Modulation of dermal microvascular endithelial cell responses to growth factors and haemostatic factors in the presence of vitronectin

In order to effect permanent closure in burns patients suffering from full thickness wounds, replacing their skin via split thickness autografting, is essential. Dermal substitutes in conjunction with widely meshed split thickness autografts (+/- cultured keratinocytes) reduce scarring at the donor and recipient sites of burns patients by reducing demand for autologous skin (both surface area and thickness), without compromising dermal delivery at the wound face. Tissue engineered products such as Integra consist of a dermal template which is rapidly remodelled to form a neodermis, at which time the temporary silicone outer layer is removed and replaced with autologous split thickness skin. Whilst provision of a thick tissue engineered dermis at full thickness burn sites reduces scarring, it is hampered by delays in vascularisation which results in clinical failure. The ultimate success of any skin graft product is dependent upon a number of basic factors including adherence, haemostasis and in the case of viable tissue grafts, success is ultimately dependent upon restoration of a normal blood supply, and hence this study. Ultimately, the goal of this research is to improve the therapeutic properties of tissue replacements, through impregnation with growth factors aimed at stimulating migration and proliferation of microvascular endothelial cells into the donor tissue post grafting. For the purpose of my masters, the aim was to evaluate the responsiveness of a dermal microvascular endothelial cell line to growth factors and haemostatic factors, in the presence of the glycoprotein vitronectin. Vitronectin formed the backbone for my hypothesis and research due to its association with both epithelial and, more specifically, endothelial migration and proliferation. Early work using a platform technology referred to as VitroGro (Tissue Therapies Ltd), which is comprised of vitronectin bound BP5/IGF-1, aided keratinocyte proliferation. I hypothesised that this result would translate to another epithelium - endothelium. VitroGro had no effect on endothelial proliferation or migration. Vitronectin increases the presence of Fibroblast Growth Factor (FGF) and Vascular Endothelial Growth Factor (VEGF) receptors, enhancing cell responsiveness to their respective ligands. So, although Human Microvascular Endothelial Cell line 1 (HMEC-1) VEGF receptor expression is generally low, it was hypothesised that exposure to vitronectin would up-regulate this receptor. HMEC-1 migration, but not proliferation, was enhanced by vitronectin bound VEGF, as well as vitronectin bound Epidermal Growth Factor (EGF), both of which could be used to stimulate microvascular endothelial cell migration for the purpose of transplantation. In addition to vitronectin's synergy with various growth factors, it has also been shown to play a role in haemostasis. Vitronectin binds thrombin-antithrombin III (TAT) to form a trimeric complex that takes on many of the attributes of vitronectin, such as heparin affinity, which results in its adherence to endothelium via heparan sulfate proteoglycans (HSP), followed by unaltered transcytosis through the endothelium, and ultimately its removal from the circulation. This has been documented as a mechanism designed to remove thrombin from the circulation. Equally, it could be argued that it is a mechanism for delivering vitronectin to the matrix. My results show that matrix-bound vitronectin dramatically alters the effect that conformationally altered antithrombin three (cATIII) has on proliferation of microvascular endothelial cells. cATIII stimulates HMEC-1 proliferation in the presence of matrix-bound vitronectin, as opposed to inhibiting proliferation in its absence. Binding vitronectin to tissues and organs prior to transplant, in the presence of cATIII, will have a profound effect on microvascular infiltration of the graft, by preventing occlusion of existing vessels whilst stimulating migration and proliferation of endothelium within the tissue.

Repair of calvarial defects with customized tissue-engineered bone grafts - I. Evaluation of osteogenesis in a three-dimensional culture system

Bone generation by autogenous cell transplantation in combination with a biodegradable scaffold is one of the most promising techniques being developed in craniofacial surgery. The objective of this combined in vitro and in vivo study was to evaluate the morphology and osteogenic differentiation of bone marrow derived mesenchymal progenitor cells and calvarial osteoblasts in a two-dimensional (2-D) and three-dimensional (3-D) culture environment (Part I of this study) and their potential in combination with a biodegradable scaffold to reconstruct critical-size calvarial defects in an autologous animal model [Part II of this study; see Schantz, J.T., et al. Tissue Eng. 2003;9(Suppl. 1):S-127-S-139; this issue]. New Zealand White rabbits were used to isolate osteoblasts from calvarial bone chips and bone marrow stromal cells from iliac crest bone marrow aspirates. Multilineage differentiation potential was evaluated in a 2-D culture setting. After amplification, the cells were seeded within a fibrin matrix into a 3-D polycaprolactone (PCL) scaffold system. The constructs were cultured for up to 3 weeks in vitro and assayed for cell attachment and proliferation using phase-contrast light, confocal laser, and scanning electron microscopy and the MTS cell metabolic assay. Osteogenic differentiation was analyzed by determining the expression of alkaline phosphatase (ALP) and osteocalcin. The bone marrow-derived progenitor cells demonstrated the potential to be induced to the osteogenic, adipogenic, and chondrogenic pathways. In a 3-D environment, cell-seeded PCL scaffolds evaluated by confocal laser microscopy revealed continuous cell proliferation and homogeneous cell distribution within the PCL scaffolds. On osteogenic induction mesenchymal progenitor cells (12 U/L) produce significantly higher (p < 0.05) ALP activity than do osteoblasts (2 U/L); however, no significant differences were found in osteocalcin expression. In conclusion, this study showed that the combination of a mechanically stable synthetic framework (PCL scaffolds) and a biomimetic hydrogel (fibrin glue) provides a potential matrix for bone tissue-engineering applications. Comparison of osteogenic differentiation between the two mesenchymal cell sources revealed a similar pattern.

Quality of life after total laparoscopic hysterectomy versus total abdominal hysterectomy for stage I endometrial cancer (LACE) : a randomised trial

Background: The two-stage Total Laparoscopic Hysterectomy (TLH) versus Total Abdominal Hysterectomy (TAH) for stage I endometrial cancer (LACE) randomised controlled trial was initiated in 2005. The primary objective of stage 1 was to assess whether TLH results in equivalent or improved QoL up to 6 months after surgery compared to TAH. The primary objective of stage 2 was to test the hypothesis that disease-free survival at 4.5 years is equivalent for TLH and TAH. Results addressing the primary objective of stage 1 of the LACE trial are presented here. Methods: The first 361 LACE participants (TAH n= 142, TLH n=190) were enrolled in the QoL substudy at 19 centres across Australia, New Zealand and Hong Kong, and 332 completed the QoL analysis. Randomisation was performed centrally and independently from other study procedures via a computer generated, web-based system (providing concealment of the next assigned treatment) using stratified permuted blocks of 3 and 6, and assigned patients with histologically confirmed stage 1 endometrioid endometrial adenocarcinoma and ECOG performance status <2 to TLH or TAH stratified by histological grade and study centre. No blinding of patients or study personnel was attempted. QoL was measured at baseline, 1 and 4 weeks (early), and 3 and 6 months (late) after surgery using the Functional Assessment of Cancer Therapy-General (FACT-G) questionnaire. The primary endpoint was the difference between the groups in QoL change from baseline at early and late time points (a 5% difference was considered clinically significant). Analysis was performed according to the intention-to-treat principle using generalized estimating equations on differences from baseline for the early and late QoL recovery. The LACE trial is registered with clinicaltrials.gov (NCT00096408) and the Australian New Zealand Clinical Trials Registry (CTRN12606000261516). Patients for both stages of the trial have now been recruited and are being followed up for disease-specific outcomes. Findings: The proportion of missing values at the 5%, 10% 15% and 20% differences in the FACT-G scale was 6% (12/190) in the TLH and 14% (20/142) in the TAH group. There were 8/332 conversions (2.4%, 7 of which were from TLH to TAH). In the early phase of recovery, patients undergoing TLH reported significantly greater improvement of QoL from baseline compared to TAH in all subscales except the emotional and social well-being subscales. Improvements in QoL up to 6 months post-surgery continued to favour TLH except for the emotional and social well-being of the FACT and the visual analogue scale of the EuroQoL five dimensions (EuroQoL-VAS). Length of operating time was significantly longer in the TLH group (138±43 mins), than in the TAH group at (109±34 mins; p=0.001). While the proportion of intraoperative adverse events was similar between the treatment groups (TAH 8/142, 5.6%; TLH 14/190, 7.4%; p=0.55), postoperatively, twice as many patients in the TAH group experienced adverse events of CTC grade 3+ than in the TLH group (33/142, 23.2% and 22/190, 11.6%, respectively; p=0.004). Postoperative serious adverse events occurred more frequently in patients who had a TAH (27/142, 19.0%) than a TLH (15/190, 7.9%) (p=0.002). Interpretation: QoL improvements from baseline during early and later phases of recovery, and the adverse event profile significantly favour TLH compared to TAH for patients treated for Stage I endometrial cancer.

Land use effects on soil carbon fractions in the southeastern United States. I. Management-intensive versus extensive grazing

Changes in grassland management intended to increase productivity can lead to sequestration of substantial amounts of atmospheric C in soils. Management-intensive grazing (MiG) can increase forage production in mesic pastures, but potential impacts on soil C have not been evaluated. We sampled four pastures (to 50 cm depth) in Virginia, USA, under MiG and neighboring pastures that were extensively grazed or bayed to evaluate impacts of grazing management on total soil organic C and N pools, and soil C fractions. Total organic soil C averaged 8.4 Mg C ha(-1) (22%) greater under MiG; differences were significant at three of the four sites examined while total soil N was greater for two sites. Surface (0-10 cm) particulate organic matter (POM) C increased at two sites; POM C for the entire depth increment (0-50 cm) did not differ significantly between grazing treatments at any of the sites. Mineral-associated C was related to silt plus clay content and tended to be greater under MiG. Neither soil C:N ratios, POM C, or POM C:total C ratios were accurate indicators of differences in total soil C between grazing treatments, though differences in total soil C between treatments attributable to changes in POM C (43%) were larger than expected based on POM C as a percentage of total C (24.5%). Soil C sequestration rates, estimated by calculating total organic soil C differences between treatments (assuming they arose from changing grazing management and can be achieved elsewhere) and dividing by duration of treatment, averaged 0.41 Mg C ha(-1) year(-1) across the four sites.

?ot I (What I?) - Re_Designing Beckett

This research explores a new approach to Beckett’s Not I, a work in which the spectator is asked to focus primarily on a human mouth suspended in space eight feet off the ground. The digital reconstitution of the x, y and z axes in Beckett's work has been retitled "?ot I". In "?ot I", the prescribed x, y and z axes of the original have been re-spatialised environmentally, physically and aurally to create an invigorated version of the text. In this work, it is primarily the reconstitution of spatial dynamics and time that are explored. An adaption and series of responses to Samuel Beckett's "Not I", first performed at Melbourne University and Deakin Motion.Lab, Deakin University, 2007

How I became the first Aboriginal person in Australia to lead a national education organisation

In this paper, Bronwyn Fredericks reflects on how, in 1997, she became the National President of the Council of Australian Postgraduate Associations Inc. (CAPA). The paper describes the election process faced by Fredericks, and identifies some of her key achievements during her time as National President. In becoming the National President, Bronwyn became the first Aboriginal person in Australia to lead a national education organisation. The story within this paper is told from the author’s autobiographical memory, drawing on the cultural, social and political context in which the story and the author were (and are) situated (Wojecki 2007). In this way the story teller reveals story lines which have not previously been articulated (Wojecki 2007). Throughout this paper, Fredericks ‘re-stories’ her experiences of leadership.

'I am smart and I am not joking': Aiming high in the middle years of schooling

In this paper, we draw on accounts from students to inform a Middle Schooling movement that has been variously described as "arrested", "unfinished" and "exhausted". We propose that if the Middle Schooling movement is to understand the changing worlds of students and develop new approaches in the middle years of schooling, then it is important to draw on the insights that individual students can provide by conducting research with "students-as-informants". The early adolescent informants to this paper report high hopes for their futures (despite their lower socio-economic surroundings), which reinforces the importance of supporting successful learner identities and highlights the role of schooling in the decline of adolescent student aspirations. However, their insights did not stop at the individual learner, with students also identifying cultural and structural constraints to reform. As such, we argue that students may be both an important resource for inquiry into individual school reform and for the Middle Schooling movement internationally.

"I'm a designer, get me out of tech class!" : closing the gap between design and technology education

The following paper explores the use of collaborative pedagogical approaches to advance foundational architectural design education, by linking design process to sustainable technology principles. After a brief discussion on architectural design education, the mentioned collaborative approach is described. This approach facilitates students’ exchange of knowledge between two courses, despite no explicit/assessable requirement to do so. The result for the students is deeper learning and a design process that is enriched through collaboration with sustainable technology. The success of this approach has been measured through questionnaires, evaluation surveys, and a comparative assessment of students common to both courses. The paper focuses on the challenges and innovations in connecting architectural design and technology education, where students are encouraged to implement lessons learnt, thereby closing the gap that these courses have traditionally represented.

‘You can interview me, but I don’t have a story’: Local Accounts of Wandering Women, Rewondering Queensland Landscapes.

This paper investigates how contemporary works of women’s travel writing are reworking canonical formations of environmental literature by presenting imaginative accounts of travel writing that are both literal and metaphorical. In this context, the paper considers how women who travel/write may intersect the spatial hybridities of travel writing and nature writing, and in doing so, create a new genre of environmental literature that is not only ecologically sensitive but gendered. As the role of female travel writers in generating this knowledge is immense but largely unexamined, this paper will investigate how a feminist geography can be applied, both critically and creatively, to local accounts of travel. It will draw on my own travels around Queensland in an attempt to explore how many female storytellers situate themselves, in and against, various discourses of mobility and morality.

“Now I know their secrets” : Kineikonic texts in the literacy classroom

Kineikonic texts – sites of the moving image – are increasingly prevalent with the rise of digital television, Web 2.0 tools, broadband Internet, and sophisticated mobile technologies. Digital practices are changing the shape of the literacy curriculum, calling for new metalanguages to describe digital and multimodal texts. This paper combines multiliteracies and functional approaches to map conventional and new textual features of a popular kineikonic text – the claymation movie. Enlivened with data from an ethnically diverse, Year 6 classroom, the author outlines filmic conventions to enable teachers and students to analyse and design movies

‘You’re a bad driver but I just made a mistake’ : attribution differences between the ‘victims’ and ‘perpetrators’ of scenario-based aggressive driving incidents

Driver aggression is an increasing concern for motorists, with some research suggesting that drivers who behave aggressively perceive their actions as justified by the poor driving of others. Thus attributions may play an important role in understanding driver aggression. A convenience sample of 193 drivers (aged 17-36) randomly assigned to two separate roles (‘perpetrators’ and ‘victims’) responded to eight scenarios of driver aggression. Drivers also completed the Aggression Questionnaire and Driving Anger Scale. Consistent with the actor-observer bias, ‘victims’ (or recipients) in this study were significantly more likely than ‘perpetrators’ (or instigators) to endorse inadequacies in the instigator’s driving skills as the cause of driver aggression. Instigators were significantly more likely attribute the depicted behaviours to external but temporary causes (lapses in judgement or errors) rather than stable causes. This suggests that instigators recognised drivers as responsible for driving aggressively but downplayed this somewhat in comparison to ‘victims’/recipients. Recipients and instigators agreed that the behaviours were examples of aggressive driving but instigators appeared to focus on the degree of intentionality of the driver in making their assessments while recipients appeared to focus on the safety implications. Contrary to expectations, instigators gave mean ratings of the emotional impact of driving aggression on recipients that were higher in all cases than the mean ratings given by the recipients. Drivers appear to perceive aggressive behaviours as modifiable, with the implication that interventions could appeal to drivers’ sense of self-efficacy to suggest strategies for overcoming plausible and modifiable attributions (e.g. lapses in judgement; errors) underpinning behaviours perceived as aggressive.