α2β1 integrin affects metastatic potential of ovarian carcinoma spheroids by supporting disaggregation and proteolysis

Background: Ovarian cancer is characterized by a wide-spread intra-abdominal metastases which represents a major clinical hurdle in the prognosis and management of the disease. A significant proportion of ovarian cancer cells in peritoneal ascites exist as multicellular aggregates or spheroids. We hypothesize that these cellular aggregates or spheroids are invasive with the capacity to survive and implant on the peritoneal surface. This study was designed to elucidate early inherent mechanism(s) of spheroid survival, growth and disaggregation required for peritoneal metastases.

Methods: In this study, we determined the growth pattern and adhesive capacity of ovarian cancer cell lines (HEY and OVHS1) grown as spheroids, using the well established liquid overlay technique, and compared them to a normal ovarian cell line (IOSE29) and cancer cells grown as a monolayer. The proteolytic capacity of these spheroids was compared with cells grown as a monolayer using a gelatin zymography assay to analyze secreted MMP-2/9 in conditioned serum-free medium. The disaggregation of cancer cell line spheroids was determined on extracellular matrices (ECM) such as laminin (LM), fibronectin (FN) and collagen (CI) and the expression of α2, α3, αv, α6 and β1 interin was determined by flow cytometric analysis. Neutralizing antibodies against α2, β1 subunits and α2β1 integrin was used to inhibit disaggregation as well as activation of MMPs in spheroids.

Results: We demonstrate that ovarian cancer cell lines grown as spheroids can sustain growth for 10 days while the normal ovarian cell line failed to grow beyond 2 days. Compared to cells grown as a monolayer, cancer cells grown as spheroids demonstrated no change in adhesion for up to 4 days, while IOSE29 cells had a 2–4-fold loss of adhesion within 2 days. Cancer cell spheroids disaggregated on extracellular matrices (ECM) and demonstrated enhanced expression of secreted pro-MMP2 as well as activated MMP2/MMP9 with no such activation of MMP's observed in monolayer cells. Flow cytometric analysis demonstrated enhanced expression of α2 and diminution of α6 integrin subunits in spheroids
versus monolayer cells. No change in the expression of α3, αv and β1 subunits was evident. Conversely, except for αv integrin, a 1.5–7.5-fold decrease in α2, α3, α6 and β1 integrin subunit expression was observed in IOSE29 cells within 2 days. Neutralizing antibodies against α2, β1 subunits and α2β1 integrin inhibited disaggregation as well as activation of
MMPs in spheroids.

Conclusion: Our results suggest that enhanced expression of α2β1 integrin may influence spheroid disaggregation and
proteolysis responsible for the peritoneal dissemination of ovarian carcinoma. This may indicate a new therapeutic target
for the suppression of the peritoneal metastasis associated with advanced ovarian carcinomas.

Influence of preexercise muscle glycogen content on transcrptional activity of metabolic and myogenic genes in well-trained humans

To determine whether preexercise muscle glycogen content influences the transcription of several early-response genes involved in the regulation of muscle growth, seven male strength-trained subjects performed one-legged cycling exercise to exhaustion to lower muscle glycogen levels (Low) in one leg compared with the leg with normal muscle glycogen (Norm) and then the following day completed a unilateral bout of resistance training (RT). Muscle biopsies from both legs were taken at rest, immediately after RT, and after 3 h of recovery. Resting glycogen content was higher in the control leg (Norm leg) than in the Low leg (435 ± 87 vs. 193 ± 29 mmol/kg dry wt; P < 0.01). RT decreased glycogen content in both legs (P < 0.05), but postexercise values remained significantly higher in the Norm than the Low leg (312 ± 129 vs. 102 ± 34 mmol/kg dry wt; P < 0.01). GLUT4 (3-fold; P < 0.01) and glycogenin mRNA abundance (2.5-fold; not significant) were elevated at rest in the Norm leg, but such differences were abolished after exercise. Preexercise mRNA abundance of atrogenes was also higher in the Norm compared with the Low leg [atrogin: 14-fold, P < 0.01; RING (really interesting novel gene) finger: 3-fold, P < 0.05] but decreased for atrogin in Norm following RT (P < 0.05). There were no differences in the mRNA abundance of myogenic regulatory factors and IGF-I in the Norm compared with the Low leg. Our results demonstrate that 1) low muscle glycogen content has variable effects on the basal transcription of select metabolic and myogenic genes at rest, and 2) any differences in basal transcription are completely abolished after a single bout of heavy resistance training. We conclude that commencing resistance exercise with low muscle glycogen does not enhance the activity of genes implicated in promoting hypertrophy.

Interaction of contractile activity and training history on mRNA abundance in skeletal muscle from trained athletes

Skeletal muscle displays enormous plasticity to respond to contractile activity with muscle from strength- (ST) and endurance-trained (ET) athletes representing diverse states of the adaptation continuum. Training adaptation can be viewed as the accumulation of specific proteins. Hence, the altered gene expression that allows for changes in protein concentration is of major importance for any training adaptation. Accordingly, the aim of the present study was to quantify acute subcellular responses in muscle to habitual and unfamiliar exercise. After 24-h diet/exercise control, 13 male subjects (7 ST and 6 ET) performed a random order of either resistance (8 x 5 maximal leg extensions) or endurance exercise (1 h of cycling at 70% peak O2 uptake). Muscle biopsies were taken from vastus lateralis at rest and 3 h after exercise. Gene expression was analyzed using real-time PCR with changes normalized relative to preexercise values. After cycling exercise, peroxisome proliferator-activated receptor- coactivator-1 (ET 8.5-fold, ST 10-fold, P < 0.001), pyruvate dehydrogenase kinase-4 (PDK-4; ET 26-fold, ST 39-fold), vascular endothelial growth factor (VEGF; ET 4.5-fold, ST 4-fold), and muscle atrophy F-box protein (MAFbx) (ET 2-fold, ST 0.4-fold) mRNA increased in both groups, whereas MyoD (3-fold), myogenin (0.9-fold), and myostatin (2-fold) mRNA increased in ET but not in ST (P < 0.05). After resistance exercise PDK-4 (7-fold, P < 0.01) and MyoD (0.7-fold) increased, whereas MAFbx (0.7-fold) and myostatin (0.6-fold) decreased in ET but not in ST. We conclude that prior training history can modify the acute gene responses in skeletal muscle to subsequent exercise.

Revisiting capital structure theory : a test of pecking order and static order trade-of model from Malaysian capital market

This study revisits the capital structure theory and test Pecking Order Hypothesis (POH) and Static Order Trade-off theory (STOT) using Malaysian Listed firms over a period from 1999 to 2002. The evidence from pecking order model suggests that the internal fund deficiency is the most important determinant that possibly explains the issuance of new debt in Malaysian capital market despite the lower predicting power.  While static trade off-model is not fit to explain the issuance of new debt issue in Malaysian capital market. This is an interesting findings that confirm the fact that Malaysian firms do not too much care about tax-shield benefit derive from employ both debt and non-debt tax-shield. The finn's size, which is used to neutralize the size effect, appears to provide some explanation for the variation in its capital structure policy choice; however asset structure and growth no evidence of static-order-trade-off is observed in Malaysian capital market.

UV-shielding ceramic nanoparticles synthesised by mechanochemcial processing

ZnO, TiO₂ and CeO₂ are known as UV-shielding ceramic materials that have advantages over organic UV absorbers for their photo-stability and non-hazardous nature to human bodies. However, they normally cause low transparency in the visible-light range due to light scattering by large particles, which is undesirable for many transparent UV-blocking applications in cosmetic and plastic industries. Light-scattering efficiency of particles can be drastically reduced by decreasing the particle sizes down below 100 nm. This paper reviews recent investigation on the synthesis of ZnO and CeO₂ nanoparticles by mechanochemical processing. The resulting particles had a significantly low degree of agglomeration, having mean particle sizes of ~ 25 nm and ~ 10 nm, respectively. The aqueous suspensions of the nanoparticles showed strong absorption in the UV-light range and high transmittance in the visible-light range. Mechanochemical processing offers the possibility of industrial-scale production of transparent UV-shielding ceramic particles for many applications.

People's war under modern conditions : an investigation into Chinese defence strategy, 1977-2049

This thesis is an investigation of the relevance of ‘people's war’ to contemporary Chinese defence policy. This loose concept has been eroded by 'modernity’, but a guerilla-based defence remains the essential theme. Prior to China's acquisition of nuclear weapons, people's war was the sole element of the state's deterrent policy, aiming to swamp an invader with China's numerical strength. In the 1980s and beyond people's war finds effectiveness through a combination of materiel of middle range technology and the traditional usage of massive manpower. People's war under modern conditions remains essentially defensive, but now incorporates 'active defence’ with accent on greater mobility. However, the central thesis of this work relates to how the traditional strategy may influence nuclear doctrine. This thesis proposes that China could abandon long-range ballistic missiles and adopt a new concept in nuclear strategy: that of, Guerilla Nuclear Warfare. Trained in guerilla tactics and equipped with battlefield nuclear weapons, this would represent the logical extension of China's people's war strategy to the new nuclear conditions associated with superpower research into space-based ballistic missile defences and which, in full deployment, could nullify a Chinese nuclear deterrent based only on 'mid-tech' delivery systems. Guerilla Nuclear Warfare, as a strategy, would involve the irregular use of locally held and controlled tactical nuclear weapons, but it would also be a method of circumventing the proposed Soviet missile defence shield by not challenging it. Guerilla Nuclear Warfare does not exist in the late 1980s, but evidence exists to suggest its development. It cannot yet be proven as the new direction but China's strategic circumstances add weight to available indications: unless the Strategic Defence programs of the established superpowers are arrested then it appears the sole option available to the Chinese for the maintenance of a nuclear deterrent in the early part of the 21st Century.

Exposure to fumes and gases during welding operations

The exposure to fumes and gases is one of the hazards associated with welding operations. Apart from research conducted on the mechanism of fume and gas formation and the relationship between fume formation rates and common welding parameters, little is known about the exposure process during welding. This research project aimed to identify the factors that influence exposure, develop an understanding of their role in the exposure process and through this understanding formulate strategies for the effective control of exposure during welding. To address these aims a literature review and an experimental program was conducted The literature review surveyed epidemiological, toxicological and exposure data. The experimental program involved three approaches, the first, an evaluation of the factors that influence exposure by assessing a metal inert gas/mild steel welding process in a workshop setting. The second approach involved the study of exposure in a controlled environment provided by a wind tunnel and simulated welding process. The final approach was to investigate workplace conditions through an assessment of exposure and control strategies in industry. The exposure to fumes and gases during welding is highly variable and frequently in excess of the health based exposure standards. Exposure is influenced by a number of a factors including the welding process, base material, arc time, electrode, arc current, arc voltage, arc length, electrode polarity, shield gas, wire-to-metal-work distance (metal inert gas), metal transfer mode, intensity of the UV radiation (ozone), the frequency of arc ignitions (ozone), thermal buoyancy generated by the arc process, ventilation (natural and mechanical), the welding environment, the position of the welder, the welders stance, helmet type, and helmet position. Exposure occurs as a result of three processes: the formation of contaminants at or around the arc region; their transport from the arc region, as influenced by the entry and thermal expansion of shield gases, the vigorous production of contaminants, thermal air currents produced by the heat of the arc process, and ventilation; and finally the entry of contaminants into the breathing zone of the welder, as influenced by the position of the welder, the welders stance, helmet type, and the helmet position. The control of exposure during welding can be achieved by several means: through the selection of welding parameters that generate low contaminant formation rates; through the limitation of arc time; and by isolating the breathing zone of the welder from the contaminant plume through the use of ventilation, welder position or the welding helmet as a physical barrier. Effective control is achieved by careful examination of the workplace, the selection of the most appropriate control option, and motivation of the workforce.

Geared equity investments : a case study of tax arbitrage down under

Geared Equity Investment (GEI) contracts are an over-the-counter financial derivative product offered by Macquarie Bank, Ltd. to individual investors in Australia and New Zealand as a managed-risk investment in local shares carrying significant tax shield benefits. Upon issuance, a geared equity contract has three stakeholders: (1) the investor; (2) the issuer; and (3) the national tax authority. We assess the value of these contracts to each stakeholder and their support for tax arbitrage. We find that the national tax authority provides a significant subsidy to GEI contracts via tax shield benefits. These benefits support investor tax arbitrage in certain cases and issuer tax arbitrage in all cases examined.

In vitro spheroids and metastatic epithelial ovarian cancer

This thesis investigated the role of cellular aggregation in the spread of epithelial ovarian cancer. Cell aggregation was studied in vitro in order to identify markers of disease progression and cell-cell adhesion. Proteomics was then used to identify additional proteins associated with cell aggregation and survival.

Multicellular spheroids in ovarian cancer metastases: Biology and pathology

Epithelial ovarian cancer (EOC) has a relatively high mortality rate (~55%). One of the presiding causes is that the current chemotherapeutic regimes are unable to achieve sustained remission, despite frequently producing a positive response at first treatment. One of the reasons that EOC is difficult to treat is that the mechanism of dissemination is unusual. EOC dissemination characteristically involves local invasion of pelvic and abdominal organs. Unlike many epithelial cancers, initial dissemination rarely requires the vasculature, although the vasculature is often implicated in the advanced stages of disease. Recently, it has become apparent that aggregates of malignant cells (spheroids) contained within malignant ascites represent a significant impediment to efficacious treatment of late stage EOC. In vivo, spheroids are present in the malignant ascites of EOC patients, while in vitro cultured spheroids are capable of tumorgenesis in vivo and display a reduced response to chemotherapeutic drugs when compared to monolayers. A major problem associated with the current generation of chemotherapy agents is that they do not address the anchorage and vascular-independent growth conditions associated with a 3-dimensional structure that has formed and/or grown in suspension. Thus, spheroid formation may represent a key component of platinum/taxanesensitive recurrence. If this is correct, a better understanding of spheroid biology may contribute to the identification of new treatment opportunities for the sustained treatment of metastatic EOC. This review article outlines the key biological features of spheroids, specifically discussing their role in EOC dissemination and chemo-response as well as providing insights into spheroid functionality.

Neighbourhood renewal in Victoria, Australia : an effective way to address social inclusion

People who live in disadvantaged communities are at increased risk of social exclusion through diminished access and quality of services, lack of opportunity and feeling powerless over decisions relating to their neighbourhoods. Neighbourhood Renewal (NR) is a Victorian State Government initiative that seeks to address this. This paper presents the findings from two individual project sites, side-by-side. Data were collected in 2004/5 and 2009 using face-to face interviewing with convenience samples of 900 NR residents across the two NR sites at each time period. A comparison group for each NR site consisted of a sample of 150 people living in the same suburb or town but outside the NR site, data were collected by telephone. Data were analysed separately for each NR project site. Findings indicate that neighbourhood renewal strategies can be effective in improving trust in government, perceptions of community participation, influence and control over community decisions and improved services. Community level strategies are valuable in addressing area-level determinants to improve social inclusion. The successes of the NR scheme support the implementation and continuation of area-specific interventions to address disadvantage and social exclusion across Victoria, Australia.

Australia's defence policy : a market-state approach?

In 2002, Professor Philip Bobbitt published 'The Shield of Achilles', in which he postulates that the nation-state is evolving into a fundamentally new form - the market state. It is my contention that Australia is evolving into something approximating Bobbitt's market-state and therefore its defence policy shuld be informed by market-state strategy.