Membrane associated transporter protein gene (SLC45A2) and the genetic basis of normal human pigmentation variation

This work is concerned with the genetic basis of normal human pigmentation variation. Specifically, the role of polymorphisms within the solute carrier family 45 member 2 (SLC45A2 or membrane associated transporter protein; MATP) gene were investigated with respect to variation in hair, skin and eye colour ― both between and within populations. SLC45A2 is an important regulator of melanin production and mutations in the gene underly the most recently identified form of oculocutaneous albinism. There is evidence to suggest that non-synonymous polymorphisms in SLC45A2 are associated with normal pigmentation variation between populations. Therefore, the underlying hypothesis of this thesis is that polymorphisms in SLC45A2 will alter the function or regulation of the protein, thereby altering the important role it plays in melanogenesis and providing a mechanism for normal pigmentation variation. In order to investigate the role that SLC45A2 polymorphisms play in human pigmentation variation, a DNA database was established which collected pigmentation phenotypic information and blood samples of more than 700 individuals. This database was used as the foundation for two association studies outlined in this thesis, the first of which involved genotyping two previously-described non-synonymous polymorphisms, p.Glu272Lys and p.Phe374Leu, in four different population groups. For both polymorphisms, allele frequencies were significantly different between population groups and the 272Lys and 374Leu alleles were strongly associated with black hair, brown eyes and olive skin colour in Caucasians. This was the first report to show that SLC45A2 polymorphisms were associated with normal human intra-population pigmentation variation. The second association study involved genotyping several SLC45A2 promoter polymorphisms to determine if they also played a role in pigmentation variation. Firstly, the transcription start site (TSS), and hence putative proximal promoter region, was identified using 5' RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). Two alternate TSSs were identified and the putative promoter region was screened for novel polymorphisms using denaturing high performance liquid chromatography (dHPLC). A novel duplication (c.–1176_–1174dupAAT) was identified along with other previously described single nucleotide polymorphisms (c.–1721C>G and c.–1169G>A). Strong linkage disequilibrium ensured that all three polymorphisms were associated with skin colour such that the –1721G, +dup and –1169A alleles were associated with olive skin in Caucasians. No linkage disequilibrium was observed between the promoter and coding region polymorphisms, suggesting independent effects. The association analyses were complemented with functional data, showing that the –1721G, +dup and –1169A alleles significantly decreased SLC45A2 transcriptional activity. Based on in silico bioinformatic analysis that showed these alleles remove a microphthalmia-associated transcription factor (MITF) binding site, and that MITF is a known regulator of SLC45A2 (Baxter and Pavan, 2002; Du and Fisher, 2002), it was postulated that SLC45A2 promoter polymorphisms could contribute to the regulation of pigmentation by altering MITF binding affinity. Further characterisation of the SLC45A2 promoter was carried out using luciferase reporter assays to determine the transcriptional activity of different regions of the promoter. Five constructs were designed of increasing length and their promoter activity evaluated. Constitutive promoter activity was observed within the first ~200 bp and promoter activity increased as the construct size increased. The functional impact of the –1721G, +dup and –1169A alleles, which removed a MITF consensus binding site, were assessed using electrophoretic mobility shift assays (EMSA) and expression analysis of genotyped melanoblast and melanocyte cell lines. EMSA results confirmed that the promoter polymorphisms affected DNA-protein binding. Interestingly, however, the protein/s involved were not MITF, or at least MITF was not the protein directly binding to the DNA. In an effort to more thoroughly characterise the functional consequences of SLC45A2 promoter polymorphisms, the mRNA expression levels of SLC45A2 and MITF were determined in melanocyte/melanoblast cell lines. Based on SLC45A2’s role in processing and trafficking TYRP1 from the trans-Golgi network to stage 2 melanosmes, the mRNA expression of TYRP1 was also investigated. Expression results suggested a coordinated expression of pigmentation genes. This thesis has substantially contributed to the field of pigmentation by showing that SLC45A2 polymorphisms not only show allele frequency differences between population groups, but also contribute to normal pigmentation variation within a Caucasian population. In addition, promoter polymorphisms have been shown to have functional consequences for SLC45A2 transcription and the expression of other pigmentation genes. Combined, the data presented in this work supports the notion that SLC45A2 is an important contributor to normal pigmentation variation and should be the target of further research to elucidate its role in determining pigmentation phenotypes. Understanding SLC45A2’s function may lead to the development of therapeutic interventions for oculocutaneous albinism and other disorders of pigmentation. It may also help in our understanding of skin cancer susceptibility and evolutionary adaptation to different UV environments, and contribute to the forensic application of pigmentation phenotype prediction.

The Second Life of urban planning? Using neogeography tools for community engagement

The majority of the world’s citizens now live in cities. Although urban planning can thus be thought of as a field with significant ramifications on the human condition, many practitioners feel that it has reached the crossroads in thought leadership between traditional practice and a new, more participatory and open approach. Conventional ways to engage people in participatory planning exercises are limited in reach and scope. At the same time, socio-cultural trends and technology innovation offer opportunities to re-think the status quo in urban planning. Neogeography introduces tools and services that allow non-geographers to use advanced geographical information systems. Similarly, is there potential for the emergence of a neo-planning paradigm in which urban planning is carried out through active civic engagement aided by Web 2.0 and new media technologies thus redefining the role of practicing planners? This paper traces a number of evolving links between urban planning, neogeography and information and communication technology. Two significant trends – participation and visualisation – with direct implications for urban planning are discussed. Combining advanced participation and visualisation features, the popular virtual reality environment Second Life is then introduced as a test bed to explore a planning workshop and an integrated software event framework to assist narrative generation. We discuss an approach to harness and analyse narratives using virtual reality logging to make transparent how users understand and interpret proposed urban designs.

Designing to maintain human agency in context-aware systems

This paper describes a series of design games, specifically aimed at exploring shifts in human agency, how they are managed, and the impact this will have on the design of future context-aware applications. The games focussed on understanding information handling issues in dental practice with participants from the University of Queensland Dental School playing an active role in the activities. Participatory design activities reveal how technology solution impact on dental practices. By finding methods of representing technological possibilities in ways which can easily be understood we enhance the contribution that dentists can make to the design process.

Designing usable ubiquitous computing

This paper demonstrates that in order to design successful ubiquitous computing, designers must consider concurrently both the end user interactions in the context of use and the sustainability of the technology and its underlying infrastructure. We describe methods used to create more useful collaboration and communication between users, designers and engineers in designing ubiquitous computing systems. We tested these methods in a real domain in an attempt to create a system that is affordable, minimally disrupts the end-user's workplace and improves human-computer interaction.

The proximal first exon architecture of the murine ghrelin gene is highly similar to its human orthologue

BACKGROUND: The murine ghrelin gene (Ghrl), originally sequenced from stomach tissue, contains five exons and a single transcription start site in a short, 19 bp first exon (exon 0). We recently isolated several novel first exons of the human ghrelin gene and found evidence of a complex transcriptional repertoire. In this report, we examined the 5' exons of the murine ghrelin orthologue in a range of tissues using 5' RACE. -----FINDINGS: 5' RACE revealed two transcription start sites (TSSs) in exon 0 and four TSSs in intron 0, which correspond to 5' extensions of exon 1. Using quantitative, real-time RT-PCR (qRT-PCR), we demonstrated that extended exon 1 containing Ghrl transcripts are largely confined to the spleen, adrenal gland, stomach, and skin. -----CONCLUSION: We demonstrate that multiple transcription start sites are present in exon 0 and an extended exon 1 of the murine ghrelin gene, similar to the proximal first exon organisation of its human orthologue. The identification of several transcription start sites in intron 0 of mouse ghrelin (resulting in an extension of exon 1) raises the possibility that developmental-, cell- and tissue-specific Ghrl mRNA species are created by employing alternative promoters and further studies of the murine ghrelin gene are warranted.

A qualitative analysis of local community communications

This paper explores how people communicate in reference to local interests and suggests information and communication technology (ICT) design for enhancement of local community networks. Qualitative data was gathered from participant observations of local community collective action and open interviews with active community members. Data analysis revealed concepts, leading to categories in relation to local interactions and interests. Design suggestions consider introducing people to local community private-strategic activity via public displays that indicate simple entry points to active participation, and creating information collections according to local community perspectives for long-term reference.

Knowledge-Based Urban Development : Planning and Applications in the Information Era

In the globalizing world, knowledge and information (and the social and technological settings for their production and communication) are now seen as keys to economic prosperity. The economy of a knowledge city creates value-added products using research, technology, and brainpower. The social benefit of knowledge-based urban development (KBUD); however, extends beyond aggregate economic growth.

Creative urban regions : harnessing urban technologies to support knowledge city initiatives

In the rapidly growing knowledge economy, the talent and creativity of those around us will be increasingly decisive in shaping economic opportunity. Creativity can be described as the ability to produce new and original ideas and things. In other words, it is any act, idea, or product that changes an existing domain or transforms an existing domain into a new one. From an economic perspective, creativity can be considered as the generation of new ideas that is the major source of innovation and new economic activities. As urban regions have become the localities of key knowledge precincts and knowledge clusters across the globe, the link between a range of new technologies and the development of ‘creative urban regions’ (CURs) has come to the fore. In this sense, creativity has become a buzz concept in knowledge-economy research and policy circles. It has spawned ‘creative milieus,’ ‘creative industries,’ ‘creative cities,’ ‘creative class,’ and ‘creative capital.’ Hence, creativity has become a key concept on the agenda of city managers, development agents, and planners as they search for new forms of urban and economic development. CURs provide vast opportunities for knowledge production and spillover, which lead to the formation of knowledge cities. Urban information and communication technology (ICT) developments support the transformation of cities into knowledge cities. This book, which is a companion volume to Knowledge-Based Urban Development: Planning and Applications in the Information Era (also published by IGI Global) focuses on some of these developments. The Forward and Afterword are written by senior respected academic researchers Robert Stimson of the University of Queensland, Australia, and Zorica Nedovic-Budic of the University of Illinois at Urbana-Champaign, USA. The book is divided into four sections, each one dealing with selected aspects of information and communication technologies and creative urban regions.

Lesser-known worlds : bridging the telematic flows with located human experience through game design

This paper represents a new theorization of the role of location-based games (LBGs) as potentially playing specific roles in peoples’ access to the culture of cities [22]. A LBG is a game that employs mobile technologies as tools for game play in real world environments. We argue that as a new genre in the field of mobile entertainment, research in this area tends to be preoccupied with the newness of the technology and its commercial possibilities. However, this overlooks its potential to contribute to cultural production. We argue that the potential to contribute to cultural production lies in the capacity of these experiences to enhance relationships between specific groups and new urban spaces. Given that developers can design LBGs to be played with everyday devices in everyday environments, what new creative opportunities are available to everyday people?

Maintaining human agency in the design on context-aware systems : Design games in a dental surgery

This paper describes a series of design games, specifically aimed at exploring shifts in human agency in order to inform the design of context-aware applications. The games focused on understanding information handling issues in dental practice with participants from a university dental school playing an active role in the activities. Participatory design activities help participants to reveal potential implicit technical resources that can be presented explicitly in technologies in order to assist humans in managing their interactions with and amidst technical systems gracefully.

Bridging technical and HCI research : creating usable ubiquitous computing

This paper describes methods used to support collaboration and communication between practitioners, designers and engineers when designing ubiquitous computing systems. We tested methods such as “Wizard of Oz” and design games in a real domain, the dental surgery, in an attempt to create a system that is: affordable; minimally disruptive of the natural flow of work; and improves human-computer interaction. In doing so we found that such activities allowed the practitioners to be on a ‘level playing ground’ with designers and engineers. The findings we present suggest that dentists are willing to engage in detailed exploration and constructive critique of technical design possibilities if the design ideas and prototypes are presented in the context of their work practice and are of a resolution and relevance that allow them to jointly explore and question with the design time. This paper is an extension of a short paper submitted to the Participatory Design Conference, 2004.

Public-private partnerships in ICT for education

A recent World Bank report notes that across the world, per capita economic growth is driven by three information and communication technology (ICT)-related factors: investments in equipment and infrastructure, investments in human capital (i.e. in education and innovation), and efficient use of labour (human resource) and capital that increases productivity (Schware 2005). These three factors have a direct impact on the provisioning of education. For one, the demand to adopt ICT-supported education services, or e-education, is outweighing the capacity of governments to adequately support education reform and expansion.

In search of a theory of integrated marketing communication

For the most part, the literature base for Integrated Marketing Communication (IMC) has developed from an applied or tactical level rather than from an intellectual or theoretical one. Since industry, practitioner and even academic studies have provided little insight into what IMC is and how it operates, our approach has been to investigate that other IMC community, that is, the academic or instructional group responsible for disseminating IMC knowledge. We proposed that the people providing course instruction and directing research activities have some basis for how they organize, consider and therefore instruct in the area of IMC. A syllabi analysis of 87 IMC units in six countries investigated the content of the unit, its delivery both physically and conceptually, and defined the audience of the unit. The study failed to discover any type of latent theoretical foundation that might be used as a base for understanding IMC. The students who are being prepared to extend, expand and enhance IMC concepts do not appear to be well-served by the curriculum we found in our research. The study concludes with a model for further IMC curriculum development.