Identification and characterisation of novel genes involved in skeletal muscle hypertrophy

There is mounting evidence in support of the view that skeletal muscle hypertrophy results from the complex and coordinated interaction of numerous signalling pathways. Well characterised components integral to skeletal muscle adaptation include the transcriptional activity of the members of the myogenic regulatory factors, numerous secreted peptide growth factors, and the regenerative potential of satellite cells. Whilst studies investigating isolated components or pathways have enhanced our current understanding of skeletal muscle hypertrophy, our knowledge of how all of these components react in concert to a common stimulus remains limited. The broad aim of this thesis was to identify and characterise novel genes involved in skeletal muscle hypertrophy. We have created a customised human skeletal muscle specific microarray which contains ∼11,000 cDNA clones derived from a normalised human skeletal muscle cDNA library as well as 270 genes with known functional roles in human skeletal muscle. The first aspect of this thesis describes the production of the microarray and evaluates the robustness and reproducibility of this analytical technique. Study one aimed to use this microarray in the identification of genes that are differentially expressed during the forced differentiation of human rhabdomyosarcoma cells, an in vitro model of skeletal muscle development. Firstly using this unique model of aberrant myogenic differentiation we aimed to identify genes with previously unidentified roles in myogenesis. Secondly, the data from this study permitted the examination of the performance of the microarray in detecting differential gene expression in a biological system. We identified several new genes with potential roles in the myogenic arrest of rhabdomyosarcoma and further characterised the expression of muscle specific genes in rhabdomyosarcoma differentiation. In study two, the molecular responses of cell cycle regulators, muscle regulatory factors, and atrophy related genes were mapped in response to a single bout of resistance exercise in human skeletal muscle. We demonstrated an increased expression of MyoD, myogenin and p21, whilst the expression of myostatin was decreased. The results of this study contribute to the existing body of knowledge on the molecular regulation skeletal muscle to a hypertrophic stimulus. In study three, the muscle samples collected in study two were analysed using the human skeletal muscle specific microarray for the identification of novel genes with potential roles in the hypertrophic process. The analysis uncovered four interesting genes (TXNIP, MLP, ASB5, FLJ 38973) that have not previously been examined in human skeletal muscle in response to resistance exercise. The functions of these genes and their potential roles in skeletal muscle are discussed. In study four, the four genes identified in study three were examined in human primary skeletal muscle cell cultures during myogenic differentiation. Human primary skeletal muscle cells were derived from the vastus lateralis muscle of 8 healthy volunteers (6 males and 2 females). Cell cultures were differentiated using serum withdrawal and serum withdrawal combined with IGF-1 supplementation. Markers of the cell proliferation, cell cycle arrest and myogenic differentiation were examined to assess the effectiveness of the differentiation stimulus. Additionally, the expressions of TXNIP, MLP, ASB5 and FLJ 38973 measured in an attempt to characterise further their roles in skeletal muscle. The expression of TXNIP changed markedly in response to both differentiation stimuli, whilst the expression of the remaining genes were not altered. Therefore it was suggested that expression of these genes might be responsive to the mechanical strain or contraction induced by the resistance exercise. In order to examine whether these novel genes responded specifically to resistance type exercise, their expression was examined following a single bout of endurance exercise. The expression of TXNIP, MLP, and FLJ 38973 remained unchanged whilst ASB5 increased 30 min following the cessation of the exercise.

User participation in the performance assessment of school buildings : a study of six primary schools in the Bellarine Peninsula region

Young children spend a significant portion of their lives at primary school. This research traces the development of school facilities in Victoria and examines the performance of six primary schools from the users' perspective. Performance assessments were carried out using participatory evaluation methods that included Touring Interviews with small groups of students aged from six to twelve years. The study found that participatory evaluation methods with both student and staff users generate significant information to improve school facilities. User comments were analysed with respect to 14 aspects of building quality and serviceability including character, thermal environment, privacy and flexibility. The study concludes that school buildings do not meet a number of key user requirements. Children expressed dissatisfaction with furniture and equipment in their classrooms and the playground, and to a lesser extent with student toilets, security of their school bags and personal privacy. Staff were dissatisfied with the provision of withdrawal areas and specialist spaces. Department of School Education facilities guidelines do not address the concerns of school users or meet user needs.

Clinical, ethical and legal considerations in the treatment of newborns with non-ketotic hyperglycinaemia

Non-ketotic hyperglycinaemia (NKH) is a devastating neurometabolic disorder leading, in its classical form, to early death or severe disability and poor quality of life in survivors. Affected neonates may need ventilatory support during a short period of respiratory depression. The transient dependence on ventilation dictates urgency in decision-making regarding withdrawal of therapy. The occurrence of patients with apparent transient forms of the disease, albeit rare, adds uncertainty to the prediction of clinical outcome and dictates that the current practice of withholding or withdrawing therapy in these neonates be reviewed. Both bioethics and law take the view that treatment decisions should be based on the best interests of the patient. The medical-ethics approach is based on the principles of non-maleficence, beneficence, autonomy and justice. The law relating to withholding or withdrawing life-sustaining treatment is complex and varies between jurisdictions. Physicians treating newborns with NKH need to provide families with accurate and complete information regarding the disease and the relative probability of possible outcomes of the neonatal presentation and to explore the extent to which family members are willing to take part in the decision making process. Cultural and religious attitudes, which may potentially clash with bioethical and juridical principles, need to be considered.

Social recognition and freedom in Axel Honneth's critical theory

The thesis examines Honneth's claim that forms of misrecognition cause undue suffering and considers the extent to which we are vulnerable to and independent of misrecognition. It formulates a sixfold classification of our responses to misrecognition - stoicism, withdrawal, conformity, reification, deconstruction, and humanism, - and thereby an alternative ethics of recognition.

Rethinking the region

This thesis explores the shift in focus of Australian regional defence and foreign policy away from Malaysia and towards Indonesia between 1965 and 1971. It finds that this shift was caused more by the consequences of Britain's military withdrawal from Malaysia and Singapore than by Suharto's accession to power.

Exercise dependence and elite Australian athletes

This thesis provides an examination of exercise dependence (EXD) among elite Australian athletes. The research identifies the main symptoms of EXD among elite athletes as excessive training, withdrawal, and continuance behaviours. EXD was found to be associated with irrational beliefs relating to training, low self esteem, and limited social support. The portfolio presents four case studies illustrating the use of solution focused brief counselling with athletes and drawing attention to the utility of a solution focused approach as an effective tool in sport psychology consultancy.

Cultural connections across time and place : Wardell's late 19th century Genazzano FCJ College Kew

The Catholic Church was profoundly affected by the 1872 Victorian Education Act, which made education secular, compulsory and free, and led to the withdrawal of state aid to religious schools. In order for the Church to run its own schools, it had to look overseas for help and invited religious teaching orders, such as the Faithful Companions of Jesus (FCJs) to set up schools in Victoria, Australia. In many instances purpose built buildings were designed by architects. William Wardell was well established in private practice in Sydney when he designed the new Convent and School, Kew, Victoria, for the FCJ Sisters, in the late 1880s. Building commenced just before the crash of Marvellous Melbourne. Less than half of the total concept of Wardell’s original plan was built. It opened for business in April 1891. Today this building forms the heart of the contemporary Genazzano FCJ College Kew. Many histories intersect in this commission. The vision for Catholic education in Victoria in the late 19th century is critical. The FCJs charism and their experience of teaching in Europe, in France, England, Ireland, Italy and Switzerland, provides a model for their work in Australia. At this time the importance of architecture to society is made manifest in education and its demands on building: if learning is valued then buildings should reflect this, for public buildings can shape morality. Wardell was trained as a Gothic Revival architect and his building participates in a broader medieval and Gothic tradition. Wardell’s original plan for this late Victorian Gothic style asymmetrical three-storeyed building, was designed to integrate a convent, school, chapel, and dormitories. This paper considers architectural history from diverse perspectives, educational, social, religious, economic and political, recognising the complexity of this project and the people who played a part in its conception and realisation.

The transnational corporation and new corporate citizenship theory : a critical analysis

A recent conceptualisation of corporate citizenship by Matten and Crane (2005) shifts focus onto the corporation's role in providing individuals with the rights they are entitled to as citizens. This expanded corporate role is depicted as filling an institutional vacuum resulting from the withdrawal of the state. Marking an innovation to the corporate citizenship literature, we devise a three-part analytical framework from political institutionalism to question the concept's ideological and empirical groundings. Incorporating a constrained game theory perspective, we use an example of the provision of Western corporate services by low-labour-cost nation-states to argue that the concept as strategy would in some circumstances exacerbate the implications of globalisation on individual citizenship rights. The analytical framework has application for research directed toward proposals to extend the reach of corporations in traditional public services and, more generally, for studies of corporate responsibilities. Future research on corporate citizenship would be strengthened in recognising, as we do, institutional incentives, constraints, decision-making modes and resources as used by the transnational corporation.

Dopamine dysregulation syndrome : implications for a dopamine hypothesis of bipolar disorder

Objective: Rational therapeutic development in bipolar is hampered by a lack of pathophysiological model. However, there is a wealth of converging data on the role of dopamine in bipolar disorder. This paper therefore examines the possibility of a dopamine hypothesis for bipolar disorder.

Method: A literature search was conducted using standard search engines Embase, PyschLIT, PubMed and MEDLINE. In addition, papers and book chapters known to the authors were retrieved and examined for further relevant articles.

Results: Collectively, in excess of 100 articles were reviewed from which approximately 75% were relevant to the focus of this paper.

Conclusion: Pharmacological models suggest a role of increased dopaminergic drive in mania and the converse in depression. In Parkinson’s disease, administration of high-dose dopamine precursors can produce a ‘maniform’ picture, which switches into a depressive analogue on withdrawal. It is possible that in bipolar disorder there is a cyclical process, where increased dopaminergic transmission in mania leads to a secondary down regulation of dopaminergic receptor sensitivity over time. This may lead to a period of decreased dopaminergic transmission, corresponding with the depressive phase, and the repetition of the cycle. This model, if verified, may have implications for rational drug development.

Variation in the gene coding for the M5 muscarinic receptor (CHRM5) influences cigarette dose but is not associated with dependence to drugs of addiction : evidence from a prospective population based cohort study of young adults

Background: The mesolimbic structures of the brain are important in the anticipation and perception of reward. Moreover, many drugs of addiction elicit their response in these structures. The M5 muscarinic receptor (M5R) is expressed in dopamine-containing neurones of the substantia nigra pars compacta and ventral tegmental area, and regulates the release of mesolimbic dopamine. Mice lacking M5R show a substantial reduction in both reward and withdrawal responses to morphine and cocaine. The CHRM5, the gene that codes for the M5R, is a strong biological candidate for a role in human addiction. We screened the coding and core promoter sequences of CHRM5 using denaturing high performance liquid chromatography to identify common polymorphisms. Additional polymorphisms within the coding and core promoter regions that were identified through dbSNP were validated in the test population. We investigated whether these polymorphisms influence substance dependence and dose in a cohort of 1947 young Australians.

Results: Analysis was performed on 815 participants of European ancestry who were interviewed at wave 8 of the cohort study and provided DNA. We observed a 26.8% increase in cigarette consumption in carriers of the rs7162140 T-allele, equating to 20.1 cigarettes per week (p=0.01). Carriers of the rs7162140 T-allele were also found to have nearly a 3-fold increased risk of developing cannabis dependence (OR=2.9 (95%CI 1.1-7.4); p=0.03).

Conclusion: Our data suggest that variation within the CHRM5 locus may play an important role in tobacco and cannabis but not alcohol addiction in European ancestry populations. This is the first study to show an association between CHRM5 and substance use in humans. These data support the further investigation of this gene as a risk factor in substance use and dependence.

Ability of predator odour exposure to elicit conditioned versus sensitised post traumatic stress disorder-like behaviours, and forebrain ΔFosB expression, in rats

At present, exposure of a rodent to the odour of a predator is one of the most common animal models of post traumatic stress disorder (PTSD). Despite this, the model remains incompletely characterized, particularly in regard to within subject assessment of major PTSD-like behaviours. In an attempt to redress this situation, we have extensively characterized the two broad categories of behaviour that are considered to characterize PTSD, that is sensitized behaviours such as social withdrawal and hypervigilance and conditioned behaviours such as avoidance of trauma linked cues. Specifically, we determined the presence and duration of both conditioned and sensitized behaviours, in the same cohort of animals, after three exposures to predator odour. Conditioned fear was assessed on the basis of inhibition of locomotor activity upon return to context 2, 7, 14, 21, and 28 days after the last odour exposure session. To assess the impact on sensitization behaviours, we monitored acoustic startle responses and social interaction behaviour 4, 9, 16, 23, and 30 days after the last exposure session. In addition to examining the behavioural consequences associated with odour exposure, we also determined the key brain regions that were activated using ΔFosB immunohistochemistry. Our results show that the two groups of behaviours thought to characterize PTSD (conditioned and sensitized) do not travel together in the predator odour model, with clear evidence of enduring changes in conditioned fear but little evidence of changes in social interaction or acoustic startle. With regard to associated patterns of activity in the brain, we observed that odour-exposed animals exhibited significantly higher numbers of FosB-positive nuclei in only the medial prefrontal cortex (mPFC), a finding that can be viewed as being consistent with the observed behavioural changes.

Down under and in between : Australian security perspectives in the 'Asian Century'

This chapter will provide an overview of Australian perspectives on the US alliance in light of ongoing and emerging challenges in the Asia-Pacific region. After a brief discussion of the motivations behind the signing of the ANZUS treaty, the first part of the chapter examines the historical context of the alliance, with a particular focus on the longstanding and ongoing tussle in Australia between independence in foreign policy making vis-à-vis broader structural constraints. While this debate has been a constant feature of the political scene in Australia, it has come into particular focus since the US withdrawal from Vietnam, which marked a turning point in Australian perspectives with regard to its own role in Asia. The collision of ideas surrounding Australian identity and Australian national interest has been reflected in policy approaches as successive governments have sought to strike a balance between the two exigencies and thus, most optimally ensure Australia’s strategic future. The chapter concludes by examining current perspectives through the lens of an ongoing debate taking place in Australian academic circles about what the rise of China means for Australia and its commitment to the US alliance, and considers options for caucus-style cooperation with fellow US allies beyond the hub-and-spokes model.

Experiencing turbulence: Asia in the Australian imaginary

Over the last century and a half the competing merits of withdrawal from and connection to Asia-and the related vocabularies of separation and engagement-have been defining themes in Australian history. Experiencing Turbulence brings together a selection of publications on Australian representation of Asia, published in various journals and books in the last ten years that follow the publication of Anxious Nation: Australia and the Rise of Asia, 1850 to 1939. Collectively they address key themes in the Australian response to Asia: survivalist anxieties, climate and race, population and immigration, empty Australia, gender and bush mythologies, and regional Identities. These essays reveal the central, often constitutive role that Asia has played in the formation of ideas of nation and identity in Australia from the late nineteenth century to the present. The collection underlines the often unpredictable character of engagement and the fluid nature of fear and fascination, proximity and distance in the Australia-Asia relationship. With the recent publication of a government White Paper on Australia in the Asian Century there is a new determination to persuade Australians that "rising Asia," turbulent though it may be, is an opportunity for Australia more than it is a threat.

Single-subject research designs and data analyses for assessing elite athletes’ conditioning

Research in conditioning (all the processes of preparation for competition) has used group research designs, where multiple athletes are observed at one or more points in time. However, empirical reports of large inter-individual differences in response to conditioning regimens suggest that applied conditioning research would greatly benefit from single-subject research designs. Single-subject research designs allow us to find out the extent to which a specific conditioning regimen works for a specific athlete, as opposed to the average athlete, who is the focal point of group research designs. The aim of the following review is to outline the strategies and procedures of single-subject research as they pertain to the assessment of conditioning for individual athletes. The four main experimental designs in single-subject research are: the AB design, reversal (withdrawal) designs and their extensions, multiple baseline designs and alternating treatment designs. Visual and statistical analyses commonly used to analyse single-subject data, and advantages and limitations are discussed. Modelling of multivariate single-subject data using techniques such as dynamic factor analysis and structural equation modelling may identify individualised models of conditioning leading to better prediction of performance. Despite problems associated with data analyses in single-subject research (e.g. serial dependency), sports scientists should use single-subject research designs in applied conditioning research to understand how well an intervention (e.g. a training method) works and to predict performance for a particular athlete.

Wave of bombings across Iraq ahead of elections

The people of Iraq have also endured bombings but on a larger scale. There are media reports saying up to thirty-one people were killed in dozens of separate explosions across different Iraqi provinces including two blasts at a checkpoint at Baghdad international airport.

This comes just days before provincial elections across Iraq, the first ballot since the withdrawal of US troops at the end of 2011. Although it is unclear who carried out the attacks different factions are increasingly stoking the fire as they vie for political power in the upcoming elections. Tawar Razaghi spoke to Senior Research Fellow, Dr Benjamin Isakhan, at the Centre for Citizenship and Globalisation at Deakin University.