Sensitivity to the MEK inhibitor E6201 in melanoma cells is associated with mutant BRAF and wildtype PTEN status

BACKGROUND: Melanoma is the most lethal form of skin cancer, but recent advances in molecularly targeted agents against the Ras/Raf/MAPK pathway demonstrate promise as effective therapies. Despite these advances, resistance remains an issue, as illustrated recently by the clinical experience with vemurafenib. Such acquired resistance appears to be the result of parallel pathway activation, such as PI3K, to overcome single-agent inhibition. In this report, we describe the cytotoxicity and anti-tumour activity of the novel MEK inhibitor, E6201, in a broad panel of melanoma cell lines (n = 31) of known mutational profile in vitro and in vivo. We further test the effectiveness of combining E6201 with an inhibitor of PI3K (LY294002) in overcoming resistance in these cell lines. RESULTS: The majority of melanoma cell lines were either sensitive (IC50 < 500 nM, 24/31) or hypersensitive (IC50 < 100 nM, 18/31) to E6201. This sensitivity correlated with wildtype PTEN and mutant BRAF status, whereas mutant RAS and PI3K pathway activation were associated with resistance. Although MEK inhibitors predominantly exert a cytostatic effect, E6201 elicited a potent cytocidal effect on most of the sensitive lines studied, as evidenced by Annexin positivity and cell death ELISA. Conversely, E6201 did not induce cell death in the two resistant melanoma cell lines tested. E6201 inhibited xenograft tumour growth in all four melanoma cell lines studied to varying degrees, but a more pronounced anti-tumour effect was observed for cell lines that previously demonstrated a cytocidal response in vitro. In vitro combination studies of E6201 and LY294002 showed synergism in all six melanoma cell lines tested, as defined by a mean combination index < 1. CONCLUSIONS: Our data demonstrate that E6201 elicits a predominantly cytocidal effect in vitro and in vivo in melanoma cells of diverse mutational background. Resistance to E6201 was associated with disruption of PTEN and activation of downstream PI3K signalling. In keeping with these data we demonstrate that co-inhibition of MAPK and PI3K is effective in overcoming resistance inherent in melanoma.

Say goodbye to the fries : graduate careers in media, cultural and communication studies

This article addresses the paucity of systematic data on graduate careers in the arts and humanities in the broader context of enduring public and policy debates about the benefits of education to society, the relation between public and private good that is derivable from education, and the specific disciplinary angle that can be brought to bear on these questions from media, cultural and communication studies. We report findings from a survey of ten years of graduates from Queensland University of Technology’s courses in media, cultural and communication studies which indicate very high employment levels and generally positive accounts of the relevance of courses to working life. A major insight that can be drawn from the research is that media, cultural and communication studies deliver capabilities, skills and orientations which are themselves strongly aligned with the kinds of transferable generic attributes which facilitate transition into the workplace.

The capsid protein of beak and feather disease virus binds to the viral DNA and is responsible for transporting the replication-associated protein into the nucleus

Circoviruses lack an autonomous DNA polymerase and are dependent on the replication machinery of the host cell for de novo DNA synthesis. Accordingly, the viral DNA needs to cross both the plasma membrane and the nuclear envelope before replication can occur. Here we report on the subcellular distribution of the beak and feather disease virus (BFDV) capsid protein (CP) and replication-associated protein (Rep) expressed via recombinant baculoviruses in an insect cell system and test the hypothesis that the CP is responsible for transporting the viral genome, as well as Rep, across the nuclear envelope. The intracellular localization of the BFDV CP was found to be directed by three partially overlapping bipartite nuclear localization signals (NLSs) situated between residues 16 and 56 at the N terminus of the protein. Moreover, a DNA binding region was also mapped to the N terminus of the protein and falls within the region containing the three putative NLSs. The ability of CP to bind DNA, coupled with the karyophilic nature of this protein, strongly suggests that it may be responsible for nuclear targeting of the viral genome. Interestingly, whereas Rep expressed on its own in insect cells is restricted to the cytoplasm, coexpression with CP alters the subcellular localization of Rep to the nucleus, strongly suggesting that an interaction with CP facilitates movement of Rep into the nucleus. Copyright © 2006, American Society for Microbiology. All Rights Reserved.

An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6-8 kb. © 2007 Springer-Verlag.

An extension to the rank constraint for improving the reliability of stereo matching algorithms using the rank transform

The rank transform is one non-parametric transform which has been applied to the stereo matching problem The advantages of this transform include its invariance to radio metric distortion and its amenability to hardware implementation. This paper describes the derivation of the rank constraint for matching using the rank transform Previous work has shown that this constraint was capable of resolving ambiguous matches thereby improving match reliability A new matching algorithm incorporating this constraint was also proposed. This paper extends on this previous work by proposing a matching algorithm which uses a dimensional match surface in which the match score is computed for every possible template and match window combination. The principal advantage of this algorithm is that the use of the match surface enforces the left�right consistency and uniqueness constraints thus improving the algorithms ability to remove invalid matches Experimental results for a number of test stereo pairs show that the new algorithm is capable of identifying and removing a large number of in incorrect matches particularly in the case of occlusions

The application of virtual reality to the understanding and treatment of schizophrenia

Virtual Reality (VR) techniques are increasingly being used in education about and in the treatment of certain types of mental illness. Research indicates VR is delivering on it's promised potential to provide enhanced training and treatment outcomes through incorporation of this high-end technology. Schizophrenia is a mental disorder affecting 1−2% of the population. A significant research project being undertaken at the University of Queensland has constructed virtual environments that reproduce the phenomena experienced by patients who have psychosis. The VR environment will allow behavioral exposure therapies to be conducted with exactly controlled exposure stimuli and an expected reduction in risk of harm. This paper reports on the work of the project, previous stages of software development and current and future educational and clinical applications of the Virtual Environments.

Mixed methods research approach to the development and review of competency standards for dietitians

Aim: Competency standards support a range of professional activities including the accreditation of university courses. Reviewing these standards is essential to ensure universities continue to produce well equipped graduates, who can meet the challenge of changing workforce requirements. This paper has two aims: a) to provide an overview of the methodological approaches utilised for compilation and review of the Competency Standards for Dietetics and b) to evaluate the Dietitians Association of Australia’s Competency Standards and capture emerging and contemporary dietetic practice. Methods: A literature review of the methods used to develop Competency Standards for dietitians in Australia, including entry level, advanced level and DAA Fellow competencies and other specific areas of competency, such as public health nutrition and nutrition education is outlined and compared to other allied health professions. The mixed methods methodology used in the most recent review is described in more detail. Results: The history of Dietetic Competency Standards development and review in Australia is compared to dietetic Competency Standards internationally and within other health professions in Australia. The political context in which these standards have been developed in Australia and which has determined their format is also discussed. The results of the most recent Competency Standards review are reported to highlight emerging practice in Australia. Conclusion: The mixed methods approach used in this review provides rich data about contemporary dietetic practice. Our view supports a planned review of all Competency Standards to ensure practice informs education and credentialling and we recommend the Dietitians Association of Australia consider this in future

Positive experiences for participants in suicide bereavement groups : a grounded theory model of elements that contribute to the process of adjusting to the loss

Grounded Theory was used to examine the experiences of 13 participants who had attended psycho-educational support groups for those bereaved by suicide. Results demonstrated core and central categories which fit well with group therapeutic factors developed by Yalom (1995) and emphasised the importance of universality, imparting information and instilling hope, catharsis and self-disclosure, and broader meaning making processes surrounding acceptance or adjustment. Participants were commonly engaged in a lengthy process of oscillating between loss oriented and restoration focused reappraisals. The functional experience of the group comprised feeling normal within the group, providing a sense of permission to feel and to express emotions and thoughts and to bestow meaning. Structural variables of information and guidance and different perspectives on the suicide and bereavement were gained from other participants, the facilitators, group content and process. Personal changes, including in relationships and in their sense of self, assisted participants to develop an altered and more positive personal narrative.

‘I always look under the bed for a man’. Needs and barriers to the expression of sexuality in residential aged care: the views of residents with and without dementia

Objectives: This qualitative study canvassed residents' perceptions of the needs and barriers to the expression of sexuality in long-term care. Methods: Sixteen residents, including five with dementia, from six aged care facilities in two Australian states were interviewed. Data were analysed using a constant comparative method. Results: Four categories describe residents' views about sexuality, their needs and barriers to its expression: ‘It still matters’; ‘Reminiscence and resignation’, ‘It's personal’, and ‘It's an unconducive environment’. Discussion: Residents, including those with dementia, saw themselves as sexual beings and with a continuing need and desire to express their sexuality. The manner in which it was expressed varied. Many barriers to sexual expression were noted, including negative attitudes of staff, lack of privacy and limited opportunities for the establishment of new relationships or the continuation of old ones. Interviewees agreed that how a resident expressed their sexuality was their business and no one else's.

Contribution of DEAF1 structural domains to the interaction with the breast cancer oncogene LMO4

The proteins LMO4 and DEAF1 contribute to the proliferation of mammary epithelial cells. During breast cancer LMO4 is upregulated, affecting its interaction with other protein partners. This may set cells on a path to tumour formation. LMO4 and DEAF1 interact, but it is unknown how they cooperate to regulate cell proliferation. In this study, we identify a specific LMO4-binding domain in DEAF1. This domain contains an unstructured region that directly contacts LMO4, and a coiled coil that contains the DEAF1 nuclear export signal (NES). The coiled coil region can form tetramers and has the typical properties of a coiled coil domain. Using a simple cell-based assay, we show that LMO4 modulates the activity of the DEAF NES, causing nuclear accumulation of a construct containing the LMO4-interaction region of DEAF1.

Responding to the 5Rs : an alternate perspective of Slowmation

This paper is a response to Hoban and Neilsen's (2010) Five Rs model for understanding how learners engage with slowmation. An alternative model (the Learning MMAEPER Model) that builds on the 5Rs model is explained in terms of its use in secondary science preservice teacher education. To probe into the surface and deep learning that can occur during the creation of a slowmation, the learning and relearning model is explored in terms of learning elements. This model can assist teachers to monitor the learning of their students and direct them to a deeper understanding of science concepts.

A multivariate approach to the identification of surrogate parameters for heavy metals in stormwater

Stormwater is a potential and readily available alternative source for potable water in urban areas. However, its direct use is severely constrained by the presence of toxic pollutants, such as heavy metals (HMs). The presence of HMs in stormwater is of concern because of their chronic toxicity and persistent nature. In addition to human health impacts, metals can contribute to adverse ecosystem health impact on receiving waters. Therefore, the ability to predict the levels of HMs in stormwater is crucial for monitoring stormwater quality and for the design of effective treatment systems. Unfortunately, the current laboratory methods for determining HM concentrations are resource intensive and time consuming. In this paper, applications of multivariate data analysis techniques are presented to identify potential surrogate parameters which can be used to determine HM concentrations in stormwater. Accordingly, partial least squares was applied to identify a suite of physicochemical parameters which can serve as indicators of HMs. Datasets having varied characteristics, such as land use and particle size distribution of solids, were analyzed to validate the efficacy of the influencing parameters. Iron, manganese, total organic carbon, and inorganic carbon were identified as the predominant parameters that correlate with the HM concentrations. The practical extension of the study outcomes to urban stormwater management is also discussed.