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.

Virtual design process : design validation tool schematic approach in car carrier

An important aspect of designing any product is validation. Virtual design process (VDP) is an alternative to hardware prototyping in which analysis of designs can be done without manufacturing physical samples. In recent years, VDP have been generated either for animation or filming applications. This paper proposes a virtual reality design process model on one of the applications when used as a validation tool. This technique is used to generate a complete design guideline and validation tool of product design. To support the design process of a product, a virtual environment and VDP method were developed that supports validation and an initial design cycle performed by a designer. The product model car carrier is used as illustration for which virtual design was generated. The loading and unloading sequence of the model for the prototype was generated using automated reasoning techniques and was completed by interactively animating the product in the virtual environment before complete design was built. By using the VDP process critical issues like loading, unloading, Australian Design rules (ADR) and clearance analysis were done. The process would save time, money in physical sampling and to large extent in complete math generation. Since only schematic models are required, it saves time in math modelling and handling of bigger size assemblies due to complexity of the models. This extension of VDP process for design evaluation is unique and was developed, implemented successfully. In this paper a Toll logistics and J Smith and Sons car carrier which is developed under author’s responsibility has been used to illustrate our approach of generating design validation via VDP.

Discussion Paper : Proposed CDIO Standard 13 - Internationalisation and Mobility

The benefits and growing need for international transparency in engineering qualifications, simple cross-credit processes, international dual awards and mechanisms to encourage student mobility, are receiving much attention around the world at present. In response, there are a number of global initiatives now looking at how these issues may be addressed, particularly in Europe, North America and Australia. CDIO has adopted 12 Standards as guiding principles for program reform and evaluation. The 12 CDIO Standards address program philosophy curriculum development, design-build experiences and workspaces, new methods of teaching and learning, faculty/academic development, and assessment and evaluation. However, none of the Standards address international qualifications nor student mobility. This discussion paper presents the underpinning case for introducing the 13th CDIO Standard, Internationalization and Mobility.

Deposition of combustion aerosols in the human respiratory tract : comparison of theoretical predictions with experimental data

Total deposition of petrol, diesel and environmental tobacco smoke (ETS) aerosols in the human respiratory tract for nasal breathing conditions was computed for 14 nonsmoking volunteers, considering the specific anatomical and respiratory parameters of each volunteer and the specific size distribution for each inhalation experiment. Theoretical predictions were 34.6% for petrol, 24.0% for diesel, and 18.5% for ETS particles. Compared to the experimental results, predicted deposition values were consistently smaller than the measured data (41.4% for petrol, 29.6% for diesel, and 36.2% for ETS particles). The apparent discrepancy between experimental data on total deposition and modeling results may be reconciled by considering the non-spherical shape of the test aerosols by diameter-dependent dynamic shape factors to account for differences between mobility-equivalent and volume-equivalent or thermodynamic diameters. While the application of dynamic shape factors is able to explain the observed differences for petrol and diesel particles, additional mechanisms may be required for ETS particle deposition, such as the size reduction upon inspiration by evaporation of volatile compounds and/or condensation-induced restructuring, and, possibly, electrical charge effects.

Addressing Challenges for Internationalisation and Mobility in Engineering Through CDIO Standards

There is a growing need for international transparency of engineering qualifications, and mechanisms to support and facilitate student mobility. In response, there are a number of global initiatives attempting to address these needs, particularly in Europe, North America and Australia. The Conceive-Design-Implement-Operate (CDIO) Initiative has a set of standards, competencies, and proficiency levels developed through a global community of practice. It is a well-structured framework in which best-practice internationalisation and student mobility can be embedded. However, the current 12 CDIO Standards do not address international qualifications or student mobility. Based on an environmental scan of global activities, the underpinning principles of best practice are identified and form the basis of the proposed 13th CDIO Standard — “Internationalization and Mobility”.

The impact of police speed enforcement practices on self-reported speeding: An exploration of the effects of visibility and mobility

Research has highlighted the relationship between vehicle speed and increased crash risk and severity. Evidence suggests that police speed enforcement, in particular speed camera operations, can be an effective tool for reducing traffic crashes. A quantitative survey of Queensland drivers (n = 852) was conducted to investigate the impact of police speed enforcement methods on self-reported speeding behaviour. Results indicate that visible enforcement was associated with significantly greater self-reported compliance than covert operations irrespective of the mobility of the approach, and the effects on behaviour were longer lasting. The mobility of operations appeared to be moderated the visibility of the approach. Specifically, increased mobility was associated with increase reported compliant behaviour, but only for covert operations, and increased longevity of reported compliant behaviour, but only for overt operations. The perceived effectiveness of various speed enforcement approaches are also analysed across a range of driving scenarios. Results are discussed in light of the small effect sizes. Recommendations for policy and future research are presented.

Walking and sitting in the Australian Antarctic Territory : mobility and imperial space

In 1995 and 1997, two major Australian expeditions travelled to Antarctica. They were the most heavily-reported Antarctican events of their two years: they were charged with the public production of Australian Antarctic spatiality. Both published exploration narratives: Don and Margie McIntyre’s Expedition Icebound generated an illustrated coffee-table book, Two Below Zero: A Year Alone in Antarctica, and the Spirit of Australia South Pole Expedition published its narrative as a video titled Walking on Ice: The History-Making Expedition to the South Pole. Yet, despite the fact that the two polar trips took place during the same period, their spatialities are markedly different. Walking on Ice is a mobile narrative of imperial exploration, while Two Below Zero is a static spatial story of colonial settlement. How polar mobility and relative immobility figure in Australia’s perceptions of, and claim to, nearly half of Antarctica is the focus of this chapter.

Mobility and safety are conflicting goals for transport policy makers when making decisions about graduated driver licensing

Policy decisions are frequently influenced by more than research results alone. This review examines one road safety countermeasure, graduated driver licensing, in three jurisdictions and identifies how the conflict between mobility and safety goals can influence policy decisions relating to this countermeasure. Evaluations from around the world of graduated driver licensing have demonstrated clear reductions in crashes for young drivers. However, the introduction of this countermeasure may be affected, both positively and negatively, by the conflict some policy makers experience between ensuring individuals remain both mobile and safe as drivers. This review highlights how this conflict in policy decision making can serve to either facilitate or hinder the introduction of graduated driver licensing systems. However, policy makers whose focus on mobility is too strong when compared with safety may be mistaken, with evidence suggesting that after a graduated driver licensing system is introduced young drivers adapt their behaviour to the new system and remain mobile. As a result, policy makers should consciously acknowledge the conflict between mobility and safety and consider an appropriate balance in order to introduce these systems. Improvements to the licensing system can then be made in an incremental manner as the balance between these two priorities change. Policy makers can achieve an appropriate balance by using empirical evidence as a basis for their decisions.

Three Carrier Ambiguity Resolutions : Generalised problems, models and solutions

In this paper, the problems of three carrier phase ambiguity resolution (TCAR) and position estimation (PE) are generalized as real time GNSS data processing problems for a continuously observing network on large scale. In order to describe these problems, a general linear equation system is presented to uniform various geometry-free, geometry-based and geometry-constrained TCAR models, along with state transition questions between observation times. With this general formulation, generalized TCAR solutions are given to cover different real time GNSS data processing scenarios, and various simplified integer solutions, such as geometry-free rounding and geometry-based LAMBDA solutions with single and multiple-epoch measurements. In fact, various ambiguity resolution (AR) solutions differ in the floating ambiguity estimation and integer ambiguity search processes, but their theoretical equivalence remains under the same observational systems models and statistical assumptions. TCAR performance benefits as outlined from the data analyses in some recent literatures are reviewed, showing profound implications for the future GNSS development from both technology and application perspectives.

Novel car carrier design : prevention of falls from heights

This article reports the details of a research on novel design in the field of semitrailer sector and discuss design by hazard prevention techniques. The novel design made addresses occupational health and safety (OHS)concerns of fall from heights. The research includes a detailed survey of national data sources to examine the fatalities caused due to fall from heights in car carriers. The study investigates OHS recommendations in Australia for semitrailer sector. Often injuries are caused due to drivers working above the 1.5 meter height for loading, unloading of the cars, moving the decks up, down, strapping the cars, and slipperly. The new design is developed using latest computer aided design and engineeing (CAD, CAE), product data management (PDM), virtual design process (VDP). The new car carrier design excels in reducing the risks of injuries to drivers and new bench mark for OHS standards. The new design has all the decks operated with hydraulics and uses unique ratchet lock mechanism (fool proof design) and loading happens at a safe working height (below 1.5 meter). All the cars are strapped on the safe working height, and then car desks operated hydraulically to transfer them to the required position. This also includes the car on the prime mover, which shuttles across from one deck to other using hydraulic and rack-pinion mechanisms. The novel design car carrier solves the problem of falls from height: next step would be to transfer this technology across other similar effected sectors.