Studying language:English in action

This book introduces key ideas and current critical debates about how English functions within its social and cultural contexts, and provides practical examples and guidance on how to approach further work in these areas. It introduces core topics of language study; language variation, pragmatics, stylistics, critical discourse analysis, language and gender and language and education. Each chapter includes case studies providing worked analysis of sample texts, suggestions for further project work and an annotated further reading section.

Intergroup relations in action: questions asked about lesbian, gay and bisexual issues in diversity training

This paper focuses on the questions which heterosexual trainees ask about lesbian, gay and bisexual (LGB) experience within diversity training about LGB issues. Drawing on a data corpus of 162 questions asked by trainees in 13 tape-recorded training sessions, questions were coded into six categories: (1) general understanding questions; (2) questions about the trainer's life, experience and practices; (3) professional practice questions; (4) questions about lesbian and gay related legislation, policies and procedures; (5) questions about specific people and projects and (6) questions about the meanings, derivations and correct use of terms and symbols. Real questions are compared with the decontexualized questions (and answers to them) that are provided in training manuals and it is demonstrated that these questions differ markedly from how questions actually get asked and how they actually get answered. Recommendations are provided for improving training and the argument made for turning towards analyses of the real world in action, especially when considering intergroup relations. Copyright © 2008 John Wiley & Sons, Ltd.

Vital democracy:a theory of democracy in action - by Frank Hendricks

Book review: Frank Hendriks, Oxford University Press, 2010, 256 pp., £47 ($85.00) (hb), ISBN-13: 9780199572786

An exploration into measurement consistency on coordinate measuring machines

In high precision industry, the measurement of geometry is often performed using coordinate measuring machines (CMMs). Measurements on CMMs can occur at many places within a long and global supply chain. In this context it is a challenge to control consistency, so that measurements are applied with appropriate levels of rigour and achieve comparable results, wherever and whenever they are performed. In this paper, a framework is outlined in which consistency is controlled through measurement strategy, such as the number and location of measurement points. The framework is put to action in a case study, demonstrating the usefulness of the approach and highlighting the dangers of imposing rigid measurement strategies across the supply chain, even if linked to standardised manufacturing processes. Potential mitigations, and the requirements for future research, are outlined.

Actionable strategy knowledge:a practice perspective

Increasingly the body of knowledge derived from strategy theory has been criticized because it is not actionable in practice, particularly under the conditions of a knowledge economy. Since strategic management is an applied discipline this is a serious criticism. However, we argue that the theory-practice question is too simple. Accordingly, this paper expands this question by outlining first the theoretical criteria under which strategy theory is not actionable, and then outlines an alternative perspective on strategy knowledge in action, based upon a practice epistemology. The paper is in three sections. The first section explains two contextual conditions which impact upon strategy theory within a knowledge economy, environmental velocity and knowledge intensity. The impact of these contextual conditions upon the application of four different streams of strategy theory is examined. The second section suggests that the theoretical validity of these contextual conditions breaks down when we consider the knowledge artifacts, such as strategy tools and frameworks, which arise from strategy research. The third section proposes a practice epistemology for analyzing strategy knowledge in action that stands in contrast to more traditional arguments about actionable knowledge. From a practice perspective, strategy knowledge is argues to be actionable as part of the everyday activities of strategizing. © 2006 Elsevier Ltd. All rights reserved.

Evaluating corporate performance:a critique of economic value added

There has been a revival of interest in economic techniques to measure the value of a firm through the use of economic value added as a technique for measuring such value to shareholders. This technique, based upon the concept of economic value equating to total value, is founded upon the assumptions of classical liberal economic theory. Such techniques have been subject to criticism both from the point of view of the level of adjustment to published accounts needed to make the technique work and from the point of view of the validity of such techniques in actually measuring value in a meaningful context. This paper critiques economic value added techniques as a means of calculating changes in shareholder value, contrasting such techniques with more traditional techniques of measuring value added. It uses the company Severn Trent plc as an actual example in order to evaluate and contrast the techniques in action. The paper demonstrates discrepancies between the calculated results from using economic value added analysis and those reported using conventional accounting measures. It considers the merits of the respective techniques in explaining shareholder and managerial behaviour and the problems with using such techniques in considering the wider stakeholder concept of value. It concludes that this economic value added technique has merits when compared with traditional accounting measures of performance but that it does not provide the universal panacea claimed by its proponents.

A contingency framework for information systems development

This research concerns information systems and information systems development. The thesis describes an approach to information systems development called Multiview. This is a methodology which seeks to combine the strengths of a number of different, existing approaches in a coherent manner. Many of these approaches are radically different in terms of concepts, philosophy, assumptions, methods, techniques and tools. Three case studies are described presenting Multiview 'in action'. The first is used mainly to expose the strengths and weaknesses of an early version of the approach discussed in the thesis. Tools and techniques are described in the thesis which aim to strengthen the approach. Two further case studies are presented to illustrate the use of this second version of Multiview. This is not put forward as an 'ideal methodology' and the case studies expose some of the difficulties and practical problems of information systems work and the use of the methodology. A more contingency based approach to information systems development is advocated using Multiview as a framework rather than a prescriptive tool. Each information systems project and the use of the framework is unique, contingent on the particular problem situation. The skills of different analysts, the backgrounds of users and the situations in which they are constrained to work have always to be taken into account in any project. The realities of the situation will cause departure from the 'ideal methodology' in order to allow for the exigencies of the real world. Multiview can therefore be said to be an approach used to explore the application area in order to develop an information system.

Signal processing for molecular and cellular biological physics:an emerging field

Recent advances in our ability to watch the molecular and cellular processes of life in action-such as atomic force microscopy, optical tweezers and Forster fluorescence resonance energy transfer-raise challenges for digital signal processing (DSP) of the resulting experimental data. This article explores the unique properties of such biophysical time series that set them apart from other signals, such as the prevalence of abrupt jumps and steps, multi-modal distributions and autocorrelated noise. It exposes the problems with classical linear DSP algorithms applied to this kind of data, and describes new nonlinear and non-Gaussian algorithms that are able to extract information that is of direct relevance to biological physicists. It is argued that these new methods applied in this context typify the nascent field of biophysical DSP. Practical experimental examples are supplied.

The mosaic test:measuring the effectiveness of colour-based image retrieval

A variety of content-based image retrieval systems exist which enable users to perform image retrieval based on colour content - i.e., colour-based image retrieval. For the production of media for use in television and film, colour-based image retrieval is useful for retrieving specifically coloured animations, graphics or videos from large databases (by comparing user queries to the colour content of extracted key frames). It is also useful to graphic artists creating realistic computer-generated imagery (CGI). Unfortunately, current methods for evaluating colour-based image retrieval systems have 2 major drawbacks. Firstly, the relevance of images retrieved during the task cannot be measured reliably. Secondly, existing methods do not account for the creative design activity known as reflection-in-action. Consequently, the development and application of novel and potentially more effective colour-based image retrieval approaches, better supporting the large number of users creating media for use in television and film productions, is not possible as their efficacy cannot be reliably measured and compared to existing technologies. As a solution to the problem, this paper introduces the Mosaic Test. The Mosaic Test is a user-based evaluation approach in which participants complete an image mosaic of a predetermined target image, using the colour-based image retrieval system that is being evaluated. In this paper, we introduce the Mosaic Test and report on a user evaluation. The findings of the study reveal that the Mosaic Test overcomes the 2 major drawbacks associated with existing evaluation methods and does not require expert participants. © 2012 Springer Science+Business Media, LLC.

Editorial overview:new protein production tools for structural biology

Full text: The idea of producing proteins from recombinant DNA hatched almost half a century ago. In his PhD thesis, Peter Lobban foresaw the prospect of inserting foreign DNA (from any source, including mammalian cells) into the genome of a λ phage in order to detect and recover protein products from Escherichia coli [ 1 and 2]. Only a few years later, in 1977, Herbert Boyer and his colleagues succeeded in the first ever expression of a peptide-coding gene in E. coli — they produced recombinant somatostatin [ 3] followed shortly after by human insulin. The field has advanced enormously since those early days and today recombinant proteins have become indispensable in advancing research and development in all fields of the life sciences. Structural biology, in particular, has benefitted tremendously from recombinant protein biotechnology, and an overwhelming proportion of the entries in the Protein Data Bank (PDB) are based on heterologously expressed proteins. Nonetheless, synthesizing, purifying and stabilizing recombinant proteins can still be thoroughly challenging. For example, the soluble proteome is organized to a large part into multicomponent complexes (in humans often comprising ten or more subunits), posing critical challenges for recombinant production. A third of all proteins in cells are located in the membrane, and pose special challenges that require a more bespoke approach. Recent advances may now mean that even these most recalcitrant of proteins could become tenable structural biology targets on a more routine basis. In this special issue, we examine progress in key areas that suggests this is indeed the case. Our first contribution examines the importance of understanding quality control in the host cell during recombinant protein production, and pays particular attention to the synthesis of recombinant membrane proteins. A major challenge faced by any host cell factory is the balance it must strike between its own requirements for growth and the fact that its cellular machinery has essentially been hijacked by an expression construct. In this context, Bill and von der Haar examine emerging insights into the role of the dependent pathways of translation and protein folding in defining high-yielding recombinant membrane protein production experiments for the common prokaryotic and eukaryotic expression hosts. Rather than acting as isolated entities, many membrane proteins form complexes to carry out their functions. To understand their biological mechanisms, it is essential to study the molecular structure of the intact membrane protein assemblies. Recombinant production of membrane protein complexes is still a formidable, at times insurmountable, challenge. In these cases, extraction from natural sources is the only option to prepare samples for structural and functional studies. Zorman and co-workers, in our second contribution, provide an overview of recent advances in the production of multi-subunit membrane protein complexes and highlight recent achievements in membrane protein structural research brought about by state-of-the-art near-atomic resolution cryo-electron microscopy techniques. E. coli has been the dominant host cell for recombinant protein production. Nonetheless, eukaryotic expression systems, including yeasts, insect cells and mammalian cells, are increasingly gaining prominence in the field. The yeast species Pichia pastoris, is a well-established recombinant expression system for a number of applications, including the production of a range of different membrane proteins. Byrne reviews high-resolution structures that have been determined using this methylotroph as an expression host. Although it is not yet clear why P. pastoris is suited to producing such a wide range of membrane proteins, its ease of use and the availability of diverse tools that can be readily implemented in standard bioscience laboratories mean that it is likely to become an increasingly popular option in structural biology pipelines. The contribution by Columbus concludes the membrane protein section of this volume. In her overview of post-expression strategies, Columbus surveys the four most common biochemical approaches for the structural investigation of membrane proteins. Limited proteolysis has successfully aided structure determination of membrane proteins in many cases. Deglycosylation of membrane proteins following production and purification analysis has also facilitated membrane protein structure analysis. Moreover, chemical modifications, such as lysine methylation and cysteine alkylation, have proven their worth to facilitate crystallization of membrane proteins, as well as NMR investigations of membrane protein conformational sampling. Together these approaches have greatly facilitated the structure determination of more than 40 membrane proteins to date. It may be an advantage to produce a target protein in mammalian cells, especially if authentic post-translational modifications such as glycosylation are required for proper activity. Chinese Hamster Ovary (CHO) cells and Human Embryonic Kidney (HEK) 293 cell lines have emerged as excellent hosts for heterologous production. The generation of stable cell-lines is often an aspiration for synthesizing proteins expressed in mammalian cells, in particular if high volumetric yields are to be achieved. In his report, Buessow surveys recent structures of proteins produced using stable mammalian cells and summarizes both well-established and novel approaches to facilitate stable cell-line generation for structural biology applications. The ambition of many biologists is to observe a protein's structure in the native environment of the cell itself. Until recently, this seemed to be more of a dream than a reality. Advances in nuclear magnetic resonance (NMR) spectroscopy techniques, however, have now made possible the observation of mechanistic events at the molecular level of protein structure. Smith and colleagues, in an exciting contribution, review emerging ‘in-cell NMR’ techniques that demonstrate the potential to monitor biological activities by NMR in real time in native physiological environments. A current drawback of NMR as a structure determination tool derives from size limitations of the molecule under investigation and the structures of large proteins and their complexes are therefore typically intractable by NMR. A solution to this challenge is the use of selective isotope labeling of the target protein, which results in a marked reduction of the complexity of NMR spectra and allows dynamic processes even in very large proteins and even ribosomes to be investigated. Kerfah and co-workers introduce methyl-specific isotopic labeling as a molecular tool-box, and review its applications to the solution NMR analysis of large proteins. Tyagi and Lemke next examine single-molecule FRET and crosslinking following the co-translational incorporation of non-canonical amino acids (ncAAs); the goal here is to move beyond static snap-shots of proteins and their complexes and to observe them as dynamic entities. The encoding of ncAAs through codon-suppression technology allows biomolecules to be investigated with diverse structural biology methods. In their article, Tyagi and Lemke discuss these approaches and speculate on the design of improved host organisms for ‘integrative structural biology research’. Our volume concludes with two contributions that resolve particular bottlenecks in the protein structure determination pipeline. The contribution by Crepin and co-workers introduces the concept of polyproteins in contemporary structural biology. Polyproteins are widespread in nature. They represent long polypeptide chains in which individual smaller proteins with different biological function are covalently linked together. Highly specific proteases then tailor the polyprotein into its constituent proteins. Many viruses use polyproteins as a means of organizing their proteome. The concept of polyproteins has now been exploited successfully to produce hitherto inaccessible recombinant protein complexes. For instance, by means of a self-processing synthetic polyprotein, the influenza polymerase, a high-value drug target that had remained elusive for decades, has been produced, and its high-resolution structure determined. In the contribution by Desmyter and co-workers, a further, often imposing, bottleneck in high-resolution protein structure determination is addressed: The requirement to form stable three-dimensional crystal lattices that diffract incident X-ray radiation to high resolution. Nanobodies have proven to be uniquely useful as crystallization chaperones, to coax challenging targets into suitable crystal lattices. Desmyter and co-workers review the generation of nanobodies by immunization, and highlight the application of this powerful technology to the crystallography of important protein specimens including G protein-coupled receptors (GPCRs). Recombinant protein production has come a long way since Peter Lobban's hypothesis in the late 1960s, with recombinant proteins now a dominant force in structural biology. The contributions in this volume showcase an impressive array of inventive approaches that are being developed and implemented, ever increasing the scope of recombinant technology to facilitate the determination of elusive protein structures. Powerful new methods from synthetic biology are further accelerating progress. Structure determination is now reaching into the living cell with the ultimate goal of observing functional molecular architectures in action in their native physiological environment. We anticipate that even the most challenging protein assemblies will be tackled by recombinant technology in the near future.