A rapid and efficient two-step gel electrophoresis method for the purification of major rye grass pollen allergens

Purified proteins are mandatory for molecular, immunological and cellular studies. However, purification of proteins from complex mixtures requires specialised chromatography methods (i.e., gel filtration, ion exchange, etc.) using fast protein liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC) systems. Such systems are expensive and certain proteins require two or more different steps for sufficient purity and generally result in low recovery. The aim of this study was to develop a rapid, inexpensive and efficient gel-electrophoresis-based protein purification method using basic and readily available laboratory equipment. We have used crude rye grass pollen extract to purify the major allergens Lol p 1 and Lol p 5 as the model protein candidates. Total proteins were resolved on large primary gel and Coomassie Brilliant Blue (CBB)-stained Lol p 1/5 allergens were excised and purified on a secondary "mini"-gel. Purified proteins were extracted from unstained separating gels and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses. Silver-stained SDS-PAGE gels resolved pure proteins (i.e., 875 μg of Lol p 1 recovered from a 8 mg crude starting material) while immunoblot analysis confirmed immunological reactivity of the purified proteins. Such a purification method is rapid, inexpensive, and efficient in generating proteins of sufficient purity for use in monoclonal antibody (mAb) production, protein sequencing and general molecular, immunological, and cellular studies.

Whisper our futures

Whisper Our Futures was an invited design proposal to produce a major public artwork for the State of Queensland’s 150th Anniversary Celebrations. It involved a network of 100 individual scrolling digital text boxes each with individual audio systems arranged together in a tessellated format. This form (specified by the originating brief) both mimicked the soaring gothic arches typical of Queensland cathedrals and was also suggestive of their stained glass windows. Each text module presented a message in both visual and audible forms for Queenslanders living 150 years hence - spoken both by the general public aw well as prominent figures. In this way the work was designed as a focus of future hope, historical reflection and inspiration to visitors to Queensland cathedrals throughout the entire year of celebrations (2009). The work was planned to premiere at Brisbane’s main Anglican Cathedral and then tour to nine other state cathedrals throughout 2009.---- Two staged proposals and budgets were invited throughout 2007. After the second successful proposal stage the State Premier and cabinet changed, ultimately leading the public art components to be dropped from the program. The proposal currently remains on file at the Queensland Premiers Office.

Enhancing an Incremental Clustering Algorithm for Web Page Collections

With the size and state of the Internet today, a good quality approach to organizing this mass of information is of great importance. Clustering web pages into groups of similar documents is one approach, but relies heavily on good feature extraction and document representation as well as a good clustering approach and algorithm. Due to the changing nature of the Internet, resulting in a dynamic dataset, an incremental approach is preferred. In this work we propose an enhanced incremental clustering approach to develop a better clustering algorithm that can help to better organize the information available on the Internet in an incremental fashion. Experiments show that the enhanced algorithm outperforms the original histogram based algorithm by up to 7.5%.

Ability of silver-impregnated contact lenses to control microbial growth and colonisation

Purpose: To examine the ability of silver nano-particles to prevent the growth of Pseudomonas aeruginosa and Staphylococcus aureus in solution or when adsorbed into contact lenses. To examine the ability of silver nano-particles to prevent the growth of Acanthamoeba castellanii. ----- ----- Methods: Etafilcon A lenses were soaked in various concentrations of silver nano-particles. Bacterial cells were then exposed to these lenses, and numbers of viable cells on lens surface or in solution compared to etafilcon A lenses not soaked in silver. Acanthamoeba trophozoites were exposed to silver nano-particles and their ability to form tracks was examined. ----- ----- Results: Silver nano-particle containing lenses reduced bacterial viability and adhesion. There was a dose-dependent response curve, with 10 ppm or 20 ppm silver showing > 5 log reduction in bacterial viability in solution or on the lens surface. For Acanthamoeba, 20 ppm silver reduced the ability to form tracks by approximately 1 log unit. ----- ----- Conclusions: Silver nanoparticles are effective antimicrobial agents, and can reduce the ability of viable bacterial cells to colonise contact lenses once incorporated into the lens.----- ----- Resumen: Objetivos: Examinar la capacidad de las nanopartículas de plata para prevenir el crecimiento de Pseudomonas aeruginosa y Staphylococcus aureus en soluciones para lentes de contacto o cuando éstas las adsorben. Examinar la capacidad de las nanopartículas de plata para prevenir el crecimiento de Acanthamoeba castellanii.----- ----- Métodos: Se sumergieron lentes etafilcon A en diversas concentraciones de nanopartículas de plata. Las células bacterianas fueron posteriormente expuestas a dichas lentes, y se compararon cantidades de células viables en la superficie de la lente o en la solución con las presentes en lentes etafilcon A que no habían sido sumergidas en plata. Trofozoítos de Acanthamoeba fueron expuestos a nanopartículas de plata y se examinó su capacidad para formar quistes.----- ----- Resultados: Las lentes que contienen nanopartículas de plata redujeron la viabilidad bacteriana y la adhesión. Hubo una curva de respuesta dependiente de la dosis, en la que 10 ppm o 20 ppm de plata mostró una reducción logarítmica > 5 en la viabilidad bacteriana tanto en la solución como en la superficie de la lente. Para Acanthamoeba, 20 ppm de plata redujeron la capacidad de formar quistes en aproximadamente 1 unidad logarítmica.----- ----- Conclusiones: Las nanopartículas de plata son agentes antimicrobianos eficaces y pueden reducir la capacidad de células bacterianas viables para colonizar las lentes de contacto una vez que se han incorporado en la lente.

A silver bullet for Australian cinema? Genre movies and the audience debate

There has been a renaissance in Australian genre cinema in recent years. Indeed, not since the 1980s have Australian genre movies across action, adventure, horror, and science-fiction among others, experienced such prominence within production, policy discourse, and industry debate. Genre movies, typically associated with commercial filmmaking and entertainment, have been identified as a strategy to improve the box-office performance of Australian feature films and to attract larger audiences. Much of this conversation has revolved around the question of whether or not genre can deliver on these high expectations and transform the unpredictable local film industry into a popular and profitable commercial production sector. However, this debate for the most part has been disconnected from analysis of Australia’s genre movie heritage in terms of their position within Australian cinema and their reception with domestic audiences, and how this correlates to contemporary trends. As this chapter argues, genre production is not a silver bullet which will single handedly improve the Australian feature film industry’s commercial performance. Genre movies have occupied, and continue to occupy, a difficult position within Australian cinema and face numerous challenges in terms of reception with national audiences, limited production scale and enterprise structures, and ongoing tensions between culture and commerce.