In vitro and in vivo MMP gene expression localisation by In Situ-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts

Background Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. In vitro, MT1-MMP (MMP-14, membrane type-1-MMP) expression is higher in more invasive human breast cancer (HBC) cell lines, whilst in vivo its expression has been associated with the stroma surrounding breast tumours. MMP-1 (interstitial collagenase) has been associated with MDA-MB-231 invasion in vitro, while MMP-3 (stromelysin-1) has been localised around invasive cells of breast tumours in vivo. As MMPs are not stored intracellularly, the ability to localise their expression to their cells of origin is difficult. Methods We utilised the unique in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR) methodology to localise the in vitro and in vivo gene expression of MT1-MMP, MMP-1 and MMP-3 in human breast cancer. In vitro, MMP induction was examined in the MDA-MB-231 and MCF-7 HBC cell lines following exposure to Concanavalin A (Con A). In vivo, we examined their expression in archival paraffin embedded xenografts derived from a range of HBC cell lines of varied invasive and metastatic potential. Mouse xenografts are heterogenous, containing neoplastic human parenchyma with mouse stroma and vasculature and provide a reproducible in vivo model system correlated to the human disease state. Results In vitro, exposure to Con A increased MT1-MMP gene expression in MDA-MB-231 cells and decreased MT1-MMP gene expression in MCF-7 cells. MMP-1 and MMP-3 gene expression remained unchanged in both cell lines. In vivo, stromal cells recruited into each xenograft demonstrated differences in localised levels of MMP gene expression. Specifically, MDA-MB-231, MDA-MB-435 and Hs578T HBC cell lines are able to influence MMP gene expression in the surrounding stroma. Conclusion We have demonstrated the applicability and sensitivity of IS-RT-PCR for the examination of MMP gene expression both in vitro and in vivo. Induction of MMP gene expression in both the epithelial tumour cells and surrounding stromal cells is associated with increased metastatic potential. Our data demonstrate the contribution of the stroma to epithelial MMP gene expression, and highlight the complexity of the role of MMPs in the stromal-epithelial interactions within breast carcinoma.

Constructions of asylum seekers and refugees in Australian political discourse

Immigration to Australia has long been the focus of negative political interest. In recent times, the proposal of exclusionary policies such as the Malaysia Deal in 2011 has fuelled further debate. In these debates, Federal politicians often describe asylum seekers and refugees as ‘illegal’, ‘queue jumpers’, and ‘boat people’. This paper investigates how the political discourse constructs asylum seekers and refugees during debates surrounding the Malaysia Deal in the Federal Parliament of Australia in 2011. Hansard Parliamentary debates were analysed to identify the underlying themes and constructions that permeate political discourse about asylum seekers and refugees. This paper argues that a dichotomous characterisation of legitimacy pervades their construction with this group constructed either as legitimate humanitarian refugees or as illegitimate ‘boat arrivals’. These constructions result in the misrepresentation of asylum seekers as illegitimate, undermining their right to protection under Australia’s laws and international obligations. This construction also represents a shift in federal political discourse from constructing asylum seekers as a border or security threat, towards an increasing preoccupation with this categorisation of people as legitimate, or illegitimate.

Making sense of purpose, direction and innovation : an embedded design led innovation case study in the Australian mining industry

The mining equipment technology services sector is driven by a reactive and user-centered design approach, with a technological focus on incremental new product development. As Australia moves out of its sustained mining boom, companies need to rethink their strategic position, to become agile to stay relevant in an enigmatic market. This paper reports on the first five months on an embedded case study within an Australian, family-owned mining manufacturer. The first author is currently engaged in a longitudinal design led innovation project, as a catalyst to guide the company’s journey to design integration. The results find that design led innovation could act as a channel for highlighting and exploring company disconnections with the marketplace and offer a customer-centric catalyst for internal change. Data collected for this study is from 12 analysed semistructured interviews, a focus group and a reflective journal, over a five-month period. This paper explores limitations to design integration, and highlights opportunities to explore and leverage entrepreneurial characteristics to stay agile, broaden innovation and future-proof through the next commodity cycle in the mining industry.

Embedded digital creatives

Digital creatives are spread widely across the economy, and growing rapidly in number. New industries such as mobile applications (apps) development are creating new occupations and services, and transforming some older ones. Digital creatives are driving innovation and economic growth in these industries and service sectors. Digital technologies and processes are becoming increasingly important in virtually all spheres of creative work, as they are across the economy. And yet there is no broad agreement either in Australia or worldwide about the constitution of the digital creative grouping, and only limited understanding of both the role of creatives in the digital economy and innovation, and of the issues and challenges they face. Who are they? Where are they working? Where are their numbers growing? The answers to these questions will help to inform the development of policy and the fostering of an environment that promotes digital content and creativity.

Embedded digital creative workers and creative services in banking

This chapter explores the roles and functions of both digital creative workers and creative services firms in an industry beyond the core creative industries: banking. The chapter focuses on the design and development of apps and mobile websites for smartphones and tablet computers, with examples drawn principally from the Australian banking sector. While it might be assumed that utility and practicality are more critical and more highly valued in apps development for financial services institutions than innovation and aesthetic design, this chapter illustrates the growing importance placed on creative work in this sector.

Cyanoacrylate glue as an alternative mounting medium for resin-embedded semithin sections

Commercially available generic Superglue (cyanoacrylate glue) can be used as an alternative mounting medium for stained resin-embedded semithin sections. It is colourless and contains a volatile, quick-setting solvent that produces permanent mounts of semithin sections for immediate inspection under the light microscope. Here, we compare the use of cyanoacrylate glue for mounting semithin sections with classical dibutyl phthalate xylene (DPX) in terms of practical usefulness, effectiveness and the quality of the final microscopic image.

The curious case of the embedded creative : creative cultural occupations outside the creative industries

This chapter is concerned with innovation that involves creative cultural occupations, but not within the creative industries. Rather, we examine the operation of cultural creative occupations that exist outside the creative industries - so-called 'embedded creatives' who work across all industry sectors (Cunningham and Higgs 2009). In doing so, we concur with Bilton (2007) that the separation of creative industries from other industries is a 'false step'. All industries must be innovative; however, they also must be able to combine both scientific and artistic creativity, and that creativity comes from the intersection of different thinking styles (Kurtzberg 2005). Moreover, we suggest that there are now detailed empirical studies, as well as a nascent theoretical base, to suggest that the transdisciplinarity which results from embedded cultural creativity is an engine of growth in the broader economy. Thus, it is relevant to both policymakers and managers. This chapter addresses the following questions: What is the role and significance of the embedded creative? Given a paucity of detailed empirical work in the area to date, what can be deduced from what extant literature there is about the nature of employment and management of these workers? And what are the practical implications of these consideration?

Distribution tap changer adjustment to improve small-scale embedded generator penetration and mitigate voltage rise

This paper addresses the voltage rise constraints that are initiated from increased renewable generation resources in low voltage distribution networks. In this paper, an approach which is able to mitigate these voltage rise constraints and allow for increased distributed generator penetration is presented. The proposed approach involves utilizing the distribution transformers static tap changer to reduce the distribution feeder voltage setpoint. The proposed approach is modeled on a generic low voltage distribution network using the PSS SINCAL© simulation software package and is also implemented in a real low voltage distribution network to verify its practicality. Results indicate that this approach can be implemented to mitigate the voltage rise constraint and increase small-scale embedded generator penetration in a high proportion of low voltage feeders while avoiding any substantial network costs.

Binding of small mono- and oligomeric integrin ligands to membrane-embedded integrins monitored by surface plasmon-enhanced fluorescence spectroscopy

We recently developed a binding assay format by incorporating native transmembrane receptors into artificial phospholipid bilayers on biosensor devices for surface plasmon resonance spectroscopy. By extending the method to surface plasmon-enhanced fluorescence spectroscopy (SPFS), sensitive recording of the association of even very small ligands is enabled. Herewith, we monitored binding of synthetic mono- and oligomeric RGD-based peptides and peptidomimetics to integrins alphavbeta3 and alphavbeta5, after having confirmed correct orientation and functionality of membrane-embedded integrins. We evaluated integrin binding of RGD multimers linked together via aminohexanoic acid (Ahx) spacers and showed that the dimer revealed higher binding activity than the tetramer, followed by the RGD monomers. The peptidomimetic was also found to be highly active with a slightly higher selectivity toward alphavbeta3. The different compounds were also evaluated in in vitro cell adhesion tests for their capacity to interfere with alphavbeta3-mediated cell attachment to vitronectin. We hereby demonstrated that the different RGD monomers were similarly effective; the RGD dimer and tetramer showed comparable IC50 values, which were, however, significantly higher than those of the monomers. Best cell detachment from vitronectin was achieved by the peptidomimetic. The novel SPFS-binding assay platform proves to be a suitable, reliable, and sensitive method to monitor the binding capacity of small ligands to native transmembrane receptors, here demonstrated for integrins.

Estimating hidden message length in binary image embedded by using boundary pixels steganography

In this paper, we propose a new steganalytic method to detect the message hidden in a black and white image using the steganographic technique developed by Liang, Wang and Zhang. Our detection method estimates the length of hidden message embedded in a binary image. Although the hidden message embedded is visually imperceptible, it changes some image statistic (such as inter-pixels correlation). Based on this observation, we first derive the 512 patterns histogram from the boundary pixels as the distinguishing statistic, then we compute the histogram difference to determine the changes of the 512 patterns histogram induced by the embedding operation. Finally we propose histogram quotient to estimate the length of the embedded message. Experimental results confirm that the proposed method can effectively and reliably detect the length of the embedded message.

Motor controller simulation and embedded implementation for a blood pump

This study was a step forward in modeling, simulation and microcontroller implementation of a high performance control algorithm for the motor of a blood pump. The rotor angle is sensed using three Hall effect sensors and an algorithm is developed to obtain better angular resolution from the three signals for better discrete-time updates of the controller. The performance of the system was evaluated in terms of actual and reference speeds, stator currents and power consumption over a range of reference speeds up to 4000 revolutions per minute. The use of fewer low cost Hall effect sensors compared to expensive high resolution sensors could reduce the cost of blood pumps for total artificial hearts.

Embedded creatives in Australian healthcare: An update

The contribution of creative occupations to Australian healthcare was examined using a mix of statistics and case studies. Creative occupations were found to be making widespread contributions in healthcare, in particular, to the development and delivery of healthcare goods and services, the initial training and ongoing professionalism of doctors and nurses and the effective functioning of healthcare buildings. Within most facets of healthcare, increasing employment over the period examined indicated that these contributions were growing. Key functions that creative activities addressed were information management and analysis and making complex information comprehensible, assisting communication and reducing psycho-social and distance-mediated barriers, and improving the efficiency and effectiveness of services.

Embedded creatives in the Australian manufacturing industry

According to 2011 Australian Census figures, embedded creative employees (creative employees not working in the core Creative Industries) make up 2 per cent (or a total of 17 635) of manufacturing industry employees. The average for all industries is 1.6 per cent. In the 2011–2012 financial year the manufacturing industry formed 7.3 per cent of Australia’s gross domestic product (GDP), contributing approximately AU$106.5 billion to the economy (Department of Industry, Innovation, Science, Research and Tertiary Education 2013). Manufacturing is central to innovation, accounting for over one-quarter of all business expenditure in R&D in 2010–2011, representing around AU$4.8 billion invested in R&D (ibid.). Facing challenges such as sustainability concerns, ever-increasing offshore production and the global financial crisis, the Australian manufacturing industry needs to remain relevant and competitive to succeed. Innovation is one way to do this. Given the contribution of the manufacturing industry to the Australian economy, and the above-average portion of embedded creatives in manufacturing, it is important to consider what exactly embedded creatives add to the industry. This chapter, inspired by the Getting Creative in Healthcare report (Pagan, Higgs and Cunningham 2008), examines the contribution of embedded creatives to innovation in the manufacturing industry via case studies and supplemental data.

Multiplexed tandem polymerase chain reaction identifies strong expression of oestrogen receptor and Her-2 from single, formalin-fixed, paraffin-embedded breast cancer sections

Aim To establish the suitability of multiplex tandem polymerase chain reaction (MT-PCR) for rapid identification of oestrogen receptor (ER) and Her-2 status using a single, formalin-fixed, paraffin-embedded (FFPE) breast tumour section. Methods Tissue sections from 29 breast tumours were analysed by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH). RNA extracted from 10μm FFPE breast tumour sections from 24 of 29 tumours (14 ER positive and 5 Her-2 positive) was analysed by MT-PCR. After establishing a correlation between IHC and/or FISH and MT-PCR results, the ER/Her-2 status of a further 32 randomly selected, archival breast tumour specimens was established by MT-PCR in a blinded fashion, and compared to IHC/FISH results. Results MT-PCR levels of ER and Her-2 showed good concordance with IHC and FISH results. Furthermore, among the ER positive tumours, MT-PCR provided a quantitative score with a high dynamic range. Threshold values obtained from this data set applied to 32 archival tumour specimens showed that tumours strongly positive for ER and/or Her-2 expression were easily identified by MT-PCR. Conclusion MT-PCR can provide rapid, sensitive and cost-effective analysis of FFPE material and may prove useful as triage to identify patients suited to endocrine or trastuzumab (Herceptin) treatment.

Constructions of cheating immune secret sharing

The work addresses the problem of cheating prevention in secret sharing. Two cheating scenarios are considered. In the first one, the cheaters always submit invalid shares to the combiner. In the second one, the cheaters collectively decide which shares are to be modified so the combiner gets a mixture of valid and invalid shares from the cheaters. The secret scheme is said to be k-cheating immune if any group of k cheaters has no advantage over honest participants. The paper investigates cryptographic properties of the defining function of secret sharing so the scheme is k-cheating immune. Constructions of secret sharing immune against k cheaters are given.