Inorganic-organic composite particle as a staged delivery platform for plasmid DNA-based biopharmaceuticals

Infectious diseases such as SARS, influenza and bird flu may spread exponentially throughout communities. In fact, most infectious diseases remain major health risks due to the lack of vaccine or the lack of facilities to deliver the vaccines. Conventional vaccinations are based on damaged pathogens, live attenuated viruses and viral vectors. If the damage was not complete, the vaccination itself may cause adverse effects. Therefore, researchers have been prompted to prepare viable replacements for the attenuated vaccines that would be more effective and safer to use. DNA vaccines are generally composed of a double stranded plasmid that includes a gene encoding the target antigen under the transcriptional directory and control of a promoter region which is active in cells. Plasmid DNA (pDNA) vaccines allow the foreign genes to be expressed transiently in cells, mimicking intracellular pathogenic infection and inducing both humoral and cellular immune responses. Currently, because of their highly evolved and specialized components, viral systems are the most effective means for DNA delivery, and they achieve high efficiencies (generally >90%), for both DNA delivery and expression. As yet, viral-mediated deliveries have several limitations, including toxicity, limited DNA carrying capacity, restricted target to specific cell types, production and packing problems, and high cost. Thus, nonviral systems, particularly a synthetic DNA delivery system, are highly desirable in both research and clinical applications.

A homogeneous surface-enhanced Raman scattering platform for ultra-trace detection of trinitrotoluene in the environment

A facile and sensitive surface-enhanced Raman scattering substrate was prepared by controlled potentiostatic deposition of a closely packed single layer of gold nanostructures (AuNS) over a flat gold (pAu) platform. The nanometer scale inter-particle distance between the particles resulted in high population of ‘hot spots’ which enormously enhanced the scattered Raman photons. A renewed methodology was followed to precisely quantify the SERS substrate enhancement factor (SSEF) and it was estimated to be (2.2 ± 0.17) × 105. The reproducibility of the SERS signal acquired by the developed substrate was tested by establishing the relative standard deviation (RSD) of 150 repeated measurements from various locations on the substrate surface. A low RSD of 4.37 confirmed the homogeneity of the developed substrate. The sensitivity of pAu/AuNS was proven by determining 100 fM 2,4,6-trinitrotoluene (TNT) comfortably. As a proof of concept on the potential of the new pAu/AuNS substrate in field analysis, TNT in soil and water matrices was selectively detected after forming a Meisenheimer complex with cysteamine.

3D cultures of prostate cancer cells cultured in a novel high-throughput culture platform are more resistant to chemotherapeutics compared to cells cultured in monolayer

Despite monolayer cultures being widely used for cancer drug development and testing, 2D cultures tend to be hypersensitive to chemotherapy and are relatively poor predictors of whether a drug will provide clinical benefit. Whilst generally more complicated, three dimensional (3D) culture systems often better recapitulate true cancer architecture and provide a more accurate drug response. As a step towards making 3D cancer cultures more accessible, we have developed a microwell platform and surface modification protocol to enable high throughput manufacture of 3D cancer aggregates. Herein we use this novel system to characterize prostate cancer cell microaggregates, including growth kinetics and drug sensitivity. Our results indicate that prostate cancer cells are viable in this system, however some non-cancerous prostate cell lines are not. This system allows us to consistently control for the presence or absence of an apoptotic core in the 3D cancer microaggregates. Similar to tumor tissues, the 3D microaggregates display poor polarity. Critically the response of 3D microaggregates to the chemotherapeutic drug, docetaxel, is more consistent with in vivo results than the equivalent 2D controls. Cumulatively, our results demonstrate that these prostate cancer microaggregates better recapitulate the morphology of prostate tumors compared to 2D and can be used for high-throughput drug testing.

Curating in uncharted territories: An examination of a programming model & communicative platform for curation in non-traditional art spaces

This thesis articulates and examines public engagement programming in an emerging, non¬-traditional site. As a practice-led research project, the creative work proposes a site responsive, engagement centric, agile model for curatorial programming that developed out of the dynamic, new media/digital, curatorial practice at QUT's Creative Industries Precinct. The model and its accompanying exegetical framework, Curating in Uncharted Territories, offer a theoretically informed approach to programming, delivering and reporting for curatorial practices in a non¬-traditional sites of public engagement. The research provides the foundation for full development of the model and the basis for further research.

Is being #instagay different from an #lgbttakeover? A cross-platform investigation of sexual and gender identity performances

This paper introduces research in progress that examines how queer women perform sexual identity across social media platforms. Applying a lens of queer theory and Actor Network Theory, it discusses women’s embodied self-representations as taking on forms that both conform to and elaborate upon the selfie genre of digital representation. Acknowledging similarities and differences across platforms, specifically between Instagram and Vine, a novel walkthrough method is introduced to identify platform characteristics that shape identity performances. This method provides insights into the role of platforms in identity performances, which can be combined with analysis of user-generated content and interviews to better understand digital media’s constraints and affordances for queer representation.

Digital Identity 3.0: The Platform for People

Developed economies are moving from an economy of corporations to an economy of people. More than ever, people produce and share value amongst themselves, and create value for corporations through co-creation and by sharing their data. This data remains in the hands of corporations and governments, but people want to regain control. Digital identity 3.0 gives people that control, and much more. In this paper we describe a concept for a digital identity platform that substantially goes beyond common concepts providing authentication services. Instead, the notion of digital identity 3.0 empowers people to decide who creates, updates, reads and deletes their data, and to bring their own data into interactions with organisations, governments and peers. To the extent that the user allows, this data is updated and expanded based on automatic, integrated and predictive learning, enabling trusted third party providers (e.g., retailers, banks, public sector) to proactively provide services. Consumers can also add to their digital identity desired meta-data and attribute values allowing them to design their own personal data record and to facilitate individualised experiences. We discuss the essential features of digital identity 3.0, reflect on relevant stakeholders and outline possible usage scenarios in selected industries.

Investigations of an electrochemical platform based on the layered MoS2-graphene and horseradish peroxidase nanocomposite for direct electrochemistry and electrocatalysis

The self-assembly of layered molybdenum disulfide–graphene (MoS2–Gr) and horseradish peroxidase (HRP) by electrostatic attraction into a novel hybrid nanomaterial (HRP–MoS2–Gr) is reported. The properties of the MoS2–Gr were characterized by X-ray diffraction (XRD), high-resolution transmission electron microscopy (TEM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). UV–vis and Fourier transform infrared spectroscopy (FT-IR) indicate that the native structure of the HRP is maintained after the assembly, implying good biocompatibility of MoS2–Gr nanocomposite. Furthermore, the HRP–MoS2–Gr composite is utilized as a biosensor, which displays electrocatalytic activity to hydrogen peroxide (H2O2) with high sensitivity (679.7 μA mM−1 cm−2), wide linear range (0.2 μM–1.103 mM), low detection limit (0.049 μM), and fast amperometric response. In addition, the biosensor also exhibits strong anti-interference ability, satisfactory stability and reproducibility. These desirable electrochemical properties are attributed to the good biocompatibility and electron transport efficiency of the MoS2–Gr composite, as well as the high loading of HRP. Therefore, this biosensor is potentially suitable for H2O2 analysis in environmental, pharmaceutical, food or industrial applications.

Multipurpose nanoporous alumina-carbon nanowall bi-dimensional nano-hybrid platform via catalyzed and catalyst-free plasma CVD

Simple, rapid, plasma-assisted synthesis of large-area arrays of vertically-aligned carbon nanowalls on highly-porous, transparent bare and gold-coated alumina membranes with the two pore sizes is reported. It is demonstrated that the complex patterns of vertically aligned nanowalls can nucleate and form different morphologies in the low-temperature plasmas. The process is stable, and the twofold change in the gas flow (10 and 20 sccm) does not noticeably influence the morphology of the nanowall pattern. Application of a thin (5 nm) gold layer to nanoporous membrane prior to the nanowall growth allows controlling the network morphology.

Lesbian, gay, bisexual, trans, and queer visibility through selfies: Comparing platform mediators across Ruby Rose’s Instagram and Vine presence

This article investigates the relationship between social media platforms and the production and dissemination of selfies in light of its implications for the visibility of lesbian, gay, bisexual, trans, and queer (LGBTQ) people. Applying an Actor Network Theory lens, two popular visual media apps, Instagram and Vine, are examined through a comparative walkthrough method. This reveals platform elements, or mediators, that can influence the conversational capacity of selfies in terms of the following: range, the variety of discourses addressed within a selfie; reach, circulation within and across publics; and salience, the strength and clarity of discourses communicated through a selfie. These mediators are illustrated through LGBTQ celebrity Ruby Rose’s Instagram selfies and Vine videos. Instagram’s use expectations encourage selfies focused on mainstream discourses of normative beauty and conspicuous consumption with an emphasis on appearance, extending through features constraining selfies’ reach and salience. In contrast, Vine’s broader use expectations enable a variety of discourses to be communicated across publics with an emphasis on creative, first-person sharing. These findings are reflected in Rose’s Instagram selfies, which mute alternative discourses of gender and sexuality through desexualized and aesthetically appealing self-representations, while Vines display her personal side, enabling both LGBTQ and heterosexual, cisgender people to identify with her without minimizing non-normative aspects of her gender and sexuality. These findings demonstrate the relevance of platforms in shaping selfies’ conversational capacity, as mediators can influence whether selfies feature in conversations reinforcing dominant discourses or in counterpublic conversations, contributing to everyday activism that challenges normative gender and sexual discourses.

Appendiceal sonography in children: A retrospective analysis as a platform for potential implementation of diagnostic categories

- Background Sonography is an important diagnostic tool in children with suspected appendicitis. Reported accuracy and appendiceal visualisation rates vary significantly, as does the management of equivocal ultrasound findings. The aim of this study was to audit appendiceal sonography at a tertiary children's hospital, and provide baseline data for a future prospective study. - Summary of work Records of children who underwent ultrasound studies for possible appendicitis between January 2008 and December 2010 were reviewed. Variables included patient demographics, sonographic appendix characteristics, and secondary signs. Descriptive statistics and analysis using ANOVA, Mann-Whitney U test, and ROC curves were performed. Mater Human Research Ethic Committee approval was granted. - Summary of results There were 457 eligible children. Using a dichotomous diagnostic model (including equivocal results), sensitivity was 89.6%, specificity 91.6%, and diagnostic yield of 40.7%. ROC curve analysis of a 6mm diameter cut-off was 0.88 AUC (95% CI 0.80 to 0.95). - Discussion and conclusions Sonography is an accurate test for acute appendicitis in children, with a high sensitivity and negative predictive value. A diameter of 6mm as an absolute cut-off in a binary model can lead to false findings. Results were compared with available literature. Recent publications propose categorising diameter1 and integrating secondary signs2 to improve accuracy and provide more meaningful results to clinicians. This study will be a benchmark for future studies with multiple diagnostic categorisation.

Improving the Flexibility of Active Grids through Web Services

Active Grids are a form of grid infrastructure where the grid network is active and programmable. These grids directly support applications with value added services such as data migration, compression, adaptation and monitoring. Services such as these are particularly important for eResearch applications which by their very nature are performance critical and data intensive. We propose an architecture for improving the flexibility of Active Grids through web services. These enable Active Grid services to be easily and flexibly configured, monitored and deployed from practically any platform or application. The architecture is called WeSPNI ('Web Services based on Programmable Networks Infrastructure'). We present the architecture together with some early experimental results on using web services to monitor data movement in an active grid.

Developing a vision for an nD modelling tool

The design of a building is a complicated process, having to formulate diverse components through unique tasks involving different personalities and organisations in order to satisfy multi-faceted client requirements. To do this successfully, the project team must encapsulate an integrated design that accommodates various social, economic and legislative factors. Therefore, in this era of increasing global competition integrated design has been increasingly recognised as a solution to deliver value to clients.----- The ‘From 3D to nD modelling’ project at the University of Salford aims to support integrated design; to enable and equip the design and construction industry with a tool that allows users to create, share, contemplate and apply knowledge from multiple perspectives of user requirements (accessibility, maintainability, sustainability, acoustics, crime, energy simulation, scheduling, costing etc.). Thus taking the concept of 3-dimensional computer modelling of the built environment to an almost infinite number of dimensions, to cope with whole-life construction and asset management issues in the design of modern buildings. This paper reports on the development of a vision for how integrated environments that will allow nD-enabled construction and asset management to be undertaken. The project is funded by a four-year platform grant from the Engineering and Physical Sciences Research Council (EPSRC) in the UK; thus awarded to a multi-disciplinary research team, to enable flexibility in the research strategy and to produce leading innovation. This paper reports on the development of a business process and IT vision for how integrated environments will allow nD-enabled construction and asset management to be undertaken. It further develops many of the key issues of a future vision arising from previous CIB W78 conferences.

A large-scale computing infrastructure for design education

Most departmental computing infrastructure reflects the state of networking technology and available funds at the time of construction, which converge in a preconceived notion of homogeneity of network architecture and usage patterns. The DMAN (Digital Media Access Network) project, a large-scale server and network foundation for the Hong Kong Polytechnic University's School of Design was created as a platform that would support a highly complex academic environment while giving maximum freedom to students, faculty and researchers through simplicity and ease of use. As a centralized multi-user computation backbone, DMAN faces an extremely hetrogeneous user and application profile, exceeding implementation and maintenance challenges of typical enterprise, and even most academic server set-ups. This paper sumarizes the specification, implementation and application of the system while describing its significance for design education in a computational context.

From concept to embodiment : challenge and strategy

Facing with the difficulty in information propagation and synthesizing from conceptual to embodiment design, this paper introduces a function-oriented, axiom based conceptual modeling scheme. Default logic reasoning is exploited for recognition and reconstitution of conceptual product geometric and topological information. The proposed product modeling system and reasoning approach testify a methodology of "structural variation design", which is verified in the implementation of a GPAL (Green Product All Life-cycle) CAD system. The GPAL system includes major enhancement modules of a mechanism layout sketching method based on fuzzy logic, a knowledge-based function-to-form mapping mechanism and conceptual form reconstitution paradigm based on default geometric reasoning. A mechanical hand design example shows a more than 20 times increase in design efficacy with these enhancement modules in the GPAL system on a general 3D CAD platform.

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.