Analysis of the NF-κB p50 dimer interface by diversity screening

An in vivo screen has been devised for NF-κB p50 activity in Escherichia coli exploiting the ability of the mammalian transcription factor to emulate a prokaryotic repressor. Active intracellular p50 was shown to repress the expression of a green fluorescent protein reporter gene allowing for visual screening of colonies expressing active p50 on agar plates. A library of mutants was constructed in which the residues Y267, L269, A308 and V310 of the dimer interface were simultaneously randomised and twenty-five novel functional interfaces were selected which repressed the reporter gene to similar levels as the wild-type protein. The leucine-269 alanine-308 core was repeatedly, but not exclusively, selected from the library whilst a diversity of predominantly non-polar residues were selected at positions 267 and 310. These results indicate that L269 and A308 may form a hot spot of interaction and allow an insight into the processes of dimer selectivity and evolution within this family of transcription factors.

Active control of car suspension systems using IDA-PBC

This paper considers the design of active control for car suspension systems using a particular form of energy-based control called Interconnection-and-Damping-Assignment Passivity-Based Control (IDA-PBC). This approach allows one to shape the kinetic and potential energy as well as modify the power flow among different components of the system by changing the interconnection and dissipative structure in a meaningful way. Different controller parameterisations are considered to design a class of controllers for active suspension systems.

Clinical dyslipidaemia is associated with changes in the lipid composition and inflammatory properties of apolipoprotein-B-containing lipoproteins from women with type 2 diabetes

The aim of this study was to use lipidomics to determine if the lipid composition of apolipoprotein-B-containing lipoproteins is modified by dyslipidaemia in type 2 diabetes and if any of the identified changes potentially have biological relevance in the pathophysiology of type 2 diabetes. VLDL and LDL from normolipidaemic and dyslipidaemic type 2 diabetic women and controls were isolated and quantified with HPLC and mass spectrometry. A detailed molecular characterisation of VLDL triacylglycerols (TAG) was also performed using the novel ozone-induced dissociation method, which allowed us to distinguish vaccenic acid (C18:1 n-7) from oleic acid (C18:1 n-9) in specific TAG species. Lipid class composition was very similar in VLDL and LDL from normolipidaemic type 2 diabetic and control participants. By contrast, dyslipidaemia was associated with significant changes in both lipid classes (e.g. increased diacylglycerols) and lipid species (e.g. increased C16:1 and C20:3 in phosphatidylcholine and cholesteryl ester and increased C16:0 [palmitic acid] and vaccenic acid in TAG). Levels of palmitic acid in VLDL and LDL TAG correlated with insulin resistance, and VLDL TAG enriched in palmitic acid promoted increased secretion of proinflammatory mediators from human smooth muscle cells. We showed that dyslipidaemia is associated with major changes in both lipid class and lipid species composition in VLDL and LDL from women with type 2 diabetes. In addition, we identified specific molecular lipid species that both correlate with clinical variables and are proinflammatory. Our study thus shows the potential of advanced lipidomic methods to further understand the pathophysiology of type 2 diabetes.

Developmentally and transgene regulated nuclear processing of primary transcripts of chalcone synthase A in petunia

The introduction of chalcone synthase A transgenes into petunia plants can result in degradation of chalcone synthase A RNAs and loss of chalcone synthase, a process called cosuppression or post-transcriptional gene silencing. Here we show that the RNA degradation is associated with changes in premRNA processing, i.e. loss of tissue specificity in transcript cleavage patterns, accumulation of unspliced molecules, and use of template-specific secondary poly(A) sites. These changes can also be observed at a lower level in leaves but not flowers of nontransgenic petunias. Based on this, a model is presented of how transgenes may disturb the carefully evolved, developmentally controlled post-transcriptional regulation of chalcone synthase gene expression by influencing the survival rate of the endogenous and their own mRNA.

An outcome evaluation of the ‘Skipper’ designated driver program

The general aim of designated driver programs is to reduce the level of drink driving by encouraging potential drink drivers to travel with a driver who has abstained from (or at least limited) consuming alcohol. Designated driver programs appear to be quite widespread around the world, however a limited number have been subject to rigorous evaluation. This paper reports results from an outcome evaluation of a designated driver program called ‘Skipper’, which was trialled in a provincial city in Queensland, Australia. The outcome evaluation included surveys three weeks prior to (baseline), four months following (1st follow-up), and 16 months following (2nd follow-up) the commencement of the trial in both the ‘intervention area’ (baseline, n = 202; 1st follow-up, n = 211; 2nd follow-up, n = 200) and a ‘comparison area’(baseline, n = 203; 1st follow-up, n = 199; 2nd follow-up, n = 201); and a comparison of random breath testing and crash data before and after the trial. The survey results indicate that awareness of the program in the intervention area was quite high four months following its introduction and that this was maintained at 16 months. The results also suggest that the ‘Skipper’ program and the related publicity had positive impacts on behaviour with an increase in the proportion of people participating in designated driver as a passenger. It is less clear, however, whether the ‘Skipper’ program impacted on other behaviours of interest, such as drink driving or involvement in alcohol-related crashes. Suggestions for further research and program improvement are discussed as well as limitations of the research.

Securing medical devices Part B : what’s available for nasogastric tubes?

This article is the second part of a two-part series examining securement options for commonly used therapeutic devices in the adult intensive care unit. Part A focused on endotracheal device securement.1 This article addresses nasogastric tube (NGT) securement options and with the aim of identifying the available range of NGT securement devices in Australia as a resource for clinicians seeking to explore options for tube stabilisation. Nasogastric feeding or gastric decompression tubes are commonly inserted via the nostril/nares. The National Pressure Ulcer Advisory Panel (NPUAP) 2011 position statement on mucosal pressure injuries, highlighted that mucosal tissues are vulnerable to pressure from devices.2 Securing of these devices sometimes leads to pressure-related injury to the internal mucosa due to difficulty visualising the mucosa and failure to reposition the nasogastric tube to relieve the pressure in a particular area.3 The nasal orifice is much smaller than the oral cavity and regular tube position changes are vital to minimise the risk of mucosal damage and ulcer development.

Analysis of unsaturated lipids by ozone-induced dissociation

Recent developments in analytical technologies have driven significant advances in lipid science. The sensitivity and selectivity of modern mass spectrometers can now provide for the detection and even quantification of many hundreds of lipids in a single analysis. In parallel, increasing evidence from structural biology suggests that a detailed knowledge of lipid molecular structure including carbon-carbon double bond position, stereochemistry and acyl chain regiochemistry is required to fully appreciate the biochemical role(s) of individual lipids. Here we review the capabilities and limitations of tandem mass spectrometry to provide this level of structural specificity in the analysis of lipids present in complex biological extracts. In particular, we focus on the capabilities of a novel technology termed ozone-induced dissociation to identify the position (s) of double bonds in unsaturated lipids and discuss its possible role in efforts to develop workflows that provide for complete structure elucidation of lipids by mass spectrometry alone: so-called top-down lipidomics. This article is part of a Special Issue entitled: Lipodomics and Imaging Mass Spectrom. (C) 2011 Elsevier B.V. All rights reserved.

A combinatorial approach to anonymous membership broadcast

A set system (X, F ) with X= {x 1,...,x m}) and F = {B1...,B n }, where B i ⊆ X, is called an (n, m) cover-free set system (or CF set system) if for any 1 ≤ i, j, k ≤ n and j ≠ k, |B i >2 |B j ∩ B k | +1. In this paper, we show that CF set systems can be used to construct anonymous membership broadcast schemes (or AMB schemes), allowing a center to broadcast a secret identity among a set of users in a such way that the users can verify whether or not the broadcast message contains their valid identity. Our goal is to construct (n, m) CF set systems in which for given m the value n is as large as possible. We give two constructions for CF set systems, the first one from error-correcting codes and the other from combinatorial designs. We link CF set systems to the concept of cover-free family studied by Erdös et al in early 80’s to derive bounds on parameters of CF set systems. We also discuss some possible extensions of the current work, motivated by different application.

Death on the front page : “Awesome things” and this thing called humanity

It has been called “the world’s worst recorded natural disaster,” and “the largest earthquake in 40 years,” galvanizing the largest global relief effort in history. For those of us involved in the discipline and/or the practice of communications, we realized that it presented a unique case study from a number of perspectives. Both the media and the public became so enraptured and enmeshed in the story of the tsunami of December 26, 2004, bringing to the fore a piece of geography and a peoples too rarely considered prior to the tragedy, that we felt compelled to examine the phenomenon. The overwhelming significance of this volume comes from its being a combination of both academic scholars and development practitioners in the field. Its poignancy becomes underscored from their wide-ranging perspectives, with 21 chapters representing some 14 different countries. Their realities provide not only credibility but also an unprecedented sensitivity to communication issues. Our approach here considers Tsunami 2004 from five communication perspectives: 1.) Interpersonal/ intercultural, 2.) Mass media, 3.) Telecommunications, 4.) Ethics, philanthropy, and development communication, and; 5.) Personal testimonies and observations. You will learn even more here about the theory and practice of disaster/crisis communication.

B-Raf mutation : a key player in molecular biology of cancer

B-Raf is one of the more commonly mutated proto-oncogenes implicated in the development of cancers. In this review, we consider the mechanisms and clinical impacts of B-Raf mutations in cancer and discuss the implications for the patient in melanoma, thyroid cancer and colorectal cancer, where B-Raf mutations are particularly common.

Strategic targeting of the PI3K–NFκB axis in cisplatin-resistant NSCLC

Chemoresistance is a major therapeutic challenge to overcome in NSCLC, in order to improve the current survival rates of <15% at 5 years. We and others have shown increased PI3K signaling in NSCLC to be associated with a more aggressive disease, and a poorer prognosis. In this study, targeted inhibition of three strategic points of the PI3K–NFκB axis was performed with the aim of exploiting vulnerabilities in cisplatin-resistant NSCLC cells. Cisplatin-resistant cell lines were previously generated through prolonged exposure to the drug. Expression of PI3K and NFκB pathway-related genes were compared between cisplatin-resistant cells and their matched parent cells using a gene expression array, qRT-PCR, DNA sequencing, western blot, and immunofluorescence. Targeted inhibition was performed using GDC-0980, a dual PI3K–mTOR inhibitor currently in Phase II clinical trials in NSCLC, and DHMEQ, an inhibitor of NFκB translocation which has been used extensively both in vitro and in vivo. Effects of the two inhibitors were assessed by BrdU proliferation assay and multiparameter viability assay. NFKBIA was shown to be 12-fold overexpressed in cisplatin-resistant cells, with no mutations present in exons 3, 4, or 5 of the gene. Corresponding overexpression of IκBα was also observed. Treatment with DHMEQ (but not GDC-0980) led to significantly enhanced effects on viability and proliferation in cisplatin-resistant cells compared with parent cells. We conclude that NFκB inhibition represents a more promising strategy than PI3K–mTOR inhibition for treatment in the chemoresistance setting in NSCLC.

Becoming an inclusive educator : applying Deleuze & Guattari to teacher education

New ways of thinking are required in teacher education to promote beginning teachers as change agents in education. Twenty years after the Salamanca Statement (UNESCO, 1994) that called for schools to provide equitable opportunities for all children, teaching practices in many classrooms are informed by the deficit view of learning. Beginning teachers need to be prepared to challenge the ideological influences that operate in schools. Deleuze and Guattari’s (1987) model of the rhizome is used to report one beginning teacher’s journey as she learnt to negotiate structural and personal obstacles to create an inclusive learning environment. Data from reflective diaries, semi-structured interviews and classroom observations highlight contextual and personal factors in one case study that contributed to the nonlinear, complex process of becoming an inclusive educator. The paper concludes by arguing the voice of beginning teachers is essential for the ongoing movement towards the creation of just, inclusive schools.