The purpose with this thesis is to investigate whether Sprite Kit is a good tool to simplify the development process for game developers when making 2D games for mobile devices. To answer this question a simple turn based strategy game has been developed with Sprite Kit. Sprite Kit is a game engine for making 2D games released by Apple. Based on the experience I got during the development I will go through and discuss some of the most important tools provided by the game engine and how they helped us to complete our game. The conclusions I reached after making a game with Sprite Kit is that the frame- work provides all the tools necessary for creating a simple 2D mobile game for iOS. Sprite Kit hides much of the lower level details and gives the game de- veloper comprehensive development support. This helps the game developer to save a lot of time and focus more on the gameplay when creating a game.

In der vorliegenden Arbeit wurden die Effekte einer frühzeitigen Zelltherapie im Langzeit in-vivo Infarktmodell studiert. Erstmals wurden dabei auch Veränderungen der kardialen -Adrenozeptoren untersucht und Zelltherapie mit einer reversiblen präinfarziösen Ischämie kombiniert. Initial wurden dafür bei 38 männlichen weißen Neuseeländer Kaninchen Knochenmarkspunktionen durchgeführt, MSC durch Kultur isoliert und 60 Minuten nach induziertem Infarkt und ohne Reperfusion in den Randbereich des Infarktgebietes injiziert. Zur Untersuchung möglicher Interaktionen zwischen Zelltherapie und Präinfarktgeschehen wurde bei einigen Tieren das Myokard durch eine kurzzeitige Präinfarktischämie präkonditioniert. Die Ergebnisse der vorliegenden Arbeit zeigen, dass auch die frühzeitige Zellinjektion ohne Reperfusion mit signifikanten Effekten auf die Kontraktilität und spezifischen sympathoadrenergen Veränderungen verbunden ist.

This thesis introduces three emerging Australian playwrights, Lally Katz, Ben Ellis and Kit Lazaroo, who are interrogating the politics of culture, identity and gender through the application of magic realism to theatre. This thesis contends that magic realist theatre offers a public site for the cultural mediation of binaries: self and other, margin and centre, life and death, western and non-western, pragmatic and spiritual. Australia, because of its history, geographical location and cultural positioning provides a fascinating case study.

Research Doctorate - Doctor of Philosophy (PhD) / Reversible protein phosphorylation plays a central role in the regulation of intracellular signalling, and is controlled by the opposing activities of protein kinases and phosphatases. Deregulation of these mechanisms can result in increased proliferation and enhanced survival, which is a hallmark feature of malignant transformation. For example, over 90% of chronic myeloid leukaemia (CML) patients express the BCR/ABL oncoprotein, which exhibits unrestrained tyrosine kinase activity. In addition, activating mutations within the receptor tyrosine kinase, c-KIT, contribute to the pathogenesis of gastrointestinal stromal tumours (GIST), systemic mastocytosis, acute myeloid leukaemia (AML), testicular seminoma and melanoma. The advent of small molecule tyrosine kinase inhibitors, such as imatinib, has revolutionised the treatment of malignancies driven by these oncogenic kinases. However, a proportion of patients are either unresponsive or develop resistance, and as such, relapse and disease progression is a major clinical problem. In order to improve the treatment outcome for these patients, a greater understanding of the signalling pathways regulated downstream of BCR/ABL and c-KIT is required. The data presented in this thesis indicates that oncogenic BCR/ABL and mutant c-KIT both require inhibition of the tumour suppressor, protein phosphatase 2A (PP2A), to induce tumourigenesis. PP2A is a large family of serine/threonine phosphatases that provide the fine control on signalling pathways by governing the rate and duration of phosphorylation. The heterotrimeric PP2A enzyme is comprised of a structural subunit (PP2A Aα and Aβ), a catalytic subunit (PP2Acα and cβ) and a regulatory subunit, which consists of three unrelated families: B55 (α, β, γ, δ), B56 (α, β, γ, δ, ε) and B" (PR72/130 / PR70/48). Binding of the regulatory subunit to the core PP2A AC dimer directs both the substrate specificity and cellular localisation of the enzyme. The combinatorial assembly of these individual components permits the formation of distinct complexes which have been implicated in numerous cellular functions such as proliferation, survival and mitosis. In particular, important roles for PP2A in various aspects of malignant transformation are beginning to emerge. Recent work demonstrates that PP2A is functionally inactivated by BCR/ABL in myeloid progenitor cells. Using the mouse myeloid progenitor cell line, FDC-P1, these observations were confirmed in the current study. Detailed investigation into the underlying mechanisms have demonstrated for the first time that active BCR/ABL increases the expression of the PP2A structural and certain regulatory subunits. This alters the PP2A holoenzyme composition and results in the abundance of complexes containing B55α and B56α. Consequently, B56γ, a known tumour suppressive subunit, appears to be simultaneously displaced. To investigate which subunits are functionally important for BCR/ABL-mediated leukaemogenesis, individual PP2A subunits were targeted with shRNA sequences in WT BCR/ABL FDC-P1 cells. Subsequent evaluation identified B56α as a key player which facilitates the leukaemic phenotype. In accordance with an increase in PP2A activity, knockdown of B56α significantly inhibited the cellular growth and reduced the clonogenic potential of BCR/ABL⁺ myeloid progenitors. Furthermore, suppression of the B56δ subunit in WT BCR/ABL FDC-P1 cells appears to delay progression through the cell cycle. Together, these findings provide new insights into the biology of PP2A and begin to define the precise mechanisms by which BCR/ABL induces leukaemogenesis via PP2A in CML. Investigation of the regulation of PP2A was also extended to the oncogenic tyrosine kinase, c-KIT. Using FDC-P1 cells expressing imatinib-sensitive (V560G) or –resistant (D816V) mutant c-KIT, this work demonstrates for the first time that constitutive activation of c-KIT impairs the activity of PP2A, and this is essential for tumourigenesis. Pharmacological reactivation of PP2A with FTY720 significantly reduced the proliferation, impaired the clonogenic potential and induced apoptosis of oncogenic c-KIT cells, whilst having no effect on empty vector controls or WT c-KIT cells stimulated with stem cell factor (SCF). These cytotoxic effects of FTY720 are mediated, in part, by the rapid dephosphorylation, and hence inactivation, of oncogenic c-KIT receptors. These promising in vitro findings were translated into an in vivo model, where the daily administration of FTY720 significantly delayed the growth of mutant c-KIT⁺ tumours. Furthermore, FTY720 markedly prevented the infiltration of D816V c-KIT tumour cells into secondary lymphoid organs, such as the spleen and bone marrow. As a result, the survival of FTY720-treated mice was significantly prolonged compared to saline-treated controls. Overall, this body of work greatly enhances our understanding of PP2A function and identifies the complex mechanisms of PP2A regulation by the oncogenic tyrosine kinases, BCR/ABL and c-KIT. Taken together, the data suggests that inhibition of PP2A may represent a general mechanism employed by constitutively active kinases to facilitate tumour growth. As such, this work supports the future application of PP2A-activating agents in a broad range of human malignancies.

Thesis(M.A.)--Auburn University, 2006. / Abstract. Vita. Includes bibliographic references.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 102-116).

Melanoblasts are the embryonic precursors of melanocytes, the pigment producing cells of the skin and hair. Melanoblasts are of key interest to developmental biologists for numerous reasons, including their ability to migrate throughout the body from a single origin in the neural crest (NC). Current methods for the study of the melanocyte lineage are limited by the heavy reliance on animal models. To challenge this, a platform of in vitro tools were designed to replace and complement current studies. A major obstacle is the transition from 2D cultures, which provide only limited behavioural information, to 3D models which are able to recapitulate the environmental conditions. 3D cultures are regularly created using tissue samples and synthetic matrices for attachment, but building a model from cell lines only has not been achieved. A co-culture model using immortalised keratinocyte (COCA) and melanoblast cell lines proved unsuitable for observing developmental processes, due to lack of movement at high cell densities, but may be practical in pigmentation research. Other methods were explored to examine melanoblast behaviour, including the use of cell derived matrices (CDMs) integrated with melanoblast cell lines, and aggregates formed by hanging drop (HD) culture. CDMs were successfully generated from the COCA line, as well as NIH3T3 fibroblasts which has been shown previously. These structures are denuded of cells to leave the deposited extracellular matrix (ECM) components intact, representative of the dermal (fibroblast) and epidermal (keratinocyte) layers of the skin. HDs were prepared from cultured melanoblast cell lines, and form tight aggregates which disseminate when plated, in a manner similar to the dissemination of cells from the NC in explant cultures. The receptor tyrosine kinase KIT and its ligand (KITL), are vital for melanoblast development. Previous study of this signalling complex has often focussed on the haematopoietic lineage and spermatogenesis, where they perform essential roles. KITL is expressed in a membrane localised form found on the surface of keratinocytes thought to promote melanoblast/melanocyte survival, and a soluble isoform found sequestered in the ECM which promotes cell migration. Cell lines expressing fluorescently tagged KIT and KITL were created to visualise their interactions using live-cell confocal imaging. Firstly, cell lines were generated to perform co-culture experiments with KIT and KITL, and we showed that these constructs are able to interact by uptake of KITL into KIT cells. Secondly, tandem fluorescent protein timers of KIT and KITL were generated which were used to observe protein kinetics. We showed that these protein timers can be manipulated using cycloheximide to block protein production, or by increasing ligand availability. These protein timers reveal that soluble KITL (sKITL) has a faster turnover than membrane bound KITL (mKITL), and that in all three proteins, there is distinct change in spatial localisation as the proteins age. Using a novel melanoblast reporter mouse, Pmel-CMN, primary mouse melanoblasts between E12.5 and E14.5 were isolated for RNA sequencing. This time period is the earliest reported for melanoblast isolation for use in gene expression analysis. We show that within this time course, there are significant changes in the RNA expression profiles, including decreasing expression of other NC cell markers, and huge increasing expression of pigmentation genes. To assess the biological relevance of using in vitro assays, cells of the immortalised melanoblast cell line, melb-a, were cultured under different conditions and examined via RNA sequencing. Results reveal differences in several areas between primary cells and those in culture, including loss of melanocyte specificity. The different tools described in this thesis provide a platform on which to study various aspects of cell behaviour, including migration, morphology and cell adhesion at both the individual cell and population levels.

Les mutations FLT3-ITD et KITD816V sont fréquemment retrouvées dans les leucémies aiguës myéloïdes où elles sont associées à un pronostic défavorable. Ces deux récepteurs à activité tyrosine kinase (RTK) mutés sont des acteurs clés de la leucémogenèse régulant la prolifération, la survie et la différenciation cellulaire. L'objectif de ce travail de thèse a été d'étudier la régulation de l'expression protéique de FLT3 en réponse aux inhibiteurs du protéasome, le rôle de l'autophagie dans les LAM KITD816V et l'impact du 2-deoxy-D-glucose sur la localisation intracellulaire des récepteurs. Les travaux réalisés ont démontré trois manières originales de cibler des cellules portant les oncogènes FLT3-ITD ou KIT en jouant sur leur dégradation, leur localisation intracellulaire et l'autophagie. / FLT3-ITD and KITD816V mutations are recurrently found in acute myeloid leukemia, where they are associated with a poor prognosis. These two Tyrosine Kinase Receptors (TKR) are involved in leukemogenesis, regulating proliferation, survival and cell differentiation. The aim of this thesis was to study the regulation of FLT3 protein expression in response to proteasome inhibitors, the role of autophagy in KITD816V-driven AML and the impact of 2-deoxy-D-glucose (2-DG) on the intracellular localization of TKRs. Our studies investigated three original ways to target cells bearing FLT3-ITD or oncogenic KIT mutations playing on their degradation, intracellular localization and autophagy.

In this thesis I aim at a design of the laboratory kit and study ways how to programme CPLD devices made by Altera company. The product is used for development and demonstration of applications in CPLD devices made by Altera company. The kit is designed for Altera programming cables and Presto (made by ASIX). Input signals are implemented by a set of switches and buttons on the board. Output states are displayed by LED diods, possibly connected to multiplex the display. The user can connect to external devices via external inputs. Thesis is also aimed at the design PCB of the laboratory kit, subsequent production, recovery and verification of compatibility ALTERA and PRESTO programmers. End of the thesis aims on working with the Quartus II design environment. In particular, it is a guide to working with templates and simulation of VHDL designs.

The hand thinks while it builds. Only by physically grappling with a material does one truly understand what it wants to become. In c o n t e m p o r a r y l a n d s c a p e architecture, there is typically a separation between the act of designing and the act of making, often causing a lack of practical knowledge of the capabilities of materials and their relationship to one another. To construct expressively means to comprehend a material's physical properties and how its process of production is revealed through repetition and exaggeration. This dissertation attempts to explore the pre-fabrication of landscapes through an iterative process of making by hand with the goal of uncovering material properties that would otherwise remain concealed. It starts with a material exploration on a detailed level, after which the resulting artefact is applied in the larger context of Pretoria. In addition, this exploration attempts to add to the very limited body of wor k c on c e r n i ng l an d s c ap e architectural tectonic theories. / Die hand dink wanneer dit bou. Slegs deur fisies met 'n materiaal te wroeg verstaan mens waarlik wat die materiaal wil word. In eietydse landskapargitektuur is daar tipies 'n verdeling tussen die daad van ontwerp en die daad van maak, wat dikwels lei tot 'n gebrek in praktiese kennis oor die geskiktheid van materiale asook hul verhouding tot mekaar. Uitdruklike konstruksie dui op 'n begrip van 'n materiaal se fisiese eienskappe en die tentoonstelling van sy produksieproses deur repetisie en oordrywing. Hierdie skripsie poog om die voorafvervaardiging van landskappe te verken deur die herhalende proses van maak met die hand, met die doel om materiaalseienskappe te ontdek wat andersins geskuil sou bly. Dit begin met 'n materiaalverkenning op 'n detail vlak, waarna die artefak toegepas word in die groter Pretoria konteks. Verder poog hierdie verkenning om by te dra tot die beperkte kennis van tektoniese teorie in landskapargitektuur. / Mini Dissertation (ML (Prof))--University of Pretoria, 2016. / Architecture / ML (Prof) / Unrestricted