Mechanical properties of interfacial films formed by lysozyme self-assembly at the air-water interface

We present the first characterization of the mechanical properties of lysozyme films formed by self-assembly at the air-water interface using the Cambridge interfacial tensiometer (CIT), an apparatus capable of subjecting protein films to a much higher level of extensional strain than traditional dilatational techniques. CIT analysis, which is insensitive to surface pressure, provides a direct measure of the extensional stress-strain behavior of an interfacial film without the need to assume a mechanical model (e.g., viscoelastic), and without requiring difficult-to-test assumptions regarding low-strain material linearity. This testing method has revealed that the bulk solution pH from which assembly of an interfacial lysozyme film occurs influences the mechanical properties of the film more significantly than is suggested by the observed differences in elastic moduli or surface pressure. We have also identified a previously undescribed pH dependency in the effect of solution ionic strength on the mechanical strength of the lysozyme films formed at the air-water interface. Increasing solution ionic strength was found to increase lysozyme film strength when assembly occurred at pH 7, but it caused a decrease in film strength at pH 11, close to the pI of lysozyme. This result is discussed in terms of the significant contribution made to protein film strength by both electrostatic interactions and the hydrophobic effect. Washout experiments to remove protein from the bulk phase have shown that a small percentage of the interfacially adsorbed lysozyme molecules are reversibly adsorbed. Finally, the washout tests have probed the role played by additional adsorption to the fresh interface formed by the application of a large strain to the lysozyme film and have suggested the movement of reversibly bound lysozyme molecules from a subinterfacial layer to the interface.

Production of Enzymatically active ketosteroid isomerase following insoluble expression in Escherichia coli

Enzymatically active Delta(5)-3-ketosteroid isomerase (KSI) protein with a C-terminus his(6)-tag was produced following insoluble expression using Escherichia coli. A simple, integrated process was used to extract and purify the target protein. Chemical extraction was shown to be as effective as homogenization at releasing the inclusion body proteins from the bacteria] cells, with complete release taking less than 20 min. An expanded bed adsorption (EBA) column utilizing immobilized metal affinity chromatography (IMAC) was then used to purify the denatured KSI-(His(6)) protein directly from the chemical extract. This integrated process greatly simplifies the recovery and purification of inclusion body proteins by removing the need for mechanical cell disruption, repeated inclusion body centrifugation, and difficult clarification operations. The integrated chemical extraction and EBA process achieved a very high purity (99%) and recovery (89%) of the KSI-(His(6)), with efficient utilization of the adsorbent matrix (9.74 mg KSI-(His(6))/mL adsorbent). Following purification the protein was refolded by dilution to obtain the biologically active protein. Seventy-nine percent of the expressed KSI-(His(6)) protein was recovered as enzymatically active protein with the described extraction, purification, and refolding process. In addition to demonstrating the operation of this intensified inclusion body process, a plate-based concentration assay detecting KSI-(His(6)) is validated. The intensified process in this work requires minimal optimization for recovering novel his-tagged proteins, and further improves the economic advantage of E. coli as a host organism. (c) 2006 Wiley Periodicals, Inc.

Career development: A project for the 21st century

This article reports of the papers present at the International Symposium 2006 'Shaping the future: connecting career development and workforce development'. The International Symposium 2006 provided an opportunity to move the project forward by considering career development in relation to the workforce development issues of human capital, labour supply, employability skills and older workers. In addition to these specific issues, it examined the broader issues of how career development services might contribute to workforce development and the career development information base needed to support public policy making. By way of background to this special issue on the International Symposium 2006, this paper briefly examines the context and the reasons behind career development's rise to a more prominent position on the public policy arena. Following this, the process of the International Symposium 2006 that resulted in the writing of the documents contained in the special issue are briefly outlined.

Going up? Women in the public sector

Abstract: Purpose – This paper aims to document women's reflections on their careers over a ten-year period to provide quantitative baseline data on which to frame follow-up in-depth interviews. The participants work in the public service in Queensland (Australia) and had been recommended for, and participated in, women in management (WIM) courses conducted in the early 1990s. Design/methodology/approach – Data were collected by means of a survey (containing closed and open items) which gathered demographic data and data related to employment history, perceptions of success and satisfaction, and the women's future career expectations. Findings – Findings revealed that the percentage of women in middle and senior management had increased over the ten-year period, although not to the extent one might have anticipated, given that the women had been targeted as high flyers by their supervisors. While not content with their classification levels (i.e. seniority), the majority of the cohort viewed their careers as being successful. Practical implications – Questions arise from this study as to why women are still “not getting to the top”. There are also policy implications for the public service concerning women's possible “reinventive contribution” and training implications associated with women only courses. Originality/value – The study is part of an Australian longitudinal study on the careers of women who attended a prestigious women-only management course in the early 1990s in Queensland. This is now becoming a study of older women.

The Pedagogical Practice of Locative Experience

We work collectively with varied locative-type projects and look to integrate our students into contemporary experience design culture. Students experience the ‘how and what’ of locative by becoming participant users, being exposed to contemporary works, and placing themselves in the role of the designer producing their own located works.