Eukaryotic expression:Developments for structural proteomics

The production of sufficient quantities of protein is an essential prelude to a structure determination, but for many viral and human proteins this cannot be achieved using prokaryotic expression systems. Groups in the Structural Proteomics In Europe (SPINE) consortium have developed and implemented high-throughput (HTP) methodologies for cloning, expression screening and protein production in eukaryotic systems. Studies focused on three systems: yeast (Pichia pastoris and Saccharomyces cerevisiae), baculovirus-infected insect cells and transient expression in mammalian cells. Suitable vectors for HTP cloning are described and results from their use in expression screening and protein-production pipelines are reported. Strategies for co-expression, selenomethionine labelling (in all three eukaryotic systems) and control of glycosylation (for secreted proteins in mammalian cells) are assessed. © International Union of Crystallography, 2006.

Structural change in apartheid era South Africa:1975-93

This paper examines the sources of structural changes in output growth of South Africa's economy over 1975-93 using a decomposition method within the inputoutput (IO) framework for analysing output changes from a demand side perspective. It decomposes output growth into private consumption, government consumption, investment and export components and also measures the impact of import substitution and changes in intermediate input use (as indicated by changes in IO coefficients). It is found that, before 1981, overall output growth was multi-components driven with all the above components contributing positively to economic growth. However, the collapse of investment demand is by far the single largest factor contributing to the economic stagnation that categorizes the post-1981 period.

Environmental scanning electron microscope imaging of vesicle systems

The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors that influence this. However, there are a few methods that study these systems in their natural hydrated state; commonly, the liposomes are visualized after drying, staining and/or fixation of the vesicles. Environmental scanning electron microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. We were the first to use ESEM to study the liposomes and niosomes, and have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses onto, or evaporates from, the sample in real-time. This provides an insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay for liposome formulation and stability.

The properties of advanced alumimium alloy systems

2XXX and 7XXX series aluminium alloys have been the accepted materials for airframe construction for many decades. However, only minor improvements in properties have been possible by the development of these alloys since the early 1970's. The constant need to reduce weight in aircraft has therefore led to a resurgence in the research for higher performance aluminium alloys. The reason for this investigation was to evaluate possible alternatives for the existing conventional aluminium alloy 2014 for aircraft wheel applications. Three new technologies in alloy development were considered: a metal matrix composite, an aluminium-lithium alloy and a powder metallurgical alloy. The basic mechanical properties of these advanced materials have already been established to an extent, but their fatigue behaviour has yet to be fully understood. The purpose of this work was to investigate the fatigue properties of the materials concerned, in both air and an aerated 3.5% NaCl solution, and compare these properties to 2014-T6. As well as the basic mechanical properties, fatigue crack propagation data is presented for all of the materials concerned. Additionally, fatigue crack initiation data is presented for the aluminium-lithium alloy and 2014. The D.C. electrical potential method was used to monitor crack growth. Of the materials investigated, the most promising was the aluminium-lithium alloy. However, short transverse properties need to be increased and the commercial cost of the material needs to be decreased before it can be considered as a direct replacement for 2014 for aircraft structural applications. It was considered that the cost of the powder metallurgical alloy would limit its further use. The metal matrix composite material proved to be unsuitable for most ambient temperature applications

Finitely coordinated models for low-temperature phases of amorphous systems

We introduce models of heterogeneous systems with finite connectivity defined on random graphs to capture finite-coordination effects on the low-temperature behaviour of finite-dimensional systems. Our models use a description in terms of small deviations of particle coordinates from a set of reference positions, particularly appropriate for the description of low-temperature phenomena. A Born-von Karman-type expansion with random coefficients is used to model effects of frozen heterogeneities. The key quantity appearing in the theoretical description is a full distribution of effective single-site potentials which needs to be determined self-consistently. If microscopic interactions are harmonic, the effective single-site potentials turn out to be harmonic as well, and the distribution of these single-site potentials is equivalent to a distribution of localization lengths used earlier in the description of chemical gels. For structural glasses characterized by frustration and anharmonicities in the microscopic interactions, the distribution of single-site potentials involves anharmonicities of all orders, and both single-well and double-well potentials are observed, the latter with a broad spectrum of barrier heights. The appearance of glassy phases at low temperatures is marked by the appearance of asymmetries in the distribution of single-site potentials, as previously observed for fully connected systems. Double-well potentials with a broad spectrum of barrier heights and asymmetries would give rise to the well-known universal glassy low-temperature anomalies when quantum effects are taken into account. © 2007 IOP Publishing Ltd.

21-residue peptide's dynamics at and between elementary structural transitions

Elementary conformational changes of the backbone of a 21-residue peptide A5(A3RA)3A are studied using molecular dynamics simulations in explicit water. The processes of the conformational transitions and the regimes of stationary fluctuations between them are investigated using minimal perturbations of the system. The perturbations consist of a few degrees rotation of the velocity of one of the systems' atoms and keep the system on the same energy surface. It is found that (i) the system dynamics is insignificantly changed by the perturbations in the regimes between the transitions; (ii) it is very sensitive to the perturbations just before the transitions that prevents the peptide from making the transitions; and (iii) the perturbation of any atom of the system, including distant water molecules is equally effective in preventing the transition. The latter implies strongly collective dynamics of the peptide and water during the transitions.

A systems approach to public sector budgeting: the case of the planning, programming, budgeting system for the health and personal social services

The aim of this thesis is to examine the specific contextual factors affecting the applicability and development of the planning, programming, budgeting system (P.P.B.S.) as a systems approach to public sector budgeting. The concept of P.P.B.S. as a systems approach to public sector budgeting will first be developed and the preliminary hypothesis that general contextual factors may be classified under political, structural and cognitive headings will be put forward. This preliminary hypothesis will be developed and refined using American and early British experience. The refined hypothesis will then be tested in detail in the case of the English health and personal social services (H.P.S.S.), The reasons for this focus are that it is the most recent, the sole remaining, and the most significant example in British central government outside of defence, and is fairly representative of non-defence government programme areas. The method of data collection relies on the examination of unpublished and difficult to obtain central government, health and local authority documents, and interviews with senior civil servants and public officials. The conclusion will be that the political constraints on, or factors affecting P.P.B.S., vary with product characteristics and cultural imperatives on pluralistic decision-making; that structural constraints vary with the degree of coincidence of programme and organisation structure and with the degree of controllability of the organisation; and finally, that cognitive constraints vary according to product characteristics, organisational responsibilities, and analytical effort.

The development, evaluation and use of freshly isolated renal proxinal tubule systems in the fischer rat

The investigation of renal pathophysiology and toxicology has traditionally been advanced by the development of increasingly defined and refined in vitro preparations. This study has sought to develop and evaluate various methods of producing pure samples of renal proximal tubules (PTs) from the Fischer rat. The introduction summarised the most common in vitro preparations together with the parameters used to monitor viability - particularly with regard to toxic events. The most prevalent isolation methods have involved the use of collagenase to produce dissociation of the cortex. However, the present study has shown that even the mildest collagenase treatment caused significant structural damage which resulted in a longevity of only 3hr in suspension. An alternative mechanical isolation technique has been developed in this study that consists of perfusion loading the renal glomeruli with Fe304 followed by disruption of the cortex by homogenisation and sequential sieving. The glomeruli are removed magnetically and the PTs then harvested by a 64μM sieve. PTs isolated in this way showed a vastly superior structural preservation over their collagenase isolated counterparts; also oxygen consumption and enzyme leakage measurements showed a longevity in excess of 6hr when incubated in a very basic medium. Attempts were then made to measure the cytosolic calcium levels in both mechanical and collagenase isolated PTs using the fluorescent calcium indicator Fura. However results were inconclusive due to significant binding of the Fura to the external PT surfaces. In conclusion, PTs prepared by the present mechanical isolation technique exhibit superior preservation and longevity compared with even the mildest collagenase isolation technique and hence appear to offer potential advantages over collagenase isolation as an in vitro renal system.

Structural limits and teachers' perspectives

This thesis is about the relationship between schooling and the economic and political structures which constrain its institutional framework. It focusses on teachers, as mediators of structural constraints, and on middle schools, as institutions which occupy a functionally transitional place between the primary and secondary traditions. In approaching the problem of linking the different perspectives of macro and micro sociologies, I argue the view that the individual mediates the contradictions between the socially cooperative processes of production and the competitive individualism which legitimates the private appropriation of wealth and income. The link is observable in the schooling process as a pattern of contradictions and tensions mediated by the rhetoric of equality of opportunity. In order to elucidate the link, the processes within the boundaried institutions must be viewed in the context of those changing tensions within the state administrative systems, which reverberate into schools as economic and political constraints. Framed within the ideology of the Flowden Report (1967), middle schooling was set within a discourse which stressed cooperative relationships rather than competitive standards. Since the mid-1970s, administrative policies have heightened the competitive battle for declining resources and attacked the Plowden ideology. Focussing the fieldwork on six middle schools in one local authority, T use an eclectic methodology to relate economic and political policies, generated in the state administrative system, to the situation in the schools between 1979 and 1981. The methodology incorporates a time dimension in order to highlight the tensions as they play upon teachers' changing definitions of the changing situation. I conclude that the intersubjective socio-cultural relations of schooling cannot be properly explained without making explicit the changing tensions in the rule/resource relationships which teachers mediate through their particular institutions.

Reactive processing of polymers: structural characterization of products by 1H and 13C NMR spectroscopy for glycidyl methacrylate grafting onto EPR in the absence and presence of a reactive comonomer

Grafted GMA on EPR samples were prepared in a Thermo-Haake internal mixer by free radical melt grafting reactions in the absence (conventional system; EPR-g-GMA(CONV)) and presence of the reactive comonomer divinyl benzene, DVB (EPR-g-GMA(DVB)). The GMA-homopolymer (poly-GMA), a major side reaction product in the conventional system, was almost completely absent in the DVB-containing system, the latter also resulted in a much higher level of GMA grafting. A comprehensive microstructure analysis of the formed poly-GMA was performed based on one-dimensional H-1 and C-13 NMR spectroscopy and the complete spectral assignments were supported by two-dimensional NMR techniques based on long range two and three bond order carbon-proton couplings from HMBC (Heteronuclear Multiple Bond Coherence) and that of one bond carbon-proton couplings from HSQC (Heteronuclear Single Quantum Coherence), as well as the use of Distortionless Enhancement by Polarization Transfer (DEPT) NMR spectroscopy. The unambiguous analysis of the stereochemical configuration of poly-GMA was further used to help understand the microstructures of the GMA-grafts obtained in the two different free radical melt grafting reactions, the conventional and comonomer-containing systems. In the grafted GMA, in the conventional system (EPR-g-GMA(CONV)), the methylene protons of the GMA were found to be sensitive to tetrad configurational sequences and the results showed that 56% of the GMA sequence in the graft is in atactic configuration and 42% is in syndiotactic configuration whereas the poly-GMA was predominantly syndiotactic. The differences in the microstructures of the graft in the conventional EPR-g-GMA(CONV) and the DVB-containing (EPR-g-GMA(DVB)) systems is also reported (C) 2009 Elsevier Ltd. All rights reserved.

Dissipative nonlinear structures in optical fiber systems II : self-similar parabolic pulses in active fibers

In this second talk on dissipative structures in fiber applications, we overview theoretical aspects of the generation, evolution and characterization of self-similar parabolic-shaped pulses in fiber amplifier media. In particular, we present a perturbation analysis that describes the structural changes induced by third-order fiber dispersion on the parabolic pulse solution of the nonlinear Schrödinger equation with gain. Promising applications of parabolic pulses in optical signal post-processing and regeneration in communication systems are also discussed.

Structural characterization of titanium-doped Bioglass using isotopic substitution neutron diffraction

Melt quenched silicate glasses containing calcium, phosphorus and alkali metals have the ability to promote bone regeneration and to fuse to living bone. Of these glasses 45S5 Bioglass® is the most widely used being sold in over 35 countries as a bone graft product for medical and dental applications; particulate 45S5 is also incorporated into toothpastes to help remineralize the surface of teeth. Recently it has been suggested that adding titanium dioxide can increase the bioactivity of these materials. This work investigates the structural consequences of incorporating 4 mol% TiO2 into Bioglass® using isotopic substitution (of the Ti) applied to neutron diffraction and X-ray Absorption Near Edge Structure (XANES). We present the first isotopic substitution data applied to melt quench derived Bioglass or its derivatives. Results show that titanium is on average surrounded by 5.2(1) nearest neighbor oxygen atoms. This implies an upper limit of 40% four-fold coordinated titanium and shows that the network connectivity is reduced from 2.11 to 1.97 for small quantities of titanium. Titanium XANES micro-fluorescence confirms the titanium environment is homogenous on the micron length scale within these glasses. Solid state magic angle spinning (MAS) NMR confirms the network connectivity model proposed. Furthermore, the results show the intermediate range order containing Na-O, Ca-O, O-P-O and O-Si-O correlations are unaffected by the addition of small quantities of TiO2 into these systems.

G-protein-coupled receptors:from structural insights to functional mechanisms

The papers resulting from the recent Biochemical Society Focused Meeting 'G-Protein-Coupled Receptors: from Structural Insights to Functional Mechanisms' held in Prato in September 2012 are introduced in the present overview. A number of future goals for GPCR (G-protein-coupled receptor) research are considered, including the need to develop biophysical and computational methods to explore the full range of GPCR conformations and their dynamics, the need to develop methods to take this into account for drug discovery and the importance of relating observations on isolated receptors or receptors expressed in model systems to receptor function in vivo. © 2013 Biochemical Society.

Dissipative nonlinear structures in optical fiber systems II:self-similar parabolic pulses in active fibers

In this second talk on dissipative structures in fiber applications, we overview theoretical aspects of the generation, evolution and characterization of self-similar parabolic-shaped pulses in fiber amplifier media. In particular, we present a perturbation analysis that describes the structural changes induced by third-order fiber dispersion on the parabolic pulse solution of the nonlinear Schrödinger equation with gain. Promising applications of parabolic pulses in optical signal post-processing and regeneration in communication systems are also discussed.

Looking under the bonnet:factors affecting student adoption of e-learning systems in Jordan

The primary questions addressed in this paper are the following: what are the factors that affect students’ adoption of an e-learning system and what are the relationships among these factors? This paper investigates and identifies some of the major factors affecting students’ adoption of an e-learning system in a university in Jordan. E-learning adoption is approached from the information systems acceptance point of view. This suggests that a prior condition for learning effectively using e-learning systems is that students must actually use them. Thus, a greater knowledge of the factors that affect IT adoption and their interrelationships is a pre-cursor to a better understanding of student acceptance of e-learning systems. In turn, this will help and guide those who develop, implement, and deliver e-learning systems. In this study, an extended version of the Technology Acceptance Model (TAM) was developed to investigate the underlying factors that influence students’ decisions to use an e-learning system. The TAM was populated using data gathered from a survey of 486 undergraduate students using the Moodle based e-learning system at the Arab Open University. The model was estimated using Structural Equation Modelling (SEM). A path model was developed to analyze the relationships between the factors to explain students’ adoption of the e-learning system. Whilst findings support existing literature about prior experience affecting perceptions, they also point to surprising group effects, which may merit future exploration.