Generating rules with predicates, terms and variables from the pruned neural networks

Artificial neural networks (ANN) have demonstrated good predictive performance in a wide range of applications. They are, however, not considered sufficient for knowledge representation because of their inability to represent the reasoning process succinctly. This paper proposes a novel methodology Gyan that represents the knowledge of a trained network in the form of restricted first-order predicate rules. The empirical results demonstrate that an equivalent symbolic interpretation in the form of rules with predicates, terms and variables can be derived describing the overall behaviour of the trained ANN with improved comprehensibility while maintaining the accuracy and fidelity of the propositional rules.

'Try hard' : Attitudes to advertising in online social networks

Advertising has recently entered many new spaces it does not fully understand. The rules that apply in traditional media do not always translate in new media environments. However, their low cost of entry and the availability of hard-to-reach target markets, such as Generation Y, make environments such as online social networking sites attractive to marketers. This paper accumulates teenage perspectives from two qualitative studies to identify attitudes towards advertising in online social network sites and develop implications for marketers seeking to advertising on social network sites.

Recombinant protein production using a Tobacco yellow dwarf virus-based episomal expression vector : control of Rep activity

Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.

Rivalry within and between strategic networks : an investigation of the United States Automotive Industry

As a consequence of the increased incidence of collaborative arrangements between firms, the competitive environment characterising many industries has undergone profound change. It is suggested that rivalry is not necessarily enacted by individual firms according to the traditional mechanisms of direct confrontation in factor and product markets, but rather as collaborative orchestration between a number of participants or network members. Strategic networks are recognised as sets of firms within an industry that exhibit denser strategic linkages among themselves than other firms within the same industry. Based on this, strategic networks are determined according to evidence of strategic alliances between firms comprising the industry. As a result, a single strategic network represents a group of firms closely linked according to collaborative ties. Arguably, the collective outcome of these strategic relationships engineered between firms suggest that the collaborative benefits attributed to interorganisational relationships require closer examination in respect to their propensity to influence rivalry in intraindustry environments. Derived in large from the social sciences, network theory allows for the micro and macro examination of the opportunities and constraints inherent in the structure of relationships in strategic networks, establishing a relational approach upon which the conduct and performance of firms can be more fully understood. Research to date has yet to empirically investigate the relationship between strategic networks and rivalry. The limited research that has been completed utilising a network rationale to investigate competitive patterns in contemporary industry environments has been characterised by a failure to directly measure rivalry. Further, this prior research has typically embedded investigation in industry settings dominated by technological or regulatory imperatives, such as the microprocessor and airline industries. These industries, due to the presence of such imperatives, are arguably more inclined to support the realisation of network rivalry, through subscription to prescribed technological standards (eg., microprocessor industry) or by being bound by regulatory constraints dictating operation within particular market segments (airline industry). In order to counter these weaknesses, the proposition guiding research - Are patterns of rivalry predicted by strategic network membership? – is embedded in the United States Light Vehicles Industry, an industry not dominated by technological or regulatory imperatives. Further, rivalry is directly measured and utilised in research, thus distinguishing this investigation from prior research efforts. The timeframe of investigation is 1993 – 1999, with all research data derived from secondary sources. Strategic networks were defined within the United States Light Vehicles Industry based on evidence of horizontal strategic relationships between firms comprising the industry. The measure of rivalry used to directly ascertain the competitive patterns of industry participants was derived from the traditional Herfindahl Index, modified to account for patterns of rivalry observed at the market segment level. Statistical analyses of the strategic network and rivalry constructs found little evidence to support the contention of network rivalry; indeed, greater levels of rivalry were observed between firms comprising the same strategic network than between firms participating in opposing network structures. Based on these results, patterns of rivalry evidenced in the United States Light Vehicle Industry over the period 1993 – 1999 were not found to be predicted by strategic network membership. The findings generated by this research are in contrast to current theorising in the strategic network – rivalry realm. In this respect, these findings are surprising. The relevance of industry type, in conjunction with prevailing network methodology, provides the basis upon which these findings are contemplated. Overall, this study raises some important questions in relation to the relevancy of the network rivalry rationale, establishing a fruitful avenue for further research.

Protein complementation assay as a display system for screening protein libraries in the intracellular environment

A wide range of screening strategies have been employed to isolate antibodies and other proteins with specific attributes, including binding affinity, specificity, stability and improved expression. However, there remains no high-throughput system to screen for target-binding proteins in a mammalian, intracellular environment. Such a system would allow binding reagents to be isolated against intracellular clinical targets such as cell signalling proteins associated with tumour formation (p53, ras, cyclin E), proteins associated with neurodegenerative disorders (huntingtin, betaamyloid precursor protein), and various proteins crucial to viral replication (e.g. HIV-1 proteins such as Tat, Rev and Vif-1), which are difficult to screen by phage, ribosome or cell-surface display. This study used the β-lactamase protein complementation assay (PCA) as the display and selection component of a system for screening a protein library in the cytoplasm of HEK 293T cells. The colicin E7 (ColE7) and Immunity protein 7 (Imm7) *Escherichia coli* proteins were used as model interaction partners for developing the system. These proteins drove effective β-lactamase complementation, resulting in a signal-to-noise ratio (9:1 – 13:1) comparable to that of other β-lactamase PCAs described in the literature. The model Imm7-ColE7 interaction was then used to validate protocols for library screening. Single positive cells that harboured the Imm7 and ColE7 binding partners were identified and isolated using flow cytometric cell sorting in combination with the fluorescent β-lactamase substrate, CCF2/AM. A single-cell PCR was then used to amplify the Imm7 coding sequence directly from each sorted cell. With the screening system validated, it was then used to screen a protein library based the Imm7 scaffold against a proof-of-principle target. The wild-type Imm7 sequence, as well as mutants with wild-type residues in the ColE7- binding loop were enriched from the library after a single round of selection, which is consistent with other eukaryotic screening systems such as yeast and mammalian cell-surface display. In summary, this thesis describes a new technology for screening protein libraries in a mammalian, intracellular environment. This system has the potential to complement existing screening technologies by allowing access to intracellular proteins and expanding the range of targets available to the pharmaceutical industry.

Making connections : creative industries networks in outer suburban locations

The role of networks and their contribution to sustaining and developing creative industries is well documented (Wittel 2001; Kong 2005; Pratt 2007). This article argues that although networks operate across geographical boundaries, particularly through the use of communication technologies, the majority of studies have focussed on the ways in which networks operate in a) specific inner-urban metropolitan regions or b) specific industries. Such studies are informed by the geographical mindset of creative city proponents such as Florida (2002) and Landry (2000) in which inner-urban precincts are seen as the prime location for creative industries activity, business development and opportunity. But what of those creative industries situated beyond the inner city? Evidence in Australia suggests there is increasing creative industries activity beyond the inner city, in outer-suburban and ex-urban areas (Gibson & Brennan-Horley 2006). This article identifies characteristics of creative industries networks in outer-suburban locations in Melbourne and Brisbane. It argues that supporting and sustaining creative industries networks in these locations may require different strategies than those applied to inner-city networks. The article thus contributes to the growing understanding of the cultural economic geography of creative industries.

Design and evaluation of an image analysis platform for low-power, low-bandwidth camera networks

We describe the design and evaluation of a platform for networks of cameras in low-bandwidth, low-power sensor networks. In our work to date we have investigated two different DSP hardware/software platforms for undertaking the tasks of compression and object detection and tracking. We compare the relative merits of each of the hardware and software platforms in terms of both performance and energy consumption. Finally we discuss what we believe are the ongoing research questions for image processing in WSNs.

Closed-form design equations for decoupling networks of small arrays

Small element spacing in compact arrays results in strong mutual coupling between the array elements. A decoupling network consisting of reactive cross-coupling elements can alleviate problems associated with the coupling. Closed-form design equations for the decoupling networks of symmetrical arrays with two or three elements are presented.

Freedom of information implications of information sharing networks for critical infrastructure protection

Protection of “critical infrastructure” has become a major issue for govern- ments worldwide. Yet in Australia, as in many other countries, including the United States, an estimated 90% of critical infrastructure is privately owned or operated commercially – in other words, critical infrastructure protection is not the exclusive domain of government. As a result, information sharing between government and the private sector has become a vitally important component of effective risk management. However, establishing effective arrangements of this kind between the public and private sector needs to take account of existing regimes of access and public disclosure which relate to government-held documents; in particular, that which is established by freedom of information (FOI) legislation. This article examines the extent to which the current Commonwealth FOI regime is likely to act as an impediment to the private sector operators of critical infrastructure participat- ing in government-operated information sharing arrangements. By examining developments in other jurisdictions, principally the United States, the article considers whether amendments to the current Australian FOI regime are necessary to ensure effective participation, consistent with the underlying object and purpose of FOI.