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.

Contrast sensitivity changes due to glaucoma and normal aging: low-spatial-frequency losses in both magnocellular and parvocellular pathways

PURPOSE: To explore the effects of glaucoma and aging on low-spatial-frequency contrast sensitivity by using tests designed to assess performance of either the magnocellular (M) or parvocellular (P) visual pathways. METHODS: Contrast sensitivity was measured for spatial frequencies of 0.25 to 2 cyc/deg by using a published steady- and pulsed-pedestal approach. Sixteen patients with glaucoma and 16 approximately age-matched control subjects participated. Patients with glaucoma were tested foveally and at two midperipheral locations: (1) an area of early visual field loss, and (2) an area of normal visual field. Control subjects were assessed in matched locations. An additional group of 12 younger control subjects (aged 20-35 years) were also tested. RESULTS: Older control subjects demonstrated reduced sensitivity relative to the younger group for the steady (presumed M)- and pulsed (presumed P)-pedestal conditions. Sensitivity was reduced foveally and in the midperiphery across the spatial frequency range. In the area of early visual field loss, the glaucoma group demonstrated further sensitivity reduction relative to older control subjects across the spatial frequency range for both the steady- and pulsed-pedestal tasks. Sensitivity was also reduced in the midperipheral location of "normal" visual field for the pulsed condition. CONCLUSIONS: Normal aging results in a reduction of contrast sensitivity for the low-spatial-frequency-sensitive components of both the M and P pathways. Glaucoma results in a further reduction of sensitivity that is not selective for M or P function. The low-spatial-frequency-sensitive channels of both pathways, which are presumably mediated by cells with larger receptive fields, are approximately equivalently impaired in early glaucoma.

Low-complexity estimation of CFO and frequency independent I/Q mismatch for OFDM systems

CFO and I/Q mismatch could cause significant performance degradation to OFDM systems. Their estimation and compensation are generally difficult as they are entangled in the received signal. In this paper, we propose some low-complexity estimation and compensation schemes in the receiver, which are robust to various CFO and I/Q mismatch values although the performance is slightly degraded for very small CFO. These schemes consist of three steps: forming a cosine estimator free of I/Q mismatch interference, estimating I/Q mismatch using the estimated cosine value, and forming a sine estimator using samples after I/Q mismatch compensation. These estimators are based on the perception that an estimate of cosine serves much better as the basis for I/Q mismatch estimation than the estimate of CFO derived from the cosine function. Simulation results show that the proposed schemes can improve system performance significantly, and they are robust to CFO and I/Q mismatch.

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.

On feasibility of regional frequency-based emergency control plans

Decentralized and regional load-frequency control of power systems operating in normal and near-normal conditions has been well studied; and several analysis/synthesis approaches have been developed during the last few decades. However in contingency and off-normal conditions, the existing emergency control plans, such as under-frequency load shedding, are usually applied in a centralized structure using a different analysis model. This paper discusses the feasibility of using frequency-based emergency control schemes based on tie-line measurements and local information available within a control area. The conventional load-frequency control model is generalized by considering the dynamics of emergency control/protection schemes and an analytic approach to analyze the regional frequency response under normal and emergency conditions is presented.

Mutation frequency of non-ESBL phenotype SENTRY (Asia-Pacific) isolates of Klebsiella pneumoniae conversion to an ESBL positive phenotype

Extended spectrum β-lactamases or ESBLs, which are derived from non-ESBL precursors by point mutation of β-lactamase genes (bla), are spreading rapidly all over the world and have caused considerable problems in the treatment of infections caused by bacteria which harbour them. The mechanism of this resistance is not fully understood and a better understanding of these mechanisms might significantly impact on choosing proper diagnostic and treatment strategies. Previous work on SHV β-lactamase gene, blaSHV, has shown that only Klebsiella pneumoniae strains which contain plasmid-borne blaSHV are able to mutate to phenotypically ESBL-positive strains and there was also evidence of an increase in blaSHV copy number. Therefore, it was hypothesised that although specific point mutation is essential for acquisition of ESBL activity, it is not yet enough, and blaSHV copy number amplification is also essential for an ESBL-positive phenotype, with homologous recombination being the likely mechanism of blaSHV copy number expansion. In this study, we investigated the mutation rate of non-ESBL expressing K. pneumoniae isolates to an ESBL-positive status by using the MSS-maximum likelihood method. Our data showed that blaSHV mutation rate of a non-ESBL expressing isolate is lower than the mutation rate of the other single base changes on the chromosome, even with a plasmid-borne blaSHV gene. On the other hand, mutation rate from a low MIC ESBL-positive (≤ 8 µg/mL for cefotaxime) to high MIC ESBL-positive (≥16 µg/mL for cefotaxime) is very high. This is because only gene copy number increase is needed which is probably mediated by homologous recombination that typically takes place at a much higher frequencies than point mutations. Using a subinhibitory concentration of novobiocin, as a homologous recombination inhibitor, revealed that this is the case.

A psychosocial approach to understanding young Australians' mobile phone behaviour

This thesis by publication contributes to our knowledge of psychological factors underlying a modern day phenomenon, young people’s mobile phone behaviour. Specifically, the thesis reports a PhD program of research which adopted a social psychological approach to explore mobile phone behaviour among young Australians aged between 15 and 24 years. A particular focus of the research program was to explore both the cognitive and behavioural aspects of young people’s mobile phone behaviour which for the purposes of this thesis is defined as mobile phone involvement. The research program comprised three separate stages which were developmental in nature, in that, the findings of each stage of the research program informed the next. The overarching goal of the program of research was to improve our understanding of the psychosocial factors influencing young people’s mobile phone behaviour. To achieve this overall goal, there were a number of aims to the research program which reflect the developmental nature of this thesis. Given the limited research into the mobile phone behaviour in Australia, the first two aims of the research program were to explore patterns of mobile phone behaviour among Australian youth and explore the social psychological factors relating to their mobile phone behaviour. Following this exploration, the research program sought to develop a measure which captures the cognitive and behavioural aspects of mobile phone behaviour. Finally, the research program aimed to examine and differentiate the psychosocial predictors of young people’s frequency of mobile phone use and their level of involvement with their mobile phone. Both qualitative and quantitative methodologies were used throughout the program of research. Five papers prepared during the three stages of the research program form the bulk of this thesis. The first stage of the research program was a qualitative investigation of young people’s mobile phone behaviour. Thirty-two young Australians participated in a series of focus groups in which they discussed their mobile phone behaviour. Thematic data analysis explored patterns of mobile phone behaviour among young people, developed an understanding of psychological factors influencing their use of mobile phones, and identified that symptoms of addiction were emerging in young people’s mobile phone behaviour. Two papers (Papers 1 and 2) emanated from this first stage of the research program. Paper 1 explored patterns of mobile phone behaviour and revealed that mobile phones were perceived as being highly beneficial to young people’s lives, with the ability to remain in constant contact with others being particularly valued. The paper also identified that symptoms of behavioural addiction including withdrawal, cognitive and behavioural salience, and loss of control, emerged in participants’ descriptions of their mobile phone behaviour. Paper 2 explored how young people’s need to belong and their social identity (two constructs previously unexplored in the context of mobile phone behaviour) related to their mobile phone behaviour. It was revealed that young people use their mobile phones to facilitate social attachments. Additionally, friends and peers influenced young people’s mobile phone behaviour; for example, their choice of mobile phone carrier and their most frequent type of mobile phone use. These papers laid the foundation for the further investigation of addictive patterns of behaviour and the role of social psychological factors on young people’s mobile behaviour throughout the research program. Stage 2 of the research program focussed on developing a new parsimonious measure of mobile phone behaviour, the Mobile Phone Involvement Questionnaire (MPIQ), which captured the cognitive and behavioural aspects of mobile phone use. Additionally, the stage included a preliminary exploration of factors influencing young people’s mobile phone behaviour. Participants (N = 946) completed a questionnaire which included a pool of items assessing symptoms of behavioural addiction, the uses and gratifications relating to mobile phone use, and self-identity and validation from others in the context of mobile phone behaviour. Two papers (Papers 3 & 4) emanated from the second stage of the research program. Paper 3 provided an important link between the qualitative and quantitative components of the research program. Qualitative data from Stage 1 indicated the reasons young people use their mobile phones and identified addictive characteristics present in young people’s mobile phone behaviour. Results of the quantitative study conducted in Stage 2 of the research program revealed the uses and gratifications relating to young people’s mobile phone behaviour and the effect of these gratifications on young people’s frequency of mobile phone use and three indicators of addiction, withdrawal, salience, and loss of control. Three major uses and gratifications: self (such as feeling good or as a fashion item), social (such as contacting friends), and security (such as use in an emergency) were found to underlie much of young people’s mobile phone behaviour. Self and social gratifications predicted young people’s frequency of mobile phone use and the three indicators of addiction but security gratifications did not. These results provided an important foundation for the inclusion of more specific psychosocial predictors in the later stages of the research program. Paper 4 reported the development of the mobile phone involvement questionnaire and a preliminary exploration of the effect of self-identity and validation from others on young people’s mobile phone behaviour. The MPIQ assessed a unitary construct and was a reliable measure amongst this cohort. Results found that self-identity influenced the frequency of young people’s use whereas self-identity and validation from others influenced their level of mobile phone involvement. These findings provided an important indication that, in addition to self factors, other people have a strong influence on young people’s involvement with their mobile phone and that mobile phone involvement is conceptually different to frequency of mobile phone use. Stage 3 of the research program empirically examined the psychosocial predictors of young people’s mobile behaviour and one paper, Paper 5, emanated from this stage. Young people (N = 292) from throughout Australia completed an online survey assessing the role of self-identity, ingroup norm, the need to belong, and self-esteem on their frequency of mobile phone use and their mobile phone involvement. Self-identity was the only psychosocial predictor of young people’s frequency of mobile phone use. In contrast, self-identity, ingroup norm, and need to belong all influenced young people’s level of involvement with their mobile phone. Additionally, the effect of self-esteem on young people’s mobile phone involvement was mediated by their need to belong. These results indicate that young people who perceive their mobile phone to be an integral part of their self-identity, who perceive that mobile phone is common amongst friends and peers, and who have a strong need for attachment to others, in some cases driven by a desire to enhance their self-esteem, are most likely to become highly involved with their mobile phones. Overall, this PhD program of research has provided an important contribution to our understanding of young Australians’ mobile phone behaviour. Results of the program have broadened our knowledge of factors influencing mobile phone behaviour beyond the approaches used in previous research. The use of various social psychological theories combined with a behavioural addiction framework provided a novel examination of young people’s mobile behaviour. In particular, the development of a new measure of mobile phone behaviour in the research program facilitated the differentiation of the psychosocial factors influencing frequency of young people’s mobile phone behaviour and their level of involvement with their mobile phone. Results of the research program indicate the important role that mobile phone behaviour plays in young people’s social development and also signals the characteristics of those people who may become highly involved with their mobile phone. Future research could build on this thesis by exploring whether mobile phones are affecting traditional social psychological processes and whether the results in this research program are generalisable to other cohorts and other communication technologies.

Characteristic analysis for multisampled digital implementation of fixed-switching-frequency closed-loop modulation of voltage-source inverter

In this paper, a fixed-switching-frequency closed-loop modulation of a voltage-source inverter (VSI), upon the digital implementation of the modulation process, is analyzed and characterized. The sampling frequency of the digital processor is considered as an integer multiple of the modulation switching frequency. An expression for the determination of the modulation design parameter is developed for smooth modulation at a fixed switching frequency. The variation of the sampling frequency, switching frequency, and modulation index has been analyzed for the determination of the switching condition under closed loop. It is shown that the switching condition determined based on the continuous-time analysis of the closed-loop modulation will ensure smooth modulation upon the digital implementation of the modulation process. However, the stability properties need to be tested prior to digital implementation as they get deteriorated at smaller sampling frequencies. The closed-loop modulation index needs to be considered maximum while determining the design parameters for smooth modulation. In particular, a detailed analysis has been carried out by varying the control gain in the sliding-mode control of a two-level VSI. The proposed analysis of the closed-loop modulation of the VSI has been verified for the operation of a distribution static compensator. The theoretical results are validated experimentally on both single- and three-phase systems.