Upregulation of matrix metalloproteinases (MMPs) in breast cancer xenografts : a major induction of stromal MMP-13

In human breast cancer (HBC), as with many carcinoma systems, most matrix metalloproteinases (MMPs) are largely expressed by the stromal cells, whereas the tumour cells are relatively silent in MMP expression. To determine the tissue source of the most relevant MMPs, we xenografted HBC cell lines and HBC tissues into the mammary fat pad (MFP) or bone of immunocompromised mice and measured the expression of human and mouse MMP-2, -9, -11, -13, membrane-type-1 MMP (MT1-MMP), MT2-MMP and MT3-MMP by species-specific real-time quantitative RT-PCR. Our data confirm a stromal origin for most tumour-associated MMPs and indicate marked and consistent upregulation of stromal (mouse) MMP-13 and MT1-MMP in all xenografts studied, irrespective of implantation in the MFP or bone environments. In addition, we show increased expression of both human MMP-13 and human MT1-MMP by the MDA-MB-231 tumour cells grown in the MFP compared to in vitro production. MMP protein and activity data confirm the upregulation of MMP mRNA production and indicate an increase in the activated MMP-2 species as a result of tumour implantation. These data directly demonstrate tumour induction of MMP production by stromal cells in both the MFP and bone environments. These xenografts are a valuable means for examining in vivo production of MMPs and suggest that MMP-13 and MT1-MMP will be relevant targets for inhibiting breast cancer progression.

Induction of tumorigenicity by galectin-3 in a nontumorigenic human breast-carcinoma cell-line

The human galectin-3 is a galactoside-binding protein of 31 kDa which functions as a receptor for glycoproteins containing poly N-acetyllactosamine side chains and as a substrate for matrix metalloproteinases-2 and -9. We studied its expression by flow cytofluorimetry, Western, Northern and Southern analyses, in five cultured human breast carcinoma cell lines previously characterized as non-tumorigenic, poorly metastatic or metastatic in nude mice. The expression of galectin-3 correlated with the reported tumorigenicity of the cells. The introduction of recombinant galectin-3 into the null expressing non-tumorigenic BT-549 cells resulted in the acquisition of anchorage-independent growth properties in alland tumorigenicity in 3/4 sense transfected cell crones. The data indicate a relationship between galectin-3 expression and malignancy of human breast carcinoma cell lines.

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.

The type I collagen induction of MT1-MMP-mediated MMP-2 activation is repressed by αVβ3 integrin in human breast cancer cells

The influence of αVβ3 integrin on MT1-MMP functionality was studied in human breast cancer cells of differing β3 integrin status. Overexpression of β3 integrin caused increased cell surface expression of αV integrin and increased cellular adhesion to extracellular matrix (ECM) substrates in BT-549, MDA-MB-231 and MCF-7 cells. β3 integrin expression also enhanced the migration of breast cancer cells on ECM substrates and enhanced collagen gel contraction. In vivo, αVβ3 cooperated with MT1-MMP to increase the growth of MCF-7 cells after orthotopic inoculation in immunocompromised mice, but had no influence on in vitro proliferation. Despite these stimulatory effects, overexpression of β3 integrin suppressed the type I collagen (Col I) induced MMP-2 activation in all breast cancer cell lines analyzed. This was also evident in extracts from the MCF-7 tumors in vivo, where MMP-2 activation was stimulated by MT1-MMP transfection, but attenuated with β3 integrin expression. Although our studies confirm important biological effects of αVβ3 integrin on enhancing cell adhesion and migration, ECM remodeling and tumor growth, β3 integrin caused reduced MMP-2 activation in response to Col I in vitro, which appears to be physiologically relevant, as it was also seen in tumor xenografts in vivo. The reduction of MMP-2 activation (and thus MT1-MMP activity) by αVβ3 in response to Col I may be important in scenarios where cells which are activated for matrix degradation need to preserve some pericellular collagen, perhaps as a substrate for cell adhesion and migration, thus maintaining a balanced level of proteolysis required for efficient tumor growth.

Adipose tissue induction in vivo

Engineering adipogenic tissue in vivo requires the concomitant induction of angiogenesis to generate a stable long-term three-dimensional construct. Histiocon-ductive tissue engineering strategies have been used. The disadvantage of using biodegradable scaffolds is a delayed angiogenic induction resulting in ischemic necrosis of the central cell population in the scaffold. We evaluated an histioinductive approach for adipose tissue engineering by combining essential key components for adipogenic induction: (1) a precursor cell source; (2) a vascular pedicle; (3) a supportive matrix, and; (4) a chamber to preserve space for the new tissue to develop. We observed concomitant adipogenic and angiogenic induction after 6 weeks in three-dimensional adipose tissue constructs.

An application of an extended theory of planned behaviour to understand drivers' compliance with the school zones speed limit in Australia and Malaysia

This research investigated individual and situational factors that influence driver compliance with the school zones speed limit in Australia and Malaysia. Despite all the countermeasures implemented in these two countries, driver compliance with the speed limit in school zones is still poor. The findings of the research provided support for the application of the Theory of Planned Behaviour to understand and therefore influence drivers' intention to comply with the school zones speed limit in both Australia and Malaysia. The research also revealed that mindfulness, a construct rarely used in road safety before, directly influenced Australian drivers' intention to comply, while habit influenced intention to comply in Malaysia. The research raised important theoretical and cross-cultural issues for future research and highlighted the need to increase the visibility of school zones with the use of more noticeable traffic control devices.

The impacts of variable speed limit on speed variation and headway distribution

This thesis investigates the impacts of variable speed limit on motorway speed variation and headway distribution. Initiative techniques of traffic flow categorisation study contribute in analysing the effects of variable speed limit on various traffic states. The project focuses on the speed harmonisation impacts within and across lanes as well as the uniformity of headway spread in the application of variable speed limit.

Condition monitoring and fault diagnosis of diesel engines using instantaneous angular speed analysis

Continuous monitoring of diesel engine performance is critical for early detection of fault developments in an engine before they materialize into a functional failure. Instantaneous crank angular speed (IAS) analysis is one of a few nonintrusive condition monitoring techniques that can be utilized for such a task. Furthermore, the technique is more suitable for mass industry deployments than other non-intrusive methods such as vibration and acoustic emission techniques due to the low instrumentation cost, smaller data size and robust signal clarity since IAS is not affected by the engine operation noise and noise from the surrounding environment. A combination of IAS and order analysis was employed in this experimental study and the major order component of the IAS spectrum was used for engine loading estimation and fault diagnosis of a four-stroke four-cylinder diesel engine. It was shown that IAS analysis can provide useful information about engine speed variation caused by changing piston momentum and crankshaft acceleration during the engine combustion process. It was also found that the major order component of the IAS spectra directly associated with the engine firing frequency (at twice the mean shaft rotating speed) can be utilized to estimate engine loading condition regardless of whether the engine is operating at healthy condition or with faults. The amplitude of this order component follows a distinctive exponential curve as the loading condition changes. A mathematical relationship was then established in the paper to estimate the engine power output based on the amplitude of this order component of the IAS spectrum. It was further illustrated that IAS technique can be employed for the detection of a simulated exhaust valve fault in this study.

Atmospheric gas plasma–induced ROS production activates TNF-ASK1 pathway for the induction of melanoma cancer cell apoptosis

Atmospheric gas plasmas (AGPs) are able to selectively induce apoptosis in cancer cells, offering a promising alternative to conventional therapies that have unwanted side effects such as drug resistance and toxicity. However, the mechanism of AGP-induced cancer cell death is unknown. In this study, AGP is shown to up-regulate intracellular reactive oxygen species (ROS) levels and induce apoptosis in melanoma but not normal melanocyte cells. By screening genes involved in apoptosis, we identify tumor necrosis factor (TNF)-family members as the most differentially expressed cellular genes upon AGP treatment of melanoma cells. TNF receptor 1 (TNFR1) antagonist-neutralizing antibody specifically inhibits AGP-induced apoptosis signal, regulating apoptosis signal-regulating kinase 1 (ASK1) activity and subsequent ASK1-dependent apoptosis. Treatment of cells with intracellular ROS scavenger N-acetyl-l-cysteine also inhibits AGP-induced activation of ASK1, as well as apoptosis. Moreover, depletion of intracellular ASK1 reduces the level of AGP-induced oxidative stress and apoptosis. The evidence for TNF-signaling dependence of ASK1-mediated apoptosis suggests possible mechanisms for AGP activation and regulation of apoptosis-signaling pathways in tumor cells.

SWIRLnet : portable anemometer network for wind speed measurements of land-falling tropical cyclones

Wind speed measurement systems are sparse in the tropical regions of Australia. Tropical cyclone wind speeds impacting communities are often ‘guestimated’ from analyzing damaged structures. A re-locatable anemometer system is required to enable measurements of wind speeds. This paper discusses design criteria of the tripods and tie down system, proposed deployment of the anemometers, instrumentation, and data logging. Preliminary assessment of the anemometer response indicates a reliable system for 1 second response, however, it is noted that the Australian building code and wind loading standard uses a moving average time of approximately 0.2 seconds for its wind speed design criteria.

Wind speed measurements of land-falling tropical cyclones using SWIRLnet, a portable anemometer network

Wind speed measurement systems are sparse in the tropical regions of Australia. Given this, tropical cyclone wind speeds impacting communities are seldom measured and often only ‘guestimated’ by analysing the extent of damage to structures. In an attempt to overcome this dearth of data, a re-locatable network of anemometers to be deployed prior to tropical cyclone landfall is currently being developed. This paper discusses design criteria of the network’s tripods and tie down system, proposed deployment of the anemometers, instrumentation and data logging. Preliminary assessment of the anemometer response indicates a reliable system for measuring the spectral component of wind with frequencies of approximately 1 Hz. This system limitation highlights an important difference between the capabilities of modern instrumentation and that of the Dines anemometer (around 0.2 seconds) that was used to develop much of the design criteria within the Australian building code and wind loading standard.

On the provable security of an efficient RSA-based pseudorandom generator

Pseudorandom Generators (PRGs) based on the RSA inversion (one-wayness) problem have been extensively studied in the literature over the last 25 years. These generators have the attractive feature of provable pseudorandomness security assuming the hardness of the RSA inversion problem. However, despite extensive study, the most efficient provably secure RSA-based generators output asymptotically only at most O(logn) bits per multiply modulo an RSA modulus of bitlength n, and hence are too slow to be used in many practical applications. To bring theory closer to practice, we present a simple modification to the proof of security by Fischlin and Schnorr of an RSA-based PRG, which shows that one can obtain an RSA-based PRG which outputs Ω(n) bits per multiply and has provable pseudorandomness security assuming the hardness of a well-studied variant of the RSA inversion problem, where a constant fraction of the plaintext bits are given. Our result gives a positive answer to an open question posed by Gennaro (J. of Cryptology, 2005) regarding finding a PRG beating the rate O(logn) bits per multiply at the cost of a reasonable assumption on RSA inversion.

Close-quarters Quadrotor flying for a pole inspection with position based visual servoing and high-speed vision

This paper presents a 100 Hz monocular position based visual servoing system to control a quadrotor flying in close proximity to vertical structures approximating a narrow, locally linear shape. Assuming the object boundaries are represented by parallel vertical lines in the image, detection and tracking is achieved using Plücker line representation and a line tracker. The visual information is fused with IMU data in an EKF framework to provide fast and accurate state estimation. A nested control design provides position and velocity control with respect to the object. Our approach is aimed at high performance on-board control for applications allowing only small error margins and without a motion capture system, as required for real world infrastructure inspection. Simulated and ground-truthed experimental results are presented.

A test rig for the condition-based maintenance application on the traction chain of very high speed trains

The preventive maintenance of traction equipment for Very High Speed Trains (VHST) nowadays is becoming very expensive owing to the high complexity and quality of these components that require high reliability. An efficient maintenance approach like the Condition-Based Maintenance (CBM) should be implemented to reduce the costs. For this purpose, an experimental full-scale test rig for the CBM of VHST traction equipment has been designed to investigate in detail failures in the main mechanical components of system, i.e. motor, bearings and gearbox. The paper describes the main characteristics of this unique test rig, able to reproduce accurately the train operating conditions, including the relative movements of the motor, the gearbox and the wheel axle. Gearbox, bearing seats and motor are equipped by accelerometers, thermocouples, torque meter and other sensors in different positions. The testing results give important information about the most suitable sensor position and type to be installed for each component and show the effectiveness of the techniques used for the signal analysis in order to identify faults of the gearbox and motor bearings.

Diagnostics of rolling element bearings for the traction system of high speed trains : experimental evidences

Monitoring of the integrity of rolling element bearings in the traction system of high speed trains is a fundamental operation in order to avoid catastrophic failures and to implement effective condition-based maintenance strategies. Diagnostics of rolling element bearings is usually based on vibration signal analysis by means of suitable signal processing techniques. The experimental validation of such techniques has been traditionally performed by means of laboratory tests on artificially damaged bearings, while their actual effectiveness in industrial applications, particularly in the field of rail transport, remains scarcely investigated. This paper will address the diagnostics of bearings taken from the service after a long term operation on a high speed train. These worn bearings have been installed on a test-rig, consisting of a complete full-scale traction system of a high speed train, able to reproduce the effects of wheel-track interaction and bogie-wheelset dynamics. The results of the experimental campaign show that suitable signal processing techniques are able to diagnose bearing failures even in this harsh and noisy application. Moreover, the most suitable location of the sensors on the traction system is also proposed.