Developing a three-dimensional in-vitro cell culture model for studying breast and prostate cancer using starPEG-heparin hydrogels

Introduction Hydrogels prepared from poly(ethylene glycol) (PEG) and maleimide-functionalized heparin provide a potential matrix for use in developing three dimensional (3D) models. We have previously demonstrated that these hydrogels support the cultivation of human umbilical vein endothelial cells (HUVECs) (1). We extend this body of work to study the ability to create an extracellular matrix (ECM)-like model to study breast and prostate cancer cell growth in 3D. Also, we investigate the ability to produce a tri-culture mimicking tumour angiogenesis with cancer spheroids, HUVECs and mesenchymal stem cells (MSC). Materials and Methods The breast cancer cell lines, MCF-7 and MDA-MB-231, and prostate cancer cell lines, LNCaP and PC3, were seeded into starPEG-heparin hydrogels and grown for 14 Days to analyse the effects of varying hydrogel stiffness on spheroid development. Resulting hydrogel constructs were analyzed via Alamar Blue assays, light microscopy, and immunofluorescence staining for cytokeratin 8/18, Ki67 and E-Cadherin. Cancer cell lines were then pre-grown in hydrogels for 5-7 days and then re-seeded into starPEG-heparin hydrogels functionalised with RGD, SDF-1, bFGF and VEGF as spheroids with HUVECs and MSC and grown for 14 days as a tri-culture in Endothelial Cell Growth Medium (ECGM; Promocell). Cell lines were also seeded as a single cell suspension into the functionalised tri-culture system. Cultures were fixed in 4% paraformaldehyde and analysed via immunostaining for Von Willebrand Factor and CD31, as well as the above mentioned markers, and observed using confocal microscopy. Results Cultures prepared in MMP-cleavable starPEG-heparin hydrogels display spheroid formation in contrast to adherent growth on tissue culture plastic. Small differences were visualised in cancer spheroid growth between different gel stiffness across the range of cell lines. Cancer cell lines were able to be co-cultivated with HUVECs and MSC. HUVEC tube formation and cancer line spheroid formation occured after 3-4 days. Interaction was visualised between tumours and HUVECs via confocal microscopy. Slightly increased interaction was seen between cancer tumours and micro-vascular tubes when seeded as single cells compared with the pre-formed spheroid approach. Further studies intend to utilise cytokine gradients to further optimise the ECM environment of in situ tumour angiogenesis. Discussion and Conclusions Our results confirm the suitability of hydrogels constructed from starPEG-heparin for HUVECs and MSC co-cultivation with cancer cell lines to study cell-cell and cell-matrix interactions in a 3D environment. This represents a step forward in the development of 3D culture models to study the pathomechanisms of breast and prostate cancer. References 1. Tsurkan MV, Chwalek K, Prokoph S, Zieris A, Levental KR, Freudenberg U, Werner C. Advanced Materials. 25, 2606-10, 2013. Disclosures The authors declare no conflicts of interest

Literacy and Digital Culture in the Early Years

In this chapter we present data drawn from observations of kindergarten children using iPads and talk with the children, their parents/guardians and teachers. We identify a continuum of practices that extends from ‘educational apps’ teaching handwriting, sight words and so forth to uses of the iPad as a device for multimodal literacy development and substantive conversation around children’s creative work. At the current time high stakes testing and the implementation of the Australian Curriculum are prompting new public and professional conversations about literacy and digital technology. The iPad is construed as both cause of and solution to problems of traditional literacy education. In this context we describe the literacies enabled by educational software available on iPads. We higlight the time constraints which bore on teachers' capacity to enact their visions of literacy education through the iPad platform and suggest ways of reflecting on responses to this constraint.

Culture in Business Process Management: How cultural values determine BPM success

There is consensus among practitioners and academics that culture is a critical factor that is able to determine success or failure of BPM initiatives. Yet, culture is a topic that seems difficult to grasp and manage. This may be the reason for the overall lack of guidance on how to address this topic in practice. We have conducted in-depth research for more than three years to examine why and how culture is relevant to BPM. In this chapter, we introduce a framework that explains the role of culture in BPM. We also present the relevant cultural values that compose a BPM culture, and we introduce a tool to examine the supportiveness of organizational cultures for BPM. Our research results provide the basis for further empirical analyses on the topic and support practitioners in the management of culture as an important factor in BPM initiatives.

Working on the edge : Exploring the creative tensions experienced by practitioners facilitating participatory media projects with refugees and asylum seekers in Australia

This paper aims to provide a contextualised and embedded exploration of how the notions of "practice" and "participation", key concepts in the study of culture and media, are manifest in an example of a complex creative project. This project aimed to engage with refugees and asylum seekers through the co-creation of cultural material and is an outcome of an? ethnographic action research (Tacchi et al. 2003) partnership involving a community development worker in a settlement support agency and a storytelling/community media researcher (the author), along with other project collaborators. The discussion of this project focuses on the role of the facilitator and illustrates the processes of orchestrating a complex project involving a series of linked stages with cumulative effect. As practitioners at this site we are working in the space where personal narratives, participatory arts and media, and the staging of intercultural, civic dialogue events, intersect. Co-creative media facilitation in these contexts involves both managing hybrid communicative spaces and (re)combining the "integrative practices" (Schatzki 1996) of a range of professional approaches and creative roles. This is liminal work, located on the boundaries of several disciplines and practices. Drawing on reflections gathered from collaborative ethnographic descriptions (Bhattacharya 2008), this paper traces moments of practitioner uncertainty that can be linked to the way "practice" and “participation” is problematised within the community cultural development field in a way that is at times an uneasy fit with conventional ways of operating in social service roles. These moments of tension also indicate where this project pushed practitioners into spaces of improvisation and new learning. Keywords: Youth, refugees, community cultural development, co-creative media facilitation, ethnographic action research, intercultural dialogue.

Negotiating indeterminacy in studio teaching: Using the zones of material and immaterial play to shape pedagogies and practices in art

The processes of studio-based teaching in visual art are often still tied to traditional models of discrete disciplines and largely immersed in skill-based learning. These approaches to training artists are also tied to an individual model of art practice that is clearly defined by the boundaries of those disciplines. This paper will explain how the open studio program at QUT can be broadly understood as an action research model of learning that ‘plays’ with the post-medium, post-studio genealogies and zones of contemporary art. This emphasises developing conceptual, contextual and formal skills as essential for engaging with and practicing in the often-indeterminate spatio-temporal sites of studio teaching. It will explore how this approach looks to Sutton-Smith’s observations on the role of play and Vygotsky’s zone of proximal development in early childhood learning as a way to develop strategies for promoting creative learning environments that are collaborative and self sustainable. Social, cultural, political and philosophical dialogues are examined as they relate to art practice with the aim of forming the shared interests, aims, and ambitions of graduating students into self initiated collectives or ARIs.

The culture of young women's drinking: Implications for developing effective policing strategies. Report to National Drug Strategy Law Enforcement Committee

Alcohol-related mortality and morbidity represents a substantial financial burden on communities across the world. Adolescence and young adulthood is a peak period for heavy episodic alcohol consumption, with over a third of all people aged 14-19 years having been at risk of acute alcoholrelated harm at least once in the previous 12 months (Australian Institute of Health and Welfare [AIHW], 2011). Excessive alcohol consumption has long been seen as a male problem; however, a gradual shift towards a social acceptance of female drunkenness has narrowed the gap in drinking quantity and style between men and women (Grucza, Bucholz, Rice, & Bierut, 2008). The presented data point to the vulnerability of women to the consequences of acute alcohol intoxication and indicate that alcohol-related offending by women is on the rise. Taken together, these findings reveal that alcohol-related harms and aggression for young women are becoming more prevalent and problematic. This report addressed these issues from a policing perspective...

Multiplicity, Embodiment and the Contemporary Dancer: Moving Identities

In recent years, the practice of contemporary dancers has altered significantly in the transition from canonical choreographic vocabularies to a proliferation of choreographic signatures within mainstream and independent dance. Dancers are often required to collaborate creatively on the formation of choreographic material, thus engaging conceptually with emerging cultural paradigms. This book explores the co-creative practice of contemporary dancers solely from the point of view of the dancer. It reveals multiple dancing perspectives, drawn from interviews, current writing and evocative accounts from inside the choreographic process, illuminating the myriad ways that dancers contribute to the production of contemporary dance culture. A key insight of the book is that a dancer's signature way of being is a 'moving identity', which incorporates past dance experience, anatomical structures and conditioned human movement as a self-in-process. The moving identity is the movement signature that the dancer forms throughout a career path.

Estimates of lost material stock of buildings and roads due to the great east Japan earthquake and tsunami

This article describes research conducted for the Japanese government in the wake of the magnitude 9.0 earthquake and tsunami that struck eastern Japan on March 11, 2011. In this study, material stock analysis (MSA) is used to examine the losses of building and infrastructure materials after this disaster. Estimates of the magnitude of material stock that has lost its social function as a result of a disaster can indicate the quantities required for reconstruction, help garner a better understanding of the volumes of waste flows generated by that disaster, and also help in the course of policy deliberations in the recovery of disaster-stricken areas. Calculations of the lost building and road materials in the five prefectures most affected were undertaken. Analysis in this study is based on the use of geographical information systems (GIS) databases and statistics; it aims to (1) describe in spatial terms what construction materials were lost, (2) estimate the amount of infrastructure material needed to rehabilitate disaster areas, and (3) indicate the amount of lost material stock that should be taken into consideration during government policy deliberations. Our analysis concludes that the material stock losses of buildings and road infrastructure are 31.8 and 2.1 million tonnes, respectively. This research approach and the use of spatial MSA can be useful for urban planners and may also convey more appropriate information about disposal based on the work of municipalities in disaster-afflicted areas.

3D cultures of prostate cancer cells cultured in a novel high-throughput culture platform are more resistant to chemotherapeutics compared to cells cultured in monolayer

Despite monolayer cultures being widely used for cancer drug development and testing, 2D cultures tend to be hypersensitive to chemotherapy and are relatively poor predictors of whether a drug will provide clinical benefit. Whilst generally more complicated, three dimensional (3D) culture systems often better recapitulate true cancer architecture and provide a more accurate drug response. As a step towards making 3D cancer cultures more accessible, we have developed a microwell platform and surface modification protocol to enable high throughput manufacture of 3D cancer aggregates. Herein we use this novel system to characterize prostate cancer cell microaggregates, including growth kinetics and drug sensitivity. Our results indicate that prostate cancer cells are viable in this system, however some non-cancerous prostate cell lines are not. This system allows us to consistently control for the presence or absence of an apoptotic core in the 3D cancer microaggregates. Similar to tumor tissues, the 3D microaggregates display poor polarity. Critically the response of 3D microaggregates to the chemotherapeutic drug, docetaxel, is more consistent with in vivo results than the equivalent 2D controls. Cumulatively, our results demonstrate that these prostate cancer microaggregates better recapitulate the morphology of prostate tumors compared to 2D and can be used for high-throughput drug testing.

Using culture to manage the transition into university: Conceptualising the dynamics of withdrawal and engagement

This conceptual paper explored the purposes of using culture in the process of coping with stress by looking how first year undergraduate students used cultural elements and activities to aid their transition into university. Results supported two key conceptualisations of the use of culture. Firstly, results indicated that students used culture either for withdrawal purposes, i.e., for escaping from the stressful situation, or for engagement purposes, i.e., for actively engaging with the stressful situation. Secondly, the results suggested three different forms of using culture to engage with stressful situations: mood management, learning, and personal interaction. While the results of the study resonate with the distinction between avoidance versus approach-oriented coping strategies that are widely explored in the stress and coping literature, they also suggest that the relationship between withdrawal and engagement might be dynamic with those two strategies serving distinct purposes in the process of coping with stress. The paper thus suggests that there is a need to develop process-oriented models of coping that would allow identifying patterns in the way people fluctuate between withdrawal and engagement that support and facilitate their personal growth and development.

The influence of organizational culture and climate on entrepreneurial intentions among research scientists

Over the past decades, universities have increasingly become involved in entrepreneurial activities. Despite efforts to embrace their ‘third mission’, universities still demonstrate great heterogeneity in terms of their involvement in academic entrepreneurship. This papers adopts an institutional perspective to understand how organizational characteristics affect research scientists’ entrepreneurial intentions. Specifically, we study the impact of university culture and climate on entrepreneurial intentions, including intentions to spin off a company, to engage in patenting or licensing and to interact with industry through contract research or consulting. Using a sample of 437 research scientists from Swedish and German universities, our results reveal that the extent to which universities articulate entrepreneurship as a fundamental element of their mission fosters research scientists’ intentions to engage in spin-off creation and intellectual property rights, but not industry-science interaction. Furthermore, the presence of university role models positively affects research scientists’ propensity to engage in entrepreneurial activities, both directly and indirectly through entrepreneurial self-efficacy. Finally, research scientists working at universities which explicitly reward people for ‘third mission’ related output show higher levels of spin-off and patenting or licensing intentions. This study has implications for both academics and practitioners, including university managers and policy makers.

Effect of constitutive material models on seismic response of two-story reinforced concrete frame

This paper focuses on the finite element (FE) response sensitivity and reliability analyses considering smooth constitutive material models. A reinforced concrete frame is modeled for FE sensitivity analysis followed by direct differentiation method under both static and dynamic load cases. Later, the reliability analysis is performed to predict the seismic behavior of the frame. Displacement sensitivity discontinuities are observed along the pseudo-time axis using non-smooth concrete and reinforcing steel model under quasi-static loading. However, the smooth materials show continuity in response sensitivity at elastic to plastic transition points. The normalized sensitivity results are also used to measure the relative importance of the material parameters on the structural responses. In FE reliability analysis, the influence of smoothness behavior of reinforcing steel is carefully noticed. More efficient and reasonable reliability estimation can be achieved by using smooth material model compare with bilinear material constitutive model.