Enterotoxigenic Escherichia coli infection of pigs: expression, extraction, purification and the adhesive properties of the K88 fimbrial adhesin

The ability of Escherichia coli to express the K88 fimbrial adhesin was satisfactorily indicated by the combined techniques of ELISA, haemagglutination and latex agglutination. Detection of expression by electron microscopy and the ability to metabolize raffinose were unsuitable. Quantitative expression of the K88 adhesin was determined by ELISA. Expression was found to vary according to the E.coli strain examined, media type and form. In general it was found that the total amount was greater, while the amount/cfu was less on agar than in broth cultures. Expression of the K88 adhesin during unshaken batch culture was related to the growth rate and was maximal during late logarithmic to early stationary phase. A combination of heat extraction, ammonium sulphate and isoelectric precipitation was found suitable for both large and small scale preparation of purified K88ab adhesin. Extraction of the K88 adhesin was sensitive to pH and it was postulated that this may affect the site of colonisation of by ETEC in vivo. Results of haemagglutination experiments were consistent with the hypothesis that the K88 receptor present on erythrocytes is composed of two elements, one responsible for the binding of K88ab and K88ac and a second responsible for the binding of the K88ad adhesin. Comparison of the haemagglutinating properties of cell-free and cell-bound K88 adhesin revealed some differences probably indicating a minor conformational change in the K88 adhesin on its isolation. The K88ab adhesin was found to bind to erythrocytes over a wide pH range (PH 4-9) and was inhibited by αK88ab and αK88b antisera. Inhibition of haemagglutination was noted with crude heparin, mannan and porcine gastric mucin, chondrosine and several hexosamines, glucosamine in particular. The most potent inhibitor of haemagglutination was n-dodecyl-β-D-glucopyranoside, one of a series of glucosides found to have inhibitory properties. Correlation between hydrophobicity of glucosides tested and degree of inhibition observed suggested hydrophobic forces were important in the interaction of the K88 adhesin with its receptor. The results of Scatchard and Hill plots indicated that binding of the K88ab adhesin to porcine enterocytes in the majority of cases is a two-step, three component system. The first K88 receptor (or site) had a K2. of 1.59x1014M-1 and a minimum of 4.3x104 sites/enterocyte. The second receptor (or site) had a K2 of 4.2x1012M-1 with a calculated 1.75x105 sites/enterocyte. Attempts to inhibit binding of cell-free K88 adhesin to porcine enterocytes by lectins were unsuccessful. However, several carbohydrates including trehalose, lactulose, galactose 1→4 mannopyranoside, chondrosine, galactosamine, stachyose and mannan were inhibitory. The most potent inhibitor was found to be porcine gastric mucin. Inhibition observed with n-octyl-α-D-glucopyranose was difficult to interpret in isolation because of interference with the assay, however, it agreed with the results of haemagglutination inhibition experiments.

Software and hardware aspects of a microprocessor controlled lathe

A small lathe has been modified to work under microprocessor control to enhance the facilities which the lathe offers and provide a wider operating range with relevant economic gains. The result of these modifications give better operating system characteristics. A system of electronic circuits have been developed, utilising the latest technology, to replace the pegboard with the associated obsolete electrical components. Software for the system includes control programmes for the implementation of the original pegboard operation and several sample machine code programmes are included, covering a wide spectrum of applications, including diagnostic testing of the control system. It is concluded that it is possible to carry out a low cost retrofit on existing machine tools to enhance their range of capabilities.

Analysis of magnetoencephaoographic data as a nonlinear dynamical system

This thesis presents the results from an investigation into the merits of analysing Magnetoencephalographic (MEG) data in the context of dynamical systems theory. MEG is the study of both the methods for the measurement of minute magnetic flux variations at the scalp, resulting from neuro-electric activity in the neocortex, as well as the techniques required to process and extract useful information from these measurements. As a result of its unique mode of action - by directly measuring neuronal activity via the resulting magnetic field fluctuations - MEG possesses a number of useful qualities which could potentially make it a powerful addition to any brain researcher's arsenal. Unfortunately, MEG research has so far failed to fulfil its early promise, being hindered in its progress by a variety of factors. Conventionally, the analysis of MEG has been dominated by the search for activity in certain spectral bands - the so-called alpha, delta, beta, etc that are commonly referred to in both academic and lay publications. Other efforts have centred upon generating optimal fits of "equivalent current dipoles" that best explain the observed field distribution. Many of these approaches carry the implicit assumption that the dynamics which result in the observed time series are linear. This is despite a variety of reasons which suggest that nonlinearity might be present in MEG recordings. By using methods that allow for nonlinear dynamics, the research described in this thesis avoids these restrictive linearity assumptions. A crucial concept underpinning this project is the belief that MEG recordings are mere observations of the evolution of the true underlying state, which is unobservable and is assumed to reflect some abstract brain cognitive state. Further, we maintain that it is unreasonable to expect these processes to be adequately described in the traditional way: as a linear sum of a large number of frequency generators. One of the main objectives of this thesis will be to prove that much more effective and powerful analysis of MEG can be achieved if one were to assume the presence of both linear and nonlinear characteristics from the outset. Our position is that the combined action of a relatively small number of these generators, coupled with external and dynamic noise sources, is more than sufficient to account for the complexity observed in the MEG recordings. Another problem that has plagued MEG researchers is the extremely low signal to noise ratios that are obtained. As the magnetic flux variations resulting from actual cortical processes can be extremely minute, the measuring devices used in MEG are, necessarily, extremely sensitive. The unfortunate side-effect of this is that even commonplace phenomena such as the earth's geomagnetic field can easily swamp signals of interest. This problem is commonly addressed by averaging over a large number of recordings. However, this has a number of notable drawbacks. In particular, it is difficult to synchronise high frequency activity which might be of interest, and often these signals will be cancelled out by the averaging process. Other problems that have been encountered are high costs and low portability of state-of-the- art multichannel machines. The result of this is that the use of MEG has, hitherto, been restricted to large institutions which are able to afford the high costs associated with the procurement and maintenance of these machines. In this project, we seek to address these issues by working almost exclusively with single channel, unaveraged MEG data. We demonstrate the applicability of a variety of methods originating from the fields of signal processing, dynamical systems, information theory and neural networks, to the analysis of MEG data. It is noteworthy that while modern signal processing tools such as independent component analysis, topographic maps and latent variable modelling have enjoyed extensive success in a variety of research areas from financial time series modelling to the analysis of sun spot activity, their use in MEG analysis has thus far been extremely limited. It is hoped that this work will help to remedy this oversight.

Development of a conceptual model of the soil-moisture-plant sub-system of the hydrological cycle

The soil-plant-moisture subsystem is an important component of the hydrological cycle. Over the last 20 or so years a number of computer models of varying complexity have represented this subsystem with differing degrees of success. The aim of this present work has been to improve and extend an existing model. The new model is less site specific thus allowing for the simulation of a wide range of soil types and profiles. Several processes, not included in the original model, are simulated by the inclusion of new algorithms, including: macropore flow; hysteresis and plant growth. Changes have also been made to the infiltration, water uptake and water flow algorithms. Using field data from various sources, regression equations have been derived which relate parameters in the suction-conductivity-moisture content relationships to easily measured soil properties such as particle-size distribution data. Independent tests have been performed on laboratory data produced by Hedges (1989). The parameters found by regression for the suction relationships were then used in equations describing the infiltration and macropore processes. An extensive literature review produced a new model for calculating plant growth from actual transpiration, which was itself partly determined by the root densities and leaf area indices derived by the plant growth model. The new infiltration model uses intensity/duration curves to disaggregate daily rainfall inputs into hourly amounts. The final model has been calibrated and tested against field data, and its performance compared to that of the original model. Simulations have also been carried out to investigate the effects of various parameters on infiltration, macropore flow, actual transpiration and plant growth. Qualitatively comparisons have been made between these results and data given in the literature.

Behavioural correlates of ocular accomodation and the autonomic nervous system

The binding issue of th is thesis was the examination of workload, induced by relinotopic and spatiotopic stimuli, on both the ocu lomotor and cardiovascular systems together with investigating the covariation between the two systems - the 'eye-heart' link. Further, the influence of refractive error on ocular accommodation and cardiovascular function was assessed. A clinical evaluation was undertaken to assess the newly available open-view infrared Shin-Nippon NVision-K 5001 optometer, its benefit being the capability to measure through pupils = 2.3 mm. Measurements of refractive error taken with the NVision-K were found to be both accurate (Difference in Mean Spherical Equivalent: 0.14 ± 0.35 D; p = 0.67) and repeatable when compared to non-cycloplegic subjective refraction. Due to technical difficulties, however, the NVision-K could not be used for the purpose of the thesis, as such, measures of accommodation were taken using the continuously recording Shin-Nippon SRW-5000 openview infrared optometer, coupled with a piezo-electric finger pulse transducer to measure pulse. Heart rate variability (HRV) was spectrally analysed to determine the systemic sympathetic and parasympathetic components of the autonomic nervous system (ANS). A large sample (n = 60), cross-sectional study showed late-onset myopes (LOMs) display less accurate responses when compared to other refractive groups at high accommodative demand levels (3 .0 0 and 4.0D). Tonic accommodation (TA) was highest in the hypermetropes, fo llowed by emmetropes and early-onset myopes while the LOM subjects demonstrated statistically significant lower levels of TA. The root-meansquare (RMS) value of the accommodative response was shown to amplify with increased levels of accommodative demand. Changes in refractive error only became significant between groups at higher demand levels (3.0 D and 4.0 D) with the LOMs showing the largest magnification in oscilIations. Examination of the stimulus-response cross-over point with the unit ratio line and TA showed a correlation between the two (r = 0.45, p = 0.001), where TA is approximately twice the dioptric value of the stimulus-response cross-over point. Investigation of the relationship between ocular accommodation and systemic ANS function demonstrated covariation between the systems. Subjects with a faster heart rate (lower heart period) tended to have a higher TA value (r = -0.27, p < 0.05). Further, an increase in accommodative demand accompanies a faster heart rate. The influence of refractive error on the cardiovascular response to changes in accommodative demand, however, was equivocal. Examination of the microfluctuations ofacconunodation demonstrated a correlation between the temporal frequency location of the accommodative high Frequency component (HFC) and the arterial pulse frequency. The correlation was present at a range of accommodative demands from 0.0 D to 4.0 D and in all four refractive groups, suggesting that the HFC was augmented by physiological factors. Examination of the effect of visual cognition on ocular accommodation and the ANS confirmed that increasing levels of cognition affect the accommodative mechanism. The accommodative response shifted away from the subject at both near and far. This shift in accommodative response accompanied a decay in the systemic parasympathetic innervation to the heart. Differences between refractive groups also existed with LOMs showing less accurate responses compared to emmetropes. This disparity, however, appeared to be augmented by the systemic sympathetic nervous system. The investigations discussed explored Ihe role of oculomotor and cardiovascular fu nction in workload enviromnents, providing evidence for a behavioural link between the cardiovascular and oculomotor systems.

A speech analysis system for use in educational research

This thesis describes work undertaken in order to fulfil a need experienced in the Department of Educational Enquiry at the University of Aston in Birmingham for speech analysis facilities suitable for use in teaching and research work within the Department. The hardware and software developed during the research project provides displays of speech fundamental frequency and intensity in real time. The system is suitable for the provision of visual feedback of these parameters of a subject's speech in a learning situation, and overcomes the inadequacies of equipment currently used for this task in that it provides a clear indication of fundamental frequency contours as the subject is speaking. The thesis considers the use of such equipment in several related fields, and the approaches that have been reported to one of the major problems of speech analysis, namely pitch-period estimation. A number of different systems are described, and their suitability for the present purposes is discussed. Finally, a novel method of pitch-period estimation is developed, and a speech analysis system incorporating this method is described. Comparison is made between the results produced by this system and those produced by a conventional speech spectrograph.

The application of robotics to the turbine blade encapsulation process

A survey of the existing state-of-the-art of turbine blade manufacture highlights two operations that have not been automated namely that of loading of a turbine blade into an encapsulation die, and that of removing a machined blade from the encapsulation block. The automation of blade decapsulation has not been pursued. In order to develop a system to automate the loading of an encapsulation die a prototype mechanical handling robot has been designed together with a computer controlled encapsulation die. The robot has been designed as a mechanical handling robot of cylindrical geometry, suitable for use in a circular work cell. It is the prototype for a production model to be called `The Cybermate'. The prototype robot is mechanically complete but due to unforeseen circumstances the robot control system is not available (the development of the control system did not form a part of this project), hence it has not been possible to fully test and assess the robot mechanical design. Robot loading of the encapsulation die has thus been simulated. The research work with regard to the encapsulation die has focused on the development of computer controlled, hydraulically actuated, location pins. Such pins compensate for the inherent positional inaccuracy of the loading robot and reproduce the dexterity of the human operator. Each pin comprises a miniature hydraulic cylinder, controlled by a standard bidirectional flow control valve. The precision positional control is obtained through pulsing of the valves under software control, with positional feedback from an 8-bit transducer. A test-rig comprising one hydraulic location pin together with an opposing spring loaded pin has demonstrated that such a pin arrangement can be controlled with a repeatability of +/-.00045'. In addition this test-rig has demonstrated that such a pin arrangement can be used to gauge and compensate for the dimensional error of the component held between the pins, by offsetting the pin datum positions to allow for the component error. A gauging repeatability of +/- 0.00015' was demonstrated. This work has led to the design and manufacture of an encapsulation die comprising ten such pins and the associated computer software. All aspects of the control software except blade gauging and positional data storage have been demonstrated. Work is now required to achieve the accuracy of control demonstrated by the single pin test-rig, with each of the ten pins in the encapsulation die. This would allow trials of the complete loading cycle to take place.

Implementation challenges of application of radio frequency identification (RFID) in warehouse

Warehouse is an essential component in the supply chain, linking the chain partners and providing them with functions of product storage, inbound and outbound operations along with value-added processes. Allocation of warehouse resources should be efficient and effective to achieve optimum productivity and reduce operational costs. Radio frequency identification (RFID) is a technology capable of providing real-time information about supply chain operations. It has been used by warehousing and logistic enterprises to achieve reduced shrinkage, improved material handling and tracking as well as increased accuracy of data collection. However, both academics and practitioners express concerns about challenges to RFID adoption in the supply chain. This paper provides a comprehensive analysis of the problems encountered in RFID implementation at warehouses, discussing the theoretical and practical adoption barriers and causes of not achieving full potential of the technology. Lack of foreseeable return on investment (ROI) and high costs are the most commonly reported obstacles. Variety of standards and radio wave frequencies are identified as source of concern for decision makers. Inaccurate performance of the RFID within the warehouse environment is examined. Description of integration challenges between warehouse management system and RFID technology is given. The paper discusses the existing solutions to technological, investment and performance RFID adoption barriers. Factors to consider when implementing the RFID technology are given to help alleviate implementation problems. By illustrating the challenges of RFID in the warehouse environment and discussing possible solutions the paper aims to help both academics and practitioners to focus on key areas constituting an obstacle to the technology growth. As more studies will address these challenges, the realisation of RFID benefits for warehouses and supply chain will become a reality.

Modelling manufacturing planning and control systems:the application of object-oriented principles and discrete-event simulation

Manufacturing planning and control systems are fundamental to the successful operations of a manufacturing organisation. 10 order to improve their business performance, significant investment is made by companies into planning and control systems; however, not all companies realise the benefits sought Many companies continue to suffer from high levels of inventory, shortages, obsolete parts, poor resource utilisation and poor delivery performance. This thesis argues that the fit between the planning and control system and the manufacturing organisation is a crucial element of success. The design of appropriate control systems is, therefore, important. The different approaches to the design of manufacturing planning and control systems are investigated. It is concluded that there is no provision within these design methodologies to properly assess the impact of a proposed design on the manufacturing facility. Consequently, an understanding of how a new (or modified) planning and control system will perform in the context of the complete manufacturing system is unlikely to be gained until after the system has been implemented and is running. There are many modelling techniques available, however discrete-event simulation is unique in its ability to model the complex dynamics inherent in manufacturing systems, of which the planning and control system is an integral component. The existing application of simulation to manufacturing control system issues is limited: although operational issues are addressed, application to the more fundamental design of control systems is rarely, if at all, considered. The lack of a suitable simulation-based modelling tool does not help matters. The requirements of a simulation tool capable of modelling a host of different planning and control systems is presented. It is argued that only through the application of object-oriented principles can these extensive requirements be achieved. This thesis reports on the development of an extensible class library called WBS/Control, which is based on object-oriented principles and discrete-event simulation. The functionality, both current and future, offered by WBS/Control means that different planning and control systems can be modelled: not only the more standard implementations but also hybrid systems and new designs. The flexibility implicit in the development of WBS/Control supports its application to design and operational issues. WBS/Control wholly integrates with an existing manufacturing simulator to provide a more complete modelling environment.

Meta level component-based framework for distributed computing applications

Adaptability for distributed object-oriented enterprise frameworks is a critical mission for system evolution. Today, building adaptive services is a complex task due to lack of adequate framework support in the distributed computing environment. In this thesis, we propose a Meta Level Component-Based Framework (MELC) which uses distributed computing design patterns as components to develop an adaptable pattern-oriented framework for distributed computing applications. We describe our novel approach of combining a meta architecture with a pattern-oriented framework, resulting in an adaptable framework which provides a mechanism to facilitate system evolution. The critical nature of distributed technologies requires frameworks to be adaptable. Our framework employs a meta architecture. It supports dynamic adaptation of feasible design decisions in the framework design space by specifying and coordinating meta-objects that represent various aspects within the distributed environment. The meta architecture in MELC framework can provide the adaptability for system evolution. This approach resolves the problem of dynamic adaptation in the framework, which is encountered in most distributed applications. The concept of using a meta architecture to produce an adaptable pattern-oriented framework for distributed computing applications is new and has not previously been explored in research. As the framework is adaptable, the proposed architecture of the pattern-oriented framework has the abilities to dynamically adapt new design patterns to address technical system issues in the domain of distributed computing and they can be woven together to shape the framework in future. We show how MELC can be used effectively to enable dynamic component integration and to separate system functionality from business functionality. We demonstrate how MELC provides an adaptable and dynamic run time environment using our system configuration and management utility. We also highlight how MELC will impose significant adaptability in system evolution through a prototype E-Bookshop application to assemble its business functions with distributed computing components at the meta level in MELC architecture. Our performance tests show that MELC does not entail prohibitive performance tradeoffs. The work to develop the MELC framework for distributed computing applications has emerged as a promising way to meet current and future challenges in the distributed environment.

Isolation, characterisation and application of novel microbial protein cross-linking enzymes

Microbial transglutaminase is favoured for use in industry over the mammalian isoform, and hence has been utilized, to great effect, as an applied biocatalyst in many industrial areas including the food and textiles industries. There are currently only a limited number of microbial TGase sources known. A number of organisms have been screened for transglutaminase activity using biochemical assays directed towards TGase catalyzed reactions (amine incorporation and peptide cross-linking assay). Of those organisms screened, TGase was identified in a number of isolates including members of the Bacillus and Streptomyces families. In addition, a protein capable of performing a TGase-like reaction was identified in the organism Pseudomonas putida that was deemed immunologically distinct from previously described TGase isoforms, though further work would be required to purify the protein responsible. The genuses Streptoverticillium and Streptomyces are known to be closely related. A number of micro-organisms relating to Streptomyces mobaraensis (formerly Streptoverticillium mobaraensis) have been identified as harboring a TGase enzyme. The exact biological role of Streptomyces TGase is not well understood, though from work undertaken here it would appear to be involved in cell wall growth. Comparison of the purified Streptomyces TGase proteins showed them to exhibit marginally different characteristics in relation to enzymatic activity and pH dependency upon comparison with Streptomyces mobaraensis TGase. In addition, TGase was identified in the organism Saccharomonospora viridis that was found to be genetically identical to that from S. mobaraensis raising questions about the enzymes dissemination in nature. TGase from S. baldaccii was found to be most diverse with respect to enzymatic characteristics whilst still retaining comparable E(y-glutamyl) lysine bond formation to S. mobaraensis TGase. As such S. baldaccii TGase was cloned into an expression vector enabling mass production of the enzyme thereby providing a viable alternative to S. mobaraensis TGase for many industrial processes.

Does internal marketing improve service quality in call centres?

Although internal marketing has been conceptually argued to improve service quality, empirical evidence in this respect has been limited. This research investigates the relationship between rewards, which are a key component of internal marketing paradigm, and service quality of frontline employees in banking call centres. In this context, the authors develop hypotheses with the help of literature review, and empirically examine the effects of various extrinsic and intrinsic rewards on service quality. The results indicate that intrinsic rewards have greater effect on service quality than extrinsic rewards, which offer managerial implications and directions for future research.

HRM systems, internal marketing and performance in Indian call centres:full research report ESRC end of award report, RES-000-22-1876

Considering the rapid growth of call centres (CCs) in India, its implications for businesses in the UK and a scarcity of research on human resource management (HRM) related issues in Indian CCs, this research has two main aims. First, to highlight the nature of HRM systems relevant to Indian call centres. Second, to understand the significance of internal marketing (IM) in influencing the frontline employees’ job-related attitudes and performance. Rewards being an important component of IM, the relationships between different types of rewards as part of an IM strategy, attitudes and performance of employees in Indian CCs will also be examined. Further, the research will investigate which type of commitment mediates the link between rewards and performance and why. The data collection will be via two phases. The first phase would involve a series of in-depth interviews with both the managers and employees to understand the functioning of CCs, and development of suitable HRM systems for the Indian context. The second phase would involve data collection through questionnaires distributed to the frontline employees and supervisors to examine the relationships among IM, employee attitudes and performance. Such an investigation is expected to contribute to development of better theory and practice.