High-throughput vectors for efficient gene silencing in plants

A major challenge in the post-genome era of plant biology is to determine the functions of all genes in the plant genome. A straightforward approach to this problem is to reduce or knockout expression of a gene with the hope of seeing a phenotype that is suggestive of its function. Insertional mutagenesis is a useful tool for this type of study but is limited by gene redundancy, lethal knockouts, non-tagged mutants, and the inability to target the inserted element to a specific gene. The efficacy of gene silencing in plants using inverted-repeat transgene constructs that encode a hairpin RNA (hpRNA) has been demonstrated by a number of groups, and has several advantages over insertional mutagenesis. In this paper we describe two improved pHellsgate vectors that facilitate rapid generation of hpRNA-encoding constructs, pHellsgate 4 allows the production of an hpRNA construct in a single step from a single polymerase chain reaction product, while pHellsgate 8 requires a two-step process via an intermediate vector. We show that these vectors are effective at silencing three endogenous genes in Arabidopsis, FLOWERING LOCUS C, PHYTOENE DESATURASE and ETHYLENE INSENSITIVE 2. We also show that a construct of sequences from two genes silences both genes.

Biological values of acupuncture and Chinese herbal medicine : impact on the life science

Editorial paper

Practical Modbus flooding attack and detection

The Modicon Communication Bus (Modbus) protocol is one of the most commonly used protocols in industrial control systems. Modbus was not designed to provide security. This paper confirms that the Modbus protocol is vulnerable to flooding attacks. These attacks involve injection of commands that result in disrupting the normal operation of the control system. This paper describes a set of experiments that shows that an anomaly-based change detection algorithm and signature-based Snort threshold module are capable of detecting Modbus flooding attacks. In comparing these intrusion detection techniques, we find that the signature-based detection requires a carefully selected threshold value, and that the anomaly-based change detection algorithm may have a short delay before detecting the attacks depending on the parameters used. In addition, we also generate a network traffic dataset of flooding attacks on the Modbus control system protocol.

WATER - a series of photographs by Louise Grayson

This body of photographic work has been created to firstly, explore a new approach to practice-led research that uses an “action genre” approach to reflective practice (Lemke) and secondly, to visually explore human interaction with the fundamental item in life - water. The first of these is based on the contention that to understand the meanings inherent in photographs we cannot look merely at the end result. It is essential to keep looking at the actions of practitioners, and the influences upon them, to determine how external influences affect the meaning potential of editorial photographs (Grayson, 2012). WATER therefore, provides an ideal platform to reflect upon the actions and influences involved in creating work within the photographic genre of photojournalism. It enables this practitioner to reflect on each stage of production to gain a better understanding of how external influences impact the narrative potential within images created. There are multi-faceted influences experienced by photographers who are creating images that, in turn, are part of constructing and presenting the narrative potential of editorial photographs. There is an important relationship between professional photographers and the technical, cultural, economic and institutional forces that impinge upon all stages of production and publication. What results is a greater understanding of technical, cultural, economic and institutional forces that impinge upon all stages of production and publication. Therefore, to understand the meanings inherent in photographs within WATER, I do not look merely at the end result. It provides a case study looking at my actions in the filed, and the influences upon me, to determine how external influences affect the meaning potential of these photographs (Grayson, 2012). As a result, this project adds to the body of scholarship around the definition of Photojournalism, how it has adapted to the current media environment and provides scope for further research into emerging new genres within editorial photography, such as citizen photojournalism. Concurrently, the photographs themselves were created to visually explore how there remains a humanistic desire to interact with the natural form of water even while living a modern cosmopolitan life around it. Taking a photojournalistic approach to exploring this phenomenon, the images were created by “capturing moments as they happened” with no posing or setting up of images. This serendipitous approach to the photographic medium provides the practitioner with at least an attempt to direct the subjectivity contained explicitly in photographs. What results is a series of images that extend the visual dialogue around the role of water within modern humanistic lifestyles and how it remains an integral part of our society’s behaviors. It captures important moments that document this relationship at this time of modern development. The resulting works were exhibited and published as part of the Head On Photo Festival, Australia's largest photo festival and the world's second largest festival in Sydney 20-24 May 2013. The WATER series of images were curated by three Magnum members; Ian Berry, Eli Reed and Chris Steele-Perkins. Magnum is a highly regarded international photographic co-operative with editorial offices in New York, London, Paris and Tokyo. There was a projection of the works as part of the official festival programme, presented to both members of the public and Sydney’s photography professionals. In addition, a sample of images from the WATER series was chosen for inclusion in the Magnum-published hardcover book. References Grayson, Louise. 2012. “Editorial photographs and patterns of practice.” Journalism Practice. Accessed: http://www.tandfonline.com/doi/abs/10.1080/17512786.2012.726836#.UbZN-L--1RQ Lemke, Jay. 1995. Textual Politics: Discourse and Social Dynamics. London: Taylor & Francis.

Academic legal research and academic research capacity enhancement working topic : "purposes and objectives of law schools beyond educating students (research, capacity building,community service and outreach)"

One of the main objectives of law schools beyond educating students is to produce viable legal research. The comments in this paper are basically confined to the Australian context, and to examine this topic effectively, it is necessary to briefly review the current tertiary research agenda in Australia. This paper argues that there is a need for recognition and support for an expanded legal research framework along with additional research training for legal academics. There also needs to be more effective methods of measuring and recognising quality in legal research. This method needs to be one that can engender respect in an interdisciplinary context.

Submission : Proposed reforms to youth justice in Queensland

The proposed reforms to the youth justice system in Queensland are premised on the assumption that offending by young people is increasing. We noted (Carrington, Dwyer, Hutchinson and Richards 2012, 8) in a recent submission about the boot camps legislation that: "Statistics suggest that this concern is not warranted. Certainly studies show that ‘rates per 100,000 juveniles in detention in Queensland have been relatively stable compared with the national trend’ (Richards 2011) and that rates of detention of child offenders have declined generally in Australia over the last three decades. Youth offending statistics are affected by the diversion options used by the police, as well as by the numbers and levels of policing, and any special strategies such as Operation Colossus in the northern part of the state. ‘Community concern’ about crime does not always reflect the true rates of crime across Queensland. Policy should be based on valid evidence, not on ‘community concern’. With stable numbers of young people being detained in Australia, the research clearly suggests that youth offending is not escalating."...

Constructing the ideal EVE online player

EVE Online, released in 2003 by CCP Games, is a space-themed Massively Multiplayer Online Game (MMOG). This sandbox style MMOG has a reputation for being a difficult game with a punishing learning curve that is fairly impenetrable to new players. This has led to the widely held belief among the larger MMOG community that “EVE players are different”, as only a very particular type of player would be dedicated to learning how to play a game this challenging. Taking a critical approach to the claim that “EVE players are different”, this paper complicates the idea that only a certain type of player capable of playing the most hardcore of games will be attracted to this particular MMOG. Instead, we argue that EVE’s “exceptionalism” is actually the result of conscious design decisions on the part of CCP games, which in turn compel particular behaviours that are continually reinforced as the norm by the game’s relatively homogenous player community.

Digital beings : an opportunity for Australian visual effects

Ongoing innovation in digital animation and visual effects technologies has provided new opportunities for stories to be visually rendered in ways never before possible. Films featuring animation and visual effects continue to perform well at the box office, proving to be highly profitable projects. The Avengers (Whedon, 2012) holds the current record for opening weekend sales, accruing as much as $207,438,708 USD and $623,357,910 USD gross at time of writing. Life of Pi (Lee, 2012) at time of writing has grossed as much as $608,791,063 USD (Box Office Mojo, 2013). With so much creative potential and a demonstrable ability to generate a large amount of revenue, the animation and visual effects industry – otherwise known as the Post, Digital and Visual Effects (PDV) industry – has become significant to the future growth and stability of the Australian film industry as a whole.

A reference station-based GNSS computing mode to support unified precise point positioning and real-time kinematic services

Currently, the GNSS computing modes are of two classes: network-based data processing and user receiver-based processing. A GNSS reference receiver station essentially contributes raw measurement data in either the RINEX file format or as real-time data streams in the RTCM format. Very little computation is carried out by the reference station. The existing network-based processing modes, regardless of whether they are executed in real-time or post-processed modes, are centralised or sequential. This paper describes a distributed GNSS computing framework that incorporates three GNSS modes: reference station-based, user receiver-based and network-based data processing. Raw data streams from each GNSS reference receiver station are processed in a distributed manner, i.e., either at the station itself or at a hosting data server/processor, to generate station-based solutions, or reference receiver-specific parameters. These may include precise receiver clock, zenith tropospheric delay, differential code biases, ambiguity parameters, ionospheric delays, as well as line-of-sight information such as azimuth and elevation angles. Covariance information for estimated parameters may also be optionally provided. In such a mode the nearby precise point positioning (PPP) or real-time kinematic (RTK) users can directly use the corrections from all or some of the stations for real-time precise positioning via a data server. At the user receiver, PPP and RTK techniques are unified under the same observation models, and the distinction is how the user receiver software deals with corrections from the reference station solutions and the ambiguity estimation in the observation equations. Numerical tests demonstrate good convergence behaviour for differential code bias and ambiguity estimates derived individually with single reference stations. With station-based solutions from three reference stations within distances of 22–103 km the user receiver positioning results, with various schemes, show an accuracy improvement of the proposed station-augmented PPP and ambiguity-fixed PPP solutions with respect to the standard float PPP solutions without station augmentation and ambiguity resolutions. Overall, the proposed reference station-based GNSS computing mode can support PPP and RTK positioning services as a simpler alternative to the existing network-based RTK or regionally augmented PPP systems.

The link between infrastructure charges and housing affordability in Australia: Where is the empirical evidence?

Developer paid charges or contributions are a commonly used infrastructure funding mechanism for local governments. However, developers claim that these costs are merely passed on to home buyers, with adverse effects to housing affordability. Despite a plethora of government reports and industry advocacy, there remains no empirical evidence in Australia to confirm or quantify this passing on effect to home buyers and hence no data for which governments to base policy decision upon. This paper examines the question of who really pays for urban infrastructure and the impact of infrastructure charges on housing affordability. It presents the findings of a number of international empirical studies that provide evidence that infrastructure charges do increase house prices. Based on international findings, and in the absence of any Australian research, then these findings suggest that if the international findings are transferable, then there is empirical evidence to support the proposition that developer paid infrastructure charges are a significant contributor to increasing house prices.

Factors associated with the isolation of Nontuberculous mycobacteria (NTM) from a large municipal water system in Brisbane, Australia

Background Nontuberculous mycobacteria (NTM) are normal inhabitants of a variety of environmental reservoirs including natural and municipal water. The aim of this study was to document the variety of species of NTM in potable water in Brisbane, QLD, with a specific interest in the main pathogens responsible for disease in this region and to explore factors associated with the isolation of NTM. One-litre water samples were collected from 189 routine collection sites in summer and 195 sites in winter. Samples were split, with half decontaminated with CPC 0.005%, then concentrated by filtration and cultured on 7H11 plates in MGIT tubes (winter only). Results Mycobacteria were grown from 40.21% sites in Summer (76/189) and 82.05% sites in winter (160/195). The winter samples yielded the greatest number and variety of mycobacteria as there was a high degree of subculture overgrowth and contamination in summer. Of those samples that did yield mycobacteria in summer, the variety of species differed from those isolated in winter. The inclusion of liquid media increased the yield for some species of NTM. Species that have been documented to cause disease in humans residing in Brisbane that were also found in water include M. gordonae, M. kansasii, M. abscessus, M. chelonae, M. fortuitum complex, M. intracellulare, M. avium complex, M. flavescens, M. interjectum, M. lentiflavum, M. mucogenicum, M. simiae, M. szulgai, M. terrae. M. kansasii was frequently isolated, but M. avium and M. intracellulare (the main pathogens responsible for disease is QLD) were isolated infrequently. Distance of sampling site from treatment plant in summer was associated with isolation of NTM. Pathogenic NTM (defined as those known to cause disease in QLD) were more likely to be identified from sites with narrower diameter pipes, predominantly distribution sample points, and from sites with asbestos cement or modified PVC pipes. Conclusions NTM responsible for human disease can be found in large urban water distribution systems in Australia. Based on our findings, additional point chlorination, maintenance of more constant pressure gradients in the system, and the utilisation of particular pipe materials should be considered.

Isolation of nontuberculous mycobacteria (NTM) from household water and shower aerosols in patients with pulmonary disease caused by NTM

It has been postulated that susceptible individuals may acquire infection with nontuberculous mycobacteria (NTM) from water and aerosol exposure. This study examined household water and shower aerosols of patients with NTM pulmonary disease. The mycobacteria isolated from clinical samples from 20 patients included M. avium (5 patients), M. intracellulare (12 patients), M. abscessus (7 patients), M. gordonae (1 patient), M. lentiflavum (1 patient), M. fortuitum (1 patient), M. peregrinum (1 patient), M. chelonae (1 patient), M. triplex (1 patient), and M. kansasii (1 patient). One-liter water samples and swabs were collected from all taps, and swimming pools or rainwater tanks. Shower aerosols were sampled using Andersen six-stage cascade impactors. For a subgroup of patients, real-time PCR was performed and high-resolution melt profiles were compared to those of ATCC control strains. Pathogenic mycobacteria were isolated from 19 homes. Species identified in the home matched that found in the patient in seven (35%) cases: M. abscessus (3 cases), M. avium (1 case), M. gordonae (1 case), M. lentiflavum (1 case), and M. kansasii (1 case). In an additional patient with M. abscessus infection, this species was isolated from potable water supplying her home. NTM grown from aerosols included M. abscessus (3 homes), M. gordonae (2 homes), M. kansasii (1 home), M. fortuitum complex (4 homes), M. mucogenicum (1 home), and M. wolinskyi (1 home). NTM causing human disease can be isolated from household water and aerosols. The evidence appears strongest for M. avium, M. kansasii, M. lentiflavum, and M. abscessus. Despite a predominance of disease due to M. intracellulare, we found no evidence for acquisition of infection from household water for this species.

Specification & design of safety technologies for complex socio-technical systems : low-cost level crossings case study

The introduction of safety technologies into complex socio-technical systems requires an integrated and holistic approach to HF and engineering, considering the effects of failures not only within system boundaries, but also at the interfaces with other systems and humans. Level crossing warning devices are examples of such systems where technically safe states within the system boundary can influence road user performance, giving rise to other hazards that degrade safety of the system. Chris will discuss the challenges that have been encountered to date in developing a safety argument in support of low-cost level crossing warning devices. The design and failure modes of level crossing warning devices are known to have a significant influence on road user performance; however, quantifying this effect is one of the ongoing challenges in determining appropriate reliability and availability targets for low-cost level crossing warning devices.

Prognostic and therapeutic relevance of FLIP and procaspase-8 overexpression in non-small cell lung cancer

Non-small cell lung carcinoma remains by far the leading cause of cancer-related deaths worldwide. Overexpression of FLIP, which blocks the extrinsic apoptotic pathway by inhibiting caspase-8 activation, has been identified in various cancers. We investigated FLIP and procaspase-8 expression in NSCLC and the effect of HDAC inhibitors on FLIP expression, activation of caspase-8 and drug resistance in NSCLC and normal lung cell line models. Immunohistochemical analysis of cytoplasmic and nuclear FLIP and procaspase-8 protein expression was carried out using a novel digital pathology approach. Both FLIP and procaspase-8 were found to be significantly overexpressed in tumours, and importantly, high cytoplasmic expression of FLIP significantly correlated with shorter overall survival. Treatment with HDAC inhibitors targeting HDAC1-3 downregulated FLIP expression predominantly via post-transcriptional mechanisms, and this resulted in death receptor- and caspase-8-dependent apoptosis in NSCLC cells, but not normal lung cells. In addition, HDAC inhibitors synergized with TRAIL and cisplatin in NSCLC cells in a FLIP- and caspase-8-dependent manner. Thus, FLIP and procaspase-8 are overexpressed in NSCLC, and high cytoplasmic FLIP expression is indicative of poor prognosis. Targeting high FLIP expression using HDAC1-3 selective inhibitors such as entinostat to exploit high procaspase-8 expression in NSCLC has promising therapeutic potential, particularly when used in combination with TRAIL receptor-targeted agents.