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.     

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.     

The antimicrobial effect of wine on Listeria innocua in a model stomach system

A model stomach, containing a food matrix and a synthetic gastric fluid, was used to study the bactericidal effect of ingested wine on Listeria innocua. Volumes of wine equivalent to the ingestion of one glass and half a bottle, led, over a period of less than 2 h, to a reduction of 3 and 4 logarithmic cycles of the initial population respectively. The influence of ethanol and organic acids, wine constituents with known antimicrobial properties, was investigated. Ethanol exhibited a higher bactericidal effect than the mixture of the main wine organic acids. When testing the organic acids separately, malic and lactic acids were found to have the strongest effect. The combination of ethanol with the organic acids acted synergistically but to a lesser extent than wine itself. The results suggest that the ingestion of wine during a meal may diminish the quantity of Listeria persisting further in the alimentary tract.     

Measurements of frequencies and spatial correlations of coherent structures in rod bundle flows

Cross-wire anemometry was used to identify and characterize coherent flow pulsations in isothermal air flow near the gap regions of a five-rod bundle with a design pitch-to-diameter ratio of 1.149 and contained in a quasi-trapezoidal duct. It was confirmed that such pulsations are quasi-periodic and contribute significantly to the velocity fluctuations across the gap. The frequency of pulsations was found to decrease with diminishing rod–wall gap size in the range between 0.015D and 0.250D, where D is the rod diameter. The pulsations in a rod–wall gap and an adjacent rod–rod gap were strongly coupled and occurred at the same frequency as one rod was displaced towards the duct wall.     

Sustaining Rural Landscapes: the role of integrated tourism

The aim of a European Union-funded research project entitled 'Supporting and Promoting Integrated Tourism in Europe's Lagging Rural Regions' (SPRITE) is to analyse and develop the potential for better-integrated tourism in the rural landscapes of Europe. Integrated tourism can be defined as that which is explicitly linked to the localities in which it takes place and, in practical terms, has clear connections with local resources, activities, products, production and service industries, and a participatory local community. Enhancing the sustainable economic potential of rural landscapes requires partnerships among rural people and the sustainable yet productive use of rural resources. A 'culture economy' approach to integrated tourism is proposed, which emphasizes the importance of local identity, the strategic commodification of resources and of place, and the importance of extra-local forces in enabling local activities. It is suggested that this approach may represent a shift beyond more traditional approaches to rural development.     

A surface distortion analysis applied to the hoop/column deployablemesh reflector antenna

A practical approach is demonstrated for the deterministic analysis of surface distortions in reflector antennas, based on a first-order approximation to the aperture field phase. Measured pattern results from the 15-m-diameter hoop/column deployable mesh reflector antenna are used to demonstrate the accuracy which can be obtained with this surface distortion analysis. The only practical limitation of the first-order approximation is determined by the slope derivations of the distorted surface from the best-fit paraboloid     

Occurrence of Lawn Care and Agricultural Pesticides in the Don River and Humber River Watersheds (1998–2002)

Between 1998 and 2002, surface water samples were collected from several sites in the Don River and Humber River watersheds, both tributaries to Lake Ontario, and analyzed for a variety of pesticides, including those used for urban lawn care. Analyses included 152 pesticide active ingredients and eight metabolites. Samples were collected during base flow periods (i.e., dry events) and rainfall events (i.e., wet events). The objectives of the study were to determine which pesticides were detectable, whether there was a difference in the detection frequency between the two watersheds and between upstream and downstream in each river, and whether precipitation influenced the frequency of detection. Eleven pesticides and one metabolite were detected in surface waters of the Don and Humber rivers or their tributaries, with approximately 72% of samples containing at least one pesticide attributable to lawn care use. The pesticides and pesticide metabolite detected in this study included 2,4-D, atrazine, bromacil, carbofuran, chlorpyrifos, cypermethrin, diazinon, dicamba, MECOPROP, metolachlor, metribuzin, and an atrazine metabolite (des-ethyl atrazine). Four pesticides exceeded federal or provincial water quality guidelines/objectives. Diazinon exceeded the provincial water quality objective in 28% of the samples taken. For the three other pesticides (atrazine, carbofuran and chlorpyrifos) exceedance of a water quality criteria occurred in less than 1% of the samples.