Controlled oxidative protein refolding using an ion-exchange column

Column-based refolding of complex and highly disulfide-bonded proteins simplifies protein renaturation at both preparative and process scale by integrating and automating a number of operations commonly used in dilution refolding. Bovine serum albumin (BSA) was used as a model protein for refolding and oxido-shuffling on an ion-exchange column to give a refolding yield of 55 % after 40 Ih incubation. Successful on-column refolding was conducted at protein concentrations of up to 10 mg/ml and refolded protein, purified from misfolded forms, was eluted directly from the column at a concentration of 3 mg/ml. This technique integrates the dithiothreitol removal, refolding, concentration and purification steps, achieving a high level of process simplification and automation, and a significant saving in reagent costs when scaled. Importantly, the current result suggests that it is possible to controllably refold disulfide-bonded proteins using common and inexpensive matrices, and that it is not always necessary to control protein-surface interactions using affinity tags and expensive chromatographic matrices. Moreover, it is possible to strictly control the oxidative refolding environment once denatured protein is bound to the ion-exchange column, thus allowing precisely controlled oxido-shuffling. (c) 2005 Elsevier B.V. All rights reserved.

Fusing subunit antigens to interleukin-2 and encapsulating them in liposomes improves their antigenicity but not their protective efficacy

Subunit vaccines commonly lack sufficient immunogenicity to stimulate a comprehensive protective immune response in vivo. We have investigated the potential of specific cytokines (interleukin-2) and particulate delivery systems (liposomes) to enhance antigenicity. Here we report that the IgG1 and IFN-gamma responses to a subunit antigen, consisting of a T and B-cell epitope from Influenza haemagglutinin, can be improved when it is both fused to interelukin-2 and encapsulated in liposomes. However, this vaccine formulation was not able to protect animals against a challenge with live Influenza A/PR/8/34 virus. The addition of more potent immune stimulators may be necessary to improve responses. (c) 2005 Elsevier Ltd. All rights reserved.

Adsorption of flexible n-alkane on graphitized thermal carbon black: analysis of adsorption isotherm by means of GCMC simulation

In this paper, we investigate the suitability of the grand canonical Monte Carlo in the description of adsorption equilibria of flexible n-alkane (butane, pentane and hexane) on graphitized thermal carbon black. Potential model of n-alkane of Martin and Siepmann (J. Phys. Chem. 102 (1998) 2569) is employed in the simulation, and we consider the flexibility of molecule in the simulation. By this we study two models, one is the fully flexible molecular model in which n-alkane is subject to bending and torsion, while the other is the rigid molecular model in which all carbon atoms reside on the same plane. It is found that (i) the adsorption isotherm results of these two models are close to each other, suggesting that n-alkane model behaves mostly as rigid molecules with respect to adsorption although the isotherm for longer chain n-hexane is better described by the flexible molecular model (ii) the isotherms agree very well with the experimental data at least up to two layers on the surface.

A study on transition of iron from active into passive state

An investigation was carried out on the transition of an iron electrode from active to passive state in a sulphuric acid solution. It was found that the active-passive transition was an auto-catalytic process in which a pre-passive film grew on the electrode surface. The growing pre-passive film had a fractal edge whose dimension was affected by the applied passivating potential and the presence of chlorides in the solution. Applying a more positive passivating potential led to a faster active-passive transition and resulted in a more irregular pre-passive film. If chlorides were introduced into the sulphuric acid solution, the active-passive transition became more rapid and the pre-passive film more irregular. Apart from the influence on the growth of the pre-passive film, the presence of chlorides in the passivating solution was found to deteriorate the stability of the final passive film. All these phenomena can be understood if the passivating iron electrode is regarded as a dissipative system. To explain these results, a fractal pre-film model is proposed in this paper. (C) 2004 Elsevier Ltd. All rights reserved.

Optofluidics enables compact tunable interferometer

An optofluidic interferometer - in which a beam propagates across an interface between fluid and air - modulates at high extinction ratios and is only microns in size.

Comparative adsorption of spherical argon and flexible n-butane in carbon slit pores - a GCMC computer simulation study

In this paper we investigate the difference between the adsorption of spherical molecule argon (at 87.3 K) and the flexible normal butane (at an equivalent temperature of 150 K) in carbon slit pores. These temperatures are equivalent in the sense that they have the same relative distances between their respective triple points and critical points. Higher equivalent temperatures are also studied (122.67 K for argon and 303 K for n-butane) to investigate the effects of temperature on the 2D-transition in adsorbed density. The Grand Canonical Monte Carlo simulation is used to study the adsorption of these two model adsorbates. Beside the longer computation times involved in the computation of n-butane adsorption, n-butane exhibits many interesting behaviors such as: (i) the onset of adsorption occurs sooner (in terms of relative pressure), (ii) the hysteresis for 2D- and 3D-transitions is larger, (iii) liquid-solid transition is not possible, (iv) 2D-transition occurs for n-butane at 150 K while it does not happen for argon except for pores that accommodate two layers of molecules, (v) the maximum pore density is about four times less than that of argon and (vi) the sieving pore width is slightly larger than that for argon. Finally another feature obtained from the Grand Canonical Monte Carlo (GCMC) simulation is the configurational arrangement of molecules in pores. For spherical argon, the arrangement is rather well structured, while for n-butane the arrangement depends very much on the pore size. (C) 2004 Elsevier B.V. All rights reserved.

Corrosion of magnesium alloys in commercial engine coolants

A number of magnesium alloys show promise as engine block materials. However, a critical issue for the automotive industry is corrosion of the engine block by the coolant and this could limit the use of magnesium engine blocks. This work assesses the corrosion performance of conventional magnesium alloy AZ91D and a recently developed engine block magnesium alloy AM-SC1 in several commercial coolants. Immersion testing, hydrogen evolution measurement, galvanic current monitoring and the standard ASTM D1384 test were employed to reveal the corrosion performance of the magnesium alloys subjected to the coolants. The results show that the tested commercial coolants are corrosive to the magnesium alloys in terms of general and galvanic corrosion. The two magnesium alloys exhibited slightly different corrosion resistance to the coolants with AZ91D being more corrosion resistant than AM-SC1. The corrosivity varied from coolant to coolant. Generally speaking. an oraganic-acid based long life coolant was less corrosive to the magnesium alloys than a traditional coolant. Among the studied commercial coolants. Toyota long, life coolant appeared to be the most promising one. In addition. it was found that potassium fluoride effectively inhibited corrosion of the magnesium alloys in the studied commercial coolants. Both general and galvanic corrosion rates were significantly decreased by addition of KF, and there were no evident side effects on the other engine block materials, such as copper, solder. brass. steel and aluminium alloys, in terms of their corrosion performance. The ASTM D 1384 test further confirmed these results and suggested that Toyota long life coolant with 1%wt KF addition is a promising coolant for magnesium engine blocks.

Why H Atom Prefers the On-Top Site and Alkali Metals Favor the Middle Hollow Site on the Basal Plane of Graphite

In this work, the different adsorption properties of H and alkali metal atoms on the basal plane of graphite are studied and compared using a density functional method on the same model chemistry level. The results show that H prefers the on-top site while alkali metals favor the middle hollow site of graphite basal plane due to the unique electronic structures of H, alkali metals, and graphite. H has a higher electronegativity than carbon, preferring to form a covalent bond with C atoms, whereas alkaline metals have lower electronegativity, tending to adsorb on the highest electrostatic potential sites. During adsorption, there are more charges transferred from alkali metal to graphite than from H to graphite.

Understanding service experience in non-profit performing arts: Implications for operations and service management

Recent research in the non-profit performing arts has shown that marketing efforts designed to increase revenue from ticket sales are not achieving the results required to sustain the performing arts. This paper applies operations management analytical techniques to the non-profit performing arts to increase understanding of operational issues and inform service management strategy. The paper takes a two-study idiographic approach. Implementing a modified version of service transaction analysis (STA), Study One describes a performing arts service from provider and customer perspectives, identifies service gaps and develops an elaborated service description incorporating both perspectives. In Study Two, building on the elaborated service description and extant research, in-depth interviews are conducted to gather thick descriptions of predictors of satisfaction, value and service quality as they relate to repurchase intention (RI). Technical, functional and critical factors required to improve organizational performance are identified. Implications for operational strategy, service design and service management theory for this context are discussed. (c) 2005 Published by Elsevier B.V.

Responding to our customers' needs: modifications to assessment in Obstetrics and Gynaecology over 20 years

Background and Objectives: This paper reports on historical changes in assessment culminating in the experience of one discipline with negotiated student feedback that has helped design and modify assessment to cater for the requirements of both students and teachers. The standard of assessment required to pass Obstetrics and Gynaecology in the four year graduate entry program in the School of Medicine at The University of Queensland, Brisbane, Australia has become less formalised and more collaborative. Changes in assessment in this discipline over the last 20 years reflect the development of an understanding of the educational principles associated with adult teaching and learning. Assessment has evolved from being teacher focussed, with questionable reliability, validity, and emphasis on outcomes, to being focussed on learning and the student. Multiplechoice examinations, combined with a collaborative approach to the reliability and validity of questions and answers and a debrief or feedback session have been found to provide an assessment format that is art acceptable measure oflearning for both teachers and students. Changes in assessment reflect a collaborative process between teachers and students based on principles of adult learning and involving negotiated student feedback. Our experience with this form of negotiated outcome for assessment is presented together with suggestions for improvement and is contrasted with assessment methods used in this department over the last 20 years. Change and refinement will continue as medical programs strive to meet the learning needs of students and assessment outcomes that are acceptable to its teachers.

Existence of complementary pairs of proportionally balanced designs

Proportionally balanced designs (pi BDs) were introduced by Gray and Matters in response to a need for the allocation of markers of the Queensland Core Skills Test to have a certain property. Subsequent papers extended the theoretical results relating to such designs and provided further instances and general constructions. This work focused on designs comprising blocks of precisely two sizes, and when each variety occurs with one of precisely two possible frequencies. Two designs based on the set V of varieties are complementary if, whenever B is a block of one, then its complement with regard to the set V is a block of the other. Here we present necessary conditions for the existence of complementary pairs of such pi BDs and provide lists of some restricted parameter sets satisfying these necessary conditions. The lists are arranged according to the number of blocks. We demonstrate that not all of these parameter sets give rise to designs. However we establish by construction of the sets of blocks that, for every feasible number of blocks less than or equal to 100, with the possible exception of 63, there exists at least one pair of complementary pi BDs. We also investigate the conditions under which the complementary design can be isomorphic to the original design, and again provide a list of feasible parameters for pairs of such designs with at most 400 blocks.

The use of internal audit by Australian companies

Purpose – The purpose of this study is to explore the voluntary use of internal audit by Australian publicly listed companies and to identify factors that lead listed companies to have an internal audit function. Design/methodology/approach – Drawing on the Institute of Internal Auditors' definition of internal auditing, the paper predicts that internal audit use is associated with factors related to risk management, strong internal controls and strong corporate governance. To test the predictions, the study combines data from a survey of listed companies with information from corporate annual reports. The paper also provides descriptive information on the use of internal audit. Findings – The results indicate that only one-third of the sample companies use internal audit. While size appears to be the dominant driver, there is also a strong association between internal audit and the level of commitment to risk management. However, the study finds only weak support for an association between the use of internal audit and strong corporate governance. Research limitations/implications – A limitation of our study is that some of the variables in the model may not be good proxies for the factors being measured. Refinement of the model and the variables used provides an opportunity for future research. Practical implications – The limited use of internal audit by Australian companies has important implications for sound corporate governance. Originality/value – This is the first study that identifies factors associated with the use of internal audit by Australian listed companies.

Mefloquine and doxycycline malaria prophylaxis in Australian soldiers in East Timor

Objectives: To describe the tolerability of mefloquine in Australian soldiers for malaria prophylaxis, including a comparison with doxycycline. Design: Open-label, prospective study and cross-sectional questionnaire and interview. Setting and participants: Two contingents of Australian soldiers, each deployed to East Timor for peacekeeping duties over a 6-month period (April 2001-October 2001 and October 2001-May 2002). Outcome measures: Withdrawals during the study; adverse events relating to mefloquine prophylaxis; willingness to use mefloquine again on deployment. Results: Of 1157 soldiers starting on mefloquine, 75 (6.5%) withdrew because of adverse responses to the drug. There were three serious adverse events of a neuropsychiatric nature, possibly relating to mefloquine. Fifty-seven per cent of soldiers using mefloquine prophylaxis reported at least one adverse event, compared with 56% using doxycycline. The most commonly reported adverse effects of both drugs were sleep disturbance, headache, tiredness and nausea. Of the 968 soldiers still taking mefloquine at the end of their deployments, 94% indicated they would use mefloquine again. Of 388 soldiers taking doxycycline prophylaxis who were deployed with the first mefloquine study contingent, 89% indicated they would use doxycycline again. Conclusions: Mefloquine was generally well tolerated by Australian soldiers and should continue to be used for those intolerant of doxycycline.