74 resultados para Domain Specific Architecture


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My perspective on the problems associated with building in bushfire prone landscapes comes from 12 years of study of the biophysical and cultural landscapes in the Great Southern Region of WA which resulted in the design and construction of the ‘Hhouse’ at Bremer Bay. The house was developed using a ‘ground up’ approach whereby I conducted a topographical survey and worked with a local botanist and a bushfire risk consultant to ascertain the level of threat that fire presented to this particular site. My intention from the outset however, was not to design a bushfire resistant house per se, but to develop a design which would place the owners in close proximity to the highly biodiverse heath vegetation of the site. I was also seeking a means—through architectural design—of linking the patterns of usage of the house with other site specific conditions related to the prevailing winds, solar orientation and seasonal change.

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The ad hoc networks are vulnerable to attacks due to distributed nature and lack of infrastructure. Intrusion detection systems (IDS) provide audit and monitoring capabilities that offer the local security to a node and help to perceive the specific trust level of other nodes. The clustering protocols can be taken as an additional advantage in these processing constrained networks to collaboratively detect intrusions with less power usage and minimal overhead. Existing clustering protocols are not suitable for intrusion detection purposes, because they are linked with the routes. The route establishment and route renewal affects the clusters and as a consequence, the processing and traffic overhead increases due to instability of clusters. The ad hoc networks are battery and power constraint, and therefore a trusted monitoring node should be available to detect and respond against intrusions in time. This can be achieved only if the clusters are stable for a long period of time. If the clusters are regularly changed due to routes, the intrusion detection will not prove to be effective. Therefore, a generalized clustering algorithm has been proposed that can run on top of any routing protocol and can monitor the intrusions constantly irrespective of the routes. The proposed simplified clustering scheme has been used to detect intrusions, resulting in high detection rates and low processing and memory overhead irrespective of the routes, connections, traffic types and mobility of nodes in the network. Clustering is also useful to detect intrusions collaboratively since an individual node can neither detect the malicious node alone nor it can take action against that node on its own.

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With the advancement of Service-Oriented Architecture in the technical and business domain, the management & engineering of services requires a thorough and systematic understanding of the service lifecycle for both business and software services. However, while service-oriented approaches acknowledge the importance of the service ecosystem, service lifecycle models are typically internally focused, paying limited attention to processes related to offering services to or using services from other actors. In this paper, we address this need by discussing the relations between a comprehensive service lifecycle approach for service management & engineering and the sourcing & purchasing of services. In particular we pay attention to the similarities and differences between sourcing business and software services, the alignment between service management & engineering and sourcing & purchasing, the role of sourcing in the transformation of an organization towards a service-oriented paradigm, the role of architectural approaches to sourcing in this transformation, and the sourcing of specific services at different levels of granularity.

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Prostate cancer is an important male health issue. The strategies used to diagnose and treat prostate cancer underscore the cell and molecular interactions that promote disease progression. Prostate cancer is histologically defined by increasingly undifferentiated tumour cells and therapeutically targeted by androgen ablation. Even as the normal glandular architecture of the adult prostate is lost, prostate cancer cells remain dependent on the androgen receptor (AR) for growth and survival. This project focused on androgen-regulated gene expression, altered cellular differentiation, and the nexus between these two concepts. The AR controls prostate development, homeostasis and cancer progression by regulating the expression of downstream genes. Kallikrein-related serine peptidases are prominent transcriptional targets of AR in the adult prostate. Kallikrein 3 (KLK3), which is commonly referred to as prostate-specific antigen, is the current serum biomarker for prostate cancer. Other kallikreins are potential adjunct biomarkers. As secreted proteases, kallikreins act through enzyme cascades that may modulate the prostate cancer microenvironment. Both as a panel of biomarkers and cascade of proteases, the roles of kallikreins are interconnected. Yet the expression and regulation of different kallikreins in prostate cancer has not been compared. In this study, a spectrum of prostate cell lines was used to evaluate the expression profile of all 15 members of the kallikrein family. A cluster of genes was co-ordinately expressed in androgenresponsive cell lines. This group of kallikreins included KLK2, 3, 4 and 15, which are located adjacent to one another at the centromeric end of the kallikrein locus. KLK14 was also of interest, because it was ubiquitously expressed among the prostate cell lines. Immunohistochemistry showed that these 5 kallikreins are co-expressed in benign and malignant prostate tissue. The androgen-regulated expression of KLK2 and KLK3 is well-characterised, but has not been compared with other kallikreins. Therefore, KLK2, 3, 4, 14 and 15 expression were all measured in time course and dose response experiments with androgens, AR-antagonist treatments, hormone deprivation experiments and cells transfected with AR siRNA. Collectively, these experiments demonstrated that prostatic kallikreins are specifically and directly regulated by the AR. The data also revealed that kallikrein genes are differentially regulated by androgens; KLK2 and KLK3 were strongly up-regulated, KLK4 and KLK15 were modestly up-regulated, and KLK14 was repressed. Notably, KLK14 is located at the telomeric end of the kallikrein locus, far away from the centromeric cluster of kallikreins that are stimulated by androgens. These results show that the expression of KLK2, 3, 4, 14 and 15 is maintained in prostate cancer, but that these genes exhibit different responses to androgens. This makes the kallikrein locus an ideal model to investigate AR signalling. The increasingly dedifferentiated phenotype of aggressive prostate cancer cells is accompanied by the re-expression of signalling molecules that are usually expressed during embryogenesis and foetal tissue development. The Wnt pathway is one developmental cascade that is reactivated in prostate cancer. The canonical Wnt cascade regulates the intracellular levels of β-catenin, a potent transcriptional co-activator of T-cell factor (TCF) transcription factors. Notably, β-catenin can also bind to the AR and synergistically stimulate androgen-mediated gene expression. This is at the expense of typical Wnt/TCF target genes, because the AR:β-catenin and TCF:β-catenin interactions are mutually exclusive. The effect of β-catenin on kallikrein expression was examined to further investigate the role of β-catenin in prostate cancer. Stable knockdown of β-catenin in LNCaP prostate cancer cells attenuated the androgen-regulated expression of KLK2, 3, 4 and 15, but not KLK14. To test whether KLK14 is instead a TCF:β-catenin target gene, the endogenous levels of β-catenin were increased by inhibiting its degradation. Although KLK14 expression was up-regulated by these treatments, siRNA knockdown of β-catenin demonstrated that this effect was independent of β-catenin. These results show that β-catenin is required for maximal expression of KLK2, 3, 4 and 15, but not KLK14. Developmental cells and tumour cells express a similar repertoire of signalling molecules, which means that these different cell types are responsive to one another. Previous reports have shown that stem cells and foetal tissues can reprogram aggressive cancer cells to less aggressive phenotypes by restoring the balance to developmental signalling pathways that are highly dysregulated in cancer. To investigate this phenomenon in prostate cancer, DU145 and PC-3 prostate cancer cells were cultured on matrices pre-conditioned with human embryonic stem cells (hESCs). Soft agar assays showed that prostate cancer cells exposed to hESC conditioned matrices had reduced clonogenicity compared with cells harvested from control matrices. A recent study demonstrated that this effect was partially due to hESC-derived Lefty, an antagonist of Nodal. A member of the transforming growth factor β (TGFβ) superfamily, Nodal regulates embryogenesis and is re-expressed in cancer. The role of Nodal in prostate cancer has not previously been reported. Therefore, the expression and function of the Nodal signalling pathway in prostate cancer was investigated. Western blots confirmed that Nodal is expressed in DU145 and PC-3 cells. Immunohistochemistry revealed greater expression of Nodal in malignant versus benign glands. Notably, the Nodal inhibitor, Lefty, was not expressed at the mRNA level in any prostate cell lines tested. The Nodal signalling pathway is functionally active in prostate cancer cells. Recombinant Nodal treatments triggered downstream phosphorylation of Smad2 in DU145 and LNCaP cells, and stably-transfected Nodal increased the clonogencity of LNCaP cells. Nodal was also found to modulate AR signalling. Nodal reduced the activity of an androgen-regulated KLK3 promoter construct in luciferase assays and attenuated the endogenous expression of AR target genes including prostatic kallikreins. These results demonstrate that Nodal is a novel example of a developmental signalling molecule that is reexpressed in prostate cancer and may have a functional role in prostate cancer progression. In summary, this project clarifies the role of androgens and changing cellular differentiation in prostate cancer by characterising the expression and function of the downstream genes encoding kallikrein-related serine proteases and Nodal. Furthermore, this study emphasises the similarities between prostate cancer and early development, and the crosstalk between developmental signalling pathways and the AR axis. The outcomes of this project also affirm the utility of the kallikrein locus as a model system to monitor tumour progression and the phenotype of prostate cancer cells.

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The function of CUB domain-containing protein 1 (CDCP1), a recently described transmembrane protein expressed on the surface of hematopoietic stem cells and normal and malignant cells of different tissue origin, is not well defined. The contribution of CDCP1 to tumor metastasis was analyzed by using HeLa carcinoma cells overexpressing CDCP1 (HeLa-CDCP1) and a high-disseminating variant of prostate carcinoma PC-3 naturally expressing high levels of CDCP1 (PC3-hi/diss). CDCP1 expression rendered HeLa cells more aggressive in experimental metastasis in immunodeficient mice. Metastatic colonization by HeLa-CDCP1 was effectively inhibited with subtractive immunization-generated, CDCP1-specific monoclonal antibody (mAb) 41-2, suggesting that CDCP1 facilitates relatively late stages of the metastatic cascade. In the chick embryo model, time- and dose-dependent inhibition of HeLa-CDCP1 colonization by mAb 41-2 was analyzed quantitatively to determine when and where CDCP1 functions during metastasis. Quantitative PCR and immunohistochemical analyses indicated that CDCP1 facilitated tumor cell survival soon after vascular arrest. Live cell imaging showed that the function-blocking mechanism of mAb 41-2 involved enhancement of tumor cell apoptosis, confirmed by attenuation of mAb 41-2–mediated effects with the caspase inhibitor z-VAD-fmk. Under proapoptotic conditions in vitro, CDCP1 expression conferred HeLa-CDCP1 cells with resistance to doxorubicin-induced apoptosis, whereas ligation of CDCP1 with mAb 41-2 caused additional enhancement of the apoptotic response. The functional role of naturally expressed CDCP1 was shown by mAb 41-2–mediated inhibition of both experimental and spontaneous metastasis of PC3-hi/diss. These findings confirm that CDCP1 functions as an antiapoptotic molecule and indicate that during metastasis CDCP1 facilitates tumor cell survival likely during or soon after extravasation.

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A line of information and information literacy research has emerged that has a strong focus on information experience. Strengthened understanding, profiling and theorising of information experience as a specific domain of interest to information researchers is required. A focus on information experience is likely to have a major influence on the field, drawing attention to interpretive and experiential forms of research.

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This thesis investigates the radically uncertain formal, business, and industrial environment of current entertainment creators. It researches how a novel communication technology, the Internet, leads to novel entertainment forms, how these lead to novel kinds of businesses that lead to novel industries; and in what way established entertainment forms, businesses, and industries are part of that process. This last aspect is addressed by focusing on one exemplary es-tablished form: movies. Using a transdisciplinary approach and a combination of historical analysis, industry interviews, and an innovative mode of ‘immersive’ textual analysis, a coherent and comprehensive conceptual framework for the creation of and re-search into a specific emerging entertainment form is proposed. That form, products based on it, and the conceptual framework describing it are all re-ferred to as Entertainment Architecture (‘entarch,’ for short). The thesis charac-terises this novel form as Internet-native transmedia entertainment, meaning it fully utilises the unique communicative characteristics of the Internet, and is spread across media. The thesis isolates four constitutive elements within Entertainment Architec-ture: story, play, ‘dance,’ and ‘glue.’ That is, entarch tells a story; offers playful interaction; invites social interaction between producer and consumer, and amongst consumers (‘dance’); and all components of it can be spread across many media, but are so well interconnected and mutually dependent that they are perceived as one product instead of many (‘glue’). This sets entarch apart from current media franchises like Star Wars or Halo, which are perceived as many products spread across many media. Entarch thus embraces the commu-nicative behaviour of Internet-native consumers instead of forcing them to de-sist from it, it harnesses the strengths of various media while avoiding some of their weaknesses, and it can sustain viable businesses. The entarch framework is an innovative contribution to scholarship that al-lows researchers to investigate this emerging entertainment form in a structured way. The thesis demonstrates this by using it to survey business models appro-priate to the entarch environment. The framework can also be used by enter-tainment creators — exemplified in the thesis by moviemakers — to delimit the room for manoeuvre available to them in a changing environment.

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Critical-sized bone defect regeneration is a remaining clinical concern. Numerous scaffold-based strategies are currently being investigated to enable in vivo bone defect healing. However, a deeper understanding of how a scaffold influences the tissue formation process and how this compares to endogenous bone formation or to regular fracture healing is missing. It is hypothesized that the porous scaffold architecture can serve as a guiding substrate to enable the formation of a structured fibrous network as a prerequirement for later bone formation. An ovine, tibial, 30-mm critical-sized defect is used as a model system to better understand the effect of the scaffold architecture on cell organization, fibrous tissue, and mineralized tissue formation mechanisms in vivo. Tissue regeneration patterns within two geometrically distinct macroscopic regions of a specific scaffold design, the scaffold wall and the endosteal cavity, are compared with tissue formation in an empty defect (negative control) and with cortical bone (positive control). Histology, backscattered electron imaging, scanning small-angle X-ray scattering, and nanoindentation are used to assess the morphology of fibrous and mineralized tissue, to measure the average mineral particle thickness and the degree of alignment, and to map the local elastic indentation modulus. The scaffold proves to function as a guiding substrate to the tissue formation process. It enables the arrangement of a structured fibrous tissue across the entire defect, which acts as a secondary supporting network for cells. Mineralization can then initiate along the fibrous network, resulting in bone ingrowth into a critical-sized defect, although not in complete bridging of the defect. The fibrous network morphology, which in turn is guided by the scaffold architecture, influences the microstructure of the newly formed bone. These results allow a deeper understanding of the mode of mineral tissue formation and the way this is influenced by the scaffold architecture. Copyright © 2012 American Society for Bone and Mineral Research.

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Tissue-specific extracellular matrix (ECM) is known to be an ideal bioscaffold to inspire the future of regenerative medicine. It holds the secret of how nature has developed such an organization of molecules into a unique functional complexity. This work exploited an innovative image processing algorithm and high resolution microscopy associated with mechanical analysis to establish a correlation between the gradient organization of cartiligous ECM and its anisotropic biomechanical response. This was hypothesized to be a reliable determinant that can elucidate how microarchitecture interrelates with biomechanical properties. Hough-Radon transform of the ECM cross-section images revealed its conformational variation from tangential interface down to subchondral region. As the orientation varied layer by layer, the anisotropic mechanical response deviated relatively. Although, results were in good agreement (Kendall's tau-b > 90%), there were evidences proposing that alignment of the fibrous network, specifically in middle zone, is not as random as it was previously thought.

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Conference curatorial outline The focus of this symposium was to question whether interior design is changing relative to local conditions, and the effect globalization has on the performance of regional, particularly Southern hemisphere identities. The intention being to understand how theory and practice is transposed to ‘distant lands’, and how ideas shift from one place to another. To this extent the symposium invited papers on the export, translation and adoption of theories and practices of interior design to differing climates, cultures, and landscapes. This process, sometimes referred to as a shift from ‘the centre to the margins’, seeks new perspectives on the adoption of European and US design ideas abroad, as well as their return to their place of origin. Papers were invited from a range of perspectives including the export of ideas/attitudes to interior spaces, history of interior spaces abroad, and the adoption of ideas/processes to new conditions. Paralleling this trafficking of ideas are broader observations about interior space that emerge through specificity of place. These include new and emerging directions and differences in our understanding of interiority; both real and virtual, and an ever-changing relationship to city, suburb and country. Keeping within the Symposium theme the intention was to examine other places, particularly on the margins of the discipline’s domain. Semantic slippage aside, there are a range of approaches that engage outside events and practices enabling a transdisciplinary practice that draws from other philosophical and theoretical frameworks. Moreover as the field expands and new territories are opened up, the virtual worlds of computer gaming, animations, and interactive environments, both rely on and produce new forms of expression. This raises questions about the extent such spaces adopt or translate existing theory and practice, that is the transposition from one area to another and their return to the discipline.

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In the face of Australia’s disaster-prone environment, architects Ian Weir and James Davidson are reconceptualising how our residential buildings might become more resilient to fire, flood and cyclone. With their first-hand experience of natural disasters, James, director of Emergency Architects Australia (EAA), and Ian, one of Australia’s few ‘bushfire architects’, discuss the ways we can design with disaster in mind. Dr Ian Weir is one of Australia’s few ‘bushfire architects’. Exploring a holistic ‘ground up’ approach to bushfire where landscape, building design and habitation patterns are orchestrated to respond to site-specific fire characteristics. Ian’s research is developed through design studio teaching at QUT and through built works in Western Australia’s fire prone forests and heathlands.

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RESEARCH BACKGROUND Enacted Cartography documents 10 years of creative research practice by Ian Weir Research Architect and was developed as standalone exhibition to support Dr Weir’s selection by the Australian Institute of Architects to represent innovative architectural practice via the Institute’s review entitled Formations: New Practices in Australian Architecture – which took the form of an exhibition and book presented in Venice, Italy for 13th International Architecture Exhibition (Venice Architecture Biennale). All works exhibited in Enacted Cartography are original works by Dr Weir and are generated either from or for the remote biodiverse landscapes of the Fitzgerald Bioregion on the south coast of Western Australia. RESEARCH CONTRIBUTION As a creative work in its own right, the Enacted Cartography exhibition makes the following contributions to knowledge: 1. Expands understandings of architectural practice by presenting a geographically-specific but multimodal form of architectural practice - wherein practitioners cross over discipline boundaries into art practice, landscape representation, website design, undergraduate university teaching and community advocacy. 2. Contributes to understandings of how such a diverse multimodal form of practice might be represented through both digital media and traditional print media in an exhibition format. 3. Expands understandings of how architectural practitioners might work within a particular place to develop a geographically-specific sense of identity, a ‘landscape of resistance’. RESEARCH SIGNIFICANCE Enacted Cartography was presented to an international audience during the 13th International Architecture Exhibition (Venice Architecture Biennale). The significance of Dr Weir’s research is evidence by his selected by the Australian Institute of Architects to represent innovation in architectural practice for the Biennale. Enacted Cartography addresses problems of national and international importance including: 1. The sustainable development of biodiverse remote landscapes; 2. The reconciliation of bushfire safety and biodiversity conservation; 3. The necessity for rethinking of architectural design methodologies to meet the complexity of landscape management and design; 4. It challenges orthodox forms of landscape representation (aerial photography, for example) which are demonstrably inadequate registrations of biophysical and cultural landscapes.

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Recent advances in the area of ‘Transformational Government’ position the citizen at the centre of focus. This paradigm shift from a department-centric to a citizen-centric focus requires governments to re-think their approach to service delivery, thereby decreasing costs and increasing citizen satisfaction. The introduction of franchises as a virtual business layer between the departments and their citizens is intended to provide a solution. Franchises are structured to address the needs of citizens independent of internal departmental structures. For delivering services online, governments pursue the development of a One-Stop Portal, which structures information and services through those franchises. Thus, each franchise can be mapped to a specific service bundle, which groups together services that are deemed to be of relevance to a specific citizen need. This study focuses on the development and evaluation of these service bundles. In particular, two research questions guide the line of investigation of this study: Research Question 1): What methods can be used by governments to identify service bundles as part of governmental One-Stop Portals? Research Question 2): How can the quality of service bundles in governmental One-Stop Portals be evaluated? The first research question asks about the identification of suitable service bundle identification methods. A literature review was conducted, to, initially, conceptualise the service bundling task, in general. As a consequence, a 4-layer model of service bundling and a morphological box were created, detailing characteristics that are of relevance when identifying service bundles. Furthermore, a literature review of Decision-Support Systems was conducted to identify approaches of relevance in different bundling scenarios. These initial findings were complemented by targeted studies of multiple leading governments in the e-government domain, as well as with a local expert in the field. Here, the aim was to identify the current status of online service delivery and service bundling in practice. These findings led to the conceptualising of two service bundle identification methods, applicable in the context of Queensland Government: On the one hand, a provider-driven approach, based on service description languages, attributes, and relationships between services was conceptualised. As well, a citizen-driven approach, based on analysing the outcomes from content identification and grouping workshops with citizens, was also conceptualised. Both methods were then applied and evaluated in practice. The conceptualisation of the provider-driven method for service bundling required the initial specification of relevant attributes that could be used to identify similarities between services called relationships; these relationships then formed the basis for the identification of service bundles. This study conceptualised and defined seven relationships, namely ‘Co-location’, ‘Resource’, ‘Co-occurrence’, ‘Event’, ‘Consumer’, ‘Provider’, and ‘Type’. The relationships, and the bundling method itself, were applied and refined as part of six Action Research cycles in collaboration with the Queensland Government. The findings show that attributes and relationships can be used effectively as a means for bundle identification, if distinct decision rules are in place to prescribe how services are to be identified. For the conceptualisation of the citizen-driven method, insights from the case studies led to the decision to involve citizens, through card sorting activities. Based on an initial list of services, relevant for a certain franchise, participating citizens grouped services according to their liking. The card sorting activity, as well as the required analysis and aggregation of the individual card sorting results, was analysed in depth as part of this study. A framework was developed that can be used as a decision-support tool to assist with the decision of what card sorting analysis method should be utilised in a given scenario. The characteristic features associated with card sorting in a government context led to the decision to utilise statistical analysis approaches, such as cluster analysis and factor analysis, to aggregate card sorting results. The second research question asks how the quality of service bundles can be assessed. An extensive literature review was conducted focussing on bundle, portal, and e-service quality. It was found that different studies use different constructs, terminology, and units of analysis, which makes comparing these models a difficult task. As a direct result, a framework was conceptualised, that can be used to position past and future studies in this research domain. Complementing the literature review, interviews conducted as part of the case studies with leaders in e-government, indicated that, typically, satisfaction is evaluated for the overall portal once the portal is online, but quality tests are not conducted during the development phase. Consequently, a research model which appropriately defines perceived service bundle quality would need to be developed from scratch. Based on existing theory, such as Theory of Reasoned Action, Expectation Confirmation Theory, and Theory of Affordances, perceived service bundle quality was defined as an inferential belief. Perceived service bundle quality was positioned within the nomological net of services. Based on the literature analysis on quality, and on the subsequent work of a focus group, the hypothesised antecedents (descriptive beliefs) of the construct and the associated question items were defined and the research model conceptualised. The model was then tested, refined, and finally validated during six Action Research cycles. Results show no significant difference in higher quality or higher satisfaction among users for either the provider-driven method or for the citizen-driven method. The decision on which method to choose, it was found, should be based on contextual factors, such as objectives, resources, and the need for visibility. The constructs of the bundle quality model were examined. While the quality of bundles identified through the citizen-centric approach could be explained through the constructs ‘Navigation’, ‘Ease of Understanding’, and ‘Organisation’, bundles identified through the provider-driven approach could be explained solely through the constructs ‘Navigation’ and ‘Ease of Understanding’. An active labelling style for bundles, as part of the provider-driven Information Architecture, had a larger impact on ‘Quality’ than the topical labelling style used in the citizen-centric Information Architecture. However, ‘Organisation’, reflecting the internal, logical structure of the Information Architecture, was a significant factor impacting on ‘Quality’ only in the citizen-driven Information Architecture. Hence, it was concluded that active labelling can compensate for a lack of logical structure. Further studies are needed to further test this conjecture. Such studies may involve building alternative models and conducting additional empirical research (e.g. use of an active labelling style for the citizen-driven Information Architecture). This thesis contributes to the body of knowledge in several ways. Firstly, it presents an empirically validated model of the factors explaining and predicting a citizen’s perception of service bundle quality. Secondly, it provides two alternative methods that can be used by governments to identify service bundles in structuring the content of a One-Stop Portal. Thirdly, this thesis provides a detailed narrative to suggest how the recent paradigm shift in the public domain, towards a citizen-centric focus, can be pursued by governments; the research methodology followed by this study can serve as an exemplar for governments seeking to achieve a citizen-centric approach to service delivery.

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This study aims to redefine spaces of learning to places of learning through the direct engagement of local communities as a way to examine and learn from real world issues in the city. This paper exemplifies Smart City Learning, where the key goal is to promote the generation and exchange of urban design ideas for the future development of South Bank, in Brisbane, Australia, informing the creation of new design policies responding to the needs of local citizens. Specific to this project was the implementation of urban informatics techniques and approaches to promote innovative engagement strategies. Architecture and Urban Design students were encouraged to review and appropriate real-time, ubiquitous technology, social media, and mobile devices that were used by urban residents to augment and mediate the physical and digital layers of urban infrastructures. Our study’s experience found that urban informatics provide an innovative opportunity to enrich students’ place of learning within the city.

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Chlamydia trachomatis is a bacterial pathogen responsible for one of the most prevalent sexually transmitted infections worldwide. Its unique development cycle has limited our understanding of its pathogenic mechanisms. However, CtHtrA has recently been identified as a potential C. trachomatis virulence factor. CtHtrA is a tightly regulated quality control protein with a monomeric structural unit comprised of a chymotrypsin-like protease domain and two PDZ domains. Activation of proteolytic activity relies on the C-terminus of the substrate allosterically binding to the PDZ1 domain, which triggers subsequent conformational change and oligomerization of the protein into 24-mers enabling proteolysis. This activation is mediated by a cascade of precise structural arrangements, but the specific CtHtrA residues and structural elements required to facilitate activation are unknown. Using in vitro analysis guided by homology modeling, we show that the mutation of residues Arg362 and Arg224, predicted to disrupt the interaction between the CtHtrA PDZ1 domain and loop L3, and between loop L3 and loop LD, respectively, are critical for the activation of proteolytic activity. We also demonstrate that mutation to residues Arg299 and Lys160, predicted to disrupt PDZ1 domain interactions with protease loop LC and strand β5, are also able to influence proteolysis, implying their involvement in the CtHtrA mechanism of activation. This is the first investigation of protease loop LC and strand β5 with respect to their potential interactions with the PDZ1 domain. Given their high level of conservation in bacterial HtrA, these structural elements may be equally significant in the activation mechanism of DegP and other HtrA family members.