Active membrane transport and receptor proteins from bacteria

A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5-35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C-terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides. ©2005 Biochemical Society.

Controlling contractual discretion

In 1993 Leggatt L.J. said in Abu Dhabi National Tanker Co. v. Product Star Shipping Ltd. (The Product Star) [1993] 1 Lloyds Rep. 397, 404: Where A and B contract with each other to confer a discretion on A, that does not render B subject to A’s uninhibited whim. In my judgment, the authorities show that not only must the discretion be exercised honestly and in good faith, but, having regard to the provisions of the contract by which it is conferred, it must not be exercised arbitrarily, capriciously or unreasonably.

Advanced liquid-liquid extraction : studies in a laboratory mixer-settler

A ten stage laboratory mixer-settler has been designed, constructed and operated with efficiencies up to 90%. The phase equilibrium data of the system acetic acid-toluene-water at different temperatures has been determined and correlated. Trials for prediction of these data have been investigated and a good agreement between the experimental data and the predictions obtained by the NRTL equation have been found. Extraction processes have been analysed. A model for determination of the time needed for a countercurrent stage-wise process to come to steady state has been derived. The experimental data was in reasonable agreement with this model. The discrete maximum principle has been applied to optimize the countercurrent extraction process and proved to be highly successful in predicting the optimum operating conditions which were confirmed by the experimental results. The temperature has proved to be a prosolvent for mass transfer in both directions but the temperature profile functioned as an anti solvent.

Protecting agents against malicious host attacks

The introduction of agent technology raises several security issues that are beyond conventional security mechanisms capability and considerations, but research in protecting the agent from malicious host attack is evolving. This research proposes two approaches to protecting an agent from being attacked by a malicious host. The first approach consists of an obfuscation algorithm that is able to protect the confidentiality of an agent and make it more difficult for a malicious host to spy on the agent. The algorithm uses multiple polynomial functions with multiple random inputs to convert an agent's critical data to a value that is meaningless to the malicious host. The effectiveness of the obfuscation algorithm is enhanced by addition of noise code. The second approach consists of a mechanism that is able to protect the integrity of the agent using state information, recorded during the agent execution process in a remote host environment, to detect a manipulation attack by a malicious host. Both approaches are implemented using a master-slave agent architecture that operates on a distributed migration pattern. Two sets of experimental test were conducted. The first set of experiments measures the migration and migration+computation overheads of the itinerary and distributed migration patterns. The second set of experiments is used to measure the security overhead of the proposed approaches. The protection of the agent is assessed by analysis of its effectiveness under known attacks. Finally, an agent-based application, known as Secure Flight Finder Agent-based System (SecureFAS) is developed, in order to prove the function of the proposed approaches.

The antecedents and consequences of e-business adoption in Malaysian manufacturing firms

Theory suggests that firms that adopt innovation share some common characteristics, just as those who do not adopt innovation and firms adopt a particular technology because the functions provided by the chosen technology fulfils their needs. Building on these arguments, this research project investigates the antecedents and consequences of e-business adoption among manufacturing firms in Malaysia. This thesis develops from the existing literature of organisational innovation adoption, information technology and strategic marketing/management. It further adds to the existing literature by using cultural-based predictors representing organisational characteristics consisting of market orientation, innovativeness and organisational learning. The study also formalises the theoretical framework of organisational-environment-technology. This study develops a new construct called technology motivation in addition to the introduction of several e-business technology scales. The results substantiate the significance of firm technology motivation in determining firm adoption of the various e-business initiatives. In addition, business environment and market orientation are found to influence firm choice of technology motivation. Meanwhile, innovativeness and organisational learning are shown to influence the magnitude of a firm’s e-business adoption. Finally, the results show that firm adoption of e-business technology does not influence organisational performance. This investigation clarifies the rationale and importance of firm technology motivation in adopting the various e-business initiatives. It also highlights the importance of having the appropriate organisational culture in ensuring a successful technology adoption.

Knowledge engineering for mental-health risk assessment and decision support

Mental-health risk assessment practice in the UK is mainly paper-based, with little standardisation in the tools that are used across the Services. The tools that are available tend to rely on minimal sets of items and unsophisticated scoring methods to identify at-risk individuals. This means the reasoning by which an outcome has been determined remains uncertain. Consequently, there is little provision for: including the patient as an active party in the assessment process, identifying underlying causes of risk, and eecting shared decision-making. This thesis develops a tool-chain for the formulation and deployment of a computerised clinical decision support system for mental-health risk assessment. The resultant tool, GRiST, will be based on consensual domain expert knowledge that will be validated as part of the research, and will incorporate a proven psychological model of classication for risk computation. GRiST will have an ambitious remit of being a platform that can be used over the Internet, by both the clinician and the layperson, in multiple settings, and in the assessment of patients with varying demographics. Flexibility will therefore be a guiding principle in the development of the platform, to the extent that GRiST will present an assessment environment that is tailored to the circumstances in which it nds itself. XML and XSLT will be the key technologies that help deliver this exibility.

Catalytic pyrolysis of rice husk for bio-oil production

Catalytic pyrolysis experiments have been carried out on Brunei rice husk (BRH) to obtain bio-oil using a fixed-bed pyrolysis rig. ZSM-5, Al-MCM-41, Al-MSU-F and Brunei rice husk ash (BRHA) were used as the catalysts for the catalytic pyrolysis experiments and comparison was done to analyse the changes in the bio-oil properties and yield. Properties of the liquid catalytic and non-catalytic bio-oil were analysed in terms of water content, pH, acid number, viscosity, density and calorific value. The bio-oil chemical composition shows that ZSM-5 increases the production of aromatic hydrocarbons and light phenols, whilst Al-MCM-41 reduces the acetic acid production. The catalytic runs increased the calorific value and water content in the bio-oil, whilst viscosity, density and acid number is decreased. © 2012 Elsevier B.V. All rights reserved.

Thermochemical characterisation of agricultural wastes from West Africa

Thermochemical characterisation of agricultural biomass wastes from West African region has been carried out and their potential use as feedstock in thermochemical conversion processes determined. Proximate, ultimate, structural compositions, calorific values, thermogravimetry (TGA) and derivative thermogravimetry (DTG) analyses were carried out on corn straw and cobs, rice straw and husks, cocoa pod, jatropha curcas and moringa olifiera seed cakes, parinari polyandra fruit shell and sugarcane bagasse. Moringa olifiera seed cakes and cocoa pods were found to contain the highest moisture contents. Rice straw was found to contain a high ash content of 45.76. wt.%. The level of nitrogen and sulphur in all the samples were very low. Rice husk was found to have the highest lignin contents while corn cob low lignin contents indicate a potential feedstock source for quality bio-oil production using thermochemical process. © 2013.

Catalytic intermediate pyrolysis of Brunei rice husk for bio-oil production

Rice husks from Brunei were subjected via intermediate pyrolysis for bio-oil production. Two main objectives were set out for this study. The application of intermediate pyrolysis on Brunei rice husk for the production of bio-oil is the main objective of this experiment. Characterisation of the rice husks was inclusive as a pre-requisite step to assess the suitability as feedstock for production of liquid fuels. Following on from the characterisation results, a temperature of 450°C was established as the optimum temperature for the production of bio-oil. A homogenous bio-oil was obtained from the pyrolysis of dry rice husk, and the physicochemical properties and chemical compositions were analysed. The second objective is the introduction of catalysts into the pyrolysis process which aims to improve the bio-oil quality, and maximise the desired liquid bio-oil properties. The incorporation of the catalysts was done via a fixed tube reactor into the pyrolysis system. Ceramic monoliths were used as the catalyst support, with montmorillonite clay as a binder to attach the catalysts onto the catalyst support. ZSM-5, Al-MCM-41, Al-MSU-F and Brunei rice husk ash (BRHA) together with its combination were adopted as catalysts. Proposed criterions dictated the selection of the best catalysts, subsequently leading to the optimisation process for bio-oil production. ZSM-5/Al-MCM-41 proved the most desirable catalyst, which increases the production of aromatics and phenols, decreased the organic acids and improved the physicochemical properties such as the pH, viscosity, density and H:C molar ratios. Variation in the ratio and positioning of both catalysts were the significant key factor for the catalyst optimisation study.

Designing multiple user perspectives and functionality for clinical decision support systems

Clinical Decision Support Systems (CDSSs) need to disseminate expertise in formats that suit different end users and with functionality tuned to the context of assessment. This paper reports research into a method for designing and implementing knowledge structures that facilitate the required flexibility. A psychological model of expertise is represented using a series of formally specified and linked XML trees that capture increasing elements of the model, starting with hierarchical structuring, incorporating reasoning with uncertainty, and ending with delivering the final CDSS. The method was applied to the Galatean Risk and Safety Tool, GRiST, which is a web-based clinical decision support system (www.egrist.org) for assessing mental-health risks. Results of its clinical implementation demonstrate that the method can produce a system that is able to deliver expertise targetted and formatted for specific patient groups, different clinical disciplines, and alternative assessment settings. The approach may be useful for developing other real-world systems using human expertise and is currently being applied to a logistics domain. © 2013 Polish Information Processing Society.

Degradation, receptor binding, insulin secreting and antihyperglycaemic actions of palmitate-derivatised native and Ala8-substituted GLP-1 analogues

The hormone glucagon-like peptide-1(7-36)amide (GLP-1) is released in response to ingested nutrients and acts to promote glucose-dependent insulin secretion ensuring efficient postprandial glucose homeostasis. Unfortunately, the beneficial actions of GLP-1 which give this hormone many of the desirable properties of an antidiabetic drug are short lived due to degradation by dipeptidylpeptidase IV (DPP IV) and rapid clearance by renal filtration. In this study we have attempted to extend GLP-1 action through the attachment of palmitoyl moieties to the E-amino group in the side chain of the LyS26 residue and to combine this modification with substitutions of the Ala 8 residue, namely Val or amino-butyric acid (Abu). In contrast to native GLP-1, which was rapidly degraded, [Lys(pal) 26]GLP-1, [Abu8,Lys(pal)26]GLP-1 and [Val8,Lys-(pal)26]GLP-1 all exhibited profound stability during 12 h incubations with DPP IV and human plasma. Receptor binding affinity and the ability to increase cyclic AMP in the clonal β-cell line BRIN-BD11 were decreased by 86- to 167-fold and 15- to 62-fold, respectively compared with native GLP-1. However, insulin secretory potency tested using BRIN-BD11 cells was similar, or in the case of [Val8,Lys(pal)26]GLP-1 enhanced. Furthermore, when administered in vivo together with glucose to diabetic (ob/ob) mice, [Lys(pal)26]GLP-1, [Abu8,Lys(pal) 26]GLP-1 and [Val8,Lys(pal) 26]GLP-1 did not demonstrate acute glucose-lowering or insulinotropic activity as observed with native GLP-1. These studies support the potential usefulness of fatty acid linked analogues of GLP-1 but indicate the importance of chain length for peptide kinetics and bioavailability. Copyright © by Walter de Gruyter.

Novel dipeptidyl peptidase IV resistant analogues of glucagon-like peptide-1(7-36)amide have preserved biological activities in vitro conferring improved glucose-lowering action in vivo

Although the incretin hormone glucagon-like peptide-1 (GLP-1) is a potent stimulator of insulin release, its rapid degradation in vivo by the enzyme dipeptidyl peptidase IV (DPP IV) greatly limits its potential for treatment of type 2 diabetes. Here, we report two novel Ala8-substituted analogues of GLP-1, (Abu8)GLP-1 and (Val8)GLP-1 which were completely resistant to inactivation by DPP IV or human plasma. (Abu8)GLP-1 and (Val8)GLP-1 exhibited moderate affinities (IC50: 4.76 and 81.1 nM, respectively) for the human GLP-1 receptor compared with native GLP-1 (IC50: 0.37 nM). (Abu8)GLP-1 and (Val8)GLP-1 dose-dependently stimulated cAMP in insulin-secreting BRIN BD11 cells with reduced potency compared with native GLP-1 (1.5- and 3.5-fold, respectively). Consistent with other mechanisms of action, the analogues showed similar, or in the case of (Val8)GLP-1 slightly impaired insulin releasing activity in BRIN BD11 cells. Using adult obese (ob/ob) mice, (Abu8 )GLP-1 had similar glucose-lowering potency to native GLP-1 whereas the action of (Val8)GLP-1 was enhanced by 37%. The in vivo insulin-releasing activities were similar. These data indicate that substitution of Ala8 in GLP-1 with Abu or Val confers resistance to DPP IV inactivation and that (Val8)GLP-1 is a particularly potent N-terminally modified GLP-1 analogue of possible use in type 2 diabetes.

The effect of slow passage in the pulse-pumped quantum dot laser

In recent years, quantum-dot (QD) semiconductor lasers attract significant interest in many practical applications due to their advantages such as high-power pulse generation because to the high gain efficiency. In this work, the pulse shape of an electrically pumped QD-laser under high current is analyzed. We find that the slow rise time of the pulsed pump may significantly affect the high intensity output pulse. It results in sharp power dropouts and deformation of the pulse profile. We address the effect to dynamical change of the phase-amplitude coupling in the proximity of the excited state (ES) threshold. Under 30ns pulse pumping, the output pulse shape strongly depends on pumping amplitude. At lower currents, which correspond to lasing in the ground state (GS), the pulse shape mimics that of the pump pulse. However, at higher currents the pulse shape becomes progressively unstable. The instability is greatest when in proximity to the secondary threshold which corresponds to the beginning of the ES lasing. After the slow rise stage, the output power sharply drops out. It is followed by a long-time power-off stage and large-scale amplitude fluctuations. We explain these observations by the dynamical change of the alpha-factor in the QD-laser and reveal the role of the slowly rising pumping processes in the pulse shaping and power dropouts at higher currents. The modeling is in very good agreement with the experimental observations. © 2014 SPIE.

Integrating service user and practitioner expertise within a web-based system for collaborative mental-health risk and safety management

Objectives: To develop a decision support system (DSS), myGRaCE, that integrates service user (SU) and practitioner expertise about mental health and associated risks of suicide, self-harm, harm to others, self-neglect, and vulnerability. The intention is to help SUs assess and manage their own mental health collaboratively with practitioners. Methods: An iterative process involving interviews, focus groups, and agile software development with 115 SUs, to elicit and implement myGRaCE requirements. Results: Findings highlight shared understanding of mental health risk between SUs and practitioners that can be integrated within a single model. However, important differences were revealed in SUs' preferred process of assessing risks and safety, which are reflected in the distinctive interface, navigation, tool functionality and language developed for myGRaCE. A challenge was how to provide flexible access without overwhelming and confusing users. Conclusion: The methods show that practitioner expertise can be reformulated in a format that simultaneously captures SU expertise, to provide a tool highly valued by SUs. A stepped process adds necessary structure to the assessment, each step with its own feedback and guidance. Practice Implications: The GRiST web-based DSS (www.egrist.org) links and integrates myGRaCE self-assessments with GRiST practitioner assessments for supporting collaborative and self-managed healthcare.

Dysregulation of placental cystathionine-β-synthase promotes fetal growth restriction

Fetal growth restriction (FGR) is characterized by the birth weight and body mass below the tenth percentile for gestational age. FGR is a major cause of perinatal morbidity and mortality and babies born with FGR are prone to develop cardiovascular diseases later in life. The underlying pathology of FGR is inadequate placental transfer of nutrients from mother to fetus, which can be caused by placental insufficiency. Hydrogen sulfide (H2S), a gaseous messenger is produced endogenously by cystathionine-lyase (Cth), cystathionine-β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (3-MST), which are present in human placenta. Recently, we demonstrated that the dysregulation of H2S/Cth pathway is associated with preeclampsia and blockade of CSE activity induces preeclampsia-like condition in pregnant mice. We hypothesized that defect in H2S pathways promote FGR and H2S donor restores fetal growth in mice where CBS or CSE activity has been compromised. Western blotting and qPCR revealed that placental CBS expressions were significantly reduced in women with FGR. ELISA analysis showed reduced placental growth factor production (PlGF) from first trimester (8–12 weeks gestation) human placental explants following inhibition of CBS activity by aminooxyacetic acid (AOA). Administration of AOA to pregnant mice had no effects on blood pressure, but caused fetal growth restriction. This was associated with reduced PlGF production. Histological analysis revealed a reduction in the placental junction zone, within which trophoblast giant cells and glycogen cells were less prominent in CBS inhibitor treated mice. These results imply that placental CBS is required for placental development and that dysregulation of CBS activity may contribute to the pathogenesis of FGR but not preeclampsia.