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.

An analysis of the images attached to referral messages in an e-mail based telemedicine system for developing countries

Little is known about the quality of the images transmitted in email telemedicine systems. The present study was designed to survey the quality of images transmitted in the Swinfen Charitable Trust email referral system. Telemedicine cases were examined for a 3 month period in 2002 and a 3 month period in 2006. The number of cases with images attached increased from 8 (38%) to 37 (53%). There were four types of images (clinical photographs, microscope pictures, notes and X-ray images) and the proportion of radiology images increased from 27 to 48%. The cases in 2002 came from four different hospitals and were associated with seven different clinical specialties. In 2006, the cases came from 19 different hospitals and 20 different specialties. The 46 cases (from both study periods) had a total of 159 attached images. The quality of the images was assessed by awarding each image a score in four categories: focus, anatomical perspective, composition and lighting. The images were scored on a five-point scale (1 = very poor to 5 =very good) by a qualified medical photographer. In comparing image quality between the two study periods, there was some evidence that the quality had reduced, although the average size of the attached images had increased. The median score for all images in 2002 was 16 (interquartile range 14-19) and the median score in 2006 was 15 (13-16). The difference was significant (P < 0.001, Mann-Whitney test).

Dominance hierarchies and health: Constructing personal zones of spiritual power and healing in modern medicine

Ethological studies of animals in groups and sociobiology indicate that hierarchies of dominance amongst some species ensure the survival of the group. When transferred to human groups, dominance hierarchies suggest a crucial role played by recasting the scope of such hierarchies of dominant and subordinate members to included hyper-dominant beings. A recognition of such beings as even more dominant than the socially dominant members of a hierarchy facilitates the empowerment of the socially subordinate members. Religious belief and practice works to establish such hyper-dominant beings (gods, goddesses, and so forth) as superior members of human groups. Doing so is a means of ensuring the survival of the species and, thus, enhancing healing and human health. The doctor-patient relationship is examined from such a point of view, with an emphasis on whether the hierarchy created by the relationship allows consideration of alternative and complementary forms of medical treatment.

Unsafe driving behaviour and four wheel drive vehicles: observational study

Objective To assess the level of compliance with the new law in the United Kingdom mandating penalties for rising a hand held mobile phone while driving, to compare compliance with this law with the one on the use of seat belts, and to compare compliance with these laws between drivers of four wheel drive vehicles and drivers of normal cars. Design Observational study with two phases-one within the grace period, the other starting one week after penalties were imposed on drivers using such telephones. Setting Three busy sites in London. Participants Drivers of 38 182 normal cars and 2944 four wheel drive vehicles. Main outcome measures Proportions of drivers seen to be using hand held mobile phones and not using seat belts. Results Drivers of four wheel drive vehicles were more likely than drivers of other cars to be seen using hand held mobile phones (8.2% v 2.0%) and not complying with the law on seat belts (19.5% v 15.0%). Levels of non-compliance with both laws were slightly higher in the penalty phase of observation, and breaking one law was associated with increased likelihood of breaking the other. Conclusions The level of non-compliance with the law on the use of hand held mobile phones by drivers in London is high, as is non-compliance with the law on seat belts. Drivers of four wheel drive vehicles were four times more likely than drivers of other cars to be seen using hand held mobile phones and slightly more likely not to comply with the law on seat belts.