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.

Benchmarking of performance of Mexican states with effective coverage

Benchmarking of the performance of states, provinces, or districts in a decentralised health system is important for fostering of accountability, monitoring of progress, identification of determinants of success and failure, and creation of a culture of evidence. The Mexican Ministry of Health has, since 2001, used a benchmarking approach based on the World Health Organization (WHO) concept of effective coverage of an intervention, which is defined as the proportion of potential health gain that could be delivered by the health system to that which is actually delivered. Using data collection systems, including state representative examination surveys, vital registration, and hospital discharge registries, we have monitored the delivery of 14 interventions for 2005-06. Overall effective coverage ranges from 54.0% in Chiapas, a poor state, to 65.1% in the Federal District. Effective coverage for maternal and child health interventions is substantially higher than that for interventions that target other health problems. Effective coverage for the lowest wealth quintile is 52% compared with 61% for the highest quintile. Effective coverage is closely related to public-health spending per head across states; this relation is stronger for interventions that are not related to maternal and child health than those for maternal and child health. Considerable variation also exists in effective coverage at similar amounts of spending. We discuss the implications of these issues for the further development of the Mexican health-information system. Benchmarking of performance by measuring effective coverage encourages decision-makers to focus on quality service provision, not only service availability. The effective coverage calculation is an important device for health-system stewardship. In adopting this approach, other countries should select interventions to be measured on the basis of the criteria of affordability, effect on population health, effect on health inequalities, and capacity to measure the effects of the intervention. The national institutions undertaking this benchmarking must have the mandate, skills, resources, and independence to succeed.

The risky business of doctoral management

Universities are under no less pressure to adopt risk management strategies than other public and private organisations. The risk management of doctoral education is a particularly important issue given that a doctorate is the highest academic qualification a university offers and stakes are high in terms of assuring its quality. However, intense risk management can interfere with the intellectual and pedagogical work which are essentially part of doctoral education. This paper seeks to understand how the culture of risk meets the culture of doctoral education and with what effect. The authors draw on sociological understandings of risk in the work of Anthony Giddens (2002) and Ulrich Beck (1992), the anthropological focus on liminality in the work of Mary Douglas (1990), and the psychological theorising of human error in the work of James Reason (1990). The paper concludes that risk consciousness brings its own risks—in particular, the potential transformation of a culture based on intellect into a culture based on compliance.

A Neglected Aspect of Business: Human and Social Capital in the Sri Lankan Tea Plantations

This paper examines idiosyncrasies of tea plantation culture and politics in relation to Sri Lankan national and popular cultural typologies, with special reference to female tea plantation workers. Tea production in Sri Lanka is heavily based on manual labour, and it is the largest industry that provides accommodation for employees and their families. In this paper, it is argued that politico-cultural production relations have dominated labour productivity in tea plantations. Ways in which female workers have been marginalized, through patriarchal politics, ethnicity, religion, education, elitism, and employment are explained. This culture of the plantation community operates negatively with respect to the management agenda. It is also argued that social capital development in tea plantations is important not only for productivity improvement, but also for reasons of political and social obligation for the nation, because migrant plantation workers have been working and living in plantations over 150 years.