IS-RT-PCR assay detection of MT-MMP in a human breast cancer cell line

The in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR) is a method that allows the direct localisation of gene expression. The method utilises the dual buffer mediated activity of the enzyme rTth DNA polymerase enabling both reverse transcription and DNA amplification. Labelled nucleoside triphosphates allow the site of expression to be labelled, rather than the PCR primers themselves, giving a more accurate localisation of transcript expression and decreased background than standard in situ hybridisation (ISH) assays. The MDA-MB-231 human breast carcinoma (HBC) cell line was assayed via the IS-RT-PCR technique, using primers encoding MT-MMP (membrane-type matrix metalloproteinase) and human β-actin. Our results clearly indicate baseline expression of MT-MMP in the relatively invasive MDA-MB-231 cell line at a signal intensity similar to the housekeeping gene β-actin, and results following induction with Concanavalin A (Con A) are consistent with our previous results obtained via Northern blotting.

High level of MT-MMP expression is associated with invasiveness of cervical cancer cells

MMP-2 (gelatinase A) has been associated with the invasive potential of many cancer cells both in vitro and in vivo. It is now becoming clear that the activation of this enzyme might be a key step in tumor invasion. This activation process has been shown to be a membrane-associated pathway inducible by various agents such as collagen type I, concanavalin A or TGF-β, but its physiological regulation is still largely unresolved. MT-MMP was recently discovered and described as a potential gelatinase-A activator. In the present study, we investigated the expression of MT-MMP (membrane-type metalloproteinase) in cervical cancer cells both in vitro and in vivo. Comparing several in vitro-transformed cervical cell lines, previously shown to display different invasive potentials, our results showed that the ability of cells to overexpress MT-MMP mRNA following ConA induction correlated with their ability to activate gelatinase A and with a highly invasive behavior. Moreover, using immunohistochemistry and in situ hybridization, we found a higher level of MT-MMP expression in invasive cervical carcinoma and lymphnode metastases compared to its expression in non-invasive CIN III lesions. Our in vivo observations also clearly demonstrated a cooperation between stromal and tumor cells for the production of MT-MMP. Taken together, our results clearly correlated high level MT-MMP expression with invasiveness, and thus suggested that MT-MMP might play a crucial role in cervical tumor invasion.

Mt Isa Shopping Precinct Survey Report : Consumer Attitudes towards Proposed Extended Trading Hours at Mt Isa

Data was collected to measure shopper’s attitudes toward the proposed Sunday and limited public holiday trading in Mt Isa. Survey questionnaires were conducted between 15th August to 17th August at Kmart Plaza, Woolworths, Miles St. and Mt Isa Plaza. In total, 300 respondents participated in the survey. Overall, the findings suggest that most respondents, especially males, couples with children and fulltime workers, embrace the proposed Sunday and limited holiday trading in Mt Isa. While there are concerns over increasing competition for smaller retailers who already trade on Sundays, a majority of respondents indicated it would suit their lifestyle, be convenient, provide more jobs, increase trade for smaller retailers within the area, reduce queues and congestion, and offer a less expensive grocery shopping. The majority of those shoppers that indicated they currently did some shopping on a Sunday reported they would continue to support smaller retailers who currently trade on Sundays and some public holidays, if changes came about. Those opposed to changes to trading hours also indicated a belief that existing trading hours were sufficient. Most people indicated the proposed extension of trading hours would not harm the community or have a negative, detrimental effect on themselves or their family. The main findings presented in the report are as follows: - 96% of respondents surveyed reported to be local, permanent residents of Mt Isa. - Residents of Mt Isa visited shopping centres and stores on average 2.4 times per week. This mirrors the average Australian shopping behaviour at 2.5 times per week (Roy Morgan Supermarket Monitor) - It was determined that Saturday was the busiest day for shopping with a majority of respondents indicating they visited stores on that day of the week. - 71% of respondents support the proposal of extended trading hours at shopping centres in Mt Isa, 25% oppose and 4% are unsure. - 87% of the respondents agreed that residents of Mt Isa should be allowed the same choice as other regional towns and cities in supporting/opposing changes to trading hours. The remaining 13% expressed a disagreement. - A larger percentage of males supported the proposal for Sunday and limited holiday trading. Of all the males surveyed, 81% were in support, 17% were opposed and 2% unsure. By contrast, 64% of female respondents support the proposal, while 31% oppose it and 5% are unsure. - The highest percentage of support exists in fulltime workers with 85% of those respondents supporting the proposal. In contrast, the lowest percentage of support was found in the non-working respondents, where 62% opposed the application. - 78% of couples living with children at home expressed support for Sunday and limited public holiday trading, while 60% of couples without children also supported the application. - Of the respondents surveyed, virtually none (less than 1%) have travelled outside of Mt Isa on a Sunday to shop. This indicates that due to the remote and isolated location of this town, residents do not have the option to travel reasonable distances in order to access a range of retailers. - 70% of the respondents often find it necessary to do some grocery shopping in Mt Isa on a Sunday. - Convenience is cited as the major reason for support (79%) followed by lifestyle (75%). - The most cited reasons for supporting ‘it would be convenient’ (81%), ‘It may create more jobs’ (77%), ‘It may reduce congestion during busy shopping periods’ (74%, and ‘It would make it easier for working families with kids’ (74%). - The most cited reasons for opposing the proposed changes are that ‘It will disadvantage smaller businesses’ (44%), ‘It is unnecessary’ (29%). - 72% of shoppers who indicated they already undertook some shopping on Sunday, indicated would continue to shop and support smaller retailers. - 98% of respondents indicated they would not change their sporting or social commitment if changes to trading hours were implemented.

Variation in parental magmas of Mt Rouse, a complex polymagmatic monogenetic volcano in the basaltic intraplate Newer Volcanics Province, southeast Australia

Monogenetic volcanoes have long been regarded as simple in nature, involving single magma batches and uncomplicated evolutions; however, recent detailed research into individual centres is challenging that assumption. Mt Rouse (Kolor) is the volumetrically largest volcano in the monogenetic Newer Volcanics Province of southeast Australia. This study presents new major, trace and Sr–Nd–Pb isotope data for samples selected on the basis of a detailed stratigraphic framework analysis of the volcanic products from Mt Rouse. The volcano is the product of three magma batches geochemically similar to Ocean–Island basalts, featuring increasing LREE enrichment with each magma batch (batches A, B and C) but no evidence of crustal contamination; the Sr–Nd–Pb isotopes define two groupings. Modelling suggests that the magmas were sourced from a zone of partial melting crossing the lithosphere–asthenosphere boundary, with batch A forming a large volume partial melt in the deep lithosphere (1.7 GPa/55.5 km); and batches B and C from similar areas within the shallow asthenosphere (1.88 GPa/61 km and 1.94 GPa/63 km, respectively). The formation and extraction of these magmas may have been due to high deformation rates in the mantle caused by edge-driven convection and asthenospheric upwelling. The lithosphere– asthenosphere boundary is important with respect to NVP volcanism. An eruption chronology involves sequential eruption of magma batches A, C and B, followed by simultaneous eruption of batches A and B. Mt Rouse is a complex polymagmatic monogenetic volcano that illustrates the complexity of monogenetic volcanism and demonstrates the importance of combining detailed stratigraphic analysis alongside systematic geochemical sampling.

Analysis and evaluation of use of the V1 and South East Freeway Bicycleway post completion of Stage C of the V1

Executive Summary: Completion of the Veloway 1 (V1) will provide a dedicated and safe route for cyclists between the Brisbane CBD and the Gateway Motorway off-ramp at Eight Mile Plains alongside the South East Motorway. The V1 is being delivered in stages and when completed will provide a dedicated 3m wide cycleway 17km in length. Two stages (D and E) remain to be constructed to complete the V1. Major trip attractors along the V1 include the Mater, Princes Alexandra and Greenslopes Hospitals, two campuses of Griffith University, Garden City shopping centre and the Australian Tax Office. This report assesses the available evidence on the impacts on cycling behaviour of the recently completed V1 Stage C. The data sources informing this review include three intercept surveys, motion activated traffic cameras and travel time surveys on the V1 and adjoining South East Freeway Bikeway (SEFB), Strava app data, and cyclist crash data along Logan Road. The key findings from the evidence are that the completed V1 Stage C has: a Attracted cyclists from Holland Park, Holland Park West, Mt Gravatt and southern parts of Tarragindi onto the V1 Stage C. b Reduced the crash exposure of pedestrians to cyclists by attracting higher speed cyclists off the adjoining SEFB onto the cycling dedicated V1 Stage C. c Reduced the potential crash exposure of cyclists to motor vehicles by attracting cyclists off Logan Road on to the V1. d Provided travel time benefits to cyclists and reduced road crossings (eight down to two). e Predominantly attracted adults commuting alone to and from work and university. The evidence shows that the two traffic crossings across Birdwood Road (required as a temporary measure until the V1 is completed) negate much of the travel time gains of the V1 Stage C compared to the adjoining SEFB for southbound cyclists. Many cyclists accessing the V1 Stage C from the south are cycling in high-volume vehicular traffic lanes to reduce their travel time along Birdwood Road, but in the process are increasing their exposure to crashes with motor vehicles. Based on these findings this report recommends that TMR: a. Continue with plans to complete the V1 Veloway b. Undertake an engineering feasibility assessment to determine the viability of constructing a section of the V1 Stage E from the intersection Weller and Birdwood Roads over Marshall Road and along Bapaume Road on the western side of the Motorway to the intersection of Bapaume and Sterculia Roads. c. In the interim, improve signage and Birdwood Road crossing points for cyclists accessing and egressing the southern end of the V1 Stage C. d. Work with Brisbane City Council to identify the safest and most practical bicycle facilities to facilitate cycle travel between Logan Road and the V1 south of Birdwood Road. e. Improve the awareness of the V1 Stage C through signage for cyclists approaching from the north with the aim of providing a better understanding of the route of the V1 to the south. f. Refine the use of motion activated traffic cameras to improve the capture rate of useable images and obtain an ongoing collection over time of V1 usage data. g. Undertake discussions with Strava, Inc. to refine the presentation of Strava data to improve visual understanding of maps showing before and after cycle route volumes along and on roads leading to the V1.