The manipulation of apoptosis-related genes to generate resistance to Fusarium wilt and water stress in banana

Bananas are susceptible to a diverse range of biotic and abiotic stresses, many of which cause serious production constraints worldwide. One of the most destructive banana diseases is Fusarium wilt caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense (Foc). No effective control strategy currently exists for this disease which threatens global banana production. Although disease resistance exists in some wild bananas, attempts to introduce resistance into commercially acceptable bananas by conventional breeding have been hampered by low fertility, long generation times and association of poor agronomical traits with resistance genes. With the advent of reliable banana transformation protocols, molecular breeding is now regarded as a viable alternative strategy to generate disease-resistant banana plants. Recently, a novel strategy involving the expression of anti-apoptosis genes in plants was shown to result in resistance against several necrotrophic fungi. Further, the transgenic plants showed increased resistance to a range of abiotic stresses. In this thesis, the use of anti-apoptosis genes to generate transgenic banana plants with resistance to Fusarium wilt was investigated. Since water stress is an important abiotic constraint to banana production, the resistance of the transgenic plants to water stress was also examined. Embryogenic cell suspensions (ECS) of two commercially important banana cultivars, Grand Naine (GN) and Lady Finger (LF), were transformed using Agrobacterium with the anti-apoptosis genes, Bcl-xL, Bcl-xL G138A, Ced-9 and Bcl- 2 3’ UTR. An interesting, and potentially important, outcome was that the use of anti-apoptosis genes resulted in up to a 50-fold increase in Agrobacterium-mediated transformation efficiency of both LF and GN cells over vector controls. Regenerated plants were subjected to a complete molecular characterisation in order to detect the presence of the transgene (PCR), transcript (RT-PCR) and gene product (Western blot) and to determine the gene copy number (Southern blot). A total of 36 independently-transformed GN lines (8 x Bcl-xL, 5 x Bcl-xL G138A, 15 x Ced-9 and 8 x Bcl-2 3’ UTR) and 41 independently-transformed LF lines (8 x Bcl-xL, 7 x BclxL G138A, 13 x Ced-9 and 13 x Bcl-2 3’ UTR) were identified. The 41 transgenic LF lines were multiplied and clones from each line were acclimatised and grown under glasshouse conditions for 8 weeks to allow monitoring for phenotypic abnormalities. Plants derived from 3 x Bcl-xL, 2 x Ced-9 and 5 x Bcl-2 3’ UTR lines displayed a variety of aberrant phenotypes. However, all but one of these abnormalities were off-types commonly observed in tissue-cultured, non-transgenic banana plants and were therefore unlikely to be transgene-related. Prior to determining the resistance of the transgenic plants to Foc race 1, the apoptotic effects of the fungus on both wild-type and Bcl-2 3’ UTR-transgenic LF banana cells were investigated using rapid in vitro root assays. The results from these assays showed that apoptotic-like cell death was elicited in wild-type banana root cells as early as 6 hours post-exposure to fungal spores. In contrast, these effects were attenuated in the root cells of Bcl-2 3’ UTR-transgenic lines that were exposed to fungal spores. Thirty eight of the 41 transgenic LF lines were subsequently assessed for resistance to Foc race 1 in small-plant glasshouse bioassays. To overcome inconsistencies in rating the internal (vascular discolouration) disease symptoms, a MatLab-based computer program was developed to accurately and reliably assess the level of vascular discolouration in banana corms. Of the transgenic LF banana lines challenged with Foc race 1, 2 x Bcl-xL, 3 x Ced-9, 2 x Bcl-2 3’ UTR and 1 x Bcl-xL G138A-transgenic line were found to show significantly less external and internal symptoms than wild-type LF banana plants used as susceptible controls at 12 weeks post-inoculation. Of these lines, Bcl-2 3’ UTR-transgenic line #6 appeared most resistant, displaying very mild symptoms similar to the wild-type Cavendish banana plants that were included as resistant controls. This line remained resistant for up to 23 weeks post-inoculation. Since anti-apoptosis genes have been shown to confer resistance to various abiotic stresses in other crops, the ability of these genes to confer resistance against water stress in banana was also investigated. Clonal plants derived from each of the 38 transgenic LF banana plants were subjected to water stress for a total of 32 days. Several different lines of transgenic plants transformed with either Bcl-xL, Bcl-xL G138A, Ced-9 or Bcl-2 3’ UTR showed a delay in visual water stress symptoms compared with the wild-type control plants. These plants all began producing new growth from the pseudostem following daily rewatering for one month. In an attempt to determine whether the protective effect of anti-apoptosis genes in transgenic banana plants was linked with reactive oxygen species (ROS)-associated programmed cell death (PCD), the effect of the chloroplast-targeting, ROS-inducing herbicide, Paraquat, on wild-type and transgenic LF was investigated. When leaf discs from wild-type LF banana plants were exposed to 10 ìM Paraquat, complete decolourisation occurred after 48 hours which was confirmed to be associated with cell death and ROS production by trypan blue and 3,3-diaminobenzidine (DAB) staining, respectively. When leaf discs from the transgenic lines were exposed to Paraquat, those derived from some lines showed a delay in decolourisation, suggesting only a weak protective effect from the transgenes. Finally, the protective effect of anti-apoptosis genes against juglone, a ROS-inducing phytotoxin produced by the causal agent of black Sigatoka, Mycosphaerella fijiensis, was investigated. When leaf discs from wild-type LF banana plants were exposed to 25 ppm juglone, complete decolourisation occurred after 48 hours which was again confirmed to be associated with cell death and ROS production by trypan blue and DAB staining, respectively. Further, TdT-mediated dUTP nick-end labelling (TUNEL) assays on these discs suggested that the cell death was apoptotic. When leaf discs from the transgenic lines were exposed to juglone, discs from some lines showed a clear delay in decolourisation, suggesting a protective effect. Whether these plants are resistant to black Sigatoka is unknown and will require future glasshouse and field trials. The work presented in this thesis provides the first report of the use of anti-apoptosis genes as a strategy to confer resistance to Fusarium wilt and water stress in a nongraminaceous monocot, banana. Such a strategy may be exploited to generate resistance to necrotrophic pathogens and abiotic stresses in other economically important crop plants.

Spatial Science News from QUT, Urban Development, November 2009.

Brief overview of topics/issues of interest end of 2009, including Spatial Science Students undertake Variety of Research Projects; labs and offices on the move again); Congratulations to Surveying Student Project- QSEA awards.

State-of-the-Art Review on Opinion Mining from Online Customers’ Feedback

Dealing with the ever-growing information overload in the Internet, Recommender Systems are widely used online to suggest potential customers item they may like or find useful. Collaborative Filtering is the most popular techniques for Recommender Systems which collects opinions from customers in the form of ratings on items, services or service providers. In addition to the customer rating about a service provider, there is also a good number of online customer feedback information available over the Internet as customer reviews, comments, newsgroups post, discussion forums or blogs which is collectively called user generated contents. This information can be used to generate the public reputation of the service providers’. To do this, data mining techniques, specially recently emerged opinion mining could be a useful tool. In this paper we present a state of the art review of Opinion Mining from online customer feedback. We critically evaluate the existing work and expose cutting edge area of interest in opinion mining. We also classify the approaches taken by different researchers into several categories and sub-categories. Each of those steps is analyzed with their strength and limitations in this paper.

Science and subjectivity

Science has been under attack in the last thirty years, and recently a number of prominent scientists have been busy fighting back. Here, an argument is presented that the `science wars' stem from an unreasonably strict adherence to the reductive method on the part of science, but that weakening this stance need not imply a lapse into subjectivity. One possible method for formalising the description of non-separable, contextually dependent complex systems is presented. This is based upon a quantum-like approach.

Continued use of process modeling grammars : the impact of individual difference factors

Process modeling grammars are used by analysts to describe information systems domains in terms of the business operations an organization is conducting. While prior research has examined the factors that lead to continued usage behavior, little knowledge has been established as to what extent characteristics of the users of process modeling grammars inform usage behavior. In this study, a theoretical model is advanced that incorporates determinants of continued usage behavior as well as key antecedent individual difference factors of the grammar users, such as modeling experience, modeling background and perceived grammar familiarity. Findings from a global survey of 529 grammar users support the hypothesized relationships of the model. The study offers three central contributions. First, it provides a validated theoretical model of post-adoptive modeling grammar usage intentions. Second, it discusses the effects of individual difference factors of grammar users in the context of modeling grammar usage. Third, it provides implications for research and practice.

Age-related differences in exercise and quality of life among breast cancer survivors

Purpose: Physical activity has become a focus of cancer recovery research as it has the potential to reduce treatment-related burden and optimize health-related quality of life (HRQoL). However, the potential for physical activity to influence recovery may be age-dependent. This paper describes physical activity levels and HRQoL among younger and older women after surgery for breast cancer and explores the correlates of physical inactivity. Methods: A population-based sample of breast cancer patients diagnosed in South-East Queensland, Australia, (n=287) were assessed once every three months, from 6 to 18 months post-surgery. The Functional Assessment of Cancer Therapy-Breast questionnaire (FACTB+4) and items from the Behavioral Risk Factor Surveillance System (BRFSS) questionnaire were used to measure HRQoL and physical activity, respectively. Physical activity was assigned metabolic equivalent task (MET) values, and categorized as < 3, 3 to 17.9 and 18+ MET-hours/weeks. Descriptive statistics, generalized linear models with age stratification (<50 years versus 50+ years), and logistic regression were used for analyses (p=0.05, two-tailed). Results: Younger women who engaged in 3 or more MET-hours/week of physical activity reported a higher HRQoL at 18 months compared to their more sedentary counterparts (p<0.05). Older women reported similar HRQoL irrespective of activity level and consistently reported clinically higher HRQoL than younger women. Increasing age, being overweight or obese, and restricting use of the treated side at six months post-surgery increased the likelihood of sedentary behavior (OR>3, p<0.05). Conclusions: Age influences the potential to observe HRQoL benefits related to physical activity participation. These results also provide relevant information for the design of exercise interventions for breast cancer survivors and highlights that some groups of women are at greater risk of long-term sedentary behavior.

From cultural studies to cultural science

This paper, first presented at a symposium on the 'past, present and future of cultural studies,' traces disciplinary changes in the study of culture from the perspective of 'cultural science', a term that was used by some of the earliest practitioners of cultural studies, including Raymond Williams. The paper goes on to describe some of the present moment, including work on the creative industries, show that a new version of cultural science is needed, based on evolutionary principles, in dialogue with the evolutionary approach in economics that was called for a century ago by Thorstein Veblen. This evolutionary turn, or 'cultural science 2.0,' it is argued, offers a radical and challenging future for cultural studies.

A Cultural Science Manifesto (in Mandarin)

We propose that a general analytic framework for cultural science can be constructed as a generalization of the generic micro meso macro framework proposed by Dopfer and Potts (2008). This paper outlines this argument along with some implications for the creative industries research agenda.

Translation of addictions science into practice : update and future directions.

Recent advances in the understanding of the genetic, neurochemical, behavioral and cultural underpinnings of addiction have led to rapid advances in the understanding of addiction as a disease. In fact, advances in basic science and the development of new pharmacological and behavioral therapies associated with them are appearing faster than can be assimilated not only by clinical researchers but practitioners and policy makers as well. Translation of science-based addictions knowledge into improved prevention, assessment and treatment, and communication of these changes to researchers and practitioners are significant challenges to the field. The general aim of this book is to summarize current and potential linkages between advances in addiction science and innovations in clinical practice. Whilst this book is primarily focused on translation, it also encompasses some scientific advances that are relevant to dissemination, and the book is itself a tool for disseminating innovative thinking. The goal is to generate interest in application opportunities from both recent research and theoretical advances.

Learning in the third space : a sociocultural perspective on learning with analogies

Research on analogies in science education has focussed on student interpretation of teacher and textbook analogies, psychological aspects of learning with analogies and structured approaches for teaching with analogies. Few studies have investigated how analogies might be pivotal in students’ growing participation in chemical discourse. To study analogies in this way requires a sociocultural perspective on learning that focuses on ways in which language, signs, symbols and practices mediate participation in chemical discourse. This study reports research findings from a teacher-research study of two analogy-writing activities in a chemistry class. The study began with a theoretical model, Third Space, which informed analyses and interpretation of data. Third Space was operationalized into two sub-constructs called Dialogical Interactions and Hybrid Discourses. The aims of this study were to investigate sociocultural aspects of learning chemistry with analogies in order to identify classroom activities where students generate Dialogical Interactions and Hybrid Discourses, and to refine the operationalization of Third Space. These aims were addressed through three research questions. The research questions were studied through an instrumental case study design. The study was conducted in my Year 11 chemistry class at City State High School for the duration of one Semester. Data were generated through a range of data collection methods and analysed through discourse analysis using the Dialogical Interactions and Hybrid Discourse sub-constructs as coding categories. Results indicated that student interactions differed between analogical activities and mathematical problem-solving activities. Specifically, students drew on discourses other than school chemical discourse to construct analogies and their growing participation in chemical discourse was tracked using the Third Space model as an interpretive lens. Results of this study led to modification of the theoretical model adopted at the beginning of the study to a new model called Merged Discourse. Merged Discourse represents the mutual relationship that formed during analogical activities between the Analog Discourse and the Target Discourse. This model can be used for interpreting and analysing classroom discourse centred on analogical activities from sociocultural perspectives. That is, it can be used to code classroom discourse to reveal students’ growing participation with chemical (or scientific) discourse consistent with sociocultural perspectives on learning.

Evaluating information systems

The book within which this chapter appears is published as a research reference book (not a coursework textbook) on Management Information Systems (MIS) for seniors or graduate students in Chinese universities. It is hoped that this chapter, along with the others, will be helpful to MIS scholars and PhD/Masters research students in China who seek understanding of several central Information Systems (IS) research topics and related issues. The subject of this chapter - ‘Evaluating Information Systems’ - is broad, and cannot be addressed in its entirety in any depth within a single book chapter. The chapter proceeds from the truism that organizations have limited resources and those resources need to be invested in a way that provides greatest benefit to the organization. IT expenditure represents a substantial portion of any organization’s investment budget and IT related innovations have broad organizational impacts. Evaluation of the impact of this major investment is essential to justify this expenditure both pre- and post-investment. Evaluation is also important to prioritize possible improvements. The chapter (and most of the literature reviewed herein) admittedly assumes a blackbox view of IS/IT1, emphasizing measures of its consequences (e.g. for organizational performance or the economy) or perceptions of its quality from a user perspective. This reflects the MIS emphasis – a ‘management’ emphasis rather than a software engineering emphasis2, where a software engineering emphasis might be on the technical characteristics and technical performance. Though a black-box approach limits diagnostic specificity of findings from a technical perspective, it offers many benefits. In addition to superior management information, these benefits may include economy of measurement and comparability of findings (e.g. see Part 4 on Benchmarking IS). The chapter does not purport to be a comprehensive treatment of the relevant literature. It does, however, reflect many of the more influential works, and a representative range of important writings in the area. The author has been somewhat opportunistic in Part 2, employing a single journal – The Journal of Strategic Information Systems – to derive a classification of literature in the broader domain. Nonetheless, the arguments for this approach are believed to be sound, and the value from this exercise real. The chapter drills down from the general to the specific. It commences with a highlevel overview of the general topic area. This is achieved in 2 parts: - Part 1 addressing existing research in the more comprehensive IS research outlets (e.g. MISQ, JAIS, ISR, JMIS, ICIS), and Part 2 addressing existing research in a key specialist outlet (i.e. Journal of Strategic Information Systems). Subsequently, in Part 3, the chapter narrows to focus on the sub-topic ‘Information Systems Success Measurement’; then drilling deeper to become even more focused in Part 4 on ‘Benchmarking Information Systems’. In other words, the chapter drills down from Parts 1&2 Value of IS, to Part 3 Measuring Information Systems Success, to Part 4 Benchmarking IS. While the commencing Parts (1&2) are by definition broadly relevant to the chapter topic, the subsequent, more focused Parts (3 and 4) admittedly reflect the author’s more specific interests. Thus, the three chapter foci – value of IS, measuring IS success, and benchmarking IS - are not mutually exclusive, but, rather, each subsequent focus is in most respects a sub-set of the former. Parts 1&2, ‘the Value of IS’, take a broad view, with much emphasis on ‘the business Value of IS’, or the relationship between information technology and organizational performance. Part 3, ‘Information System Success Measurement’, focuses more specifically on measures and constructs employed in empirical research into the drivers of IS success (ISS). (DeLone and McLean 1992) inventoried and rationalized disparate prior measures of ISS into 6 constructs – System Quality, Information Quality, Individual Impact, Organizational Impact, Satisfaction and Use (later suggesting a 7th construct – Service Quality (DeLone and McLean 2003)). These 6 constructs have been used extensively, individually or in some combination, as the dependent variable in research seeking to better understand the important antecedents or drivers of IS Success. Part 3 reviews this body of work. Part 4, ‘Benchmarking Information Systems’, drills deeper again, focusing more specifically on a measure of the IS that can be used as a ‘benchmark’3. This section consolidates and extends the work of the author and his colleagues4 to derive a robust, validated IS-Impact measurement model for benchmarking contemporary Information Systems (IS). Though IS-Impact, like ISS, has potential value in empirical, causal research, its design and validation has emphasized its role and value as a comparator; a measure that is simple, robust and generalizable and which yields results that are as far as possible comparable across time, across stakeholders, and across differing systems and systems contexts.

Discovery and characterization of IGFBP-mediated endocytosis in the human retinal pigment epithelial cell line ARPE-19

Insulin-like growth factor binding proteins (IGFBPs) are prime regulators of IGF-action in numerous cell types including the retinal pigment epithelium (RPE). The RPE performs several functions essential for vision, including growth factor secretion and waste removal via a phagocytic process mediated in part by vitronectin (Vn). In the course of studying the effects of IGFBPs on IGF-mediated VEGF secretion and Vn-mediated phagocytosis in the RPE cell line ARPE-19, we have discovered that these cells avidly ingest synthetic microspheres (2.0 μm diameter) coated with IGFBPs. Given the novelty of this finding and the established role for endocytosis in mediating IGFBP actions in other cell types, we have explored the potential role of candidate cell surface receptors. Moreover, we have examined the role of key IGFBP structural motifs, by comparing responses to three members of the IGFBP family (IGFBP-3, IGFBP-4 and IGFBP-5) which display overlapping variations in primary structure and glycosylation status. Coating of microspheres (FluoSpheres®, sulfate modified polystyrene filled with a fluorophore) was conducted at 37 °C for 1 h using 20 μg/mL of test protein, followed by extensive washing. Binding of proteins was confirmed using a microBCA assay. The negative control consisted of microspheres treated with 0.1% bovine serum albumin (BSA), and all test samples were post-treated with BSA in an effort to coat any remaining free protein binding sites, which might otherwise encourage non-specific interactions with the cell surface. Serum-starved cultures of ARPE-19 cells were incubated with microspheres for 24 h, using a ratio of approximately 100 microspheres per cell. Uptake of microspheres was quantified using a fluorometer and was confirmed visually by confocal fluorescence microscopy. The ARPE-19 cells displayed little affinity for BSA-treated microspheres, but avidly ingested large quantities of those pre-treated with Vn (ANOVA; p < 0.001). Strong responses were also observed towards recombinant formulations of non-glycosylated IGFBP-3, glycosylated IGFBP-3 and glycosylated IGFBP-5 (all p < 0.001), while glycosylated IGFBP-4 induced a relatively minor response (p < 0.05). The response to IGFBP-3 was unaffected in the presence of excess soluble IGFBP-3, IGF-I or Vn. Likewise, soluble IGFBP-3 did not induce uptake of BSA-treated microspheres. Antibodies to either the transferrin receptor or type 1 IGF-receptor displayed slight inhibitory effects on responses to IGFBPs and Vn. Heparin abolished responses to Vn, IGFBP-5 and non-glycosylated IGFBP-3, but only partially inhibited the response to glycosylated IGFBP-3. Our results demonstrate for the first time IGFBP-mediated endocytosis in ARPE-19 cells and suggest roles for the IGFBP-heparin-binding domain and glycosylation status. These findings have important implications for understanding the mechanisms of IGFBP actions on the RPE, and in particular suggest a role for IGFBP-endocytosis.

Age-related maculopathy : linking aetiology and pathophysiological changes to the ischaemia hypothesis

Age-related maculopathy (ARM) has remained a challenging topic with respect to its aetiology, pathomechanisms, early detection and treatment since the late 19th century when it was first described as its own entity. ARM was previously considered an inflammatory disease, a degenerative disease, a tumor and as the result of choroidal hemodynamic disturbances and ischaemia. The latter processes have been repeatedly suggested to have a key role in its development and progression. In vivo experiments under hypoxic conditions could be models for the ischaemic deficits in ARM. Recent research has also linked ARM with gene polymorphisms. It is however unclear what triggers a person's gene susceptibility. In this manuscript, a linking hypothesis between aetiological factors including ischaemia and genetics and the development of early clinicopathological changes in ARM is proposed. New clinical psychophysical and electrophysiological tests are introduced that can detect ARM at an early stage. Models of early ARM based upon hemodynamic, photoreceptor and post-receptoral deficits are described and the mechanisms by which ischaemia may be involved as a final common pathway are considered. In neovascular age-related macular degeneration (neovascular AMD), ischaemia is thought to promote release of vascular endothelial growth factor (VEGF) which induces chorioretinal neovascularisation. VEGF is critical in the maintenance of the healthy choriocapillaris. In the final section of the manuscript the documentation of the effect of new anti-VEGF treatments on retinal function in neovascular AMD is critically viewed.

Post-buckling Strength of LiteSteel Beams in Shear

This paper presents the details of experimental and numerical studies on the shear behaviour of a recently developed, cold-formed steel beam known as LiteSteel Beam (LSB). The LSB sections are produced by a patented manufacturing process involving simultaneous cold-forming and electric resistance welding. It has a unique shape of a channel beam with two rectangular hollow flanges. Recent research has demonstrated the presence of increased shear capacity of LSBs due to the additional fixity along the web to flange juncture, but the current design rules ignore this effect. Therefore they were modified by including a higher elastic shear buckling coefficient. In the present study, the ultimate shear capacity results obtained from the experimental and numerical studies of 10 different LSB sections were compared with the modified shear capacity design rules. It was found that they are still conservative as they ignore the presence of post-buckling strength. Therefore the design rules were further modified to include the available post-buckling strength. Suitable design rules were also developed under the direct strength method format. This paper presents the details of this study and the results including the final design rules for the shear capacity of LSBs.