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.

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.

Low cost localisation for agricultural robotics

This paper presents a pose estimation approach that is resilient to typical sensor failure and suitable for low cost agricultural robots. Guiding large agricultural machinery with highly accurate GPS/INS systems has become standard practice, however these systems are inappropriate for smaller, lower-cost robots. Our positioning system estimates pose by fusing data from a low-cost global positioning sensor, low-cost inertial sensors and a new technique for vision-based row tracking. The results first demonstrate that our positioning system will accurately guide a robot to perform a coverage task across a 6 hectare field. The results then demonstrate that our vision-based row tracking algorithm improves the performance of the positioning system despite long periods of precision correction signal dropout and intermittent dropouts of the entire GPS sensor.

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.

Laser-camera data discrepancies and reliable perception in outdoor robotics

This work aims to promote integrity in autonomous perceptual systems, with a focus on outdoor unmanned ground vehicles equipped with a camera and a 2D laser range finder. A method to check for inconsistencies between the data provided by these two heterogeneous sensors is proposed and discussed. First, uncertainties in the estimated transformation between the laser and camera frames are evaluated and propagated up to the projection of the laser points onto the image. Then, for each pair of laser scan-camera image acquired, the information at corners of the laser scan is compared with the content of the image, resulting in a likelihood of correspondence. The result of this process is then used to validate segments of the laser scan that are found to be consistent with the image, while inconsistent segments are rejected. Experimental results illustrate how this technique can improve the reliability of perception in challenging environmental conditions, such as in the presence of airborne dust.

Creating synergy in teacher education through robotics-based STEM activities

Robotics has created opportunities for educators to teach concepts across Science, Technology, Engineering, and Mathematics (STEM). This is one of the reasons robotics is becoming increasingly common in primary and secondary classrooms in Australia. To enable pre-service teachers to design engaging STEM activities that incorporate these technologies, robotics is part of the teaching program in the primary education degree at Queensland University of Technology (QUT). A number of pre-service teachers also choose to extend their abilities by implementing robotics activities on field studies, in schools on a voluntary basis, and in outreach activities such as the Robotics@QUT project. The Robotics@QUT project is a support network developed to build professional knowledge and capacity of classroom teachers in schools from a low SES area, engaging in robotics-based STEM activities. Professional Development (PD) workshops are provided to teachers in order to build their knowledge and confidence in implementing robotics activities in their classrooms, loan kits are provided, and pre-service teacher visits arranged to provide the teachers with on-going support. A key feature of the project is the partnerships developed between the teachers and the pre-service teachers involved in the project. The purpose of this study was to ascertain how the teachers in the project perceived the value of the PD workshops and the pre-service teachers’ involvement and what the benefits of the involvement in the project were for the pre-service teachers. Seventeen teachers completed a five-point (1-5) likert scale questionnaire regarding their involvement in the Robotics@QUT project. Teachers’ responses on the value of the project and the pre-service teacher support highlighted the benefits of the partnerships formed and provided insights into the value of the support provided by the pre-service teachers. This paper also describes one pre-service teacher’s experience with the project and the perceived benefits from being involved.

A collaborative robotics engagement project

This paper reports on an evaluation of a collaborative robotics engagement project involving teachers from local schools and an academic from Queensland University of Technology (QUT). Engaged community projects are aimed at building stronger relationships between universities and their local communities (Sandman, Williams & Abrams, 2009). This partnership leads to mutually beneficial outcomes, builds community capacity, and can focus on aspirations and access to higher education for school students (Scull & Cuthill, 2010). The Robotics@QUT project aimed to build a partnership between local teachers and the university in order to provide students from a low SES area opportunity to engage in robotics-based Science, Technology, Engineering, and Mathematics (STEM) activities. Students from low SES regions are underrepresented at university and less likely to pursue studies in these fields (Bradley, Noonan, Nugent, & Scales, 2008). Having teachers who provide engaging STEM activities is an important motivating factor for students to enjoy STEM and do well in STEM subjects (Tytler, Osborne Williams Tytler & Clark, 2008).

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.

Robotics for sustainable broad-acre agriculture

This paper describes the development of small low-cost cooperative robots for sustainable broad-acre agriculture to increase broad-acre crop production and reduce environmental impact. The current focus of the project is to use robotics to deal with resistant weeds, a critical problem for Australian farmers. To keep the overall system affordable our robot uses low-cost cameras and positioning sensors to perform a large scale coverage task while also avoiding obstacles. A multi-robot coordinator assigns parts of a given field to individual robots. The paper describes the modification of an electric vehicle for autonomy and experimental results from one real robot and twelve simulated robots working in coordination for approximately two hours on a 55 hectare field in Emerald Australia. Over this time the real robot 'sprayed' 6 hectares missing 2.6% and overlapping 9.7% within its assigned field partition, and successfully avoided three obstacles.

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.