Genetic Complementation to Identify DNA Elements That Influence Complement Resistance in Leishmania chagasi

Past studies showed that Leishmania spp. promastigotes exhibit differential sensitivity to complement mediated lysis (CML) during development in vitro and in vivo. Leishmania chagasi promastigotes in cultures during logarithmic and stationary growth phases are CML-sensitive or CML-resistant when exposed to human serum, respectively, but only in cultures recently initiated with parasites from infected animals; serially passaged cultures become constitutively CML-sensitive regardless of growth phase. Building on these observations, a genetic screen was conducted to identify novel complement resistance factors of L. chagasi. A cosmid library containing genomic DNA was transfected into a promastigote line previously subjected to \u3e50 serial passages. Selection with human serum for CML resistance yielded 12 transfectant clones. Cosmids isolated from 7 of these clones conferred CML resistance when transfected into an independent, high-passage promastigote culture; at 12% human serum, the mean survival of transfectants was 37% (±11.6%), and that of control transfectants was about 1%. Inserts within the 7 cosmids were unique. Determination of the complete DNA sequence for 1 cosmid indicated that its 32-kilobase insert was 89% identical (overall) to a 31-kilobase region of Leishmania major chromosome 36, which is predicted to encode 6 genes, all of which encode hypothetical proteins

Performance evaluation of rail trackbed stiffness: pre and post stabilisation

Excessive deflection of a rail in response to axle loading can lead to discomfort for passengers and increased wear of both railway structures and trains. These oscillations are often caused by poor trackbed stiffness which may be due to either soft subgrade and/or contaminated ballast. A variety of trackbed stabilisation (TBS) techniques are available to remediate soft subgrades and increase the safety of tracks within the railway network. Traditional TBS methods require track removal, which is expensive, disruptive and often inefficient maintenance works. Micro-piling, using screw piles installed between sleepers, is an innovative low disruption TBS technique. This paper investigates the performance of a soft subgrade and contaminated ballast section of rail line in the UK, before and after screw pile TBS. Pre and post remediation, a computer vision-based system was used to measure rail vertical deflections during train passages and then analysed to quantify the trackbed stiffness. Additionally, 3D finite element models are created and validated by the site measurements. The finite element models are used to simulate a range of different scenarios exploring how changes to the TBS piling layout and/or further works, such as ballast improvement could add further improvements or design efficiencies. Site measurements show TBS reduced rail deflection by 20–30%, indicating that micro-piling is an effective technique for soft subgrades. The finite element analysis revealed the efficiency of micro-piling is highly dependent on the conditions of ballast, strength of the ground at the pile toe, and the pile arrangement. When the aforementioned are optimised the rail deflection could be reduced to approximately 50% of the pre TBS condition

Adapting to the digital age: a narrative approach

The article adopts a narrative inquiry approach to foreground informal learning and exposes a collection of stories from tutors about how they adapted comfortably to the digital age. We were concerned that despite substantial evidence that bringing about changes in pedagogic practices can be difficult, there is a gap in convincing approaches to help in this respect. In this context, this project takes a “bottom-up” approach and synthesises several life-stories into a single persuasive narrative to support the process of adapting to digital change. The project foregrounds the small, every-day motivating moments, cultural features and environmental factors in people's diverse lives which may have contributed to their positive dispositions towards change in relation to technology enhanced learning. We expect that such narrative approaches could serve to support colleagues in other institutions to warm up to ever-changing technological advances

Democratic cultural policy : democratic forms and policy consequences

The forms that are adopted to give practical meaning to democracy are assessed to identify what their implications are for the production of public policies in general and cultural policies in particular. A comparison of direct, representative, democratic elitist and deliberative versions of democracy identifies clear differences between them in terms of policy form and democratic practice. Further elaboration of these differences and their consequences are identified as areas for further research

Researching trust in the police and trust in justice: a UK perspective

This paper describes the immediate and more distant origins of a programme of comparative research that is examining cross-national variations in public trust in justice and in the police. The programme is built around a module of the fifth European Social Survey, and evolved from a study funded by the European Commission. The paper describes the conceptual framework within which we are operating – developed in large measure from theories of procedural justice. It reviews some of the methodological issues raised by the use of sample surveys to research issues of public trust in the police, public perceptions of institutional legitimacy and compliance with the law. Finally it gives a flavour of some of the early findings emerging from the programme

Characterization of DNA Sequences that Confer Complement Resistance in \u3ci\u3eLeishmania chagasi\u3c/i\u3e

Serial passage of axenically cultured Leishmania chagasi promastigotes results in a progressive diminution in resistance to complement-mediated lysis (CML), whereas high CML resistance is seen in infectious metacyclic promastigotes from the sandfly vector as well as metacyclic-like promastigotes within low-passage cultures at stationary growth phase. As we previously reported, in a screen seeking to identify novel genes involved in CML resistance: (1) a genomic cosmid library derived from DNA of CML-resistant L. chagasi promastigotes was transfected into highpassage (constitutively CML-sensitive) L. chagasi promastigotes; (2) transformants were screened for acquisition of CML-resistance; (3) multiple cosmid-transfectants exhibited partial CML resistance; and (4) the sequence for one of the cosmids (Cosmid 51) was determined. This report extends the analysis of Cosmid 51, and identifies by deletion analysis a subregion of the cosmid insert that is critical to the CML-resistance phenotype of Cosmid 51 transformants. We also report the sequence determination and initial CML-resistance activity of another cosmid that also confers partial resistance to CML