Construction of cDNA libraries from trifluoroacetic acid-solvated amphibian skin secretions: molecular cloning of multiple bombinin-like peptide precursor transcripts from a library of yellow-bellied toad (Bombina variegata) secretion

Amphibian skin secretions are renowned as complex mixtures of bioactive peptides many of which are analogues of endogenous regulatory peptides. While skin secretions can be obtained non-invasively for peptidome analysis, parallel studies on the granular gland transcriptome required specimen sacrifice. The aim of the present study was to analyse archived skin secretions to determine the robustness of bioactive peptide precursor-encoding polyadenylated mRNAs in an attempt to extract maximum molecular information from rare samples. A range of solvated skin secretion samples were examined after lyophilisation for their potential to generate viable and comprehensive cDNA libraries based upon polyadenylated mRNA capture and amplification/cloning using appropriate commercial kits. Here we present unequivocal data that the granular gland transcriptome persists in a PCR amenable format even after storage for as long as 12 years in 0.1%(v/v) aqueous trifluoroacetic acid (TFA). We used a pooled skin secretion sample (2 ml) from the yellow-bellied toad, Bombina variegata (n = 14), containing the equivalent of 5 mg/ml of lyophilised skin secretion, that had been used in part for peptide isolation purposes in 1998 and had been stored at - 20 °C since that time. In the first cloning experiment, 12 different bombinin-like peptide precursor cDNAs were cloned encoding 17 different bombinins, the majority of which were novel. Subsequently, bombesin and bradykinin-related peptide precursor transcripts have been cloned successfully. These data illustrate the unexpected stability/longevity of the transcriptome in these secretions — a finding with implications for both this field of research and for the wider field of molecular biology.

A Multicenter Blinded Study to Evaluate KRAS Mutation Testing Methodologies in the Clinical Setting

Evidence that activating mutations of the KRAS oncogene abolish the response to anti-epidermal growth factor receptor therapy has revolutionized the treatment of advanced colorectal cancer. This has resulted in the urgent demand for KRAS mutation testing in the clinical setting to aid choice of therapy. The Am of this study was to evaluate six different KRAS mutation detection methodologies on two series of primary colorectal cancer samples. Two series of 80 frozen and 74 formalin-fixed paraffin-embedded tissue samples were sourced and DNA was extracted at a central site before distribution to seven different testing sites. KRAS mutations in codons 12 and 13 were assessed by using single strand conformation polymorphism analysis, pyrosequencing, high resolution melting analysis, dideoxy sequencing, or the commercially available TIB Molbiol (Berlin, Germany) or DxS Diagnostic innovations (Manchester, UK) kits. in frozen tissue samples, concordance in KRAS status (defined as consensus in at least five assays) was observed in 66/80 (83%) cases. In par-affin tissue, concordance was 46/74 (63%) if all assays were considered or 71/74 (96%) using the five best performing assays. These results demonstrate that a variety of detection methodologies are suitable and provide comparable results for KRAS mutation analysis of clinical samples. (J Mol Diagn 2009, 11:543-552; DOI: 10.2353/jmoldx.2009.090057)

Development and validation of dry reagent time-resolved fluoroimmuno assays for zeranol and alpha-zearalenol to assist in distinguishing zeranol abuse from Fusarium spp. toxin contamination in bovine urine

Zeranol, an oestrogenic growth promoter in food animals, is banned within the European Union (EU). However, commercially available immunoassay kits for zeranol cross-react with toxins formed by naturally occurring Fusarium spp. fungi, leading to false-positive screening results. This paper describes the validation of a specificity enhanced, rapid dry reagent time-resolved fluoroimmunoassay (TR-FIA) for zeranol (recovery 99%, limit of detection 1.3 ng ml(-1)) demonstrating that up to 150 ng ml(-1) of Fusarium spp. toxins in urine do not lead to false-positive results. This assay will assist EU Member States to implement Council Directive 961 23\EC, which requires states to monitor for potential abuses of zeranol. A similar TR-FIA for the Fusarium spp. toxin a-zearalenol, using the same sample extract, is also described (recovery 68%, limit of detection 5.6 ng ml(-1)). Only the addition of diluted sample extract is required to perform these dry-reagent TR-FIAs, the results being available within 1 h of extract application. The EU-funded project 'Natural Zeranol' (FAIR5-CT97-3443) will use these fluoroimmunoassays to screen bovine urine in four Member States to gather data on the seasonality of Fusarium spp. toxin contamination of urine and the incidence of zeranol screening test positives.

How Does Justice Sound? Enunciative Modalities, Law’s Logos and Woman’s Voice

In the throes of her mimetic exposure of the lie of phallocratic discursive unity in 'Speculum of the Other Woman', Irigaray paused on the impossibility of woman’s voice and remarked that ‘it [was] still better to speak only in riddles, allusions, hints, parables.’ Even if asked to clarify a few points. Even if people plead that they just don’t understand. After all, she said, ‘they never have understood.’ (Irigaray 1985, 143).

That the law has never understood a uniquely feminine narrative is hardly controversial, but that this erasure continues to have real and substantive consequences for justice is a reality that feminists have been compelled to remain vigilant in exposing. How does the authority of the word compound law’s exclusionary matrix? How does law remain impervious to woman’s voice and how might it hear woman’s voice? Is there capacity for a dialogic engagement between woman, parler femme, and law?

This paper will explore these questions with particular reference to the experience of women testifying to trauma during the rape trial. It will argue that a logically linked historical genealogy can be traced through which law has come to posit itself as an originary discourse by which thinking is very much conflated with being, or in other terms, law is conflated with justice. This has consequences both for women’s capacity to speak or represent the harm of rape to law, but also for law’s ability to ‘hear’ woman’s voice and objectively adjudicate in cases of rape. It will suggest that justice requires law acknowledge the presence of two distinct and different subjects and that this must be done not only at the symbolic level but also at the level of the parole, syntax and discourse.

Response of farmland biodiversity to the introduction of bioenergy crops: effects of local factors and surrounding landscape context

The recent growth in bioenergy crop cultivation, stimulated by the need to implement measures to reduce net CO emissions, is driving major land-use changes with consequences for biodiversity and ecosystem service provision. Although the type of bioenergy crop and its associated management is likely to affect biodiversity at the local (field) scale, landscape context and its interaction with crop type may also influence biodiversity on farms. In this study, we assessed the impact of replacing conventional agricultural crops with two model bioenergy crops (either oilseed rape Brassica napus or Miscanthus × giganteus) on vascular plant, bumblebee, solitary bee, hoverfly and carabid beetle richness, diversity and abundance in 50 sites in Ireland. We assessed whether within-field biodiversity was also related to surrounding landscape structure. We found that local- and landscape-scale variables correlated with biodiversity in these agricultural landscapes. Overall, the differences between the bioenergy crops and the conventional crops on farmland biodiversity were mostly positive (e.g. higher vascular plant richness in Miscanthus planted on former conventional tillage, higher solitary bee abundance and richness in Miscanthus and oilseed rape compared with conventional crops) or neutral (e.g. no differences between crop types for hoverflies and bumblebees). We showed that these crop type effects were independent of (i.e. no interactions with) the surrounding landscape composition and configuration. However, surrounding landscape context did relate to biodiversity in these farms, negatively for carabid beetles and positively for hoverflies. Although we conclude that the bioenergy crops compared favourably with conventional crops in terms of biodiversity of the taxa studied at the field scale, the effects of large-scale planting in these landscapes could result in very different impacts. Maintaining ecosystem functioning and the delivery of ecosystem services will require a greater understanding of impacts at the landscape scale to ensure the sustainable development of climate change mitigation measures.

Plasma IL-6 levels correlate with clinical and ultrasound measures of arthritis in patients with systemic lupus erythematosus

ObjectiveThe objective of this paper is to elucidate the role of specific cytokines in lupus (SLE) arthritis.MethodsFifty SLE and 40 RA patients had an ultrasound (US) scan of their hand as per standardized protocols. US scores were expressed per joint and as a total 'US activity' score, (sum of power Doppler (PD) and grey-scale synovial hypertrophy scores in all joints) and a total erosion score. SLE disease activity was assessed (BILAG and SELENA-SLEDAI). Plasma levels of IL-6, TNF-alpha and BLyS were measured using sandwich ELISA kits (Quantikine kits, R & D).ResultsOn the basis of the US results SLE patients were divided into three groups: erosive arthritis (n?=?20), non-erosive arthritis (n?=?18) and those with a normal US scan (n?=?12). Across the SLE groups plasma IL-6 levels correlated with CRP (p?

Who was Andrea? Writing Oneself as a Feminist Icon

This article explores the construction of Andrea Dworkin as a public persona, or a ‘feminist icon’, revered by some and demonized by others. It argues that in both her fiction and non-fiction, Dworkin engaged in a process of writing herself as an exceptional woman, a ‘feminist militant’ as she describes herself in the subheading of her 2002 memoir, Heartbreak. The article illustrates Dworkin’s autobiographical logic of exceptionalism by comparing the story told in Heartbreak to the story of Dworkin’s major novel, Mercy, which features a heroine, Andrea, who shares Dworkin’s name and significant biographical details. While Dworkin has insisted that Mercy is not an autobiographical novel, the author undertakes a reading here of Mercy as the story of Dworkin if she had not become the feminist icon of her own and others’ construction. In Mercy, Andrea unsuccessfully attempts to escape the silent, victimized status that Dworkin has insistently argued is imposed upon women. In her repeated victimization, Andrea functions for Dworkin as an ‘everywoman’ who both embodies Dworkin’s world-view and highlights how Dworkin’s own biography exists in tension with some of her central assumptions about women, gender and contemporary society.

Experimental Evaluations of Elementary Science Programs: A Best-Evidence Synthesis

This article presents a systematic review of research on the achievement outcomes of all types of approaches to teaching science in elementary schools. Study inclusion criteria included use of randomized or matched control groups, a study duration of at least 4 weeks, and use of achievement measures independent of the experimental treatment. A total of 23 studies met these criteria. Among studies evaluating inquiry-based teaching approaches, programs that used science kits did not show positive outcomes on science achievement measures (weighted ES=+0.02 in 7 studies), but inquiry-based programs that emphasized professional development but not kits did show positive outcomes (weighted ES=+0.36 in 10 studies). Technological approaches integrating video and computer resources with teaching and cooperative learning showed positive outcomes in a few small, matched studies (ES=+0.42 in 6 studies). The review concludes that science teaching methods focused on enhancing teachers’ classroom instruction throughout the year, such as cooperative learning and science-reading integration, as well as approaches that give teachers technology tools to enhance instruction, have significant potential to improve science learning.

Restorative Justice and Sex Offending

This essay examines the origins and uses of restorative justice with sexual offending and the contemporary challenges and controversies surrounding this. It charts the range of ad hoc initiatives which have sought to apply a restorative form of intervention with violent or sexual offending from first time and ‘acquaintance’ rape as well as young sexual abusers to high risk sexual offenders in the form of circles of support and accountability. Such schemes are often presented as a counter to the failings of retributive forms of justice and are premised on Braithwaite’s (1989) notion of ‘reintegrative shaming’ that seek to reintegrate offenders into the community. Critics of restorative justice traditionally put forward a number of core objectives when restorative justice is applied to serious forms of offending such as sexual offending. The essay also sets out and seeks to counter these principal challenges and how they may be overcome. For the most part, however, restorative justice has failed to reach its potential as a fully fledged sentencing rationale in being applied as a mainstream response to a wide range of offending including that at the higher end of the spectrum. The essay also seeks to examine barriers to restorative justice within contemporary penal policy and to highlight some of the most controversial applications of the restorative paradigm including those related to clergy sexual abuse. It concludes by offering some thoughts on the future of restorative justice as a mainstream responses to serious forms of offending.

Characterisation of UK and Irish Cold-Pressed Rapeseed Oils in Relation to Quality Indices and Constituents.

Cold-pressed rapeseed oil (CPRSO) is produced when seeds from an oilseed rape crop are mechanically crushed whilst at a low temperature. CPRSO’s popularity is rapidly expanding and is now produced in most Northern European countries including both N.Ireland and ROI. The CPRSO industry is still relatively new and therefore not as widely researched as other high quality oils. Fifteen CPRSO from The UK, Ireland and France were examined to determine characteristic differences between the oils. Two samples of extra-virgin olive oil and two samples of refined rapeseed oil were also included in the investigation to assess performance against market competitors. The antioxidant potential of the oils was assessed using the ABTS and DPPH radical scavenging assays. Both unexpectedly showed that refined rapeseed oil had the highest potential whilst there was significant difference between many of the CPRSO’s. The acid value (ACOS method Cd 3d-63) ranged widely from 0.47-3.41. To predict the stability during storage, an accelerated oxidation test was carried out where the oils were placed in an oven (60°C) and peroxide value was monitored. The results showed extra-virgin olive oil underwent the least oxidation during the trial. The refined rapeseed oil suffered the worst levels of oxidation whilst the CPRSO’s performed similarly but with some variation. Fatty acid composition was investigated with GC-MS and some of the major fatty acids were found to differ significantly between producers. Minor compound analysis was achieved with extraction and identification through HPLC. All results are critically discussed and compared to relevant published studies.

“How Can a Woman Who Has Been Raped Be Believed?”: Andrea Dworkin, Sexual Violence and the Ethics of Belief

In June 2000, Andrea Dworkin, an American feminist activist and author, published an account of being raped in a Paris hotel room a year earlier. The story was met with widespread disbelief, including from feminist readers. This article explores the reasons for this disbelief, asking how and why narratives of rape are granted – or denied – truth status by their readers. The article argues for understanding the conferral of belief as a narrative transaction involving the actions of both narrator and reader. It posits that Dworkin was widely seen as an unreliable narrator but argues that for ideologically charged narratives such as rape narratives judgements of reliability and belief inevitably draw upon the normative standpoint of the reader. I suggest that there are opposing criteria for establishing the truth of rape narratives; a ‘factual’ or legal model, which sees rape narratives as requiring scrutiny, and an ‘experiential’ model, located within certain strands of feminist politics, which emphasises the ethical importance of believing women’s narratives. The article finishes with a consideration of the place of belief within an ethics of reading and reception of rape narratives.

Beyond Criminal Justice: An anthology of abolitionist papers presented to conferences of the European Group for the Study of Deviance and Social Control over the past decade

Beyond Criminal Justice presents a vision of a future without brutal, authoritarian and repressive penal regimes. Many of the papers brought together here have been unavailable for more than two decades. Their republication indicates not only their continuing theoretical importance to abolitionist studies but also how they provide important insights into the nature and legitimacy of criminal processes in the here and now. Contributors highlight the human consequences of the harms of imprisonment, evidencing the hurt, injury and damage of penal incarceration across a number of different countries in Europe. Focusing on penal power and prisoner contestation to such power, the moral and political crises of imprisonment are laid bare. The contributors to Beyond Criminal Justice explore the urgent need for a coherent, rational and morally and politically sophisticated theoretical basis for penal abolitionism. Advocating a utopian imagination and at the same time practical solutions already implemented in countries around Europe - alongside grappling with controversial debates such as abolitionist responses to rape and sexual violence - the book steps outside of common sense assumptions regarding 'crime', punishment and 'criminal justice'. Beyond Criminal Justice will be of interest to students of criminology, zemiology, sociology, penology and critical legal studies as well as anyone interested in rethinking the problem of 'crime' and challenging the logic of the penal rationale.

Propagation Characteristics of Groove Gap Waveguide Below and Above Cutoff

Recently, gap waveguides have been shown as a potential alternative to conventional waveguides in the millimeterwave band. Until now, Groove Gap Waveguide (GGW) has been studied through direct correspondence with rectangular waveguide with the same physical dimensions. However there have been observed differences in the above cutoff propagation characteristics between these two waveguide types. Furthermore, the behavior of GGW below cutoff remains unknown. This work presents a discussion of both below and above cutoff propagation characteristics of GGW, and introduces a simple model that explains the observed GGW behavior and establishes well its propagation characteristics. Two TRL calibration kits have been manufactured, and the measurements have good agreement with the proposed analysis model results.

Violence against women in the Irish War of Independence, 1919-1921

This chapter examines the nature and extent of violence experienced by women in Ireland during the Irish War of Independence (1919-1921) at the hands of both the Crown forces and the Irish Republican Army. It argues that targetted killings of women by either side was rare. The most common forms of such violence can be categorised as physical, gendered (cutting of hair) and psychological (intimidation and the killing of male relatives). It argues that there was a difference between gendered and sexual crime, the latter of which appears to have been very uncommon. A considerable part of the chapter uses theoretical literature on violence against women in conflict zones to explain why sexual violence was uncommon, arguing that neither side had much to gain from its employment, that the Crown forces were aware of the damage it could do to Britain's international reputation and that the terror tactics adopted by the Crown forces were sufficient to achieve their ends without resorting to rape. In regard to the IRA, the absence of any evidence of rape or sexual assault being perpetrated could be attributable to their Catholicism, reliance on support from the community, the efforts of the first Dáil to achieve foreign recognition of the Republic and the role of Cumann na mBan women in the guerrilla conflict. The historiography of women in the Irish revolution is also analysed.

A microbiological study of Papillon-Lefévre syndrome in two patients

AIM: To analyse the microflora of subgingival plaque from patients with Papillon-Lefévre syndrome (PLS), which is a very rare disease characterised by palmar-plantar hyperkeratosis with precocious periodontal destruction.

METHODS: Bacterial isolates were identified using a combination of commercial identification kits, traditional laboratory tests, and gas liquid chromatography. Some isolates were also subjected to partial 16S rDNA sequencing. Plaque samples were also assayed for the presence of Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in a quantitative enzyme linked immunosorbent assay (ELISA) using monoclonal antibodies.

RESULTS: The culture results showed that most isolates were capnophilic and facultatively anaerobic species-mainly Capnocytophaga spp and Streptococcus spp. The latter included S. constellatus, S. oralis, and S. sanguis. Other facultative bacteria belonged to the genera gemella, kingella, leuconostoc, and stomatococcus. The aerobic bacteria isolated were species of neisseria and bacillus. Anaerobic species included Prevotella intermedia, P. melaninogenica, and P. nigrescens, as well as Peptostreptococcus spp. ELISA detected P gingivalis in one patient in all sites sampled, whereas A. actinomycetemcomitans was detected in only one site from the other patient. Prevotella intermedia was present in low numbers.

CONCLUSIONS: Patients with PLS have a very complex subgingival flora including recognised periodontal pathogens. However, no particular periodontopathogen is invariably associated with PLS.