Purification of the cardiac sarcoplasmic reticulum membrane protein phospholamban from recombinant Escherichia coli

Phospholamban (PLN) was expressed in Escherichia coli as a protein fusion with glutathione S-transferase (GST). GST-PLN was mostly present in the insoluble protein fraction and accounted for approximately 50% of total insoluble protein. Attempts to suppress inclusion body formation or to use GST as an affinity-purification tag failed. A successful purification method is based on preparative SDS/PAGE and electrodialysis. From 1 g cells we typically purified 13.5 mg fusion protein with a PLN content of 2.8 mg. We genetically inserted an enterokinase (EK) protease site just in front of the PLN sequence and demonstrated the proteolytical liberation of PLN from the carrier protein. The approach described represents a substantial advancement in PLN expression and purification.     

Perspectives on Design and Innovation

A series of studies of top European and other firms has revealed patterns of design management associated with commercial success. Firms that invest resources and professional expertise in product and industrial design in traditional and new industries have been commercially more successful than firms that pay less attention to these aspects of design. As an industry matures there is a shift in emphasis from design associated with technological innovation, to designs supporting technical improvements, and then to supporting user needs, fashion and product variants. These issues are illustrated through the history of the evolution of the bicycle.     

A comparison of CCS tandem overload controls

In this paper we analyse issues associated with overloads in tandem switching systems in Common Channel Signalling (CCS) networks. In particular, we examine the need for, and effectiveness of, Automatic Congestion Control (ACC), a CCS mechanism which allows a congested switching system to inform adjacent CCS nodes of its level of overload, and subsequently controls the rate at which traffic is sent to the congested node. We demonstrate the critical need for end-offices and tandem switching systems to implement some form of ACC if network performance is to be maintained during congestion, especially in tandem exchanges. In addition, we find that the presently defined ACC should be expanded to allow more levels of congestion to be reported to adjacent nodes, so that a more effective, finer grained control is produced.     

A delta transform approach to loop gain-phase shaping design of robust digital control systems

This paper addresses the existence of loop gain-phase shaping (LGPS) solutions for the design of robust digital control systems for SISO, minimum-phase, continuous-time processes with parametric uncertainty. We develop the frequency response properties of LGPS for discrete-time systems using the Δ-transform, a transform method that applies to both continuous-time and discrete-time systems. A theorem is presented which demonstrates that for reasonable specifications there always exists a sampling period such that the robust digital control problem has a solution. Finally, we offer a procedure for estimating the maximum feasible sampling period for LGPS solutions to robust digital control problems.     

Guide to conducting a psychological autopsy.

Much of the writing on the psychological autopsy is reviewed. Several writers' opinions about what should be included in a comprehensive behavioral analysis conducted after an individual's death are compiled in an Appendix. Several additional and new areas to be covered in a psychological autopsy are included. The primary emphasis is on the provision of a practical guide for the working professional. Twenty-four separate factors, ranging from reconstruction of events that occurred before the death to the deceased's family death history, are presented. Some legal considerations are examined briefly in order to acquaint the professional with the significant impact that the results can have on family benefits and criminal prosecution. (PsycINFO Database Record (c) 2010 APA, all rights reserved)