Thiamine plays a critical role in the acid tolerance of Listeria monocytogenes

Understanding the molecular basis of acid tolerance in the food-borne pathogen Listeria monocytogenes is important as this property contributes to survival in the food-chain and enhances survival within infected hosts. The aim of this study was to identify genes contributing to acid tolerance in L. monocytogenes using transposon mutagenesis and subsequently to elucidate the physiological role of these genes in acid tolerance. One mutant harboring a Tn917 insertion in the thiT gene (formerly lmo1429), which encodes a thiamine (vitamin B1) uptake system, was found to be highly sensitive to acid. The acid-sensitive phenotype associated with loss of this gene was confirmed with an independently isolated mutant, from which the thiT gene was deleted (ΔthiT). Cells of both wild-type and ΔthiT mutant that were thiamine depleted were found to be significantly more acid sensitive than control cultures. Thiamine-depleted cultures failed to produce significant concentrations of acetoin, consistent with the known thiamine dependence of acetolactate synthase, an enzyme required for acetoin synthesis from pyruvate. As acetoin synthesis is a proton-consuming process, we suggest that the acid sensitivity observed in thiamine-depleted cultures may be owing to an inability to produce acetoin.

The AcrAB-TolC efflux system of Salmonella enterica serovar Typhimurium plays a role in pathogenesis

The ability of an isogenic set of mutants of Salmonella enterica serovar Typhimurium L354 (SL1344) with defined deletions in genes encoding components of tripartite efflux pumps, including acrB, acrD, acrF and tolC, to colonize chickens was determined in competition with L354. In addition, the ability of L354 and each mutant to adhere to, and invade, human embryonic intestine cells and mouse monocyte macrophages was determined in vitro. The tolC and acrB knockout mutants were hyper-susceptible to a range of antibiotics, dyes and detergents; the tolC mutant was also more susceptible to acid pH and bile and grew more slowly than L354. Complementation of either gene ablated the phenotype. The tolC mutant poorly adhered to both cell types in vitro and was unable to invade macrophages. The acrB mutant adhered, but did not invade macrophages. In vivo, both the acrB mutant and the tolC mutant colonized poorly and did not persist in the avian gut, whereas the acrD and acrF mutant colonized and persisted as well as L354. These data indicate that the AcrAB-TolC system is important for the colonization of chickens by S. Typhimurium and that this system has a role in mediating adherence and uptake into target host cells.

Stylised worlds: Colour Separation Overlay in BBC television plays of the 1970s

This essay aims to understand and interrogate the use of Colour Separation Overlay (CSO) as a mode of experimental production and aesthetic innovation in television drama in the 1970s. It sets out to do this by describing, accounting for and evaluating CSO as a production technique, considering the role of key production personnel, and analysing four specific BBC productions. Deploying methodologies of archival research, practitioner interview, and close textual analysis, the essay also delivers a significant reassessment of the role of the producer and designer in the conceptualisation and realisation of small-screen dramatic fiction.

Chaos, culture and fantasy: The television plays of Howard Schuman

The single plays of American ex-pat playwright Howard Schuman produced for British television between 1973 and 1983 have received little critical attention. Written in a distinctly un-British madcap, non-naturalistic and often pulpy 'B movie' style, they centre around caricatured, hysterical and/or camp characters and make frequent references to popular culture. This article provides a general survey of Schuman's plays and analyses his sensibility as a screenwriter, drawing extensively on material from interviews with the writer. The article's particular focus is how and why different cultural forms including music, film and theatre are used and referred to in Schuman's plays, and how this conditions the plays' narrative content and visual and aural form. It also considers the reception of Schuman's plays and their status as non-naturalistic dramas that engage heavily with American pop culture, within the context of British drama. Finally, it explores the writer's relationship to style and aesthetics, and considers how his written works have been enhanced through creative design decisions, comparing his directions (in one of his scripts) with the realized play to reflect on the use of key devices.

Playful language alternation in an online discussion forum: the example of digital code plays

This paper explores the linguistic practice of digital code plays in an online discussion forum, used by the community of English-speaking Germans living in Britain. By adopting a qualitative approach of Computer-Mediated Discourse Analysis, the article examines the ways in which these bilinguals deploy linguistic and other semiotic resources on the forum to co-construct humorous code plays. These performances occur in the context of negotiating language norms and are based on conscious manipulations of both codes, English and German. They involve play with codes at three levels: play with forms, meanings, and frames. Although, at first sight, such alternations appear to be used mainly for a comic effect, there is more to this than just humour. By mixing both codes at all levels, the participants deliberately produce aberrant German ‘polluted’ with English and, in so doing, dismantle the ideology of language purity upheld by the purist movement. The deliberate character of this type of code alternation demonstrates heightened metalinguistic awareness as well as creativity and criticality. By exploring the practice of digital code plays, the current study contributes to the growing body of research on networked multilingualism as well as to practices associated with translanguaging, poly- and metrolingualism.

Performing right: legal constraints and Beckett’s plays on BBC television

Drawing on BBC archival documentation, this article outlines how BBC television versions of Beckett’s plays were affected by copyright. Rights to record and broadcast original drama for the screen differ from those governing adaptations of existing theatre plays. Rights can be assigned for specific territories and periods of time, and are negotiated and traded via complex contractual agreements. Examining how Beckett’s agents and the BBC dealt with rights sheds new light on the history of his work on television.

How to watch television? Pedagogy and paedocracy in Beckett’s television plays

This article responds to scholarship on Beckett’s television plays that regards them as positive interventions which encourage the viewer to reconsider the conventions of the medium, and that raise the cultural standards of television drama. In making claims about how the plays address and educate their viewers, critical approaches shift between conceptions of audience. This analysis of Beckett’s plays on British television reconsiders their aesthetic strategies, their relationship with television culture, and the dominant assumptions of critical writing about them by examining the parallel between conceptions of the audience and conceptions of the child in writing about television and Beckett’s television plays.

The United States in Southern Africa: Chester Crocker's linkage strategy 1981-1988

Chester Crocker was appointed as Reagan's Assistant Secretary of State for African Affairs in 1981. He had criticised the inconsistencies of US African policy and proposed a renewed emphasis on the balance between America's global interests with specific regional priorities. While the focus of Congressmen, journalists and public opinion centred on the issue of apartheid, it was the Namibian War of Independence (South African Border War) that initially drew the attention of the Reagan administration, and it was the resolution of this war that remained the priority for the US government in this region throughout Crocker's time in office.

Exploring the role the brand plays in the choice of charity by UK volunteers

Charities need to understand why volunteers choose one brand rather than another in order to attract more volunteers to their organisation. There has been considerable academic interest in understanding why people volunteer generally. However, this research explores the more specific question of why a volunteer chooses one charity brand rather than another. It builds on previous conceptualisations of volunteering as a consumption decision. Seen through the lens of the individual volunteer, it considers the under-researched area of the decision-making process. The research adopts an interpretivist epistemology and subjectivist ontology. Qualitative data was collected through depth interviews and analysed using both Means-End Chain (MEC) and Framework Analysis methodology. The primary contribution of the research is to theory: understanding the role of brand in the volunteer decision-making process. It identifies two roles for brand. The first is as a specific reason for choice, an ‘attribute’ of the decision. Through MEC, volunteering for a well-known brand connects directly through to a sense of self, both self-respect but also social recognition by others. All four components of the symbolic consumption construct are found in the data: volunteers choose a well-known brand to say something about themselves. The brand brings credibility and reassurance, it reduces the risk and enables the volunteer to meet their need to make a difference and achieve a sense of accomplishment. The second closely related role for brand is within the process of making the volunteering decision. Volunteers built up knowledge about the charity brands from a variety of brand touchpoints, over time. At the point of decision-making that brand knowledge and engagement becomes relevant, enabling some to make an automatic choice despite the significant level of commitment being made. The research identifies four types of decision-making behaviour. The research also makes secondary contributions to MEC methodology and to the non-profit context. It concludes within practical implications for management practice and a rich agenda for future research.

Mannose-binding lectin complement pathway plays a key role in complement activation by Paracoccidioides brasiliensis

Paracoccidioides brasiliensis (Pb) is a dimorphic fungal pathogen that causes paracoccidioidomycosis the most severe deep mycosis from South America Although cell mediated immunity is considered the most efficient protective mechanism against Pb infection mechanisms of innate immunity are poorly defined Herein we investigated the interaction of the complement system with high and low virulence isolates of Pb We demonstrated that Pb18 a high virulence Pb Isolate when incubated with normal human serum (NHS) induces consumption of hemolytic complement and when immobilized promotes binding of C4b C3b and C5b-C9 Both low virulence (Pb265) and high virulence (Pb18) isolates consumed C4 C3 and mannose-binding learn (MBL) of MBL-sufficient but not of MBL-deficient serum as revealed by deposition of residual C4 C3 and MBL on immune complexes and mannan However higher complement components consumption was observed with Pb265 as compared with Pb18 The suggested relationship between low virulence and significant complement activation properties of Pb isolates was confirmed by the demonstration that virulence attenuation of Pb 18 results in acquisition of the ability to activate complement Conversely reactivation of attenuated Pb18 results in loss of the ability to activate complement Our results demonstrate for the first time that Pb yeasts activate the complement system by the lectin pathway and there is an Inverse correlation between complement activating ability and Pb virulence These differences could exert an influence on Innate immunity and severity of the disease developed by infected hosts (C) 2010 Elsevier Ltd All rights reserved

5-Lipoxygenase plays a role in the control of parasite burden and contributes to oxidative damage of erythrocytes in murine Chagas` disease

Chagas` disease is accompanied by severe anemia and oxidative stress, which may contribute to mortality. In this study, we investigated the role of 5-lipoxygenase (5-LO) in the control of parasitism and anemia associated with oxidative damage of erythrocytes in experimental Trypanosoma cruzi infection. Wild-type C57BL/6, 129Sv mice treated or not with nordihydroguaiaretic acid (NDGA, 5-LO inhibitor), mice lacking the 5-LO enzyme gene (5-LO(-/-)) and inducible nitric oxide synthase gene (iNOS(-/-)) were infected with the Y strain of T cruzi. impairment of 5-LO resulted in increased numbers of trypomastigote forms in the blood and amastigote forms in the heart of infected mice. We assessed oxidative stress in erythrocytes by measuring oxygen uptake, induction time and chemiluminescence following treatment with tert-butyl hydroperoxide (TBH). Our results show that 5-LO metabolites increased lipid peroxidation levels in erythrocytes during the early phase of murine T cruzi infection. NDGA treatment reduced oxidative damage of erythrocytes in C57BL/6 T cruzi-infected mice but not in C57BL/6 iNOS-/- infected mice, showing that the action of NDGA is dependent on endogenous nitric oxide (NO). In addition, our results show that 5-LO metabolites do not participate directly in the development of anemia in infected mice. We conclude that 5-LO products may not only play a major role in controlling heart tissue parasitism, i.e., host resistance to acute infection with T cruzi in vivo, but in the event of an infection also play an important part in erythrocyte oxidative stress, an NO-dependent effect. (C) 2009 Elsevier B.V. All rights reserved.