968 resultados para Garnier, Louise Bary, 1836-
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2015
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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.
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Objective: To assess extent of coder agreement for external causes of injury using ICD-10-AM for injury-related hospitalisations in Australian public hospitals. Methods: A random sample of 4850 discharges from 2002 to 2004 was obtained from a stratified random sample of 50 hospitals across four states in Australia. On-site medical record reviews were conducted and external cause codes were assigned blinded to the original coded data. Code agreement levels were grouped into the following agreement categories: block level, 3-character level, 4-character level, 5th-character level, and complete code level. Results: At a broad block level, code agreement was found in over 90% of cases for most mechanisms (eg, transport, fall). Percentage disagreement was 26.0% at the 3-character level; agreement for the complete external cause code was 67.6%. For activity codes, the percentage of disagreement at the 3-character level was 7.3% and agreement for the complete activity code was 68.0%. For place of occurrence codes, the percentage of disagreement at the 4-character level was 22.0%; agreement for the complete place code was 75.4%. Conclusions: With 68% agreement for complete codes and 74% agreement for 3-character codes, as well as variability in agreement levels across different code blocks, place and activity codes, researchers need to be aware of the reliability of their specific data of interest when they wish to undertake trend analyses or case selection for specific causes of interest.
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This study explores teenager perceptions towards advertising in the online social networking environment. The future of online social networking sites is dependant upon the continued support of advertisers in this new medium, which is linked to the acceptance of advertising on these sites by their targeted audience. This exploratory study used the qualitative research methods of focus groups and in-depth personal interviews to gain insights from the teenager participants. The literature review in Chapter Two examined the previous research into advertising theories, consumer attitudes and issues such as advertising avoidance, advertising as a service and trust and privacy in the online social networking environment. The teenage consumer was also examined as were the influences of social identity theory. From this literature review eleven propositions were formed which provided a structure to the analysis of the research. Chapter Three outlined the multi-method research approach of using focus groups and in-depth interviews. The key findings were outlined in Chapter Four and Chapter Five provides discussion regarding these findings and the implications for theory and advertising practice. The main findings from this study suggest that teenagers have very high levels of advertising avoidance and are sceptical towards advertising on their online social networking sites. They have an inherent distrust of commercial messages in the online social networking environment; however they are extremely trusting with the information that they disclose online. They believe that if their site is classified as private, then the information disclosed on this site is not accessible to anyone. The study explores the reasons behind these views. This research has resulted in the identification of seven motivations behind online social networking use. A new model of advertising avoidance in the online social networking environment is also presented and discussed. This model makes a contribution towards filling the gap in available research on online social networking sites and advertising perception. The findings of this study have also resulted in the identification of the characteristics of online social networking sites as an advertising medium. The newness of online social networking sites coupled with the enthusiastic adoption of online social networking by the teenage demographic means that this exploratory study will be of interest to both academics and practitioners alike.
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Treatment resistors, that is children whose reading skills have not responded to sustained periods of appropriate intervention, have long been the concern of parents, educators, clinicians and researchers. Rudolph Flesch's book of 1955, Why Johnny Can't Read and What You Can Do About It, sparked the "great (and largely continuing) debate" about appropriate instructional methods for teaching reading.
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Among the students in Australian classrooms who are experiencing learning difficulties are increasing numbers of children who have been diagnosed with Asperger's syndrome. Although the general cognitive and language abilities of these students are comparable with most of their peers, they experience significant difficulties with social communication, social interactions and social-emotional/behavioural functioning. Despite indications that there are features inherent in Asperger's syndrome that are likely to have a negative effect on the development of advanced literacy skills, studies to date have primarily focused on social-emotional/behavioural challenges. Without effective literacy skills, however, students' access to educational and career opportunities may be curtailed. This article reviews features of Asperger's syndrome that appear to have a negative impact upon the development of advanced literacy skills and suggests ways in which inclusive classroom teachers could support the development of their learners.
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Think back to your teaching experiences over the past four years. Were there a number of children or adolescents in your classes who seemed to have learning difficulties? What area of their learning seemed to be of greatest concern for these students? Did most of these struggling learners have difficulty with reading? Was expressing their thoughts in writing also a challenge? Did some of these students also have difficulty in listening to and following directions? And did some of these students seem hesitant when asked to speak to the class?
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Theories that inform pedagogical practices have positioned young children as innocent, pre-political and egocentric. This paper draws from an action research study that investigates the impact of “transformative storytelling”, where stories purposefully crafted to counter metanarratives, revealed the impact of human greed with one class of children aged five to six years of age. Derrida’s notion of “cinders” provided a concept for investigating the traces or imprints the language of story left behind, amidst the children’s comments and actions, enabling the possibilities of the history of these “cinders” (that is what informed these comments and actions) to be noticed. Readings of some of the children’s responses suggest that children aged five and six years can engage in political discourse through the provocation of “transformative storytelling”, and that their engagement demonstrated the consideration of others through critical awareness and intersubjectivity. These early readings raise questions regarding curriculum content and pedagogical practices in early years education and the validity of ongoing educational goals that incorporate critical awareness and intersubjectivity to equip students with communitarian strategies to counter the individualistic outlook of neoliberalist societies.
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This study is an inquiry into the professional identity constructions of early childhood educators, where identity is conceptualised as social and contextual. Through a genealogical analysis of narratives of four Queensland early childhood teachers, the thesis renders as problematic universal and fixed notions of what it is to be an early childhood professional. The data are the four teachers’ professional life history narratives recounted through a series of conversational interviews with each participant. As they spoke about professionalism and ethics, these teachers struggled to locate themselves as professionals, as they drew on a number of dominant discourses available to them. These dominant discourses were located and mapped through analysis of the participants’ talk about relationships with parents, colleagues and authorities. Genealogical analysis enabled multiple readings of the ways in which the participants’ talk held together certainties and uncertainties, as they recounted their experiences and spoke of early childhood expertise, relational engagement and ethics. The thesis concludes with suggestions for ways to support early childhood teachers and pre-service teachers to both engage with and resist normative processes and expectations of professional identity construction. In so doing, multiple and contextual opportunities can be made available when it comes to being professional and ‘doing’ ethics. The thesis makes an argument for new possibilities for thinking and speaking professional identities that include both certainty and uncertainty, comfort and discomfort, and these seemingly oppositional terms are held together in tension, with an insistence that both are necessary and true. The use of provocations offers tools through which pre-service teachers, teachers and teacher educators can access new positions associated with certainties and uncertainties in professional identities. These new positions call for work that supports experiences of ‘de-comfort’ – that is, experiences that encourage early childhood educators to step away from the comfort zones that can become part of expertise, professional relationships and ethics embedded within normative representations of what it is to be an early childhood professional.