifrån ett destinationslojalitetsperspektiv. Vidare har undersökts hur DMO:n Visit Stockholms faktiska marknadsföring och kommunikation till utländska följare förs på digitala plattformar. För att nå denna kunskap utfördes en dokumentstudie av Visit Stockholm samt Svensk Turism, därefter ägde tre intervjuer rum med representanter från dessa två organisationer. Till sist utfördes även en netnografisk undersökning av Visit Stockholms officiella hemsida, samt deras konton på Facebook och Instagram. Resultaten visade att det ej i dagsläget finns några uttalade formella strategier kring destinationslojalitet, men att det finns en medvenhet kring frågan och att det troligtvis kommer att bli mer aktuellt i framtiden. Utöver detta har empiriska fynd gjorts som visar på att det faktiskt förs marknadsföring och kommunikation på sociala medier som kan kopplas till destinationslojalitet. En utmaning för Stockholm inför framtiden blir att ytterligare ta till vara på de möjligheter som de digitala kanalerna kan erbjuda. Ett aktivt arbete kommer att krävas inför skapandet av nya relationer till följare och besökare, samt bibehållandet av existerande relationer.

This thesis explores the forces of change in lives and landscapes that have altered the Lamosangu-to-Everest route in northeastern Nepal and shows how a transect in photographs and conversations across the east-central Himalaya allows us insight and a greater understanding into the processes and consequences of this change. Three forces of change over the last twenty-five years dominated discussions with local informants: the rise of the "People's War"-Nepal's Maoist Insurgency beginning in 1996; the Democratic Revolution of 1990; and dependence on tourism for livelihood after the establishment of Sagarmatha National Park in 1976. Understanding the cultural-historical context for these forces is necessary to understand the concerns of today's residents living along the Lamosangu-to-Mount Everest Base Camp transect. The visual and ethnographic evidence discussed in this thesis takes a larger role than strict analysis of conspicuous large-scale land use change and I hope the comparative 200 I images will be used as benchmarks for future research as well as for further exploration into the ways people and place have been represented.

ITC/USA 2011 Conference Proceedings / The Forty-Seventh Annual International Telemetering Conference and Technical Exhibition / October 24-27, 2011 / Bally's Las Vegas, Las Vegas, Nevada / A novel type-II hybrid automatic repeat request (HARQ) algorithm using adaptive modulations and bit-interleaved coded modulation (BICM) is presented. The algorithm uses different optimized puncturing patterns for different transmissions of the same data packet. The proposed approach exploits mapping diversity through BICM with iterative decoding. The modulation order is changed in each transmission to keep the number of symbols transmitted constant. We present new bit error rate and frame error rate analytical results for the proposed technique showing good agreement with simulation results. We compare the throughput performance of our proposed HARQ technique with a reference HARQ technique that uses different mapping arrangements but keeps the modulation order fixed. By using optimized puncturing patterns and adaptive modulations, our method provides significantly better throughput performance over the reference HARQ method in the whole signalto- noise ratio (SNR) range, and achieves a gain of 12 dB in the medium SNR region.

Flooding of residential property is a real and growing phenomenon in the UK causing short and long-term detriment of various kinds to its victims. The issue of potential decrease in value of those properties which are located on the floodplain, though much discussed in the media, has received scant attention in the UK research literature. An extensive literature survey has revealed a need for methodological innovation in the field of temporal impact of flooding and the inadequacy of the current paradigms for inclusion of insurance into flood modelling. A wide-ranging review of data sources, including discussion with industry experts, has identified the requirement to generate primary data on the availability and cost of flood insurance. A novel framework has been developed for this research. This framework is an extension of the recent research in flood modelling and incorporates ideas from the wider house price analysis literature. Data collected via a questionnaire survey of householders has been combined with secondary data on property prices and flood designation in order to attribute any loss in property value to the correct vector of underlying flood status. The output from this study makes a contribution to the understanding of the impact of flooding on house prices, allowing for better valuation advice. Empirical findings are that the understandable concerns of residential property owners at risk of flooding regarding long term loss of property value are largely unfounded. Price discounts are observed for some recently flooded areas but they are temporary Improved appreciation of the impact of claims and flood risk on the cost of insurance has also emerged. The insurance market was not found to be instrumental in reducing the price of property. The output from the study also makes a methodological contribution in extending concepts relating to the relationship between flooding, insurance and house prices. This development is anticipated to facilitate refinement and updating of the empirical findings with reduced effort in the light of future events.

Repeat track analysis is one tool that can be used to derive parameters describing elevation changes from elevation data collected from a satellite with a near-repeat groundtrack. While initially developed to study ocean topography, it was then applied to ice sheet data. This study expands upon that previous research by testing the method’s ability to estimate parameters using different amounts of data, different grid sizes and types, and different elevation models containing different parameters to be estimated. In all cases, ICESat-derived elevations were used as input data, as ICESat has a near-repeat groundtrack with extensive coverage of the Greenland and Antarctica ice sheets. Results were compared using the differences between modeled and ICESat-derived elevations, correlation of estimated parameters to known physical features, and differences between known and estimated parameter values for simulated elevation data. It was found that there should be data from at least as many distinct time periods (or, in the case of ICESat, laser campaigns) as parameters being estimated, grids centered on and aligned with the reference groundtrack should be used, and that elevation models containing a constant elevation change rate, slopes, an initial elevation at the grid center, and annual terms should be used. Crossover analysis is a different method to determine elevation change rate with elevation data and serves as an independent verification of the repeat track analysis method. It was found that the hdot values determined from crossover and repeat track analyses agreed to within 5 cm/yr in most areas of the ice sheets, with differences greater than 40 cm/yr along the coasts. While repeat track analysis provides greater coverage than crossover analysis, it is uncertain which method provides the most accurate results. / text

The expansion of gene-specific trinucleotide repeats is responsible for a growing list of human disorders, including myotonic dystrophy type 1 (DM1). Repeat instability for most of these disorders, including DM1, is characterized by complex patterns of inherited and ongoing tissue-specific instability and pathogenesis. While the mechanistic basis behind the unique locus-specific instability of trinucleotide repeats is currently unknown, DNA metabolic processes are likely to play a role. My thesis involves investigating the contribution of DNA replication to the trinucleotide instability of myotonic dystrophy type 1. Herein I have designed an in vivo primate model system, based on the SV40 replication system, to assess the contribution of DNA replication to DM1 repeat instability. This system allows the assessment, under controlled conditions, and manipulation of variables that may affect replication-associated repeat instability, under a primate cellular system. Using the SV40 model system, I not only confirmed previous observations that repeat length and replication direction affect repeat instability, but also for the first time determined that the location of the replication origin relative to the repeat tract plays an important role in repeat instability. This novel observation allowed for the development of a fork-shift model of repeat instability, in which cis-elements adjacent to the repeat tract affect replication, in turn altering the propensity for repeat instability. To further my study of DNA replication in DM1 repeat instability, I have mapped the origin of replication adjacent to the DM1 locus in human patient cells and the tissues of DM1 transgenic mice actively undergoing repeat instability. The position of the replication origins adjacent to the repeat tract at the DM1 locus places several known cis-elements, including CTCF binding sites, in a position to alter replication as predicted by the fork-shift model. My analysis of the CTCF sites showed them capable of altering replication and repeat instability at the DM1 locus. Taken together these results suggest that the placement of replication origins, repeat tracts and cis-elements, may mark trinucleotide repeat tracts, such as the DM1, for locus-, tissue- and development-specific replication-associated repeat instability.