Toward biomimetic materials in bone regeneration : functional behavior of mesenchymal stem cells on a broad spectrum of extracellular matrix components

Bone defect treatments can be augmented by mesenchymal stem cell (MSC) based therapies. MSC interaction with the extracellular matrix (ECM) of the surrounding tissue regulates their functional behavior. Understanding of these specific regulatory mechanisms is essential for the therapeutic stimulation of MSC in vivo. However, these interactions are presently only partially understood. This study examined in parallel, for the first time, the effects on the functional behavior of MSCs of 13 ECM components from bone, cartilage and hematoma compared to a control protein, and hence draws conclusions for rational biomaterial design. ECM components specifically modulated MSC adhesion, migration, proliferation, and osteogenic differentiation, for example, fibronectin facilitated migration, adhesion, and proliferation, but not osteogenic differentiation, whereas fibrinogen enhanced adhesion and proliferation, but not migration. Subsequently, the integrin expression pattern of MSCs was determined and related to the cell behavior on specific ECM components. Finally, on this basis, peptide sequences are reported for the potential stimulation of MSC functions. Based on the results of this study, ECM component coatings could be designed to specifically guide cell functions.

Neo-Dandy

Neo-Dandy was a practice-led research project that explored histories of a quintessential men’s and womenswear garment from across the ages — the formal white dress shirt. The aim was to generate a body of radically new mens’ shirts that, whilst incorporating characteristics normally associated with womenswear, would remain acceptable to male wearers. A detailed study identified a broad spectrum of historical design approaches, ranging from the orthodox man’s shirt to the many variations of the women’s blouse. Within this spectrum a threshold was discovered where the men’s shirt morphed into the woman’s blouse — a ‘design moment’ that appeared to typify the dandy figure (a fashion character who subversively confronts dress norms of their day). The research analysed thousands of archive catwalk images from leading contemporary menswear designers, and of these, only a small number tampered appreciably with the men’s white dress shirt — suggesting a new realm of possibility for fashion design innovation. This led to the creation of a new body of work labelled ‘Neo-Dandy’. Sixty ‘concept shirts’ were produced, with differing styles and varying degrees of detailing, that fitted the brief of being acceptable to male wearers, eminently ‘wearable’ and on a threshold position between menswear and womenswear. These designs were each tested, documented, and assessed in their capacity to evolve the Neo-Dandy aesthetic. Based on these outcomes, a list of key design principles for achieving this aesthetic was identified to assist designers in further evolving this style. The creative work achieved substantial public acclaim with the ‘Neo Dandy Collection’ winning a prestigious Design Institute of Australia Award (Lifestyle category) and being one of four finalists in the prestigious overall field for design excellence. It was subsequently curated into three major Brisbane exhibitions — the ARC Biennial, at Artisan Gallery and the industry leader, the Mercedes Benz Fashion Festival. The collection was also exhibited at the Queensland Art Gallery.

Neo-Dandy

Neo-Dandy was a practice-led research project that explored histories of a quintessential men’s and womenswear garment from across the ages — the formal white dress shirt. The aim was to generate a body of radically new mens’ shirts that incorporated characteristics normally associated with womenswear, whist remaining acceptable to male wearers. A detailed study identified a broad spectrum of historical design approaches, ranging from the orthodox man’s shirt to the many variations of the women’s blouse. Within this spectrum a threshold was discovered where the men’s shirt morphed into the woman’s blouse — a ‘design moment’ that appeared to typify the dandy figure (a fashion character who subversively confronts dress norms of their day). The research analysed thousands of archive catwalk images from leading contemporary menswear designers, and of these, only a small number tampered appreciably with the men’s white dress shirt — suggesting a new realm of possibility for fashion design innovation. This led to the creation of a new body of work labelled ‘Neo-Dandy’. Sixty ‘concept shirts’ were produced, with differing styles and varying degrees of detailing, that fitted the brief of being acceptable to male wearers, eminently ‘wearable’ and on a threshold position between menswear and womenswear. These designs were each tested, documented, and assessed in their capacity to evolve the Neo-Dandy aesthetic. Based on these outcomes, a list of key design principles for achieving this aesthetic was identified to assist designers in further evolving this style. The creative work achieved substantial public acclaim with the ‘Neo Dandy Collection’ winning a prestigious Design Institute of Australia Award (Lifestyle category) and being one of four finalists in the prestigious overall field for design excellence. It was subsequently curated into three major Brisbane exhibitions — the ARC Biennial, at Artisan Gallery and the industry leader, the Mercedes Benz Fashion Festival. The collection was also exhibited at the Queensland Art Gallery.

Development of a Rep-inducible, BBTV-based expression system in banana

Banana bunchy top is regarded as the most important viral disease of banana, causing significant yield losses worldwide. The disease is caused by Banana bunchy top virus (BBTV), which is a circular ssDNA virus belonging to the genus Babuvirus in the family Nanoviridae. There are currently few effective control strategies for this and other ssDNA viruses. “In Plant Activation” (InPAct) is a novel technology being developed at QUT for ssDNA virus-activated suicide gene expression. The technology exploits the rolling circle replication mechanism of ssDNA viruses and is based on a unique “split” gene design such that suicide gene expression is only activated in the presence of the viral Rep. This PhD project aimed to develop a BBTV-based InPAct system as a suicide gene strategy to control BBTV. The BBTV-based InPAct vector design requires a BBTV intergenic region (IR) to be embedded within an intron in the gene expression cassette. To ensure that the BBTV IR would not interfere with intron splicing, a TEST vector was initially generated that contained the entire BBTV IR embedded within an intron in a β-glucuronidase (GUS) expression vector. Transient GUS assays in banana embryogenic cell suspensions indicated that cryptic intron splice sites were present within the IR. Transcript analysis revealed two cryptic intron splice sites in the Domain III sequence of the CR-M within the IR. Removal of the CR-M from the TEST vector resulted in an enhancement of GUS expression suggesting that the cryptic intron splice sites had been removed. An InPAct GUS vector was subsequently generated that contained the modified BBTV IR, with the CR-M (minus Domain III) repositioned within the InPAct cassette. Using transient histochemical and fluorometric GUS assays in banana embryogenic cells, the InPAct GUS vector was shown to be activated in the presence of the BBTV Rep. However, the presence of both BBTV Rep and Clink was shown to have a deleterious effect on GUS expression suggesting that these proteins were cytotoxic at the levels expressed. Analysis of replication of the InPAct vectors by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector through the nicking/ligation activity of BBTV Rep. However, Rep-mediated episomal replicons, indicative of rolling circle replication of the released circularised cassettes, were not observed. The inability of the InPAct cassette to be replicated was further investigated. To examine whether the absence of Domain III of the CR-M was responsible, a suite of modified BBTV-based InPAct GUS vectors was constructed that contained the CR-M with the inclusion of Domain III, the CR-M with the inclusion of Domain III and additional upstream IR sequence, or no CR-M. Analysis of replication by Southern hybridisation revealed that neither the presence of Domain III, nor the entire CR-M, had an effect on replication levels. Since the InPAct cassette was significantly larger than the native BBTV genomic components (approximately 1 kb), the effect of InPAct cassette size on replication was also investigated. A suite of size variant BBTV-based vectors was constructed that increased the size of a replication competent cassette to 1.1 kbp through to 2.1 kbp.. Analysis of replication by Southern hybridisation revealed that an increase in vector size above approximately 1.5 - 1.7 kbp resulted in a decrease in replication. Following the demonstration of Rep-mediated release, circularisation and expression from the InPAct GUS vector, an InPAct vector was generated in which the uidA reporter gene was replaced with the ribonuclease-encoding suicide gene, barnase. Initially, a TEST vector was generated to assess the cytotoxicity of Barnase on banana cells. Although transient assays revealed a Barnase-induced cytotoxic effect in banana cells, the expression levels were sub-optimal. An InPAct BARNASE vector was generated and tested for BBTV Rep-activated Barnase expression using transient assays in banana embryogenic cells. High levels of background expression from the InPAct BARNASE vector made it difficult to accurately assess Rep-activated Barnase expression. Analysis of replication by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector but no Rep-mediated episomal replicons indicative of rolling circle replication of the released circularised cassettes were again observed. Despite the inability of the InPAct vectors to replicate to enable high level gene expression, the InPAct BARNASE vector was assessed in planta for BBTV Rep-mediated activation of Barnase expression. Eleven lines of transgenic InPAct BARNASE banana plants were generated by Agrobacterium-mediated transformation and were challenged with viruliferous Pentalonia nigronervosa. At least one clonal plant in each line developed bunchy top symptoms and infection was confirmed by PCR. No localised lesions were observed on any plants, nor was there any localised GUS expression in the one InPAct GUS line challenged with viruliferous aphids. The results presented in this thesis are the first study towards the development of a BBTV-based InPAct system as a Rep-activatable suicide gene expression system to control BBTV. Although further optimisation of the vectors is necessary, the preliminary results suggest that this approach has the potential to be an effective control strategy for BBTV. The use of iterons within the InPAct vectors that are recognised by Reps from different ssDNA plant viruses may provide a broad-spectrum resistance strategy against multiple ssDNA plant viruses. Further, this technology holds great promise as a platform technology for the molecular farming of high-value proteins in vitro or in vivo through expression of the ssDNA virus Rep protein.

The MESH book : landscape / infrastructure

Resulting from a series of student-run 'Edge' conferences that have been held in Australia and New Zealand (beginning at RMIT in 1983), The Mesh Book is a collection of essays grouped into themes of Invisible Infrastructures (systems of belief), Immanent Infrastructures (natural systems) and Present Infrastructures (roads and services). Ranging from esoteric discussions to analytical case studies, the book assembles a broad spectrum of ideas on the landscape within the context of Australia and a contemporary study of place.

Simulation of the nutrient supply in fracture healing

The healing process for bone fractures is sensitive to mechanical stability and blood supply at the fracture site. Most currently available mechanobiological algorithms of bone healing are based solely on mechanical stimuli, while the explicit analysis of revascularization and its influences on the healing process have not been thoroughly investigated in the literature. In this paper, revascularization was described by two separate processes: angiogenesis and nutrition supply. The mathematical models for angiogenesis and nutrition supply have been proposed and integrated into an existing fuzzy algorithm of fracture healing. The computational algorithm of fracture healing, consisting of stress analysis, analyses of angiogenesis and nutrient supply, and tissue differentiation, has been tested on and compared with animal experimental results published previously. The simulation results showed that, for a small and medium-sized fracture gap, the nutrient supply is sufficient for bone healing, for a large fracture gap, non-union may be induced either by deficient nutrient supply or inadequate mechanical conditions. The comparisons with experimental results demonstrated that the improved computational algorithm is able to simulate a broad spectrum of fracture healing cases and to predict and explain delayed unions and non-union induced by large gap sizes and different mechanical conditions. The new algorithm will allow the simulation of more realistic clinical fracture healing cases with various fracture gaps and geometries and may be helpful to optimise implants and methods for fracture fixation.

Frontiers in Materials Science and Technology

This special issue aims to provide up-to-date knowledge and the latest scientific concepts and technological developments in the processing, characterization, testing, mechanics, modeling and applications of a broad range of advanced materials. The many contributors, from Denmark, Germany, UK, Iran, Saudi Arabia, Malaysia, Japan, the People’s Republic of China, Singapore, Taiwan, USA, New Zealand and Australia, present a wide range of topics including: nanomaterials, thin films and coatings, metals and alloys, composite materials, materials processing and characterization, biomaterials and biomechanics, and computational materials science and simulation. The work will therefore be of great interest to a broad spectrum of researchers and technologists.

Generative Music editorial

An issue on generative music in Contemporary Music Review allows space to explore many of these controversies, and to explore the rich algorithmic scene in contemporary practice, as well as the diverse origins and manifestations of such a culture. A roster of interesting exponents from both academic and arts practice backgrounds are involved, matching the broad spectrum of current work. Contributed articles range from generative algorithms in live systems, from live coding to interactive music systems to computer games, through algorithmic modelling of longer-term form, evolutionary algorithms, to interfaces between modalities and mediums, in algorithmic choreography. A retrospective on the intensive experimentation into algorithmic music and sound synthesis at the Institute of Sonology in the 1960s and 70s creates a complementary strand, as well as an open paper on the issues raised by open source, as opposed to proprietary, software and operating systems, with consequences in the creation and archiving of algorithmic work.

The experience of emotional expression in the context of social relations for people diagnosed with schizophrenia : a phenomenological explication

The experience of emotional expression in the context of social relations is not well understood for people diagnosed with schizophrenia. Early phenomenological research on the experience of people diagnosed with schizophrenia traditionally focussed on self experience in isolation from others, with later research explicating isolated aspects of self experience in relation to others. The current research aimed to focus on the progressive experience of emotional expression of people diagnosed with schizophrenia in relation to others over 12 months, in order to gain a broad spectrum of experience. This study involved unstructured interviews with 7 participants, an average of 4 times each, over a period of 12 months. Due to the unstructured nature of the interviews, a great breadth of experience was explicated. From the interviews there emerged 6 themes grouped together as a transition into, and 5 themes grouped together as a recovery from, symptoms associated with a diagnosis of schizophrenia. Special significance was given to the theme of relational confusion as an experience that provides an understanding of the relationship between social stressors and personal characteristics with responses that are associated with a diagnosis of schizophrenia. It was suggested that participants experienced themselves, including their distancing and isolating responses, as a part of a social system. The breadth of experiences that emerged afforded a framework of experiences within which prior phenomenological research findings on static moments of experience have been located. A more meaningful understanding of the transitioning into and recovery from the experiences associated with a diagnosis of schizophrenia will afford advances in mental health practice.

Exercise, appetite, and energy balance: The interactions between energy expenditure and intake, and the implications for weight management

This book disseminates current information pertaining to the modulatory effects of foods and other food substances on behavior and neurological pathways and, importantly, vice versa. This ranges from the neuroendocrine control of eating to the effects of life-threatening disease on eating behavior. The importance of this contribution to the scientific literature lies in the fact that food and eating are an essential component of cultural heritage but the effects of perturbations in the food/cognitive axis can be profound. The complex interrelationship between neuropsychological processing, diet, and behavioral outcome is explored within the context of the most contemporary psychobiological research in the area. This comprehensive psychobiology- and pathology-themed text examines the broad spectrum of diet, behavioral, and neuropsychological interactions from normative function to occurrences of severe and enduring psychopathological processes

Nuclear localization of NPR1 is required for activation of PR gene expression

Systemic acquired resistance (SAR) is a broad-spectrum resistance in plants that involves the upregulation of a battery of pathogenesis-related (PR) genes. NPR1 is a key regulator in the signal transduction pathway that leads to SAR. Mutations in NPR1 result in a failure to induce PR genes in systemic tissues and a heightened susceptibility to pathogen infection, whereas overexpression of the NPR1 protein leads to increased induction of the PR genes and enhanced disease resistance. We analyzed the subcellular localization of NPR1 to gain insight into the mechanism by which this protein regulates SAR. An NPR1–green fluorescent protein fusion protein, which functions the same as the endogenous NPR1 protein, was shown to accumulate in the nucleus in response to activators of SAR. To control the nuclear transport of NPR1, we made a fusion of NPR1 with the glucocorticoid receptor hormone binding domain. Using this steroid-inducible system, we clearly demonstrate that nuclear localization of NPR1 is essential for its activity in inducing PR genes.

Re-conceptualising and re-positioning Australian library and information science education for the 21st century [Final Report 2011]

How can Australian library and information science (LIS) education produce, in a sustainable manner, the diverse supply of graduates with the appropriate attributes to develop and maintain high quality professional practice in the rapidly changing 21st century? This report presents the key findings of a project that has examined this question through research into future directions for LIS education in Australia. Titled Re-conceptualising and re-positioning Australian library and information science education for the twenty-first century, the purpose of the project was to establish a consolidated and holistic picture of the Australian LIS profession, and identify how its future education and training can be mediated in a cohesive and sustainable manner. The project was undertaken with a team of 12 university and vocational LIS educators from 11 institutions around Australia between November 2009 and December 2010. Collectively, these eleven institutions represented the broad spectrum and diversity of LIS education in Australia, and enabled the project to examine education for the information profession in a holistic and synergistic manner. Participating institutions in the project included Queensland University of Technology (Project Leader), Charles Sturt University, Curtin University of Technology, Edith Cowan University, Monash University, RMIT University, University of Canberra, University of South Australia, University of Tasmania, University of Technology Sydney and Victoria University. The inception and need for the project was motivated by a range of factors. From a broad perspective several of these factors relate to concerns raised at national and international levels regarding problems with education for LIS. In addition, the motivation and need for the project also related to some unique challenges that LIS education faces in the Australian tertiary education landscape. Over recent years a range of responses to explore the various issues confronting LIS education in Australia have emerged at local and national levels however this project represented the first significant investment of funding for national research in this area. In this way, the inception of the project offered a unique opportunity and powerful mechanism through which to bring together key stakeholders and inspire discourse concerning future education for the profession. Therefore as the first national project of its kind, its intent has been to provide foundation research that will inform and guide future directions for LIS education and training in Australia. The primary objective of the project was to develop a Framework for the Education of the Information Professions in Australia. The purpose of this framework was to provide evidence based strategic recommendations that would guide Australia’s future education for the information professions. Recognising the three major and equal players in the education process the project was framed around three areas of consideration: LIS students, the LIS workforce and LIS educators. Each area of consideration aligned to a research substudy in the project. The three research substudies were titled Student Considerations, Workforce Planning Considerations and Tertiary Education Considerations. The Students substudy provided a profile of LIS students and an analysis of their choices, experiences and expectations in regard to LIS education and their graduate destinations. The Workforce substudy provided an overview and analysis of the nature of the current LIS workforce, including a focus on employer expectations and employment opportunities and comment on the core and elective skill, knowledge and attitudes of current and future LIS professionals. Finally the Tertiary Education substudy provided a profile of LIS educators and an analysis of their characteristics and experiences including the key issues and challenges. In addition it also explored current national and international trends and priorities impacting on LIS education. The project utilised a Community Based Participatory Research (CBPR) approach. This approach involves all members of the community in all aspects of the project. It recognised the unique strengths and perspectives that community members bring to the process. For this project ‘community’ comprised of all individuals who have a role in, or a vested interest in, LIS education and included LIS educators, professionals, employers, students and professional associations. Individuals from these sub-groups were invited to participate in a range of aspects of the project from design through to implementation and evaluation. A range of research methodologies were used to consider the many different perspectives of LIS education, including employers and recruiters, professional associations, students, graduates and LIS teaching staff. Data collection involved a mixed method approach of questionnaires, focus groups, semi-structured interviews and environmental scans. An array of approaches was selected to ensure that broadest possible access to different facets of the information profession would be achieved. The main findings and observations from each substudy have highlighted a range of challenges for LIS education that need to be addressed. These findings and observations have grounded the development of the Framework for the Education of the Information Professions in Australia. The framework presents eleven recommendations to progress the national approach to LIS education and guide Australia’s future education for the information professions. The framework will be used by the LIS profession, most notably its educators, as strategic directions for the future of LIS education in Australia. Framework for the Education of the Information Professions in Australia: Recommendation 1: It is recommended that a broader and more inclusive vocabulary be adopted that both recognises and celebrates the expanding landscape of the field, for example ‘information profession’, ‘information sector’, ‘information discipline’ and ‘information education’. Recommendation 2: It is recommended that a self-directed body composed of information educators be established to promote, support and lead excellence in teaching and research within the information discipline. Recommendation 3: It is recommended that Australia’s information discipline continue to develop excellence in information research that will raise the discipline’s profile and contribute to its prominence within the national and international arena. Recommendation 4: It is recommended that further research examining the nature and context of Australia’s information education programs be undertaken to ensure a sustainable and relevant future for the discipline. Recommendation 5: It is recommended that further research examining the pathways and qualifications available for entry into the Australian information sector be undertaken to ensure relevance, attractiveness, accessibility and transparency. Recommendation 6: It is recommended that strategies are developed and implemented to ensure the sustainability of the workforce of information educators. Recommendation 7: It is recommended that a national approach to promoting and marketing the information profession and thereby attracting more students to the field is developed. Recommendation 8: It is recommended that Australia’s information discipline continues to support a culture of quality teaching and learning, especially given the need to accommodate a focus on the broader information landscape and more flexible delivery options. Recommendation 9: It is recommended that strategies are developed that will support and encourage collaboration between information education within the higher education and VET sectors. Recommendation 10: It is recommended that strategies and forums are developed that will support the information sector working together to conceptualise and articulate their professional identity and educational needs. Recommendation 11: It is recommended that a research agenda be established that will identify and prioritise areas in which further development or work is needed to continue advancing information education in Australia. The key findings from this project confirm that a number of pressing issues are confronting LIS education in Australia. Left unaddressed these issues will have significant implications for the future of LIS education as well as the broader LIS profession. Consequently creating a sustainable and cohesive future can only be realised through cooperation and collaboration among all stakeholders including those with the capacity to enact radical change in university and vocational institutions. Indeed the impending adoption and implementation of the project’s recommendations will fundamentally determine whether Australian LIS education is assured both for the present day and into the future.

A human capital framework : designed for convergence

The 21" century business environment is dominated by unprecedented change across a broad spectrum of social, economic, technological and cultural factors (Nowotny, Scott & Gibbons 2001). Among these, two broad trends -economic globalisation and rising knowledge intensity (Hart 2006)have come to distinguish organisational life. Under the weight of these transformational influences, the developed world, it seems, has arrived at a transformational moment. The far-reaching effects of the global financial crisis and its shadowy twin: the threat of a double dip recession, continue to exert an unsteadying influence on global and corporate finances. Growth in developed economies has slumped, share prices have declined, the market value of corporations has slipped and unemployment rates, in the vast majority of developed economies, have risen. Gross domestic product (GDP) growth has retreated from the strong growth experienced in the late 1990s to negative growth in 2009 and a sluggish and unsteady recovery in 2010. In response, the reach of Government in terms of its participation in markets has been extended, bringing with it the need to transition to new governance and regulatory arrangements. Ongoing concerns regarding the pace and sustainability of the recovery remains a front-of-mind concern with bailouts, buybacks, borrowings and BP dominating news services: 'We are witnessing the reweaving of the social, political and economic fabric that binds our planet, with long-term consequences that are as or more profound than those of the industrial era' (Tapscott & Williams 2006, p. 59).

Clusterin facilitates COMMD1 and I-kB degradation to enhance NF-kB activity in prostate cancer cells

Secretory clusterin (sCLU) is a stress-activated, cytoprotective chaperone that confers broad-spectrum cancer treatment resistance, and its targeted inhibitor (OGX-011) is currently in phase II trials for prostate, lung, and breast cancer. However, the molecular mechanisms by which sCLU inhibits treatment-induced apoptosis in prostate cancer remain incompletely defined. We report that sCLU increases NF-κB nuclear translocation and transcriptional activity by serving as a ubiquitin-binding protein that enhances COMMD1 and I-κB proteasomal degradation by interacting with members of the SCF-βTrCP E3 ligase family. Knockdown of sCLU in prostate cancer cells stabilizes COMMD1 and I-κB, thereby sequestrating NF-κB in the cytoplasm and decreasing NF-κB transcriptional activity. Comparative microarray profiling of sCLU-overexpressing and sCLU-knockdown prostate cancer cells confirmed that the expression of many NF-κB–regulated genes positively correlates with sCLU levels. We propose that elevated levels of sCLU promote prostate cancer cell survival by facilitating degradation of COMMD1 and I-κB, thereby activating the canonical NF-κB pathway.

Viruses of banana in East Africa

Bananas are one of the world's most important food crops, providing sustenance and income for millions of people in developing countries and supporting large export industries. Viruses are considered major constraints to banana production, germplasm multiplication and exchange, and to genetic improvement of banana through traditional breeding. In Africa, the two most important virus diseases are bunchy top, caused by Banana bunchy top virus (BBTV), and banana streak disease, caused by Banana streak virus (BSV). BBTV is a serious production constraint in a number of countries within/bordering East Africa, such as Burundi, Democratic Republic of Congo, Malawi, Mozambique, Rwanda and Zambia, but is not present in Kenya, Tanzania and Uganda. Additionally, epidemics of banana streak disease are occurring in Kenya and Uganda. The rapidly growing tissue culture (TC) industry within East Africa, aiming to provide planting material to banana farmers, has stimulated discussion about the need for virus indexing to certify planting material as virus-free. Diagnostic methods for BBTV and BSV have been reported and, for BBTV, PCR-based assays are reliable and relatively straightforward. However for BSV, high levels of serological and genetic variability and the presence of endogenous virus sequences within the banana genome complicate diagnosis. Uganda has been shown to contain the greatest diversity in BSV isolates found anywhere in the world. A broad-spectrum diagnostic test for BSV detection, which can discriminate between endogenous and episomal BSV sequences, is a priority. This PhD project aimed to establish diagnostic methods for banana viruses, with a particular focus on the development of novel methods for BSV detection, and to use these diagnostic methods for the detection and characterisation of banana viruses in East Africa. A novel rolling-circle amplification (RCA) method was developed for the detection of BSV. Using samples of Banana streak MY virus (BSMYV) and Banana streak OL virus (BSOLV) from Australia, this method was shown to distinguish between endogenous and episomal BSV sequences in banana plants. The RCA assay was used to screen a collection of 56 banana samples from south-west Uganda for BSV. RCA detected at least five distinct BSV isolates in these samples, including BSOLV and Banana streak GF virus (BSGFV) as well as three BSV isolates (Banana streak Uganda-I, -L and -M virus) for which only partial sequences had been previously reported. These latter three BSV had only been detected using immuno-capture (IC)-PCR and thus were possible endogenous sequences. In addition to its ability to detect BSV, the RCA protocol was also demonstrated to detect other viruses within the family Caulimoviridae, including Sugar cane bacilliform virus, and Cauliflower mosaic virus. Using the novel RCA method, three distinct BSV isolates from both Kenya and Uganda were identified and characterised. The complete genome of these isolates was sequenced and annotated. All six isolates were shown to have a characteristic badnavirus genome organisation with three open reading frames (ORFs) and the large polyprotein encoded by ORF 3 was shown to contain conserved amino acid motifs for movement, aspartic protease, reverse transcriptase and ribonuclease H activities. As well, several sequences important for expression and replication of the virus genome were identified including the conserved tRNAmet primer binding site present in the intergenic region of all badnaviruses. Based on the International Committee on Taxonomy of Viruses (ICTV) guidelines for species demarcation in the genus Badnavirus, these six isolates were proposed as distinct species, and named Banana streak UA virus (BSUAV), Banana streak UI virus (BSUIV), Banana streak UL virus (BSULV), Banana streak UM virus (BSUMV), Banana streak CA virus (BSCAV) and Banana streak IM virus (BSIMV). Using PCR with species-specific primers designed to each isolate, a genotypically diverse collection of 12 virus-free banana cultivars were tested for the presence of endogenous sequences. For five of the BSV no amplification was observed in any cultivar tested, while for BSIMV, four positive samples were identified in cultivars with a B-genome component. During field visits to Kenya, Tanzania and Uganda, 143 samples were collected and assayed for BSV. PCR using nine sets of species-specific primers, and RCA, were compared for BSV detection. For five BSV species with no known endogenous counterpart (namely BSCAV, BSUAV, BSUIV, BSULV and BSUMV), PCR was used to detect 30 infections from the 143 samples. Using RCA, 96.4% of these samples were considered positive, with one additional sample detected using RCA which was not positive using PCR. For these five BSV, PCR and RCA were both useful for identifying infected samples, irrespective of the host cultivar genotype (Musa A- or B-genome components). For four additional BSV with known endogenous counterparts in the M. balbisiana genome (BSOLV, BSGFV, BSMYV and BSIMV), PCR was shown to detect 75 infections from the 143 samples. In 30 samples from cultivars with an A-only genome component there was 96.3% agreement between PCR positive samples and detection using RCA, again demonstrating either PCR or RCA are suitable methods for detection. However, in 45 samples from cultivars with some B-genome component, the level of agreement between PCR positive samples and RCA positive samples was 70.5%. This suggests that, in cultivars with some B-genome component, many infections were detected using PCR which were the result of amplification of endogenous sequences. In these latter cases, RCA or another method which discriminates between endogenous and episomal sequences, such as immuno-capture PCR, is needed to diagnose episomal BSV infection. Field visits were made to Malawi and Rwanda to collect local isolates of BBTV for validation of a PCR-based diagnostic assay. The presence of BBTV in samples of bananas with bunchy top disease was confirmed in 28 out of 39 samples from Malawi and all nine samples collected in Rwanda, using PCR and RCA. For three isolates, one from Malawi and two from Rwanda, the complete nucleotide sequences were determined and shown to have a similar genome organisation to previously published BBTV isolates. The two isolates from Rwanda had at least 98.1% nucleotide sequence identity between each of the six DNA components, while the similarity between isolates from Rwanda and Malawi was between 96.2% and 99.4% depending on the DNA component. At the amino acid level, similarities in the putative proteins encoded by DNA-R, -S, -M, - C and -N were found to range between 98.8% to 100%. In a phylogenetic analysis, the three East African isolates clustered together within the South Pacific subgroup of BBTV isolates. Nucleotide sequence comparison to isolates of BBTV from outside Africa identified India as the possible origin of East African isolates of BBTV.