Human SULT1A genes: Cloning and activity assays of the SULT1A promoters

The three human SULT1A sulfotransferase enzymes are closely related in amino acid sequence (>90%), yet differ in their substrate preference and tissue distribution. SULT1A1 has a broad tissue distribution and metabolizes a range of xenobiotics as well as endogenous substrates such as estrogens and iodothyronines. While the localization of SULT1A2 is poorly understood, it has been shown to metabolize a number of aromatic amines. SULT1A3 is the major catecholamine sulfonating form, which is consistent with it being expressed principally in the gastrointestinal tract. SULT1A proteins are encoded by three separate genes, located in close proximity to each other on chromosome 16. The presence of differential 5′-untranslated regions identified upon cloning of the SULT1A cDNAs suggested the utilization of differential transcriptional start sites and/or differential splicing. This chapter describes the methods utilized by our laboratory to clone and assay the activity of the promoters flanking these different untranslated regions found on SULT1A genes. These techniques will assist investigators in further elucidating the differential mechanisms that control regulation of the human SULT1A genes. They will also help reveal how different cellular environments and polymorphisms affect the activity of SULT1A gene promoters.

Using context and preferences to implement self-adapting pervasive computing applications

Applications that exploit contextual information in order to adapt their behaviour to dynamically changing operating environments and user requirements are increasingly being explored as part of the vision of pervasive or ubiquitous computing. Despite recent advances in infrastructure to support these applications through the acquisition, interpretation and dissemination of context data from sensors, they remain prohibitively difficult to develop and have made little penetration beyond the laboratory. This situation persists largely due to a lack of appropriately high-level abstractions for describing, reasoning about and exploiting context information as a basis for adaptation. In this paper, we present our efforts to address this challenge, focusing on our novel approach involving the use of preference information as a basis for making flexible adaptation decisions. We also discuss our experiences in applying our conceptual and software frameworks for context and preference modelling to a case study involving the development of an adaptive communication application.

Structure and activity in assessing antioxidant activity in vitro and in vivo:a critical appraisal illustrated with the flavonoids

Structure–activity relationships are indispensable to identify the most optimal antioxidants. The advantages of in vitro over in vivo experiments for obtaining these relationships are, that the structure is better defined in vitro, since less metabolism takes place. It is also the case that the concentration, a parameter that is directly linked to activity, is more accurately controlled. Moreover, the reactions that occur in vivo, including feed-back mechanisms, are often too multi-faceted and diverse to be compensated for during the assessment of a single structure–activity relationship. Pitfalls of in vitro antioxidant research include: (i) by definition, antioxidants are not stable and substantial amounts of oxidation products are formed and (ii) during the scavenging of reactive species, reaction products of the antioxidants accumulate. Another problem is that the maintenance of a defined concentration of antioxidants is subject to processes such as oxidation and the formation of reaction products during the actual antioxidant reaction, as well as the compartmentalization of the antioxidant and the reactive species in the in vitro test system. So determinations of in vitro structure-activity relationships are subject to many competing variables and they should always be evaluated critically. (c) 2005 Published by Elsevier B.V.

The role of context and timeframe in moderating relationships within the theory of planned behaviour

This study examined the moderating effect of context and timeframe on the predictive ability of Theory of Planned Behaviour (TPB) constructs. Three hundred and eighty-three students completed TPB measures either in a campus bar or a library and were randomly allocated to one of three timeframe conditions: tonight, tomorrow or next week. There was a threeway interaction such that the subjective norms of participants in a bar were more predictive of their intentions to binge drink that night, whereas thesubjective norms of participants in a library were less predictive of intentions to binge drink that night. This research provides empirical evidence that ignoring context may result in underestimation of the importance of normative factors in binge drinking. It also suggests that other research utilising the TPB needs to take greater account of the impact of context of data collection, which has been neglected to date.

The heteromeric 5-HT3A/B receptor:the effect of 5-HT3B subunits on receptor structure and ligand recognition

The 5-HT3 receptors are members of the cys-loop family of ligand-gated ion channels. Two functional subtypes are known, the homomeric 5HT3A and the heteromeric 5HT3A/B receptors, which exhibit distinct biophysical characteristics but are difficult to differentiate pharmacologically. Atomic force microscopy has been used to determine the stoichiometry and architecture of the heteromeric 5HT3A/B receptor. Each subunit was engineered to express a unique C-terminal epitope tag, together with six sequential histidine residues to facilitate nickel affinity purification. The 5-HT3 receptors, ectopically expressed in HEK293 cells, were solubilised, purified and decorated with antibodies to the subunit specific epitope tags. Imaging of individual receptors by atomic force microscopy revealed a pentameric arrangement of subunits in the order BBABA, reading anti-clockwise when viewed from the extracellular face. Homology models for the heteromeric receptor were then constructed using both the electron microscopic structure of the nicotinic acetylcholine receptor, from Torpedo marmorata, and the X-ray crystallographic structure of the soluble acetylcholine binding protein, from Lymnaea stagnalis, as templates. These homology models were used, together with equivalent models constructed for the homomeric receptor, to interpret mutagenesis experiments designed to explore the minimal recognition differences of both the natural agonist, 5-HT, and the competitive antagonist, granisetron, for the two human receptor subtypes. The results of this work revealed that the 5-HT3B subunit residues within the ligand binding site, for both the agonist and antagonist, are accommodating to conservative mutations. They are consistent with the view that the 5-HT3A subunit provides the principal and the 5-HT38 subunit the complementary recognition interactions at the binding interface.

Solid-supported boronic acid conjugates for sugar and glycopeptide recognition

The Fmoc synthetic strategy was employed to synthesise two identical combinatorial peptide libraries on a hydrophilic PEG-PS resin. One library was appended with boronic acid moieties at two positionally-fixed locations. Successful inclusion of the boronic acid units was confirmed using a novel UV fluorescent colorimetric assay employing carminic acid as the dye compound. A study of the effect had by the resin-bound peptides bearing boronic acid groups on the binding characteristics of vancomycin, a medically relevant antibiotic glycoprotein, was conducted. In all, 132 library compounds were tested for their binding affinity with vancomycin, via immobilisation of the glycopeptide onto the solid support through hydrogen bonding or complexation with the boronic acid moieties. Subsequent cleavage via acidolysis afforded vancomycin containing solutions which were quantified by growth inhibition of methicillin susceptible Staphylococcus aureus. Comparison of the diameters of the resultant zones of inhibition and those produced by vancomycin of known concentrations afforded a means of calculating the vancomycin concentration of the cleavage solutions, and thereby determining the binding affinity of vancomycin to each peptide sequence. Five peptide sequences and twenty one of the peptidyl-boronic acid sequences showed zones of inhibition, demonstrating their reversible affinity for vancomycin. Three peptide sequences showed zones of inhibition in both libraries. The presence of boronic acid was therefore shown to impart, enhance, detract and remove the affinity of vancomycin to a range of resin-bound peptide sequences.

A comparison of microphone and speech recognition engine efficacy for mobile data entry

The research presented in this paper is part of an ongoing investigation into how best to incorporate speech-based input within mobile data collection applications. In our previous work [1], we evaluated the ability of a single speech recognition engine to support accurate, mobile, speech-based data input. Here, we build on our previous research to compare the achievable speaker-independent accuracy rates of a variety of speech recognition engines; we also consider the relative effectiveness of different speech recognition engine and microphone pairings in terms of their ability to support accurate text entry under realistic mobile conditions of use. Our intent is to provide some initial empirical data derived from mobile, user-based evaluations to support technological decisions faced by developers of mobile applications that would benefit from, or require, speech-based data entry facilities.

Peripheral vision and pattern recognition : a review

We summarize the various strands of research on peripheral vision and relate them to theories of form perception. After a historical overview, we describe quantifications of the cortical magnification hypothesis, including an extension of Schwartz's cortical mapping function. The merits of this concept are considered across a wide range of psychophysical tasks, followed by a discussion of its limitations and the need for non-spatial scaling. We also review the eccentricity dependence of other low-level functions including reaction time, temporal resolution, and spatial summation, as well as perimetric methods. A central topic is then the recognition of characters in peripheral vision, both at low and high levels of contrast, and the impact of surrounding contours known as crowding. We demonstrate how Bouma's law, specifying the critical distance for the onset of crowding, can be stated in terms of the retinocortical mapping. The recognition of more complex stimuli, like textures, faces, and scenes, reveals a substantial impact of mid-level vision and cognitive factors. We further consider eccentricity-dependent limitations of learning, both at the level of perceptual learning and pattern category learning. Generic limitations of extrafoveal vision are observed for the latter in categorization tasks involving multiple stimulus classes. Finally, models of peripheral form vision are discussed. We report that peripheral vision is limited with regard to pattern categorization by a distinctly lower representational complexity and processing speed. Taken together, the limitations of cognitive processing in peripheral vision appear to be as significant as those imposed on low-level functions and by way of crowding.

Business and social entrepreneurs in the UK:gender, context and commitment

Purpose – The purpose of this paper is to investigate what sort of people become social entrepreneurs, and in what way they differ from business entrepreneurs. More importantly, to investigate in what socio-economic context entrepreneurial individuals are more likely to become social than business entrepreneurs. These questions are important for policy because there has been a shift from direct to indirect delivery of many public services in the UK, requiring a professional approach to social enterprise. Design/methodology/approach – Evidence is presented from the Global Entrepreneurship Monitor (GEM) UK survey based upon a representative sample of around 21,000 adults aged between 16 and 64 years interviewed in 2009. The authors use logistic multivariate regression techniques to identify differences between business and social entrepreneurs in demographic characteristics, effort, aspiration, use of resources, industry choice, deprivation, and organisational structure. Findings – The results show that the odds of an early-stage entrepreneur being a social rather than a business entrepreneur are reduced if they are from an ethnic minority, if they work ten hours or more per week on the venture, and if they have a family business background; while they are increased if they have higher levels of education and if they are a settled in-migrant to their area. While women social entrepreneurs are more likely than business entrepreneurs to be women, this is due to gender-based differences in time commitment to the venture. In addition, the more deprived the community they live in, the more likely women entrepreneurs are to be social than business entrepreneurs. However, this does not hold in the most deprived areas where we argue civic society is weakest and therefore not conducive to support any form of entrepreneurial endeavour based on community engagement. Originality/value – The paper's findings suggest that women may be motivated to become social entrepreneurs by a desire to improve the socio-economic environment of the community in which they live and see social enterprise creation as an appropriate vehicle with which to address local problems.