Functional and Structural Diversification of the Anguimorpha Lizard Venom System

Venom has only been recently discovered to be a basal trait of the Anguimorpha lizards. Consequently, very little is known about the timings of toxin recruitment events, venom protein molecular evolution, or even the relative physical diversifications of the venom system itself. A multidisciplinary approach was used to examine the evolution across the full taxonomical range of this similar to 130 million-year-old clade. Analysis of cDNA libraries revealed complex venom transcriptomes. Most notably, three new cardioactive peptide toxin types were discovered (celestoxin, cholecystokinin, and YY peptides). The latter two represent additional examples of convergent use of genes in toxic arsenals, both having previously been documented as components of frog skin defensive chemical secretions. Two other novel venom gland-overexpressed modified versions of other protein frameworks were also recovered from the libraries (epididymal secretory protein and ribonuclease). Lectin, hyaluronidase, and veficolin toxin types were sequenced for the first time from lizard venoms and shown to be homologous to the snake venom forms. In contrast, phylogenetic analyses demonstrated that the lizard natriuretic peptide toxins were recruited independently of the form in snake venoms. The de novo evolution of helokinestatin peptide toxin encoding do-mains within the lizard venom natriuretic gene was revealed to be exclusive to the helodermatid/anguid subclade. New isoforms were sequenced for cysteine-rich secretory protein, kallikrein, and phospholipase A 2 toxins. Venom gland morphological analysis revealed extensive evolutionary tinkering. Anguid glands are characterized by thin capsules and mixed glands, serous at the bottom of the lobule and mucous toward the apex. Twice, independently this arrangement was segregated into specialized serous protein-secreting glands with thick capsules with the mucous lobules now distinct (Heloderma and the Lanthanotus/Varanus clade). The results obtained highlight the importance of utilizing evolution-based search strategies for biodiscovery and emphasize the largely untapped drug design and development potential of lizard venoms. Molecular & Cellular Proteomics 9:2369-2390, 2010.

Confined site construction: A qualitative review of issues regarding health and safety on-site

The construction industry is inherently hazardous, with a significant number of accidents and incidents occurring, particularly on confined construction sites. This research identifies, clarifies and tabulates the various managerial health and safety issues encountered on confined construction sites, based on a qualitative approach, to aid in the management of the complex health and safety concerns. The methodology is based on qualitative research incorporating case studies, interviews, causal loop diagrams and mind mapping. The key findings in the managerial issues in the management of health and safety on confined construction sites can be summarised as follows; (1) A lack of space, (2) Increased management of site personnel, (3) Overcrowding of the work place. The implication for the industry is that due to the sustained development of urban centres on a global scale, coupled with the increasing complexity of architectural designs, the majority of on-site project management professionals are faced with the onerous task of completing often intricate designs within a limited spatial environment, under strict health and safety parameters. The value of such research is to aid management professionals successfully identify the various managerial issues highlighted, resulting in the successful management of health and safety on a confined construction site.

Confined Site Construction: A qualitative investigation of critical issues affecting management of Health and Safety

The construction industry is inherently risky, with a significant number of accidents and disasters occurring, particularly on confined construction sites. This research investigates and identifies the various issues affecting successful management of health and safety in confined construction sites. The rationale is that identifying the issues would assist the management of health and safety particularly in inner city centres which are mostly confined sites. Using empiricism epistemology, the methodology was based on qualitative research approach by means of multiple case studies in three different geographical locations of Ireland, UK and USA. Data on each case study were collected through individual interviews and focus group discussion with project participants. The findings suggest that three core issues are the underlying factors affecting management of health and safety on confined construction sites. It include, (i) lack of space, (ii) problem of co-ordination and management of site personnel, and (iii) overcrowding of workplace. The implication of this is that project teams and their organisations should see project processes from a holistic point of view, as a unified single system, where quick intervention in solving a particular issue should be the norm, so as not to adversely affect interrelated sequence of events in project operations. Proactive strategies should be devised to mitigate these issues and may include detail project programming, space management, effective constructability review and efficient co-ordination of personnel, plant and materials among others. The value of this research is to aid management and operation of brownfield sites by identifying issues impacting on health and safety management in project process.

The Interview:An Ethnographic Approach

What are new interview methods and practices in our new 'interview society' and how do they relate to traditional social science research? This volume interrogates the interview as understood, used - and under-used - by anthropologists. It puts the interview itself in the hotseat by exploring the nature of the interview, interview techniques, and illustrative cases of interview use.

What is a successful and representative interview? How are interviews best transcribed and integrated into our writing? Is interview knowledge production safe, ethical and representative? And how are interviews used by anthropologists in their ethnographic practice?

This important volume leads the reader from an initial scrutiny of the interview to interview techniques and illustrative case studies. It is experimental, innovative, and covers in detail matters such as awkwardness, silence and censorship in interviews that do not feature in general interview textbooks. It will appeal to social scientists engaged in qualitative research methods in general, and anthropology and sociology students using interviews in their research and writing in particular.

Functional peptidomics of amphibian skin secretion: A novel Kunitz-type chymotrypsin inhibitor from the African hyperoliid frog, Kassina senegalensis

Amphibian skin secretions are, for the most part, complex peptidomes. While many peptide components have been biologically- and structurally-characterised into discrete "families", some of which are analogues of endogenous vertebrate regulatory peptides, a substantial number are of unique structure and unknown function. Among the components of these secretory peptidomes is an array of protease inhibitors. Inhibitors of trypsin are of widespread occurrence in different taxa and are representative of many established structural classes, including Kunitz, Kazal and Bowman-Birk. However, few protease inhibitors with activity against other specific proteases have been described from this source. Here we report for the first time, the isolation and structural characterisation of an inhibitor of chymotrypsin of Kunitz-type from the skin secretion of the African hyperoliid frog, Kassina senegalensis. To this end, we employed a functional peptidomic approach. This scheme involves fractionation of the peptidome, functional end-point screening, structural characterisation of resultant actives followed by molecular cloning of biosynthetic precursor-encoding cDNA(s). The novel mature and active polypeptide identified consisted of 62 amino acid residues (average molecular mass 6776.24 Da), of which 6 were positionally-conserved cysteines. The P(1) position within the active site was occupied by a phenylalanyl residue. Bioinformatic analysis of the sequence using BLAST, revealed a structural similarity to Kunitz-type chymotrypsin inhibitors from other organisms, ranging from silkworms to snakes.

Synthesis and SAR of novel, non-MPEP chemotype mGluR5 NAMs identified by functional HTS

This Letter describes the discovery and SAR of three novel series of mGluR5 non-competitive antagonists/negative allosteric modulators (NAMs) not based on manipulation of an MPEP/MTEP chemotype identified by a functional HTS approach. This work demonstrates fundamentally new mGluR5 NAM chemotypes with submicromolar potencies, and further examples of a mode of pharmacology 'switch' to provide PAMs with a non-MPEP scaffold. (C) 2009 Elsevier Ltd. All rights reserved.

Synthesis and SAR of novel, 4-(phenylsulfamoyl)phenylacetamide mGlu(4) positive allosteric modulators (PAMs) identified by functional high-throughput screening (HTS)

Herein we disclose the synthesis and SAR of a series of 4-(phenylsulfamoyl)phenylacetamide compounds as mGlu(4) positive allosteric modulators (PAMs) that were identified via a functional HTS. An iterative parallel approach to these compounds culminated in the discovery of VU0364439 (11) which represents the most potent (19.8 nM) mGlu(4) PAM reported to date. (C) 2010 Elsevier Ltd. All rights reserved.

A Systems Biology Approach Identifies SART1 as a Novel Determinant of Both 5-Fluorouracil and SN38 Drug Resistance in Colorectal Cancer

Chemotherapy response rates for advanced colorectal cancer remain disappointingly low, primarily because of drug resistance, so there is an urgent need to improve current treatment strategies. To identify novel determinants of resistance to the clinically relevant drugs 5-fluorouracil (5-FU) and SN38 (the active metabolite of irinotecan), transcriptional profiling experiments were carried out on pretreatment metastatic colorectal cancer biopsies and HCT116 parental and chemotherapy-resistant cell line models using a disease-specific DNA microarray. To enrich for potential chemoresistance-determining genes, an unsupervised bioinformatics approach was used, and 50 genes were selected and then functionally assessed using custom-designed short interfering RNA(siRNA) screens. In the primary siRNA screen, silencing of 21 genes sensitized HCT116 cells to either 5-FU or SN38 treatment. Three genes (RAPGEF2, PTRF, and SART1) were selected for further analysis in a panel of 5 colorectal cancer cell lines. Silencing SART1 sensitized all 5 cell lines to 5-FU treatment and 4/5 cell lines to SN38 treatment. However, silencing of RAPGEF2 or PTRF had no significant effect on 5-FU or SN38 sensitivity in the wider cell line panel. Further functional analysis of SART1 showed that its silencing induced apoptosis that was caspase-8 dependent. Furthermore, silencing of SART1 led to a downregulation of the caspase-8 inhibitor, c-FLIP, which we have previously shown is a key determinant of drug resistance in colorectal cancer. This study shows the power of systems biology approaches for identifying novel genes that regulate drug resistance and identifies SART1 as a previously unidentified regulator of c-FLIP and drug-induced activation of caspase-8. Mol Cancer Ther; 11(1); 119-31. (C) 2011 AACR.

Notes from Some Small Countries: A Study of the 'New Punitiveness' in Ireland, Scotland and New Zealand

In recent years there has been a remarkable surge of interest in the concept of punitiveness in theoretical criminology. Accounts serve to emphasise rupture over continuity, drawing attention to the increased focus on managerialism, risk and expressive penal policies in countries such as England and the US. Criticisms of these accounts have drawn attention to the weak empirical base for such assertions and the continued relevance of local cultural, historical and political conditions in mediating the effect of more punitive trends. In light of the relative neglect of smaller jurisdictions in this literature it was decided to locate these debates in three small common law jurisdictions, namely, Ireland, Scotland and New Zealand over the period 1976-2006 with a view to assessing the empirical evidence for penal change. This was done using a broader definition of punitiveness than normally employed incorporating indices relating to the ‘front end’ (eg police powers) as well as the ‘back end’ (eg prison and probation) of the criminal justice system. Data were collected on the three case studies using a multi-method approach involving examination of extensive quantitative data, interviews with key criminal justice stakeholders and documentary analysis. The data provide some support for the ‘new punitiveness’ thesis in these countries through a pattern of increased legislative activity aimed at controlling violent and sexual offenders and significant increases in the lengths of sentences imposed. However, analysis of qualitative data and a larger number of variables reveals distinctly different patterns of punitiveness over the thirty year period in the three countries. It is argued that the study holds important lessons for comparative criminology into the ‘new punitiveness’. There is a need for qualitative as well as quantitative data; for multiple rather than singular indices across a wide range of areas (juvenile justice, prison conditions, etc); and for ‘front end’ as well as ‘back end’ indices.

Managing i-branding to create brand equity

Purpose: Given the emergent nature of i-branding as an academic field of study and a lack of applied research output, the aim of this paper is to explain how businesses manage i-branding to create brand equity.

Design/methodology/approach: Within a case-study approach, seven cases were developed from an initial sample of 20 food businesses. Additionally, utilising secondary data, the analysis of findings introduces relevant case examples from other industrial sectors.

Findings: Specific internet tools and their application are discussed within opportunities to create brand equity for products classified by experience, credence and search characteristics. An understanding of target customers will be critical in underpinning the selection and deployment of relevant i-branding tools. Tools facilitating interactivity – machine and personal – are particularly significant.

Research limitations/implications: Future research positioned within classification of goods constructs could provide further contributions that recognise potential moderating effects of product/service characteristics on the development of brand equity online. Future studies could also employ the i-branding conceptual framework to test its validity and develop it further as a means of explaining how i-branding can be managed to create brand equity.

Originality/value: While previous research has focused on specific aspects of i-branding, this paper utilises a conceptual framework to explain how diverse i-branding tools combine to create brand equity. The literature review integrates fragmented literature around a conceptual framework to produce a more coherent understanding of extant thinking. The location of this study within a classification of goods context proved critical to explaining how i-branding can be managed.

A normative approach to the criminalisation of cartel activity

Although cartel behaviour is almost universally (and rightly) condemned, it is not clear why cartel participants deserve the full wrath of the criminal law and its associated punishment. To fill this void, I develop a normative (or principled) justification for the criminalisation of conduct characteristic of ‘hard core’ cartels. The paper opens with a brief consideration of the rhetoric commonly used to denounce cartel activity, eg that it ‘steals from’ or ‘robs’ consumers. To put the discussion in context, a brief definition of ‘hard core’ cartel behaviour is provided and the harms associated with this activity are identified. These are: welfare losses in the form of appropriation (from consumer to producer) of consumer surplus, the creation of deadweight loss to the economy, the creation of productive inefficiency (hindering innovation of both products and processes), and the creation of so-called X-inefficiency. As not all activities which cause harm ought to be criminalised, a theory as to why certain harms in a liberal society can be criminalised is developed. It is based on JS Mill's harm to others principle (as refined by Feinberg) and on a choice of social institutions using Rawls's ‘veil of ignorance.’ The theory is centred on the value of individual choice in securing one's own well-being, with the market as an indispensable instrument for this. But as applied to the harm associated with cartel conduct, this theory shows that none of the earlier mentioned problems associated with this activity provide sufficient justification for criminalisation. However, as the harm from hard core cartel activity strikes at an important institution which permits an individual's ability to secure their own well-being in a liberal society, criminalisation of hard core cartel behaviour can have its normative justification on this basis.

Functional Complexity: The source of value in biodiversity.

Biodiversity may be seen as a scientific measure of the complexity of a biological system, implying an information basis. Complexity cannot be directly valued, so economists have tried to define the services it provides, though often just valuing the services of 'key' species. Here we provide a new definition of biodiversity as a measure of functional information, arguing that complexity embodies meaningful information as Gregory Bateson defined it. We argue that functional information content (FIC) is the potentially valuable component of total (algorithmic) information content (AIC), as it alone determines biological fitness and supports ecosystem services. Inspired by recent extensions to the Noah's Ark problem, we show how FIC/AIC can be calculated to measure the degree of substitutability within an ecological community. Establishing substitutability is an essential foundation for valuation. From it, we derive a way to rank whole communities by Indirect Use Value, through quantifying the relation between system complexity and the production rate of ecosystem services. Understanding biodiversity as information evidently serves as a practical interface between economics and ecological science. © 2012 Elsevier B.V.

Determining trophic niche width: An experimental test of the stable isotope approach

Determining the trophic niche width of an animal population and the relative degree to which a generalist population consists of dietary specialists are long-standing problems of ecology. It has been proposed that the variance of stable isotope values in consumer tissues could be used to quantify trophic niche width of consumer populations. However, this promising idea has not yet been rigorously tested. By conducting controlled laboratory experiments using model consumer populations (Daphnia sp., Crustacea) with controlled diets, we investigated the effect of individual- and population-level specialisation and generalism on consumer d C mean and variance values. While our experimental data follow general expectations, we extend current qualitative models to quantitative predictions of the dependence of isotopic variance on dietary correlation time, a measure for the typical time over which a consumer changes its diet. This quantitative approach allows us to pinpoint possible procedural pitfalls and critical sources of measurement uncertainty. Our results show that the stable isotope approach represents a powerful method for estimating trophic niche widths, especially when taking the quantitative concept of dietary correlation time into account. © 2012 The Authors.

Quantitative and qualitative trapping of arsines deployed to assess loss of volatile arsenic from paddy soil

Arsenic volatilization in the environment is thought to be an important pathway for transfer from terrestrial pools to the atmosphere. However, this phenomenon is not well characterized due to inherent sampling issues in trapping, quantifying and qualifying these arsine gases; including arsine (AsH(3)), monomethyl arsine (MeAsH(2)), dimethyl arsine (Me(2)AsH) and trimethyl arsine (TMAs). To quantify and qualify arsines in air we developed a novel technique based on silver nitrate impregnated silica gel filled tubes. The method was characterized by measuring the recovery of trapped arsines after elution of this chemo-trap with hot boiling diluted nitric acid. Results from three separate experiments, measured by ICP-MS, showed that the method is reproducible and quantitative. Arsine species recovery ranged from 80.1 to 95.6%, with limit of detection as low as 3.8 ng per chemo-trap tube. Moreover, HPLC-ICP-MS analysis of hot boiling water eluted traps showed that the corresponding oxy ions of the arsines were formed with the As-C bonds of the molecule intact, hence, allowing qualification of trapped arsine species. A microcosm study examining volatile arsenic evolution from field contaminated Bangladeshi paddy soils (24.2 mg/kg arsenic) was used to show the application of silver nitrate chemo-trapping approach. Traps were placed on the inlet and the outlet of microcosms containing the soils that were either (cattle derived) manured or not, or flooded or not, in a factorial design. The headspace was purged with air at a flow rate of 12 mL/min. Results showed that as much as 320 ng of arsenic (0.014% of total soil content) could be emitted in a 3 week period for manured and flooded soils and that TMAs was the dominant species evolved, with lesser quantities of Me(2)AsH. No volatile arsenic evolution was observed for nonmanured treatments, and arsine release from the nonflooded, manured treatment was much less than the flooded treatment.

The influence of evidence-based medicine training on decision-making in relation to over-the-counter medicines: a qualitative study

Objectives
To explore the role of evidence of effectiveness when making decisions about over-the-counter (OTC) medication and to ascertain whether evidence-based medicine training raised awareness in decision-making. Additionally, this work aimed to complement the findings of a previous study because all participants in this current study had received training in evidence-based medicine (unlike the previous participants).

Methods
Following ethical approval and an e-mailed invitation, face-to-face, semi-structured interviews were conducted with newly registered pharmacists (who had received training in evidence-based medicine as part of their MPharm degree) to discuss the role of evidence of effectiveness with OTC medicines. Interviews were recorded and transcribed verbatim. Following transcription, all data were entered into the NVivo software package (version 8). Data were coded and analysed using a constant comparison approach.

Key findings
Twenty-five pharmacists (7 males and 18 females; registered for less than 4 months) were recruited and all participated in the study. Their primary focus with OTC medicines was safety; sales of products (including those that lack evidence of effectiveness) were justified provided they did no harm. Meeting patient expectation was also an important consideration and often superseded evidence. Despite knowledge of the concept, and an awareness of ethical requirements, an evidence-based approach was not routinely implemented by these pharmacists. Pharmacists did not routinely utilize evidence-based resources when making decisions about OTC medicines and some felt uncomfortable discussing the evidence-base for OTC products with patients.

Conclusions
The evidence-based medicine training that these pharmacists received appeared to have limited influence on OTC decision-making. More work could be conducted to ensure that an evidence-based approach is routinely implemented in practice