Quantification of gammaH2AX foci in response to ionising radiation

DNA double-strand breaks (DSBs), which are induced by either endogenous metabolic processes or by exogenous sources, are one of the most critical DNA lesions with respect to survival and preservation of genomic integrity. An early response to the induction of DSBs is phosphorylation of the H2A histone variant, H2AX, at the serine-139 residue, in the highly conserved C-terminal SQEY motif, forming gammaH2AX(1). Following induction of DSBs, H2AX is rapidly phosphorylated by the phosphatidyl-inosito 3-kinase (PIKK) family of proteins, ataxia telangiectasia mutated (ATM), DNA-protein kinase catalytic subunit and ATM and RAD3-related (ATR)(2). Typically, only a few base-pairs (bp) are implicated in a DSB, however, there is significant signal amplification, given the importance of chromatin modifications in DNA damage signalling and repair. Phosphorylation of H2AX mediated predominantly by ATM spreads to adjacent areas of chromatin, affecting approximately 0.03% of total cellular H2AX per DSB(2,3). This corresponds to phosphorylation of approximately 2000 H2AX molecules spanning approximately 2 Mbp regions of chromatin surrounding the site of the DSB and results in the formation of discrete gammaH2AX foci which can be easily visualized and quantitated by immunofluorescence microscopy(2). The loss of gammaH2AX at DSB reflects repair, however, there is some controversy as to what defines complete repair of DSBs; it has been proposed that rejoining of both strands of DNA is adequate however, it has also been suggested that re-instatement of the original chromatin state of compaction is necessary(4-8). The disappearence of gammaH2AX involves at least in part, dephosphorylation by phosphatases, phosphatase 2A and phosphatase 4C(5,6). Further, removal of gammaH2AX by redistribution involving histone exchange with H2A.Z has been implicated(7,8). Importantly, the quantitative analysis of gammaH2AX foci has led to a wide range of applications in medical and nuclear research. Here, we demonstrate the most commonly used immunofluorescence method for evaluation of initial DNA damage by detection and quantitation of gammaH2AX foci in gamma-irradiated adherent human keratinocytes(9)

Quantitation of gammaH2AX foci in tissue samples

DNA double-strand breaks (DSBs) are particularly lethal and genotoxic lesions, that can arise either by endogenous (physiological or pathological) processes or by exogenous factors, particularly ionizing radiation and radiomimetic compounds. Phosphorylation of the H2A histone variant, H2AX, at the serine-139 residue, in the highly conserved C-terminal SQEY motif, forming γH2AX, is an early response to DNA double-strand breaks1. This phosphorylation event is mediated by the phosphatidyl-inosito 3-kinase (PI3K) family of proteins, ataxia telangiectasia mutated (ATM), DNA-protein kinase catalytic subunit and ATM and RAD3-related (ATR)2. Overall, DSB induction results in the formation of discrete nuclear γH2AX foci which can be easily detected and quantitated by immunofluorescence microscopy2. Given the unique specificity and sensitivity of this marker, analysis of γH2AX foci has led to a wide range of applications in biomedical research, particularly in radiation biology and nuclear medicine. The quantitation of γH2AX foci has been most widely investigated in cell culture systems in the context of ionizing radiation-induced DSBs. Apart from cellular radiosensitivity, immunofluorescence based assays have also been used to evaluate the efficacy of radiation-modifying compounds. In addition, γH2AX has been used as a molecular marker to examine the efficacy of various DSB-inducing compounds and is recently being heralded as important marker of ageing and disease, particularly cancer3. Further, immunofluorescence-based methods have been adapted to suit detection and quantitation of γH2AX foci ex vivo and in vivo4,5. Here, we demonstrate a typical immunofluorescence method for detection and quantitation of γH2AX foci in mouse tissues.

Youth off the Streets "Our Place, Walgett Youth and Young Families Project" Project Evaluation.

The Project: • YOTS is a major youth specific agency established in 1991. It is a non-denominational, non-discriminatory and not-for-profit organisation, providing a wide range of services and offering a full continuum of care. It seeks to build on the strengths and positive aspects of marginalised young people and communities. • The 'Our Place, Walgett Youth and Young Families Project' further develops an existing YOTS capacity to provide services to Aboriginal young people. • The project adopted an action-research and community development model in which YOTS worked in partnership with the Youth Sub-committee of the Walgett Interagency. • Specific goals/objectives of the program were to: Coordinate youth and young family activities in partnership with local services and the community to build self-esteem, pride, resilience, motivation and skills; Contribute to the prevention and reduction of homelessness, unstable and unsafe housing and disruptive mobility (Walgett/Redfern) in youth and young families; Increase and improve collaborative engagement between youth and family focused services; and, research, adapt and implement Australian and international best-practice homelessness prevention/reduction initiatives to contribute to new models of practice relevant to rural and regional areas. • The project centred around an out-reach model that focused on providing a safe space with relevant structured activities coordinated by YOTS youth and family workers. Through community and service provider consultation, it was proposed that local services could coordinate strategies and activities and run them, where possible, from the centre, providing ease of access in a safe and supportive context. • Specific activities included: Implementing regular meetings with the stakeholders and community representatives; Developing a Terms of Reference for YOTS presence in the Walgett community; Undertaking a community consultation prior to finalising program activities; Implementing a range of recreational activities (sports, music, arts and crafts) early on in the activity; Implementing young family support initiatives; implementing a volunteering program, including volunteer support to young families through intergenerational volunteering; running a series of Culture and Healing Camps in partnership with local Elders and other services; Running a series of Music Camps; Providing alternative education support and referrals in partnership with local schools; Researching, identifying and adapting other best-practice models.

Unsupervised online learning of condition-invariant images for place recognition

This paper presents an online, unsupervised training algorithm enabling vision-based place recognition across a wide range of changing environmental conditions such as those caused by weather, seasons, and day-night cycles. The technique applies principal component analysis to distinguish between aspects of a location’s appearance that are condition-dependent and those that are condition-invariant. Removing the dimensions associated with environmental conditions produces condition-invariant images that can be used by appearance-based place recognition methods. This approach has a unique benefit – it requires training images from only one type of environmental condition, unlike existing data-driven methods that require training images with labelled frame correspondences from two or more environmental conditions. The method is applied to two benchmark variable condition datasets. Performance is equivalent or superior to the current state of the art despite the lesser training requirements, and is demonstrated to generalise to previously unseen locations.

Insurance and transport law : controlling insurance contract terms : Section 54 of the Insurance Contracts Act – compliance, recovery and accountability

Commonwealth legislation covering insurance contracts contains numerous provisions designed to control the operation and effect of terms in life and general insurance contracts. For example, the Life Insurance Act 1995 (Cth) contains provisions regulating the consequences attendant upon incorrect statements in proposals [1] and non-payment of premiums, [2] provides that an insurer may only exclude liability in the case of suicide if it has made express provision for such contingency in its policy, [3] and severely restricts the efficacy of conditions as to war risks. [4] The Insurance Contracts Act 1984 (Cth) is even more intrusive and has a major impact upon contractual provisions in the general insurance field. It is beyond the scope of this note to explore all of these provisions in any detail but examples of controls and constraints imposed upon the operation and effect of contractual provisions include the following. A party is precluded from relying upon a provision in a contract of insurance if such reliance would amount to a failure to act with the utmost good faith. [5] Similarly, a policy provision which requires differences or disputes arising out of the insurance to be submitted to arbitration is void, [6] unless the insurance is a genuine cover for excess of loss over and above another specified insurance. [7] Similarly clause such as conciliation clauses, [8] average clauses, [9] and unusual terms [10] are given qualified operation. [11] However the provision in the Insurance Contracts Act that has the greatest impact upon, and application to, a wide range of insurance clauses and claims is s 54. This section has already generated a significant volume of case law and is the focus of this note. In particular this note examines two recent cases. The first, Johnson v Triple C Furniture and Electrical Pty Ltd [2012] 2 Qd R 337, (hereafter the Triple C case), is a decision of the Queensland Court of Appeal; and the second, Matthew Maxwell v Highway Hauliers Pty Ltd [2013] WASCA 115, (hereafter the Highway Hauliers case), is a decision of the Court of Appeal in Western Australia. This latter decision is on appeal to the High Court of Australia. The note considers too the decision of the New South Wales Court of Appeal in Prepaid Services Pty Ltd v Atradius Credit Insurance NV [2013] NSWCA 252 (hereafter the Prepaid Services case).These cases serve to highlight the complex nature of s 54 and its application, as well as the difficulty in achieving a balance between an insurer and an insured's reasonable expectations.

Natural convection heat transfer in a baffled triangular enclosure

To reduce the natural convection heat loss from enclosures many researchers used convection suppression devices in the past. In this study a single baffle is used under the top tip to investigate numerically the natural convection heat loss in an attic shaped enclosure which is a cost effective approach. The case considered here is one inclined wall of the enclosure is uniformly heated while the other inclined wall is uniformly cooled with adiabatic bottom wall. The finite volume method has been used to discretize the governing equations, with the QUICK scheme approximating the advection term. The diffusion terms are discretized using central-differencing with second order accuracy. A wide range of governing parameters are studied (Rayleigh number, aspect ratio, baffle length etc.). It is observed that the heat transfer due to natural convection in the enclosure reduces when the baffle length is increased. Effects of other parameters on heat transfer and flow field are described in this study.

Onset of transition in mixed convection of a lid-driven trapezoidal enclosure filled with water-Al2O3 nanofluid

A numerical study is carried out to investigate the transition from laminar to chaos in mixed convection heat transfer inside a lid-driven trapezoidal enclosure. In this study, the top wall is considered as isothermal cold surface, which is moving in its own plane at a constant speed, and a constant high temperature is provided at the bottom surface. The enclosure is assumed to be filled with water-Al2O3 nanofluid. The governing Navier–Stokes and thermal energy equations are expressed in non-dimensional forms and are solved using Galerkin finite element method. Attention is paid in the present study on the pure mixed convection regime at Richandson number, Ri = 1. The numerical simulations are carried out over a wide range of Reynolds (0.1 ≤ Re ≤ 103) and Grashof (0.01 ≤ Gr ≤ 106) numbers. Effects of the presence of nanofluid on the characteristics of mixed convection heat transfer are also explored. The average Nusselt numbers of the heated wall are computed to demonstrate the influence of flow parameter variations on heat transfer. The corresponding change of flow and thermal fields is visualized from the streamline and the isotherm contour plots.

Integrated marketing communications

Organisations continually use integrated marketing communications to achieve a competitive advantage and meet their marketing objectives. This 4th edition of Integrated Marketing Communications emphasises digital and interactive marketing, the most dynamic and crucial components to a successful IMC campaign today. Incorporating the most up to date theories and practice, the text clearly explains and demonstrates how to best select and co-ordinate all of a brand’s marketing communications elements to effectively engage the target market. Chapters adopt an integrative approach to examine marketing communications from both a consumer’s and marketer’s perspective. A wide range of local and global examples include: Spotify, Pandora, Coca-Cola, Pepsi, Woolworths, Nike, KFC, Victoria Bitter, Tigerair and Air New Zealand. Each new copy of the text also offers 12 month access to wealth of student on-line revision and learning tools: CourseMate Express + Search me! marketing. Unique to the text is a series of end of chapter local videos showing students how key objectives in IMC theory are applied by real businesses.

Collaboration to improve evacuation centre operations in Queensland

Purpose The purpose of this paper is to provide a case study of two organisations working in evacuation centres which overcame challenges to develop a constructive relationship, resulting in improved outcomes for disaster-affected people. A wide range of services for disaster-affected communities are provided as part of emergency sheltering. Collaboration between agencies providing services is essential, but sometimes challenging. Design/methodology/approach A wide range of services for disaster-affected communities are provided as part of emergency sheltering. Collaboration between agencies providing services is essential, but sometimes challenging. The purpose of this paper is to provide a case study of two organisations working in evacuation centres which overcame challenges to develop a constructive relationship, resulting in improved outcomes for disaster-affected people. Findings The Preferred Sheltering Practices provides an ongoing anchor for Australian Red Cross and Environmental Health Australia (EHA) (Queensland) Inc.’s relationship and has led to other tangible benefits such as involvement in each other’s events and trainings. The relationship has become embedded in each organisation’s day-to-day business ensuring the relationship’s sustainability beyond individual staff movements. Originality/value This case study provides an example of how collaboration can be achieved between two organisations with seemingly different mandates to improve the response for disaster-affected communities.

Microalgae: An alternative source of biodiesel for the compression ignition (CI) engine

This thesis is a comprehensive study of microalgae biodiesel for the compression ignition engine. It examines microalgae growing conditions, the extraction process and physiochemical properties with a wide range of microalgae species. It also evaluates microalgae biodiesel with regards to engine performance and emission characteristics and explains the difficulties and potentiality of microalgae as a biodiesel. In doing so, an extensive analysis of different extraction methods and engine testing was conducted and a comprehensive study on microalgae biodiesel is presented.

Injection velocity control of thermoplastic injection molding via a double controller scheme

Injection velocity has been recognized as a key variable in thermoplastic injection molding. Its closed-loop control is, however, difficult due to the complexity of the process dynamic characteristics. The basic requirements of the control system include tracking of a pre-determined injection velocity curve defined in a profile, load rejection and robustness. It is difficult for a conventional control scheme to meet all these requirements. Injection velocity dynamics are first analyzed in this paper. Then a novel double-controller scheme is adopted for the injection velocity control. This scheme allows an independent design of set-point tracking and load rejection and has good system robustness. The implementation of the double-controller scheme for injection velocity control is discussed. Special techniques such as profile transformation and shifting are also introduced to improve the velocity responses. The proposed velocity control has been experimentally demonstrated to be effective for a wide range of processing conditions.

The role of university research centres in building road safety capacity : implications for China

In recent decades, highly motorised countries, such as Australia, have witnessed significant improvements in population health through reductions in fatalities and injuries from road traffic crashes. In Australia, concerted efforts have been made to reduce the road trauma burden since road fatalities reached their highest level in in the early 1970s. Since that time, many improvements have been made drawing on various disciplines to reduce the trauma burden (e.g., road and vehicle design, road user education, traffic law enforcement practices and enforcement technologies). While road fatalities have declined significantly since the mid-1970s, road trauma remains a serious public health concern in Australia. China has recently become the largest car market in the world (Ma, Li, Zhou, Duan, & Bishai, 2012). This rapid motorisation has been accompanied by substantial expansion of the road network as well as a large road trauma burden. Road traffic injuries are a major cause of death in China, reported as accounting for one third of all injury-deaths between 2002 and 2006 (Ma et al., 2012). In common with Australia, China has experienced a reported decline in fatalities since 2002 (see Hu, Wen & Baker, 2008). However, there remains a strong need for action in this area as rates of motorisation continue to climb in China. In Australia, a wide range of organisations have contributed to the improvements in road safety including government agencies, professional organisations, advocacy groups and research centres. In particular, Australia has several highly regarded and multi-disciplinary, university-based research centres that work across a range of road safety fields, including engineering, intelligent transportation systems, the psychology of road user behaviour, and traffic law enforcement. Besides conducting high-quality research, these centres fulfil an important advocacy role in promoting safer road use and facilitating collaborations with government and other agencies, at both the national and international level. To illustrate the role of these centres, an overview will be provided of the Centre for Accident Research and Road Safety-Queensland (CARRS-Q), which was established in 1996 and has gone on to become a recognised world-leader in road safety and injury prevention research. The Centre’s research findings are used to provide evidence-based recommendations to government and have directly contributed to promoting safer road use in Australia. Since 2006, CARRS-Q has also developed strong collaborative links with various universities and organisations in China to assist in building understanding, connections and capacity to assist in reducing the road trauma burden. References Hu, G., Wen, M., Baker, T. D., & Baker, S. P. (2008). Road-traffic deaths in China, 1985–2005: threat and opportunity. Injury Prevention, 14, 149-153. Ma, S., Li, Q., Zhou, M., Duan, L., & Bishai, D. (2012). Road Traffic Injury in China: A Review of National Data Sources. Traffic Injury Prevention, 13(S1), 57-63.

Interventions to control nosocomial infections : study designs and statistical issues

There is a wide range of potential study designs for intervention studies to decrease nosocomial infections in hospitals. The analysis is complex due to competing events, clustering, multiple timescales and time-dependent period and intervention variables. This review considers the popular pre-post quasi-experimental design and compares it with randomized designs. Randomization can be done in several ways: randomization of the cluster [intensive care unit (ICU) or hospital] in a parallel design; randomization of the sequence in a cross-over design; and randomization of the time of intervention in a stepped-wedge design. We introduce each design in the context of nosocomial infections and discuss the designs with respect to the following key points: bias, control for nonintervention factors, and generalizability. Statistical issues are discussed. A pre-post-intervention design is often the only choice that will be informative for a retrospective analysis of an outbreak setting. It can be seen as a pilot study with further, more rigorous designs needed to establish causality. To yield internally valid results, randomization is needed. Generally, the first choice in terms of the internal validity should be a parallel cluster randomized trial. However, generalizability might be stronger in a stepped-wedge design because a wider range of ICU clinicians may be convinced to participate, especially if there are pilot studies with promising results. For analysis, the use of extended competing risk models is recommended.

Assessment of the short-term success of a translocation of lesser short-tailed bats Mystacina tuberculata

Translocation is a powerful tool that has been used in the conservation of a wide range of taxa. However, few translocations of bats have been attempted and we know of no successes. The few translocations which have been attempted have either failed due to dispersal from the release site or have not been monitored sufficiently to determine the cause of failure. We assessed the short-term success of a translocation of lesser short-tailed bats Mystacina tuberculata by the New Zealand Department of Conservation, where 3 release methods were used to minimise dispersal or mortality: bats were juveniles, were maintained in captivity at the release site, and were provided with supplementary food and roosts following release. Success was assessed by determining if founders remained at the release site and maintained condition (weight). Recapture showed that at least 9 of the 20 bats remained at the release site 232 d after release. There was weak evidence that bats lost weight, although final weights were comparable to those of bats from a natural population. However, all bats caputred 8 mo after release had damaged, infected ears and some were balding. The problem was treated but recurred, and bats were returned to captivity. Our results are th first to demonstrate that translocated bats can remain at their release site and survive. However, disease may be an issue in future translocations.

Translocation of bats as a conservation strategy : previous attempts and potential problems

Translocation is an increasingly popular conservation tool from which a wide range of taxa have benefited. However, to our knowledge, bats have not been translocated successfully. Bats differ behaviourally, morphologically and physiologically from the taxa for which translocation the- ory has been developed, so existing guidelines may not be directly transferable. We review previous translocations of bats and discuss characteristics of bats that may require special consideration dur- ing translocation. Their vagility and homing ability, coloniality, roost requirements, potential ability to transmit diseases, susceptibility to anthropomorphic impacts, and cryptic nature have implications for establishing populations, effects of these populations on the release site, and ability to monitor translocation success following release. We hope that our discussion of potential problems will be able to supplement the existing, more generic guidelines to provide a starting point for the planning of bat translocations.