High-level expression of Rhodotorula gracilis d-amino acid oxidase in Pichia pastoris

By combining gene design and heterologous over-expression of Rhodotorula gracilis D-amino acid oxidase (RgDAO) in Pichia pastoris, enzyme production was enhanced by one order of magnitude compared to literature benchmarks, giving 350 kUnits/l of fed-batch bioreactor culture with a productivity of 3.1 kUnits/l h. P. pastoris cells permeabilized by freeze-drying and incubation in 2-propanol (10% v/v) produce a highly active (1.6 kUnits/g dry matter) and stable oxidase preparation. Critical bottlenecks in the development of an RgDAO catalyst for industrial applications have been eliminated.

Identification and characterisation of F3GT1 and F3GGT1, two glycosyltransferases responsible for anthocyanin biosynthesis in red-fleshed kiwifruit (Actinidia chinensis)

Much of the diversity of anthocyanins is due to the action of glycosyltransferases, which add sugar moieties to anthocyanidins. We identified two glycosyltransferases, F3GT1 and F3GGT1, from red-fleshed kiwifruit (Actinidia chinensis) that perform sequential glycosylation steps. Red-fleshed genotypes of kiwifruit accumulate anthocyanins mainly in the form of cyanidin 3-O-xylo-galactoside. Genes in the anthocyanin and flavonoid biosynthetic pathway were identified and shown to be expressed in fruit tissue. However, only the expression of the glycosyltransferase F3GT1 was correlated with anthocyanin accumulation in red tissues. Recombinant enzyme assays in vitro and in vivo RNA interference (RNAi) demonstrated the role of F3GT1 in the production of cyanidin 3-O-galactoside. F3GGT1 was shown to further glycosylate the sugar moiety of the anthocyanins. This second glycosylation can affect the solubility and stability of the pigments and modify their colour. We show that recombinant F3GGT1 can catalyse the addition of UDP-xylose to cyanidin 3-galactoside. While F3GGT1 is responsible for the end-product of the pathway, F3GT1 is likely to be the key enzyme regulating the accumulation of anthocyanin in red-fleshed kiwifruit varieties.

Environmental regulation of leaf colour in red 35S : PAP1 Arabidopsis thaliana

High-temperature, low-light (HTLL) treatment of 35S:PAP1 Arabidopsis thaliana over-expressing the PAP1 (Production of Anthocyanin Pigment 1) gene results in reversible reduction of red colouration, suggesting the action of additional anthocyanin regulators. High-performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LCMS) and Affimetrix®-based microarrays were used to measure changes in anthocyanin, flavonoids, and gene expression in response to HTLL. HTLL treatment of control and 35S:PAP1 A. thaliana resulted in a reversible reduction in the concentrations of major anthocyanins despite ongoing over-expression of the PAP1 MYB transcription factor. Twenty-one anthocyanins including eight cis-coumaryl esters were identified by LCMS. The concentrations of nine anthocyanins were reduced and those of three were increased, consistent with a sequential process of anthocyanin degradation. Analysis of gene expression showed down-regulation of flavonol and anthocyanin biosynthesis and of transport-related genes within 24 h of HTLL treatment. No catabolic genes up-regulated by HTLL were found. Reductions in the concentrations of anthocyanins and down-regulation of the genes of anthocyanin biosynthesis were achieved by environmental manipulation, despite ongoing over-expression of PAP1. Quantitative PCR showed reduced expression of three genes (TT8, TTG1 and EGL3) of the PAP1 transcriptional complex, and increased expression of the potential transcriptional repressors AtMYB3, AtMYB6 and AtMYBL2 coincided with HTLL-induced down-regulation of anthocyanin biosynthesis. HTLL treatment offers a model system with which to explore anthocyanin catabolism and to discover novel genes involved in the environmental control of anthocyanins.

STI-GMaS : an open-source environment for simulation of sexually-transmitted infections

Background Sexually-transmitted pathogens often have severe reproductive health implications if treatment is delayed or absent, especially in females. The complex processes of disease progression, namely replication and ascension of the infection through the genital tract, span both extracellular and intracellular physiological scales, and in females can vary over the distinct phases of the menstrual cycle. The complexity of these processes, coupled with the common impossibility of obtaining comprehensive and sequential clinical data from individual human patients, makes mathematical and computational modelling valuable tools in developing our understanding of the infection, with a view to identifying new interventions. While many within-host models of sexually-transmitted infections (STIs) are available in existing literature, these models are difficult to deploy in clinical/experimental settings since simulations often require complex computational approaches. Results We present STI-GMaS (Sexually-Transmitted Infections – Graphical Modelling and Simulation), an environment for simulation of STI models, with a view to stimulating the uptake of these models within the laboratory or clinic. The software currently focuses upon the representative case-study of Chlamydia trachomatis, the most common sexually-transmitted bacterial pathogen of humans. Here, we demonstrate the use of a hybrid PDE–cellular automata model for simulation of a hypothetical Chlamydia vaccination, demonstrating the effect of a vaccine-induced antibody in preventing the infection from ascending to above the cervix. This example illustrates the ease with which existing models can be adapted to describe new studies, and its careful parameterisation within STI-GMaS facilitates future tuning to experimental data as they arise. Conclusions STI-GMaS represents the first software designed explicitly for in-silico simulation of STI models by non-theoreticians, thus presenting a novel route to bridging the gap between computational and clinical/experimental disciplines. With the propensity for model reuse and extension, there is much scope within STI-GMaS to allow clinical and experimental studies to inform model inputs and drive future model development. Many of the modelling paradigms and software design principles deployed to date transfer readily to other STIs, both bacterial and viral; forthcoming releases of STI-GMaS will extend the software to incorporate a more diverse range of infections.

Patterns of diversity in fatty acid composition in the Australian groundnut germplasm collection

Knowledge of the amounts and types of fatty acids in groundnut oil is beneficial, particularly from a nutritional standpoint. Germplasm evaluation data for fatty acid composition on 819 accessions of groundnut (Arachis hypogaea L.) from the Australian Tropical Field Crops Genetic Resource Centre, Biloela, Queensland were examined. Data for eight quantitative fatty acid descriptors have been documented. Statistical assessment, via methods of pattern analysis, summarised and described the patterns of variation in fatty acid composition of the groundnut accessions in the Australian germplasm collection. Presentation of the results from principal components analysis and hierarchical cluster analysis using a biplot was shown to be a very useful interpretative tool. Such a biplot enables a simultaneous examination of the relationships among all the accessions and the fatty acids. Unlike that information available via database searches, the results from contribution analysis together with the biplot provide a global picture of the diversity available for use in plant breeding programs. The use of standardised data for eight fatty acids, compared to using three specific fatty acids, provided a better description of the total diversity available because it remains relevant with possible changes in the nutritional preferences for fatty acids. Fatty acid composition was found to vary in relation to the branching pattern of the accessions. This pattern is generally indicative of the botanical types of groundnuts; Virginia (alternate) compared to Spanish and Valencia (sequential) botanical types.

Universal codes from switching strategies

We discuss algorithms for combining sequential prediction strategies, a task which can be viewed as a natural generalisation of the concept of universal coding. We describe a graphical language based on Hidden Markov Models for defining prediction strategies, and we provide both existing and new models as examples. The models include efficient, parameterless models for switching between the input strategies over time, including a model for the case where switches tend to occur in clusters, and finally a new model for the scenario where the prediction strategies have a known relationship, and where jumps are typically between strongly related ones. This last model is relevant for coding time series data where parameter drift is expected. As theoretical contributions we introduce an interpolation construction that is useful in the development and analysis of new algorithms, and we establish a new sophisticated lemma for analysing the individual sequence regret of parameterised models.

Follow the leader if you can, hedge if you must

Follow-the-Leader (FTL) is an intuitive sequential prediction strategy that guarantees constant regret in the stochastic setting, but has poor performance for worst-case data. Other hedging strategies have better worst-case guarantees but may perform much worse than FTL if the data are not maximally adversarial. We introduce the FlipFlop algorithm, which is the first method that provably combines the best of both worlds. As a stepping stone for our analysis, we develop AdaHedge, which is a new way of dynamically tuning the learning rate in Hedge without using the doubling trick. AdaHedge refines a method by Cesa-Bianchi, Mansour, and Stoltz (2007), yielding improved worst-case guarantees. By interleaving AdaHedge and FTL, FlipFlop achieves regret within a constant factor of the FTL regret, without sacrificing AdaHedge’s worst-case guarantees. AdaHedge and FlipFlop do not need to know the range of the losses in advance; moreover, unlike earlier methods, both have the intuitive property that the issued weights are invariant under rescaling and translation of the losses. The losses are also allowed to be negative, in which case they may be interpreted as gains.

Characterising and predicting cyber attacks using the Cyber Attacker Model Profile (CAMP)

Purpose Ethnographic studies of cyber attacks typically aim to explain a particular profile of attackers in qualitative terms. The purpose of this paper is to formalise some of the approaches to build a Cyber Attacker Model Profile (CAMP) that can be used to characterise and predict cyber attacks. Design/methodology/approach The paper builds a model using social and economic independent or predictive variables from several eastern European countries and benchmarks indicators of cybercrime within the Australian financial services system. Findings The paper found a very strong link between perceived corruption and GDP in two distinct groups of countries – corruption in Russia was closely linked to the GDP of Belarus, Moldova and Russia, while corruption in Lithuania was linked to GDP in Estonia, Latvia, Lithuania and Ukraine. At the same time corruption in Russia and Ukraine were also closely linked. These results support previous research that indicates a strong link between been legitimate economy and the black economy in many countries of Eastern Europe and the Baltic states. The results of the regression analysis suggest that a highly skilled workforce which is mobile and working in an environment of high perceived corruption in the target countries is related to increases in cybercrime even within Australia. It is important to note that the data used for the dependent and independent variables were gathered over a seven year time period, which included large economic shocks such as the global financial crisis. Originality/value This is the first paper to use a modelling approach to directly show the relationship between various social, economic and demographic factors in the Baltic states and Eastern Europe, and the level of card skimming and card not present fraud in Australia.

Parallel authentication and public-key encryption

A parallel authentication and public-key encryption is introduced and exemplified on joint encryption and signing which compares favorably with sequential Encrypt-then-Sign (ɛtS) or Sign-then-Encrypt (Stɛ) schemes as far as both efficiency and security are concerned. A security model for signcryption, and thus joint encryption and signing, has been recently defined which considers possible attacks and security goals. Such a scheme is considered secure if the encryption part guarantees indistinguishability and the signature part prevents existential forgeries, for outsider but also insider adversaries. We propose two schemes of parallel signcryption, which are efficient alternative to Commit-then-Sign-and- Encrypt (Ct&G3&S). They are both provably secure in the random oracle model. The first one, called generic parallel encrypt and sign, is secure if the encryption scheme is semantically secure against chosen-ciphertext attacks and the signature scheme prevents existential forgeries against random-message attacks. The second scheme, called optimal parallel encrypt. and sign, applies random oracles similar to the OAEP technique in order to achieve security using encryption and signature components with very weak security requirements — encryption is expected to be one-way under chosen-plaintext attacks while signature needs to be secure against universal forgeries under random-plaintext attack, that is actually the case for both the plain-RSA encryption and signature under the usual RSA assumption. Both proposals are generic in the sense that any suitable encryption and signature schemes (i.e. which simply achieve required security) can be used. Furthermore they allow both parallel encryption and signing, as well as parallel decryption and verification. Properties of parallel encrypt and sign schemes are considered and a new security standard for parallel signcryption is proposed.

Effect of Coulomb blockade, gold resistance, and thermal expansion on the electrical resistance of ultrathin gold films

Controlling the electrical resistance of granular thin films is of great importance for many applications, yet a full understanding of electron transport in such films remains a major challenge. We have studied experimentally and by model calculations the temperature dependence of the electrical resistance of ultrathin gold films at temperatures between 2 K and 300 K. Using sputter deposition, the film morphology was varied from a discontinuous film of weakly coupled meandering islands to a continuous film of strongly coupled coalesced islands. In the weak-coupling regime, we compare the regular island array model, the cotunneling model, and the conduction percolation model with our experimental data. We show that the tunnel barriers and the Coulomb blockade energies are important at low temperatures and that the thermal expansion of the substrate and the island resistance affect the resistance at high temperatures. At low temperatures our experimental data show evidence for a transition from electron cotunneling to sequential tunneling but the data can also be interpreted in terms of conduction percolation. The resistivity and temperature coefficient of resistance of the meandering gold islands are found to resemble those of gold nanowires. We derive a simple expression for the temperature at which the resistance changes from non-metal-like behavior into metal-like behavior. In the case of strong island coupling, the total resistance is solely determined by the Ohmic island resistance.

Hydrogen in plasma-nanofabrication : selective control of nanostructure heating and passivation

The possibility of independent control of the surface fluxes of energy and hydrogen-containing radicals, thus enabling selective control of the nanostructure heating and passivation, is demonstrated. In situ energy flux measurements reveal that even a small addition of H2 to low-pressure Ar plasmas leads to a dramatic increase in the energy deposition through H recombination on the surface. The heat release is quenched by a sequential addition of a hydrocarbon precursor while the surface passivation remains effective. Such selective control offers an effective mechanism for deterministic control of the growth shape, crystallinity, and density of nanostructures in plasma-aided nanofabrication. © 2010 American Institute of Physics.

The role of the posterior superior temporal sulcus in audiovisual processing

In this study we investigate previous claims that a region in the left posterior superior temporal sulcus (pSTS) is more activated by audiovisual than unimodal processing. First, we compare audiovisual to visual-visual and auditory-auditory conceptual matching using auditory or visual object names that are paired with pictures of objects or their environmental sounds. Second, we compare congruent and incongruent audiovisual trials when presentation is simultaneous or sequential. Third, we compare audiovisual stimuli that are either verbal (auditory and visual words) or nonverbal (pictures of objects and their associated sounds). The results demonstrate that, when task, attention, and stimuli are controlled, pSTS activation for audiovisual conceptual matching is 1) identical to that observed for intramodal conceptual matching, 2) greater for incongruent than congruent trials when auditory and visual stimuli are simultaneously presented, and 3) identical for verbal and nonverbal stimuli. These results are not consistent with previous claims that pSTS activation reflects the active formation of an integrated audiovisual representation. After a discussion of the stimulus and task factors that modulate activation, we conclude that, when stimulus input, task, and attention are controlled, pSTS is part of a distributed set of regions involved in conceptual matching, irrespective of whether the stimuli are audiovisual, auditory-auditory or visual-visual.

Audit fees and asset securitization risks

The thesis investigates “where were the auditors in asset securitizations”, a criticism of the audit profession before and after the onset of the global financial crisis (GFC). Asset securitizations increase audit complexity and audit risks, which are expected to increase audit fees. Using US bank holding company data from 2003 to 2009, this study examines the association between asset securitization risks and audit fees, and its changes during the global financial crisis. The main test is based on an ordinary least squares (OLS) model, which is adapted from the Fields et al. (2004) bank audit fee model. I employ a principal components analysis to address high correlations among asset securitization risks. Individual securitization risks are also separately tested. A suite of sensitivity tests indicate the results are robust. These include model alterations, sample variations, further controls in the tests, and correcting for the securitizer self-selection problem. A partial least squares (PLS) path modelling methodology is introduced as a separate test, which allows for high intercorrelations, self-selection correction, and sequential order hypotheses in one simultaneous model. The PLS results are consistent with the main results. The study finds significant and positive associations between securitization risks and audit fees. After the commencement of the global financial crisis in 2007, there was an increased focus on the role of audits on asset securitization risks resulting from bank failures; therefore I expect that auditors would become more sensitive to bank asset securitization risks after the commencement of the crisis. I find that auditors appear to focus on different aspects of asset securitization risks during the crisis and that auditors appear to charge a GFC premium for banks. Overall, the results support the view that auditors consider asset securitization risks and market changes, and adjust their audit effort and risk considerations accordingly.

Essays in the political economy of voting and reforms

This thesis presents four essays in the political economy of elections and reforms. The first study exploits discontinuities around school entry cut-off dates to show that early childhood conditions can impact the probability to become a top-flight politician. The second study provides empirical estimates of the effect of sequential voting on turnout and bandwagon voting outside the laboratory. The third work describes a novel nonparametric strategy to identify tactical voting patterns directly from balloting results using British election data. Finally, a study is put forward that examines the political feasibility of reforms.

A review of modern computational algorithms for Bayesian optimal design

Bayesian experimental design is a fast growing area of research with many real-world applications. As computational power has increased over the years, so has the development of simulation-based design methods, which involve a number of algorithms, such as Markov chain Monte Carlo, sequential Monte Carlo and approximate Bayes methods, facilitating more complex design problems to be solved. The Bayesian framework provides a unified approach for incorporating prior information and/or uncertainties regarding the statistical model with a utility function which describes the experimental aims. In this paper, we provide a general overview on the concepts involved in Bayesian experimental design, and focus on describing some of the more commonly used Bayesian utility functions and methods for their estimation, as well as a number of algorithms that are used to search over the design space to find the Bayesian optimal design. We also discuss other computational strategies for further research in Bayesian optimal design.