The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2

Activation of the cyclin-dependent protein kinases p34cdc2 and p33cdk2 requires binding with a cyclin partner and phosphorylation on the first threonine residue in the sequence THEVVTLWYRAPE. We present evidence that this threonine residue, number 160 in p33cdk2, can be specifically phosphorylated by a cdc2-related protein kinase from Xenopus oocytes called p40MO15. Binding to cyclin A and phosphorylation of this threonine are both required to activate fully the histone H1 kinase activity of p33cdk2. In cell extracts, a portion of p40MO15 is found in a high molecular weight complex that is considerably more active than a lower molecular weight form. Wild-type MO15 protein expressed in bacteria does not possess kinase activity, but acquires p33cdk2-T160 kinase activity after incubation with cell extract and ATP. We conclude that p40MO15 corresponds to CAK (cdc2/cdk2 activating kinase) and speculate that, like p33cdk2 and p34cdc2, p40MO15 requires activation by phosphorylation and association with a companion subunit.     

Benchmarking of electricity networks: Practical problems with its use for regulation

Benchmarking is a technique derived for management purposes, intended to identify outliers in any population as possible problems for resolution. Several regulatory agencies have tried to use benchmarking to define the degree of inefficiency of regulated companies, by reference to some target or frontier. This paper identifies the main problem inherent in trying to adapt a management technique to a regulatory purpose – namely that it requires the exercise of subjective judgements. The resulting lack of predictability and objectivity is not conducive to the provision of efficient regulatory incentives.This paper explains where subjective judgement enters into regulatory applications of benchmarking, which is not always apparent. The choice of model and selection of data sets are two obvious areas, but the main problem arises over the assumption that any costs not explained by the model must be due to inefficiency. Such assumptions are simply unsupported by evidence and lead to cost targets (or cost reduction targets defined over several years) that are little better than subjective guesses. There are other, superior ways to set cost reduction targets, based on long-term trends in total factor productivity for the regulated sector, which are used in the US and in other countries. However, some regulators feel obliged to use benchmarking and the paper concludes with recommendations as to how benchmarking should support further and more objective investigations into the costs of regulated utilities.     

Over-production of 5-enolpyruvylshikimate-3-phosphate synthase in Escherichia coli: use of the T7 promoter

5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, the productof the Escherichia coli aroA gene, has been overproduced inE.coli BL21(DE3) under the control of the T7 gene 10 promoterand ribosome binding site, to a level of {small tilde}50% oftotal cell protein. EPSP synthase is the primary target of thepost-emergence herbicide, glyphosate, commonly known as Roundup^TM.A simple two step purification is described, which results in99% pure homogeneous protein (as determined by PAGE). The integrityof the protein has been compared with previously characterizedprotein from .E.coli AB2829(pKD501) by determination of itskinetic parameters, N-terminal protein and DNA sequences, aminoacid analysis and ¹³C-NMR spectroscopy. This new overproducingstrain readily provides the gram quantities of highly pure proteinrequired for NMR studies of the active site and the developmentof novel time-resolved solid-state NMR techniques currentlyunderway in this laboratory.     

Automatic detection of microaneurysms in colour fundus images for diabetic retinopathy screening

Regular eye screening is essential for the early detection and treatment of the diabetic retinopathy. This paper presents a novel automatic screening system for diabetic retinopathy that focuses on the detection of the earliest visible signs of retinopathy, which are microaneurysms. Microaneurysms are small dots on the retina, formed by ballooning out of a weak part of the capillary wall. The detection of the microaneurysms at an early stage is vital, and it is the first step in preventing the diabetic retinopathy. The paper first explores the existing systems and applications related to diabetic retinopathy screening, with a focus on the microaneurysm detection methods. The proposed decision support system consists of an automatic acquisition, screening and classification of diabetic retinopathy colour fundus images, which could assist in the detection and management of the diabetic retinopathy. Several feature extraction methods and the circular Hough transform have been employed in the proposed microaneurysm detection system, alongside the fuzzy histogram equalisation method. The latter method has been applied in the preprocessing stage of the diabetic retinopathy eye fundus images and provided improved results for detecting the microaneurysms.     

Transient analysis using quasiperiodic forcing functions

The unit step is expressed as a convolution of frequency-modulated sinusoids. The response of a system to the step defined in this manner is then examined, and a method is indicated whereby transient information may be extracted.     

FieldMAP: A Spatiotemporal Field Monitoring Application Prototyping Framework

The fundamental aim of monitoring is to identify abnormalities in the observed phenomena and allow inference of the likely cause. Faced with the common problems of spatially irregular sensor distribution and intermittent sensor measurement availability, key to fulfilling the monitoring aim is filling in the spatiotemporal gaps in the data. While wireless sensor networks (WSNs) technology, combined with microelectromechanical systems availability potentially offer sensing solutions for a variety of application domains, in the context of monitoring applications a conceptual shift is needed from currently available, point-measurement-based ldquosense-and-sendrdquo systems toward the provision of phenomena field representations, in real time, enabling effective visualization of the spatiotemporal patterns. This paper argues the case for a generic, rapid prototyping framework for end-to-end sensing systems that support the approach of providing field representations for visualization. A formal approach to framework development was taken, ensuring that resulting instrumentation systems are well specified. Both the framework development and its evaluation are linked to the full cycle of requirements setting, design, and deployment of a prototype instrumentation system for aerospace applications-specifically, health monitoring of a gas turbine engine. The field monitoring application prototyping (FieldMAP) framework supports multimodal sensing, provides a number of opportunities for data processing and information extraction, caters for monitoring of the instrumentation health, offers a modular field-mapping design component and allows for real-time phenomena visualization, data and information logging, and postanalysis. Experience with the FieldMAP has shown that sophisticated and robust prototypes can be developed in a short period of time.     

Scanning transmission electron microscopy mass analysis of fibrillin-containing microfibrils from foetal elastic tissues

We have applied scanning transmission electron microscopy to intact native fibrillin-containing microfibrils isolated from foetal bovine elastic tissues in order to derive new insights into microfibril organisation. This technique provides quantitative data on the mass per unit length and axial mass distribution of unstained, unshadowed macromolecules. Scanning transmission electron microscopy of microfibrils from aorta, skin and nuchal ligament revealed that the beads corresponded to peaks of mass and the interbead regions to troughs of mass. These major features of axial mass distribution were characteristic of all microfibrils examined. Tissue-specific and age-dependent variations in mass were identified in microfibrils that were structurally comparable by rotary shadowing electron microscopy. Increased microfibril mass correlated with increasing gestational age. The additional mass was associated predominantly at, or close to, the bead. Some microfibril populations exhibited pronounced assymetry in their axial mass distribution. These data indicate that intact native microfibrillar assemblies from developing elastic tissues are heterogeneous in composition. Loss of mass following chondroitinase ABC or AC lyase treatment confirmed the presence of chondroitin sulphate in nuchal ligament microfibrillar assemblies.     

Impact of horizontal and vertical heterogeneities on retrievals using multiangle microwave brightness temperature data

This paper investigates the impact of heterogeneity at the land surface on geophysical parameters retrieved from multiangle microwave brightness temperature data, such as would be obtained from the Soil Moisture and Ocean Salinity (SMOS) mission. Synthetic brightness temperature data were created using the Common Land (land surface) Model, coupled with a microwave emission model and set within the framework of the North American Land Data Assimilation System (NLDAS). Soil moisture, vegetation optical depth, and effective physical temperature were retrieved using a multiobjective calibration routine similar to the proposed SMOS retrieval algorithm for a typical on-axis range of look angles. The impact of heterogeneity both in the near-surface profiles of soil moisture and temperature and in the land cover on the accuracy of the retrievals was examined. There are significant errors in the retrieved parameters over regions with steep gradients in the near-surface soil moisture profile. These errors are approximately proportional to the difference in the soil water content between the top (at 0.7 cm) and second layer (at 2.7 cm) of the land surface model. The errors resulting from heterogeneity in the land cover are smaller and increase nonlinearly with increasing land-surface heterogeneity (represented by the standard deviation of the optical depth within the pixel). The most likely use of retrieved soil moisture is through assimilation into an LDAS for improved initiation of weather and climate models. Given that information on the soil moisture profile is already available within the LDAS, the error in the retrieved soil moisture as a result of the near-surface profile can be corrected for. The potential errors as a result of land-surface heterogeneity can also be assessed for use in the assimilation process.     

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