Platelet-derived endothelial cell growth factor expression and angiogenesis in cervical intraepithelial neoplasia and squamous cell carcinoma of the cervix

Growth and metastatic spread of invasive carcinoma depends on angiogenesis, the formation of new blood vessels. Platelet-derived endothelial cell growth factor (PD-ECGF) is an angiogenic growth factor for a number of solid tumors, including lung, bladder, colorectal, and renal cell cancer. Cervical intraepithelial neoplasia (CIN) is the precursor to squamous cell cervical carcinoma (SCC). Mean vessel density (MVD) increases from normal cervical tissue, through low- and high-grade CIN to SCC. We evaluated PD-ECGF immunoreactivity and correlated its expression with MVD in normal, premalignant, and malignant cervical tissue. PD-ECGF expression was assessed visually within the epithelial tissues and scored on the extent and intensity of staining. MVD was calculated by counting the number of vessels positive for von Willebrand factor per unit area subtending normal or CIN epithelium or within tumor hotspots for SCC. Cytoplasmic and/or nuclear PD-ECGF immunoreactivity was seen in normal epithelium. PD-ECGF expression significantly increased with histologic grade from normal, through low- and high-grade CIN, to SCC (P < .02). A progressive significant increase in the microvessel density was also seen, ranging from a mean of 28 vessels for normal tissue to 57 for SCC (P < .0005). No correlation was found between PD-ECGF expression and MVD (P = .45). We conclude that PD-ECGF expression and MVD increase as the cervix transforms from a normal to a malignant phenotype. PD-ECGF is thymidine phosphorylase, a key enzyme in the activation of fluoropyrimidines, including 5-fluorouracil. Evaluation of PD-ECGF thymidine phosphorylase expression may be important in designing future chemotherapeutic trials in cervical cancer. Copyright (C) 2000 by W.B. Saunders Company.

Satiety responsiveness in toddlerhood predicts energy intake and weight status at four years of age

The aim of this study was to examine whether maternal-report of child eating behaviour at two years predicted self-regulation of energy intake and weight status at four years. Using an ‘eating in the absence of hunger’ paradigm, children’s energy intake (kJ) from a semi-standardized lunch meal and a standardized selection of snacks were measured. Participants were 37 mother-child dyads (16 boys, Median child age = 4.4 years, Inter-quartile range = 3.7-4.5 years) recruited from an existing longitudinal study (NOURISH randomised controlled trial). All participants were tested in their own home. Details of maternal characteristics, child eating behaviours (at age two years) reported by mothers on a validated questionnaire, and measured child height and weight (at age 3.5-4 years) were sourced from existing NOURISH trial data. Correlation and partial correlation analyses were used to examine longitudinal relationships. Satiety responsiveness and Slowness in eating were inversely associated with energy intake of the lunch meal (partial r = -.40, p =.023, and partial r = -.40, p = .023) and the former was also negatively associated with BMI-for-age Z score (partial r = -.42, p = .015). Food responsiveness and Enjoyment of food were not related to energy intake or BMI Z score. None of the eating behaviours were significantly associated with energy intake of the snacks (i.e., eating in the absence of hunger). The small and predominantly ‘healthy weight’ sample of children may have limited the ability to detect some hypothesized effects. Nevertheless, the study provides evidence for the predictive validity of two eating behaviours and future research with a larger and more diverse sample should be able to better evaluate the predictive validity of other children’s early eating behaviour styles.

School children’s personal exposure to ultrafine particles in the urban environment

There has been considerable scientific interest in personal exposure to ultrafine particles (UFP). In this study, the inhaled particle surface area doses and dose relative intensities in the tracheobronchial and alveolar regions of lungs were calculated using the measured 24-hour UFP time series of school children personal exposures for each recorded activity. Bayesian hierarchical modelling was used to determine mean doses and dose intensities for the various microenvironments. Analysis of measured personal exposures for 137 participating children from 25 schools in the Brisbane Metropolitan Area showed similar trends for all the participating children. Bayesian regression modelling was performed to calculate the daily proportion of children's total doses at different microenvironments. The proportion of alveolar doses in the total daily dose for \emph{home}, \emph{school}, \emph{commuting} and \emph{other} were 55.3\%, 35.3\%, 4.5\% and 5.0\%, respectively, with the \emph{home} microenvironment contributing a majority of children's total daily dose. Children's mean indoor dose was never higher than the outdoor's at any of the schools, indicating there were no persistent indoor particle sources in the classrooms during the measurements. Outdoor activities, eating/cooking at home and commuting were the three activities with the highest dose intensities. Personal exposure was more influenced by the ambient particle levels than immediate traffic.

Gene silencing: Fleshing out the bones

Genetic studies are revealing the pathway for RNA-mediated gene silencing. Short RNA molecules are the key, giving sequence specificity for RNA degradation and mediating communication within and between cells; these short RNAs are common to transcriptional and post-transcriptional silencing pathways. The expression of transgenes in plants varies between independent transformants and there are many examples where the transgenic trait is not expressed, or disappears in subsequent generations, despite the presence of the transgene. This loss of a trait, but not of the transgene, has become known as gene silencing and can take two forms, transcriptional or post-transcriptional. As their names imply, transcriptional gene silencing occurs when a transgene is not transcribed, whereas in post-transcriptional gene silencing, the transgene mRNA is produced but degraded before it is translated (reviewed in [1]). Both forms of silencing seem to be the result of inherent mechanisms for protecting plants against mobile or invading DNA — for example, transposable elements or the T-DNA of Agrobacterium — or RNA viruses. Plants are not alone in their capacity for transgene silencing; both forms of silencing occur in flies and fungi, where it is known as RIP or quelling, while nematodes exhibit post-transcriptional silencing, generally referred to as RNA interference (RNAi). A clearer picture of the mechanisms and relationships of the different types of transgene silencing is beginning to emerge from a number of recent studies [2], [3], [4], [5], [6], [7] and [8]. Some of these studies [2], [3], [4] and [5] have enhanced our understanding of the steps within the post-transcriptional silencing pathway, and others [6], [7] and [8] have demonstrated that the two forms of silencing may be mechanistically linked.

HMM relative entropy rate concepts for vision-based aircraft manoeuvre detection

Machine vision is emerging as a viable sensing approach for mid-air collision avoidance (particularly for small to medium aircraft such as unmanned aerial vehicles). In this paper, using relative entropy rate concepts, we propose and investigate a new change detection approach that uses hidden Markov model filters to sequentially detect aircraft manoeuvres from morphologically processed image sequences. Experiments using simulated and airborne image sequences illustrate the performance of our proposed algorithm in comparison to other sequential change detection approaches applied to this application.

Consistent HMM parameter estimation using Kerridge inaccuracy rates

In this paper, we propose a novel online hidden Markov model (HMM) parameter estimator based on Kerridge inaccuracy rate (KIR) concepts. Under mild identifiability conditions, we prove that our online KIR-based estimator is strongly consistent. In simulation studies, we illustrate the convergence behaviour of our proposed online KIR-based estimator and provide a counter-example illustrating the local convergence properties of the well known recursive maximum likelihood estimator (arguably the best existing solution).

A bioengineered 3D ovarian cancer model for the assessment of peptidase-mediated enhancement of spheroid growth and intraperitoneal spread

Cancer-associated proteases promote peritoneal dissemination and chemoresistance in malignant progression. In this study, kallikrein-related peptidases 4, 5, 6, and 7 (KLK4-7)-cotransfected OV-MZ-6 ovarian cancer cells were embedded in a bioengineered three-dimensional (3D) microenvironment that contains RGD motifs for integrin engagement to analyze their spheroid growth and survival after chemotreatment. KLK4-7-cotransfected cells formed larger spheroids and proliferated more than controls in 3D, particularly within RGD-functionalized matrices, which was reduced upon integrin inhibition. In contrast, KLK4-7-expressing cell monolayers proliferated less than controls, emphasizing the relevance of the 3D microenvironment and integrin engagement. In a spheroid-based animal model, KLK4-7-overexpression induced tumor growth after 4 weeks and intraperitoneal spread after 8 weeks. Upon paclitaxel administration, KLK4-7-expressing tumors declined in size by 91% (controls: 87%) and showed 90% less metastatic outgrowth (controls: 33%, P<0.001). KLK4-7-expressing spheroids showed 53% survival upon paclitaxel treatment (controls: 51%), accompanied by enhanced chemoresistance-related factors, and their survival was further reduced by combination treatment of paclitaxel with KLK4/5/7 (22%, P=0.007) or MAPK (6%, P=0.006) inhibition. The concomitant presence of KLK4-7 in ovarian cancer cells together with integrin activation drives spheroid formation and proliferation. Combinatorial approaches of paclitaxel and KLK/MAPK inhibition may be more efficient for late-stage disease than chemotherapeutics alone as these inhibitory regimens reduced cancer spheroid growth to a greater extent than paclitaxel alone.

Molecular dynamics investigation on shearing between osteopontin and hydroxyapatite in biological materials

Bone, a hard biological material, possesses a combination of high stiffness and toughness, even though the main basic building blocks of bone are simply mineral platelets and protein molecules. Bone has a very complex microstructure with at least seven hierachical levels. This unique material characteristic attracts great attention, but the deformation mechanisms in bone have not been well understood. Simulation at nano-length scale such as molecular dynamics (MD) is proven to be a powerful tool to investigate bone nanomechanics for developing new artificial biological materials. This study focuses on the ultra large and thin layer of extrafibrillar protein matrix (thickness = ~ 1 nm) located between mineralized collagen fibrils (MCF). Non-collagenous proteins such as osteopontin (OPN) can be found in this protein matrix, while MCF consists mainly of hydroxyapatite (HA) nanoplatelets (thickness = 1.5 – 4.5 nm). By using molecular dynamics method, an OPN peptide was pulled between two HA mineral platelets with water in presence. Periodic boundary condition (PBC) was applied. The results indicate that the mechanical response of OPN peptide greatly depends on the attractive electrostatics interaction between the acidic residues in OPN peptide and HA mineral surfaces. These bonds restrict the movement of OPN peptide, leading to a high energy dissipation under shear loading.

Innovation through action research in environmental education : from project to praxis

This thesis is a work-in-progress that articulates my research journey based on the development of a curriculum innovation in environmental education. This journey had two distinct, but intertwined phases: action research based fieldwork, conducted collaboratively, to create a whole school approach to environmental education curriculum planning; and a phase of analysis and reflection based on the emerging findings, as I sought to create personal "living educational theory" about change and innovation. A key stimulus for the study was the perceived theory-practice gap in environmental education, which is often presented in the literature as a criticism of teachers for failing to achieve the values and action objectives of critical environmental education. Hence, many programs and projects are considered to be superficial and inconsequential in terms of their ability to seriously address environmental issues. The intention of this study was to work with teachers in a project that would be an exemplar of critical environmental education. This would be in the form of a whole school "learnscaping" curriculum in a primary school whereby the schoolgrounds would be utilised for interdisciplinary critical environment education. Parallel with the three cycles of action research in this project, my research objectives were to identify and comment upon the factors that influence the generation of successful educational innovation. It was anticipated that the project would be a collaboration involving me, as researcher-facilitator, and many of the teachers in the school as active participants. As the project proceeded through its action cycles, however, it became obvious that the goal of developing a critical environmental education curriculum, and the use of highly participatory processes, were unrealistic. Institutional and organisational rigidities in education generally, teachers' day-to-day work demands, and the constant juggle of work, family and other responsibilities for all participants acted as significant constraints. Consequently, it became apparent that the learnscaping curriculum would not be the hoped-for exemplar. Progress was slow and, at times, the project was in danger of stalling permanently. While the curriculum had some elements of critical environmental education, these were minor and not well spread throughout the school. Overall, the outcome seemed best described as a "small win"; perhaps just another example of the theory-practice gap that I had hoped this project would bridge. Towards the project's end, however, my continuing reflection led to an exploration of chaos/complexity theory which gave new meaning to the concept of a "small win". According to this theory, change is not the product of linear processes applied methodically in purposeful and diligent ways, but emerges from serendipitous events that cannot be planned for, or forecast in advance. When this perspective of change is applied to human organisations - in this study, a busy school - the context for change is recognised not as a stable, predictable environment, but as a highly complex system where change happens all the time, cannot be controlled, and no one can be really sure where the impacts might lead. This so-called "butterfly effect" is a central idea of this theory where small changes or modifications are created - the effects of which are difficult to know, let alone determine - and which can have large-scale impacts. Allied with this effect is the belief that long term developments in an organisation that takes complexity into account, emerge by spontaneous self-organising evolution, requiring political interaction and learning in groups, rather than systematic progress towards predetermined goals or "visions". Hence, because change itself and the contexts of change are recognised as complex, chaos/complexity theory suggests that change is more likely to be slow and evolutionary - cultural change - rather than fast and revolutionary where the old is quickly ushered out by radical reforms and replaced by new structures and processes. Slow, small-scale changes are "normal", from a complexity viewpoint, while rapid, wholesale change is both unlikely and unrealistic. Therefore, the frustratingly slow, small-scale, imperfect educational changes that teachers create - including environmental education initiatives - should be seen for what they really are. They should be recognised as successful changes, the impacts of which cannot be known, but which have the potential to magnify into large-scale changes into the future. Rather than being regarded as failures for not meeting critical education criteria, "small wins" should be cause for celebration and support. The intertwined phases of collaborative action research and individual researcher reflection are mirrored in the thesis structure. The first three chapters, respectively, provide the thesis overview, the literature underpinning the study's central concern, and the research methodology. Chapters 4, 5, and 6 report on each of the three action research cycles of the study, namely Laying the Groundwork, Down to Work!, and The Never-ending Story. Each of these chapters presents a narrative of events, a literature review specific to developments in the cycle, and analysis and critique of the events, processes and outcomes of each cycle. Chapter 7 provides a synthesis of the whole of the study, outlining my interim propositions about facilitating curriculum change in schools through action research, and the implications of these for environmental education.

The Brisbane Northbank Urban Film Studio : screening of shortlisted films

Research Statement: In this research project film groups of 4-5 students under my direction produced a 3-5 minute urban film that explored the Brisbane Northbank, and which would become the basis for an urban proposal and design of a small film studio for independent filmmakers in the site. The theoretical premise was that a film studio does not simply produce movies, it creates urban effects all around it and acts as a vortex of cultural activity and social life. For this modest facility where the cinema goes out into the street, the city itself becomes the studio. Students were called to observe the historical problematics of technique, image and effect that arise in the cinema, and to apply these to their own urban-film practice. A panel of judges working in film and architecture shortlisted the 12 best films in 2010 and a major public film screening event took place at the Tribal Cinema. The Shortlisted films today form a permanent "exhibit" in YouTube. The research project was funded by the Queensland University of Technology, School of Design and received accolades from film faculty in the Creative Industries Faculty. The diverse body of work that emanated from the screening contributed a unique analysis of the Northbank to Brisbane.

Meta-analysis of individual patient data from randomized trials of chemotherapy plus cetuximab as first-line treatment for advanced non-small cell lung cancer

OBJECTIVES: Four randomized phase II/III trials investigated the addition of cetuximab to platinum-based, first-line chemotherapy in patients with advanced non-small cell lung cancer (NSCLC). A meta-analysis was performed to examine the benefit/risk ratio for the addition of cetuximab to chemotherapy. MATERIALS AND METHODS: The meta-analysis included individual patient efficacy data from 2018 patients and individual patient safety data from 1970 patients comprising respectively the combined intention-to-treat and safety populations of the four trials. The effect of adding cetuximab to chemotherapy was measured by hazard ratios (HRs) obtained using a Cox proportional hazards model and odds ratios calculated by logistic regression. Survival rates at 1 year were calculated. All applied models were stratified by trial. Tests on heterogeneity of treatment effects across the trials and sensitivity analyses were performed for all endpoints. RESULTS: The meta-analysis demonstrated that the addition of cetuximab to chemotherapy significantly improved overall survival (HR 0.88, p=0.009, median 10.3 vs 9.4 months), progression-free survival (HR 0.90, p=0.045, median 4.7 vs 4.5 months) and response (odds ratio 1.46, p<0.001, overall response rate 32.2% vs 24.4%) compared with chemotherapy alone. The safety profile of chemotherapy plus cetuximab in the meta-analysis population was confirmed as manageable. Neither trials nor patient subgroups defined by key baseline characteristics showed significant heterogeneity for any endpoint. CONCLUSION: The addition of cetuximab to platinum-based, first-line chemotherapy for advanced NSCLC significantly improved outcome for all efficacy endpoints with an acceptable safety profile, indicating a favorable benefit/risk ratio.

The epidemiology of basal cell carcinoma

Basal cell carcinoma (BCC) is a skin cancer of particular importance to the Australian community. Its rate of occurrence is highest in Queensland, where 1% to 2% of people are newly affected annually. This is an order of magnitude higher than corresponding incidence estimates in European and North American populations. Individuals with a sun-sensitive complexion are particularly susceptible because sun exposure is the single most important causative agent, as shown by the anatomic distribution of BCC which is in general consistent with the levels of sun exposure across body sites. A distinguishing feature of BCC is the occurrence of multiple primary tumours within individuals, synchronously or over time, and their diagnosis and treatment costs contribute substantially to the major public health burden caused by BCC. A primary knowledge gap about BCC pathogenesis however was an understanding of the true frequency of multiple BCC occurrences and their body distribution, and why a proportion of people do develop more than one BCC in their life. This research project sought to address this gap under an overarching research aim to better understand the detailed epidemiology of BCC with the ultimate goal of reducing the burden of this skin cancer through prevention. The particular aim was to document prospectively the rate of BCC occurrence and its associations with constitutional and environmental (solar) factors, all the while paying special attention to persons affected by more than one BCC. The study built on previous findings and recent developments in the field but set out to confirm and extend these and propose more adequate theories about the complex epidemiology of this cancer. Addressing these goals required a new approach to researching basal cell carcinoma, due to the need to account for the phenomenon of multiple incident BCCs per person. This was enabled by a 20 year community-based study of skin cancer in Australians that provided the methodological foundation for this thesis. Study participants were originally randomly selected in 1986 from the electoral register of all adult residents of the subtropical township of Nambour in Queensland, Australia. On various occasions during the study, participants were fully examined by dermatologists who documented cumulative photodamage as well as skin cancers. Participants completed standard questionnaires about skin cancer-related factors, and consented to have any diagnosed skin cancers notified to the investigators by regional pathology laboratories in Queensland. These methods allowed 100% ascertainment of histologically confirmed BCCs in this study population. 1339 participants had complete follow-up to the end of 2007. Statistical analyses in this thesis were carried out using SAS and SUDAAN statistical software packages. Modelling methods, including multivariate logistic regressions, allowed for repeated measures in terms of multiple BCCs per person. This innovative approach gave new findings on two levels, presented in five chapters as scientific papers: 1. Incidence of basal cell carcinoma multiplicity and detailed anatomic distribution: longitudinal study of an Australian population The incidence of people affected multiple times by BCC was 705 per 100,000 person years compared to an incidence rate of people singly affected of 935 per 100,000 person years. Among multiply and singly affected persons alike, site-specific BCC incidence rates were far highest on facial subsites, followed by upper limbs, trunk, and then lower limbs 2. Melanocytic nevi and basal cell carcinoma: is there an association? BCC risk was significantly increased in those with forearm nevi (Odds Ratios (OR) 1.43, 95% Confidence Intervals (CI) 1.09-1.89) compared to people without forearm nevi, especially among those who spent their time mainly outdoors (OR 1.6, 95%CI 1.1-2.3) compared to those who spent their time mainly indoors. Nevi on the back were not associated with BCC. 3. Clinical signs of photodamage are associated with basal cell carcinoma multiplicity and site: a 16-year longitudinal study Over a 16-year follow-up period, 58% of people affected by BCC developed more than one BCC. Among these people 60% developed BCCs across different anatomic sites. Participants with high numbers of solar keratoses, compared to people without solar keratoses, were most likely to experience the highest BCC counts overall (OR 3.3, 95%CI 1.4-13.5). Occurrences of BCC on the trunk (OR 3.3, 95%CI 1.4-7.6) and on the limbs (OR 3.7, 95%CI 2.0-7.0) were strongly associated with high numbers of solar keratoses on these sites. 4. Occurrence and determinants of basal cell carcinoma by histological subtype in an Australian community Among 1202 BCCs, 77% had a single growth pattern and 23% were of mixed histological composition. Among all BCCs the nodular followed by the superficial growth patterns were commonest. Risk of nodular and superficial BCCs on the head was raised if 5 or more solar keratoses were present on the face (OR 1.8, 95%CI 1.2-2.7 and OR 4.5, 95%CI 2.1-9.7 respectively) and similarly on the trunk in the presence of multiple solar keratoses on the trunk (OR 4.2, 95%CI 1.5-11.9 and OR 2.2, 95%CI 1.1-4.4 respectively). 5. Basal cell carcinoma and measures of cumulative sun exposure: an Australian longitudinal community-based study Dermal elastosis was more likely to be seen adjacent to head and neck BCCs than trunk BCCs (p=0.01). Severity of dermal elastosis increased on each site with increasing clinical signs of cutaneous sun damage on that site. BCCs that occurred without perilesional elastosis per se, were always found in an anatomic region with signs of photodamage. This thesis thus has identified the magnitude of the burden of multiple BCCs. It does not support the view that people affected by more than one BCC represent a distinct group of people who are prone to BCCs on certain body sites. The results also demonstrate that BCCs regardless of site, histology or order of occurrence are strongly associated with cumulative sun exposure causing photodamage to the skin, and hence challenge the view that BCCs occurring on body sites with typically low opportunities for sun exposure or of the superficial growth pattern are different in their association with the sun from those on typically sun-exposed sites, or nodular BCCs, respectively. Through dissemination in the scientific and medical literature, and to the community at large, these findings can ultimately assist in the primary and secondary prevention of BCC, perhaps especially in high-risk populations.

Improved molecular tools for sugar cane biotechnology

Sugar cane is a major source of food and fuel worldwide. Biotechnology has the potential to improve economically-important traits in sugar cane as well as diversify sugar cane beyond traditional applications such as sucrose production. High levels of transgene expression are key to the success of improving crops through biotechnology. Here we describe new molecular tools that both expand and improve gene expression capabilities in sugar cane. We have identified promoters that can be used to drive high levels of gene expression in the leaf and stem of transgenic sugar cane. One of these promoters, derived from the Cestrum yellow leaf curling virus, drives levels of constitutive transgene expression that are significantly higher than those achieved by the historical benchmark maize polyubiquitin-1 (Zm-Ubi1) promoter. A second promoter, the maize phosphonenolpyruvate carboxylate promoter, was found to be a strong, leaf-preferred promoter that enables levels of expression comparable to Zm-Ubi1 in this organ. Transgene expression was increased approximately 50-fold by gene modification, which included optimising the codon usage of the coding sequence to better suit sugar cane. We also describe a novel dual transcriptional enhancer that increased gene expression from different promoters, boosting expression from Zm-Ubi1 over eightfold. These molecular tools will be extremely valuable for the improvement of sugar cane through biotechnology.

An improved chemically inducible gene switch that functions in the monocotyledonous plant sugar cane

Chemically inducible gene switches can provide precise control over gene expression, enabling more specific analyses of gene function and expanding the plant biotechnology toolkit beyond traditional constitutive expression systems. The alc gene expression system is one of the most promising chemically inducible gene switches in plants because of its potential in both fundamental research and commercial biotechnology applications. However, there are no published reports demonstrating that this versatile gene switch is functional in transgenic monocotyledonous plants, which include some of the most important agricultural crops. We found that the original alc gene switch was ineffective in the monocotyledonous plant sugar cane, and describe a modified alc system that is functional in this globally significant crop. A promoter consisting of tandem copies of the ethanol receptor inverted repeat binding site, in combination with a minimal promoter sequence, was sufficient to give enhanced sensitivity and significantly higher levels of ethanol inducible gene expression. A longer CaMV 35S minimal promoter than was used in the original alc gene switch also substantially improved ethanol inducibility. Treating the roots with ethanol effectively induced the modified alc system in sugar cane leaves and stem, while an aerial spray was relatively ineffective. The extension of this chemically inducible gene expression system to sugar cane opens the door to new opportunities for basic research and crop biotechnology.