In der vorliegenden Arbeit wurden Cluster- und QM/MM-Einbettungsmodelle für geträgerte Vanadiumoxidkatalysatoren auf Siliziumdioxid mit Hilfe von DFT-Rechnungen untersucht. Es wurden Strukturen, Schwingungen und die Stabilität gegenüber Wasser und Sauerstoff unter Reaktionsbedingungen betrachtet. Ferner wurde die Reaktivität bei der Oxidativen Dehydrierung (ODH) von n-Propan zu n-Propen und der Oxidation von Methanol zu Formaldehyd untersucht. Zur Durchführung der Einbettungsrechnung wurde ein angepasstes Shell-Model-Potential verwendet. Bezüglich der Schwingungen zeigte sich, dass sich der Einfluss der Einbettung hauptsächlich auf die Kopplung von Schwingungen beschränkt, aber die Frequenzen kaum verändert werden. Die lokale Struktur um das Vanadiumatom beeinflusst die Vanadylschwingung wenig. Die Stabilitätsberechnungen haben gezeigt, dass die untersuchten Modelle unter Reaktionsbedingungen vorliegen und hydroxylierte Spezies keine Bedeutung haben. Bei der Untersuchung der beiden Reaktionen wird für die Cluster- und die QM/MM-Einbettungsrechnungen jeweils derselbe Mechanismus gefunden. Der Vorteil der Einbettungsrechnungen besteht darin, dass die lokale Struktur um das aktive Zentrum variiert werden kann. Bei der ODH von Propan hat die lokale Struktur einen eher geringen Einfluss auf die Reaktionsenergien, denn die Reaktion verläuft hauptsächlich am Vanadylsauerstoff. Der Übergangszustand des geschwindigkeitsbestimmenden Schrittes liegt bei der Einbettungsrechnung dennoch energetisch höher. Grund ist eine sterische Hinderung durch die Hydroxylgruppen der Oberfläche. Bei der Oxidation von Methanol ist der Einfluss der lokalen Struktur größer, denn die Reaktion verläuft sowohl über den Vanadylsauerstoff als auch über die Brückensauerstoffatome zum Trägermaterial. Für beide Reaktionen wird ein Einfluss der Vanadiumbeladung auf die Reaktionsenergien gefunden. Bei höherer Beladung werden die Reaktionen exothermer. / In this work cluster models and models for QM/MM-embedding for supported vanadia catalysts on silica were studied with help of DFT-calculations. The structures, vibrations and stability towards water and oxygen under reaction conditions were examined. Furthermore the reactivities towards the oxidative dehydrogenation (ODH) of n-propane to n-propene and the oxidation of methanol to formaldehyde were tested. For the embedding an adapted shell-model-potential was employed. Regarding the vibrations it was shown that the influence of the embedding lies mostly in the coupling of vibrations and not in their frequencies. The local structure surrounding the vanadium atom has only minor influence. The stability calculations have shown that the tested model systems exist under reaction conditions while hydroxylated species do not exist. When studying the reactivity of the two reactions always the same mechanism is found for cluster and embedded calculations. The benefit of the embedded calculations is the possibility to vary the local structure surrounding the active center. For the ODH of propane the local structure has only small impact on reaction energies because the reaction takes mainly place at the vanadyl oxygen. The transition state of the rate determining step nevertheless is energetically higher in the embedded calculations due to steric hindrance caused by the hydroxyl groups of the surface. The impact of local structure is more important for the oxidation of methanol since in this case vanadyl oxygen and bridging oxygens to the support are involved in the reaction. For both reactions an influence on reaction energies of the vanadia loading is found. With more loading the reactions becomes more exothermic.

Rett Syndrome (RTT) is an X-linked genetic disorder and a major cause of intellectual disability in girls. Mutations in the methyl-CpG binding protein 2 (MECP2) gene, are the primary cause of the disorder. Despite the dominant neurological phenotypes that characterise RTT, MECP2 is expressed ubiquitously throughout the body and a number of peripheral phenotypes such as growth retardation (reduced height and weight), skeletal deformities (scoliosis/kyphosis), reduced bone mass and low energy fractures are also common yet under-reported clinical features of the disorder. In order to explore whether MeCP2 protein deficiency results in altered structural and functional properties of bone and to test the potential reversibility of any such defects, I have conducted series of histological, imaging and biomechanical tests of bone using an accurate genetic (functional knockout) mouse model of RTT. Initial experiments using a GFP reporter mouse line demonstrated the presence of MeCP2 in bone cells and the effective silencing on the gene in functional knockout mice. Different aspects of the study were conducted in different types of bone tissues that were especially suited for individual assays. For instance, biomechanical three point bending tests were conducted in long bone (femur) whilst trabecular geometry measures were measured in spinal vertebrae. Both hemizygous Mecp2stop/y male mice in which Mecp2 is silenced in all cells and female Mecp2stop/+ mice in which Mecp2 is silenced in ~50% of cells as a consequence of random X-chromosome inactivation (XCI), revealed, lighter and smaller long bones and significant reductions in cortical bone mechanical properties (~ 39.5% reduction in stiffness, 31% reduction in ultimate load and 37% reduction in Young’s modulus respectively in Mecp2stop/y male mice; %) and material properties (microhardess reduced 12.3% in Mecp2stop/y male mice and 14% inMecp2stop/+ female mice) as compared to age wild type control mice. Micro structural analysis conducted using µCT also revealed a significant reduction in cortical (54% reduction in cortical thickness, 30% in bone volume, 20% in total area, and 38% in marrow area) and trabecular (~30% in trabecular thickness) bone parameters as compared to age matched wild-type controls MeCP2-deficent mice. Histological analysis using Sirius red staining as a marker of collagen revealed a ~25% reduction in collagen content in MeCP2 deficient mice as compared to age matched wild type controls. In experiments designed to establish the potential for reversal of MeCP2-related deficits, unsilencing of Mecp2 in adult mice by tamoxifen-induced and cre-mediated excision of a stop cassette located at the endogenous Mecp2 locus (male; Mecp2stop/y, CreER and female; Mecp2+/stop, CreER), resulted in a restoration of biomechanical properties towards the wild-type levels. Specifically, Male Mecp2stop/y, CreER mice displayed improvement in mechanical properties (stiffness 40%, ultimate load 10%, young’s modulus 61% and micro hardness 12%) and structural bone parameter (trabecular thickness 80%) as compared to Mecp2stop/y male mice. Female Mecp2+/stop, CreER, displayed a significant improvement (19%) in microhardess measures as compared to Mecp2 deficient mice. Overall, the results of my studies show that MeCP2-deficiency results in overt, but potentially reversible, alterations in the biomechanical integrity of bone and highlights the importance of targeting skeletal phenotypes in considering the development of pharmacological and gene-based therapies for Rett Syndrome.

Pulmonary arterial hypertension (PAH) is a progressive and debilitating disease characterised by increases in pulmonary vasoconstriction and excessive remodelling of the pulmonary arteries. Together, these processes lead to sustained elevations in pulmonary arterial pressure, right heart failure and eventual death if left untreated. Despite the number and variety of treatment options available, the survival rate in incident and prevalent cases of PAH remains poor. Therefore, a better understanding of the pathobiology of PAH is required to generate novel therapeutic approaches with improved efficiency in patients. In PAH there is a well described gender bias. Women are consistently reported to represent up to 75% of the total PAH population; however, the reasons for this female predominance remain unclear. Recently, estrogen has been implicated as a major risk factor, for example, elevated estrogen levels and alterations in estrogen metabolism are closely correlated with PAH development in females. The role of testosterone in PAH is currently under investigated. Effects of estrogen are mediated through two classical estrogen receptors (ER)-α and –β, or the novel G-protein-coupled estrogen receptor (GPER). Expression of all of these receptors is identified in pulmonary vasculature, including in smooth muscle and endothelial cells. The role they play in PAH pathogenesis in females is largely undetermined. Given the diverse effects of estrogen described in the pulmonary vasculature during PAH, for example, proliferative effects in pulmonary artery smooth muscle cells (PASMCs), we hypothesised that estrogen receptors play an integral role in PAH in females. To examine this, we used both translational and experimental studies to characterise ERs in PAH. Chronic hypoxic male and female mice, and mice over-expressing the serotonin transporter (SERT+ mice), which demonstrate female susceptibility, were used to investigate the effects of an ERα antagonist in vivo. GPER knockout mice were also investigated in chronic hypoxia. In situ and in vitro studies in human PASMCs with ER agonists and antagonists added clinical relevance to our findings. In addition, testosterone manipulation was investigated in male mice by castration in vivo. Immunohistochemistry, immunoblotting and qRT-PCR analysis demonstrated that ERα was increased in PASMCs and pulmonary arteries from female PAH patients and chronic hypoxic mice, respectively. On the other hand, ERβ was decreased in PAH and hypoxia. It was also observed that females expressed higher levels of ERα in PAH compared to males whereas ERβ was lower in females. PAH was assessed by measuring right ventricular systolic pressure (RVSP), right ventricular hypertrophy (RVH) and pulmonary vascular remodelling and muscularisation. Chronic hypoxia induced-pulmonary hypertension (PH) was attenuated in female mice dosed with the ERα antagonist MPP, shown by marked reductions in RVSP and pulmonary vascular remodelling. Hypoxic male mice remained unaffected by MPP treatment. Spontaneous PH and chronic hypoxia induced-PH observed in female SERT+ mice were reversed by treatment with MPP. Immunoblotting and qRT-PCR analysis revealed that the possible mechanism involved in the beneficial effect of MPP in females in vivo involved restoring the dysfunctional bone morphogenetic protein receptor-2 (BMPR2) axis observed in PAH. This effect was only observed in female mice. In addition, chronic hypoxia induced- PH in male and female mice was unaffected by GPER deletion. Expression of GPER between female non-PAH controls and PAH patients was unchanged. In isolated human PASMCs estrogen induced proliferation was inhibited by MPP, but not PHTPP or G15, an ERβ and GPER antagonist, respectively. The ERα agonist, PPT stimulated proliferation of human PASMCs. Both estrogen and PPT induced proliferation was dependent on downstream PI3K/Akt and ERK MAPK activity. In males, testosterone deprivation by surgical castration had no effect on chronic-hypoxia induced PH. RVSP, RVH and pulmonary vascular remodelling were unchanged in hypoxic castrated mice relative to sham controls. Testosterone levels, assessed by enzyme linked immunosorbent assay (ELISA) demonstrated no effects of hypoxia on plasma testosterone levels. Testosterone levels were approximately halved by castration. qRT-PCR analysis showed that in mouse lung there were also no difference in expression of the androgen receptor (AR) and 5α-reducatse, the testosterone metabolising enzyme. Testosterone had no effect on proliferation of human PASMCs, although its primary metabolite, dihydrotestosterone (DHT), stimulated proliferation in a dose-dependent manner. In summary of these findings, we have identified an ERα-dependent mechanism of PAH in females, but not in males. ERα is noticeably increased in female human PASMCs from PAH patients compared to male PAH patients. Additionally, ERα activation in female human PASMCs leads to proliferation driven by PI3K/Akt and ERK MAPK activation. Treatment with an ERα antagonist attenuated the development of chronic hypoxia induced-PH in females but not males, and reversed PH in SERT+ female mice. We demonstrate that the mechanism attributed to the beneficial effect of MPP in vivo involved restoration of the dysfunctional BMPR2 signalling axis. Our results suggest that increased ERα expression may drive PAH development in females. Furthermore, we demonstrate that ERα does not play a key role in the development of hypoxia induced-PH in male mice. In addition we conclude that testosterone does not contribute to chronic hypoxic-PH observed in males. We suggest that altered local synthesis and metabolism in the lung and right ventricle may however, facilitate progression of established PAH in males and worsening survival rates. Overall, our results provide evidence for ERα in PAH development and implicate targeting ERs as a novel therapeutic target in PAH treatment.

Previous studies have identified stem cell populations in articular cartilage using colony forming assays and mesenchymal stem cell (MSC) marker expression. The specificity of classical MSC markers for isolation of stem cells within articular cartilage is insufficient, with large and highly variable quantities being reported in the literature. This study has demonstrated, for the first time, a panel of stem cell markers specific for articular cartilage-derived stem cells (ACSC). ACSCs were isolated, quantified and cultured from healthy and OA joints. Stem cells were clonally-derived cell lines that proliferated beyond 50 population doublings whilst maintaining a phenotype, and demonstrated tri-lineage potential. We discovered that OA cartilage had a two-fold increase in stem cell number, consisting of two divergent stem cell sub-populations. These divergent populations varied in proliferative capacity with only 50% of stem cells from the OA joint capable of extended proliferation in vitro. Using transcriptomic next generation sequencing of culture-expanded chondrocytes and ACSCs we successfully identified differentially expressed genes and a panel of novel markers of cartilage-specific stem cells. Novel markers were validated using qPCR and protein labelling and, were specifically expressed in ACSCs, with no expression in the culture-expanded full-depth chondrocytes. Using immunofluorescence for novel stem cell markers we found articular cartilage-derived stem cells are localised within the transitional zone in normal cartilage and the superficial zone in OA cartilage. OA cartilage was found to contain a 2-fold increase in stem cells using immunofluorescence. Subsequently, we used the panel of novel markers and fluorescent active cell sorting to isolate a sub-population from full-depth cartilage with stem cell characteristics. These cells were plastic adherent, clonogenic, with proliferative capacity greater than 50PD and displayed tri-lineage potential, therefore meeting all criteria for classification as a MSC population. The use of specific markers to isolate ACSCs will allow for further characterisation of stem cells, including a more in-depth understanding of the mechanisms of proliferation, differentiation and degeneration within articular cartilage.

The fundamental corneal properties of mechanical rigidity, maintenance of curvature and optical transparency result from the specific organisation of collagen fibrils in the corneal stroma. The exact arrangement of stromal collagen is currently unknown but several structural models have been proposed. The purpose of the present study is to investigate inconsistencies between current x‐ray derived structural models of the cornea and optically derived birefringence data. Firstly, the thesis reviews the current understanding of corneal structure, particularly in relation to corneal birefringence. It also reviews and develops the different analytical approaches used to model optical biaxial behaviour, particularly as applied to predict corneal optical phase retardation. The second part develops a novel technique of elliptic polarization biomicroscopy (EPB), enabling study of corneal birefringence in vivo. Using EPB, the pattern of corneal retardation is recorded for a range of human subjects. This dataset is then used to investigate both central and peripheral corneal birefringence as well as the corneal microstructure. A key finding is that the central parts of the cornea exhibit a retardation pattern compatible with a negative biaxial crystal, whereas the peripheral corneal regions do not. Furthermore, within the central regions of the cornea, orthogonal confocal conic fibrillar structures are identified which resemble the analytically derived contours of equal refractive index of an ideal negative biaxial crystal. The third part of this work presents a synthesis of previous published experimental, anatomical and theoretical findings and the experimental results presented in this thesis. Based on these findings, a novel corneal structural model is proposed that comprises overlapping spherical elliptic structural units. Finally, ensuing biomechanical and clinical consequences of the spherical elliptic structural model and of the EPB technique are discussed including their potential diagnostic and surgical applications.

Stereophotogrammetric imaging systems produce representations of surfaces (two-dimensional manifolds in three-dimensional space) through triangulations of a large number of estimated surface points. Traditional forms of analysis of these surfaces are based on point locations (manually marked anatomical landmarks) as described in Chapter 1. An advanced application of these types of landmarks will be thoroughly examined in Chapter 2 through the concept of Ghost Imaging. The results of this chapter necessitated a reliability study of stereophotogrammetric imaging systems which is discussed in Chapter 3. Given the results of the reliability study, an investigation info new definitions of landmarks and facial shape description is undertaken in Chapter 4. A much richer representation is expressed by the curves which track the ridges and valleys of the dense surface and by the relatively smooth surface patches which lie between these curves. New automatic methods for identifying anatomical curves and the resulting full surface representation, based on shape index, curvature, smoothing techniques, warping, and bending energy, are described. Chapter 5 discussed new and extended tools of analysis that are necessary for this richer representation of facial shape. These methods will be applied in Chapter 6 to different shape objects, including the human face, mussel shells, and computational imaging comparisons. Issues of sexual dimorphism (differences in shapes between males and females), change in shape with age, as well as pre- and post-facial surgical intervention will be explored. These comparisons will be made using new methodological tools developed specifically for the new curve and surface identification method. In particular, the assessment of facial asymmetry and the questions involved in comparing facial shapes in general, at both the individual and the group level, will also be considered. In Chapter 7, Bayesian methods are explored to determine further ways in which to understand and compare human facial features. In summary, this thesis shows a novel method of curve and full facial mesh identification that is used, successfully, in pilot case studies of multiple types of surfaces. It then shows a novel proof of principle for using Bayesian methods to create a fully automatic process in facial shape characterisation. In order to view this thesis in full, please view in Adobe Reader.

EU's 2007 enlargement by Bulgaria and Romania is evaluated by applying a simple macroeconomic integration model able to encompass as many of the theoretically predicted integration effects possible. The direct integration effects of Bulgaria and Romania spill-over to EU15, including Austria and the 10 new member states of the 2004 EU enlargement. The pattern of the integration effects is qualitatively similar to those of EU's 2004 enlargement by 10 new member states. Bulgaria and Romania gain much more from EU accession than the incumbents in the proportion of 20:1. In the medium-run up to 2020, Bulgaria and Romania can expect a sizable overall integration gain, amounting to additional ½ percentage point real GDP growth per annum. Within the incumbent EU member states Austria will gain somewhat more (+0.05%) than the average of EU15 (+0.02%) and the 10 new EU member states (+0.01%), which joined the EU in 2004. (author's abstract) / Series: EI Working Papers / Europainstitut

Regression models for de facto currency regime classification are complemented by inferential techniques for tracking the stability of exchange rate regimes. Several structural change methods are adapted to these regressions: tools for assessing the stability of exchange rate regressions in historical data (testing), in incoming data (monitoring) and for determining the breakpoints of shifts in the exchange rate regime (dating). The tools are illustrated by investigating the Chinese exchange rate regime after China gave up on a fixed exchange rate to the US dollar in 2005 and to track the evolution of the Indian exchange rate regime since 1993. / Series: Research Report Series / Department of Statistics and Mathematics

xv Abstract The aim of this thesis is to investigate the accurate stabilization energy and binding free energy in various non-covalent complexes spanned from small organic molecules to biomolecules. Non-covalent interactions such as H-bonds, π...π stacking and halogen bonds are mainly responsible for understanding of most biological processes, such as small molecule interactions with surface, protein-ligand binding in the cell machinery, etc. In the thesis, different non-covalent complexes such as graphene…electron donor- acceptor complexes, DNA base pair interaction with silica surface, etc, were investigated. The reference stabilization energies were calculated at ab initio level, e.g., CCSD(T)/CBS method wherever possible. On the other hand, more approximated scaled MP2 method (MP2.5/CBS/6-31G*(0.25)) is taken as reference instead of CCSD(T)/CBS due to the size of the complexes. Further, the DFT and MM energies were also tested towards the reference one. The knowledge of non- covalent interaction is required for rationalizing of any association processes in nature which requires accurate description of the free energy change. The state-of- the-art molecular dynamics simulation in full atomic scale and biased metadynamics free energy method is used for binding free energy calculations. The well tempered...

How do we know where objects are in the environment and how do we use this information to guide our actions? Recovering the three-dimensional (3D) structure of our surroundings from the two-dimensional retinal input received from the eyes is a computationally challenging task and depends on the brain processing and combining ambiguous sources of sensory information (cues) to depth. This thesis combines psychophysical and computational techniques to gain further insight into (i) which cues the brain uses for perceptual judgments of depth and motion-in-depth; and (ii) the processes underlying the combination of the information from these cues into a single percept of depth. The first chapter deals with the question which sources of information the visual system uses to estimate the time remaining until an approaching object will hit us; a problem that is complicated by the fact that the variable of interest (time) is highly correlated to other perceptual variables that may be used (e.g. distance). Despite these high correlations we show that the visual system recovers a temporal estimate, rather than using one or more of its covariates. In the second chapter I ask how extra-retinal signals (changes in the convergence angles of the eyes) contribute to estimates of 3D speed. Traditionally, extra-retinal signals are reputed to be a poor indicator of 3D motion. Using techniques to isolate extra-retinal signals to changes in vergence, we show that judgments of 3D speed are best explained on the basis that the visual system computes a weighted average of retinal and extra-retinal signals. The third and fourth chapters investigate how the visual system combines binocular and monocular cues to depth in judgments of relative depth and the speed of 3D motion. In chapter three I show that differences in retinal size systematically affect the perceived disparityde defined depth between two unfamiliar targets, so that a target with a larger retinal size is seen as closer than a target with a smaller retinal size at the same disparity-defined distance. This perceptual bias increases as the retinal size ratio between the targets is increased but remains constant as the absolute sizes of the targets change concurrently while keeping the retinal size ratio constant. In addition, bias increases as the absolute distance to both targets increases. I propose that these findings can be explained on the basis that the visual system attempts to optimally combine disparity with retinal size cues (or in the case of 3D motion: changing disparity information with looming cues), but assumes that both objects are of equal size while they are not. In chapter 4 these findings are extended to 3D motion: physically larger unfamiliar targets are reported to approach faster than a smaller target moving at the same speed at the same distance. These findings cannot be explained on the basis of observers' use of a biased perceived distance, caused by differences in the retinal size (as found in chapter 3). I conclude that, in line with contemporary theories of visual perception, the brain solves the puzzle of 3D perception by combining all available sources of visual information in an optimal manner, even though this may lead to inaccuracies in the final estimate of depth.