EdU, a new thymidine analogue for labelling proliferating cells in the nervous system

The standard method of labelling proliferating cells uses the thymidine analogue, bromodeoxyuridine (BrdU), which incorporates into the DNA during S-phase of the cell cycle. A disadvantage of this method is that the immunochemical processing requires pre-treatment of the cells and tissue with heat or acid to reveal the antigen. This pre-treatment reduces reliability of the method and degrades the specimen, reducing the ability for multiple immuno-fluorescence labelling at high resolution. We report here the utility of a novel thymidine analogue, ethynyl deoxyuridine (EdU), detected with a fluorescent azide via the “click” chemistry reaction (the Huisgen 1,3-dipolar cycloaddition reaction of an organic azide to a terminal acetylene). The detection of EdU requires no heat or acid treatment and the incorporated EdU is covalently conjugated to fluorescent probe. The reaction is quick and compatible with fluorescence immunochemistry and other fluorescent probes. We show here that EdU is non-toxic in vitro and in vivo and can be used in place of BrdU to label cells during neurogenesis and the progeny identified at least 30 days later. The fluorescent labelling of EdU, markedly improves the detection of proliferating cells and allows concurrent high resolution fluorescence immunochemistry.

Reimagining Creativity : Critically, Ethically, and Practically

True creativity sits in stark contrast with global trends to standardise education systems. It is, though, an attractive semantic and conjures up all sorts of positives. However none of the positive potentials of the creativity push are likely to materialise while process-oriented bureaucratic understandings of creativity dominate curriculum policy and development. Creativity requires pedagogy to forego substantial levels of control. It also requires ethical content because “creative” is an empty epithet, applying equally to the creation of beautiful music as it does to the creation of a nuclear weapon. In responding to Kapitzke and Hay, this chapter outlines the stark contradictions embedded in the creativity push.

Tomorrow's Technologies

Australian research and technological solutions are now being applied throughout the world.

Improved healing of large segmental defects in the rat femur by reverse dynamization in the presence of bone morphogenetic protein-2

Background Large segmental defects in bone do not heal well and present clinical challenges. This study investigated modulation of the mechanical environment as a means of improving bone healing in the presence of bone morphogenetic protein (BMP)-2. Although the influence of mechanical forces on the healing of fractures is well established, no previous studies, to our knowledge, have described their influence on the healing of large segmental defects. We hypothesized that bone-healing would be improved by initial, low-stiffness fixation of the defect, followed by high-stiffness fixation during the healing process. We call this reverse dynamization. Methods A rat model of a critical-sized femoral defect was used. External fixators were constructed to provide different degrees of stiffness and, importantly, the ability to change stiffness during the healing process in vivo. Healing of the critical-sized defects was initiated by the implantation of 11 mg of recombinant human BMP (rhBMP)-2 on a collagen sponge. Groups of rats receiving BMP-2 were allowed to heal with low, medium, and high-stiffness fixators, as well as under conditions of reverse dynamization, in which the stiffness was changed from low to high at two weeks. Healing was assessed at eight weeks with use of radiographs, histological analysis, microcomputed tomography, dual x-ray absorptiometry, and mechanical testing. Results Under constant stiffness, the low-stiffness fixator produced the best healing after eight weeks. However, reverse dynamization provided considerable improvement, resulting in a marked acceleration of the healing process by all of the criteria of this study. The histological data suggest that this was the result of intramembranous, rather than endochondral, ossification. Conclusions Reverse dynamization accelerated healing in the presence of BMP-2 in the rat femur and is worthy of further investigation as a means of improving the healing of large segmental bone defects. Clinical Relevance These data provide the basis of a novel, simple, and inexpensive way to improve the healing of critical-sized defects in long bones. Reverse dynamization may also be applicable to other circumstances in which bonehealing is problematic.

Toward a sustainable culture of peer partnership

This project is a two phase design working in partnership with five universities to develop, implement and systematically embed a distributive leadership model that aims to embed peer partnership (review, development) within the culture of teaching and learning excellence. This presentation will posit a ‘prototype’ peer review leadership model based on ongoing research that brings together both the fundamentals of peer review with the broader importance of context and persons. It will be argued that essential to teaching development is a need to address not only the implementation of peer partnership programs but also strategies to influence and change both the contexts of teaching and the advantages for colleagues. Peer review as a strategy to develop excellence in teaching needs to be considered from a holistic perspective encompassing all elements of the teaching environment. The emphasis is on working to foster the type of conditions needed for leadership and change to begin and be sustained. The work has implications for policy, research, leadership development and student outcomes and has potential application world-wide. Phase 1 has collected focus interview and questionnaire data to inform the research and is being analysed using a thematic qualitative approach and statistical analysis Evidence is emerging currently as the project is ongoing

Developing a culture of peer review of teaching through a distributive leadership approach

Teaching is a core function of higher education and must be effective if it is to provide students with learning experiences that are stimulating, challenging and rewarding Obtaining feedback on teaching is indispensable to enhancing the quality of learning design, facilitating personal and/or professional development and maximising student learning outcomes. Peer review of teaching has the potential to improve the quality of teaching at tertiary level, by encouraging critical reflection on teaching, innovation in teaching practice and scholarship of teaching at all academic levels. However, embedding peer review within the culture of teaching and learning is a significant challenge that requires sustained commitment from senior leadership as well as those in leadership roles within local contexts.

Overview of the INEX 2008 Ad Hoc Track

This paper gives an overview of the INEX 2008 Ad Hoc Track. The main goals of the Ad Hoc Track were two-fold. The first goal was to investigate the value of the internal document structure (as provided by the XML mark-up) for retrieving relevant information. This is a continuation of INEX 2007 and, for this reason, the retrieval results are liberalized to arbitrary passages and measures were chosen to fairly compare systems retrieving elements, ranges of elements, and arbitrary passages. The second goal was to compare focused retrieval to article retrieval more directly than in earlier years. For this reason, standard document retrieval rankings have been derived from all runs, and evaluated with standard measures. In addition, a set of queries targeting Wikipedia have been derived from a proxy log, and the runs are also evaluated against the clicked Wikipedia pages. The INEX 2008 Ad Hoc Track featured three tasks: For the Focused Task a ranked-list of nonoverlapping results (elements or passages) was needed. For the Relevant in Context Task non-overlapping results (elements or passages) were returned grouped by the article from which they came. For the Best in Context Task a single starting point (element start tag or passage start) for each article was needed. We discuss the results for the three tasks, and examine the relative effectiveness of element and passage retrieval. This is examined in the context of content only (CO, or Keyword) search as well as content and structure (CAS, or structured) search. Finally, we look at the ability of focused retrieval techniques to rank articles, using standard document retrieval techniques, both against the judged topics as well as against queries and clicks from a proxy log.

There is less to this argument than meets the eye

In this article the authors continue the Artificial Intelligence and the law debate begun with Moles' 1991 article. In it the authors answer the latest criticisms made by Moles and others as they explain and argue the case for the practical benefits to be gained by AI systems involving the law.

Cough and exhaled nitric oxide levels : what happens with exercise?

Cough associated with exertion is often used as a surrogate marker of asthma. However, to date there are no studies that have objectively measured cough in association with exercise in children. Our primary aim was to examine whether children with a pre-existing cough have an increase in cough frequency during and post-exercise. We hypothesized that children with any coughing illness will have an increase in cough frequency post-exercise regardless of the presence of exercise-induced broncho-constriction (EIB) or atopy. In addition, we hypothesized that Fractional exhaled nitric oxide (FeNO) levels decreases post-exercise regardless of the presence of EIB or atopy. Children with chronic cough and a control group without cough undertook an exercise challenge, FeNO measurements and a skin prick test, and wore a 24-h voice recorder to objectively measure cough frequency. The association between recorded cough frequency, exercise, atopy, and presence of EIB was tested. We also determined if the change in FeNO post exercise related to atopy or EIB. Of the 50 children recruited (35 with cough, 15 control), 7 had EIB. Children with cough had a significant increase in cough counts (median 7.0, inter-quartile ranges, 0.5, 24.5) compared to controls (2.0, IQR 0, 5.0, p = 0.028) post-exercise. Presence of atopy or EIB did not influence cough frequency. FeNO level was significantly lower post-exercise in both groups but the change was not influenced by atopy or EIB. Cough post-exertion is likely a generic response in children with a current cough. FeNO level decreases post-exercise irrespective of the presence of atopy or EIB. A larger study is necessary confirm or refute our findings.

Artefact ecologies : supporting embodied meeting practices with distance access

Frameworks such as activity theory, distributed cognition and structuration theory, amongst others, have shown that detailed study of contextual settings where users work (or live) can help the design of interactive systems. However, these frameworks do not adequately focus on accounting for the materiality (and embodiment) of the contextual settings. Within the IST-EU funded AMIDA project (Augmented Multiparty Interaction with Distance Access) we are looking into supporting meeting practices with distance access. Meetings are inherently embodied in everyday work life and that material artefacts associated with meeting practices play a critical role in their formation. Our eventual goal is to develop a deeper understanding of the dynamic and embodied nature of meeting practices and designing technologies to support these. In this paper we introduce the notion of "artefact ecologies" as a conceptual base for understanding embodied meeting practices with distance access. Artefact ecologies refer to a system consisting of different digital and physical artefacts, people, their work practices and values and lays emphasis on the role artefacts play in embodiment, work coordination and supporting remote awareness. In the end we layout our plans for designing technologies for supporting embodied meeting practices within the AMIDA project.

The cost of changing your mind!

A recent decision of the Queensland Civil and Administrative Tribunal dealt with the liability of a purchaser to pay a termination penalty where a contract for the purchase of a residential property was terminated during the ‘cooling-off’ period. The decision is Lucy Cole Prestige Properties Broadbeach Pty Ltd ATF Gaindri FT Trust t/as Lucy Cole Prestige Properties Broadbeach Pty Ltd v Kastrissios [2013] QCAT 653.

Analysing the Prospects of a Successful Application Under s 180 of the Property Law Act 1974 (Qld)

Section 180 of the Property Law Act 1974 (Qld) makes provision for an applicant to seek a statutory right of user over a neighbour’s property where such right of use is reasonably necessary in the interests of effective use in any reasonable manner of the dominant land. In recent years, the Queensland courts have been confronted with a number of such applications. Litigation has also been common in New South Wales which has a statutory provision in largely similar terms. This article seeks to identify those factors that have underpinned successful applications, the obstacles that an applicant may encounter and the considerations that have guided the courts when considering the associated issues of compensation and costs.

MCF7/LCC2: A 4-hydroxytamoxifen resistant human breast cancer variant that retains sensitivity to the steroidal antiestrogen ICI 182,780

The development of resistance to the antiestrogen tamoxifen occurs in a high percentage of initially responsive patients. We have developed a new model in which to investigate acquired resistance to triphenylethylenes. A stepwise in vitro selection of the hormone-independent human breast cancer variant MCF-7/LCC1 against 4-hydroxytamoxifen produced a stable resistant population designated MCF7/LCC2. MCF7/LCC2 cells retain levels of estrogen receptor expression comparable to the parental MCF7/LCC1 and MCF-7 cells. Progesterone receptor expression remains estrogen inducible in MCF7/LCC2 cells, although to levels significantly lower than observed in MCF-7 and MCF7/LCC1 cells. MCF7/ LCC2 cells form tumors in ovariectomized nude mice without estrogen supplementation, and these tumors are tamoxifen resistant but can be tstrogen stimulated. Significantly, MCF7/LCC2 cells have retained sensitivity to the steroidal antiestrogen ICI 182,780. These data suggest that some breast cancer patients who acquire resistance to tamoxifen may not develop cross-resistance to treatment with steroidal antiestrogens.

An acute growth factor treatment that preserves function after spinal cord contusion injury

Inflammation of the spinal cord after traumatic spinal cord injury leads to destruction of healthy tissue. This “secondary degeneration” is more damaging than the initial physical damage and is the major contributor to permanent loss of functions. In our previous study we showed that combined delivery of two growth factors, vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), significantly reduced secondary degeneration after hemi-section injury of the spinal cord in the rat. Growth factor treatment reduced the size of the lesion cavity at 30d compared to control animals and further reduced the cavity at 90d in treated animals while in control animals the lesion cavity continued to increase in size. Growth factor treatment also reduced astrogliosis and reduced macroglia/macrophage activation around the injury site. Treatment with individual growth factors alone had similar effects to control treatments. The present study investigated whether growth factor treatment would improve locomotor behaviour after spinal contusion injury, a more relevant preclinical model of spinal cord injury. The growth factors were delivered for the first 7d to the injury site via osmotic minipump. Locomotor behaviour was monitored at 1-28d after injury using the BBB score and at 30d using automated gait analysis. Treated animals had BBB scores of 18; Control animals scored 10. Treated animals had significantly reduced lesion cavities and reduced macroglia/macrophage activation around the injury site. We conclude that growth factor treatment preserved spinal cord tissues after contusion injury, thereby allowing functional recovery. This treatment has the potential to significantly reduce the severity of human spinal cord injuries.