In vitro and in vivo studies on protease-activated receptor-2

Protease-activated receptor-2 (PAR2) is a G protein coupled receptor (GPCR) that is activated by proteolytic cleavage of its amino terminal domain by trypsin-like serine proteases. Cleavage of this receptor exposes a neoepitope, termed the tethered ligand (TL), which binds intramolecularly within the receptor to stimulate signal transduction via coupled G proteins. PAR2-mediated signal transduction is also experimentally stimulated by hexapeptides (agonist peptides; APs) that are homologous to the TL sequence. Due to the irreversible nature of PAR2 proteolysis, downstream signal transduction is tightly regulated. Following activation, PAR2 is rapidly uncoupled from downstream signalling by the post-translational modifications phosphorylation and ubiquination which facilitate interactions with â- arrestin. This scaffolding protein couples PAR2 to the internalisation machinery initiating its desensitisation and trafficking through the early and late endosomes followed by receptor degradation. PAR2 is widely expressed in mammalian tissues with key roles for this receptor in cardiovascular, respiratory, nervous and musculoskeletal systems. This receptor has also been linked to pathological states with aberrant expression and signalling noted in several cancers. In prostate cancer, PAR2 signalling induces migration and proliferation of tumour derived cell lines, while elevated receptor expression has been noted in malignant tissues. Importantly, a role for this receptor has also been suggested in prostate cancer bone metastasis as coexpression of PAR2 and a proteolytic activator has been demonstrated by immunohistochemical analysis. Based on these data, the primary focus of this project has been on two aspects of PAR2 biology. The first is characterisation of cellular mechanisms that regulate PAR2 signalling and trafficking. The second aspect is the role of this receptor in prostate cancer bone metastasis. In addition, to permit these studies, it was first necessary to evaluate the specificity of the commercially available anti-PAR2 antibodies SAM11, C17, N19 and H99. The evaluation of the four commercially available antibodies was assessed using four techniques: immunoprecipitation; Western blot analysis; immunofluorescence; and flow cytometry. These approaches demonstrated that three of the antibodies efficiently detect ectopically expressed PAR2 by each of these techniques. A significant finding from this study was that N19 was the only antibody able to specifically detect N-glycosylated endogenous PAR2 by Western blot analysis. This analysis was performed on lysates from prostate cancer derived cell lines and tissue derived from wildtype and PAR2 knockout mice. Importantly, further evaluation demonstrated that this antibody also efficiently detects endogenous PAR2 at the cell surface by flow cytometry. The anti-PAR2 antibody N19 was used to explore the in vitro role of palmitoylation, the post-translational addition of palmitate, in PAR2 signalling, trafficking, cell surface expression and desensitization. Significantly, use of the palmitoylation inhibitor 2-bromopalmitate indicated that palmitate addition is important in trafficking of PAR2 endogenously expressed by prostate cancer cell lines. This was supported by palmitate labelling experiments using two approaches which showed that PAR2 stably expressed by CHO cells is palmitoylated and that palmitoylation occurs on cysteine 361. Another key finding from this study is that palmitoylation is required for optimal PAR2 signalling as Ca2+ flux assays indicated that in response to trypsin agonism, palmitoylation deficient PAR2 is ~9 fold less potent than wildtype receptor with a reduction of about 33% in the maximum signal induced via the mutant receptor. Confocal microscopy, flow cytometry and cell surface biotinylation analyses demonstrated that palmitoylation is required for efficient cell surface expression of PAR2. Importantly, this study also identified that palmitoylation of this receptor within the Golgi apparatus is required for efficient agonist-induced rab11amediated trafficking of PAR2 to the cell surface. Interestingly, palmitoylation is also required for receptor desensitization, as agonist-induced â-arrestin recruitment and receptor degradation were markedly reduced in CHO-PAR2-C361A cells compared with CHO-PAR2 cells. Collectively, these data provide new insights on the life cycle of PAR2 and demonstrate that palmitoylation is critical for efficient signalling, trafficking, cell surface localization and degradation of this receptor. This project also evaluated PAR2 residues involved in ligand docking. Although the extracellular loop (ECL)2 of PAR2 is known to be required for agonist-induced signal transduction, the binding pocket for receptor agonists remains to be determined. In silico homology modelling, based on a crystal structure for the prototypical GPCR rhodopsin, and ligand docking were performed to identify PAR2 transmembrane (TM) amino acids potentially involved in agonist binding. These methods identified 12 candidate residues that were mutated to examine the binding site of the PAR2 TL, revealed by trypsin cleavage, as well as of the soluble ligands 2f-LIGRLO-NH2 and GB110, which are both structurally based on the AP SLIGRLNH2. Ligand binding was evaluated from the impact of the mutated residues on PAR2-mediated calcium mobilisation. An important finding from these experiments was that mutation of residues Y156 and Y326 significantly reduced 2f-LIGRLO-NH2 and GB110 agonist activity. L307 was also important for GB110 activity. Intriguingly, mutation of PAR2 residues did not alter trypsin-induced signalling to the same extent as for the soluble agonists. The reason for this difference remains to be further examined by in silico and in vitro experimentation and, potentially, crystal structure studies. However, these findings identified the importance of TM domains in PAR2 ligand docking and will enhance the design of both PAR2 agonists and potentially agents to inhibit signalling (antagonists). The potential importance of PAR2 in prostate cancer bone metastasis was examined using a mouse model. In patients, prostate cancer bone metastases cause bone growth by disrupting bone homeostasis. In an attempt to mimic prostate cancer growth in bone, PAR2 responsive 22Rv1 prostate cancer cells, which form mixed osteoblastic and osteolytic lesions, were injected into the proximal aspect of mouse tibiae. A role for PAR2 was assessed by treating these mice with the recently developed PAR2 antagonist GB88. As controls, animals bearing intra-tibial tumours were also treated with vehicle (olive oil) or the prostate cancer chemotherapeutic docetaxel. The effect of these treatments on bone was examined radiographically and by micro-CT. Consistent with previous studies, 22Rv1 tumours caused osteoblastic periosteal spicule formation and concurrent osteolytic bone loss. Significantly, blockade of PAR2 signalling reduced the osteoblastic and osteolytic phenotype of 22Rv1 tumours in bone. No bone defects were detected in mice treated with docetaxel. These qualitative data will be followed in the future by quantitative micro-CT analysis as well as histology and histomorphometry analysis of already collected tissues. Nonetheless, these preliminary experiments highlight a potential role for PAR2 in prostate cancer growth in bone. In summary, in vitro studies have defined mechanisms regulating PAR2 activation, downstream signalling and trafficking and in vivo studies point to a potential role for this receptor in prostate cancer bone metastasis. The outcomes of this project are that a greater understanding of the biology of PAR2 may lead to the development of strategies to modulate the function of this receptor in disease.

Re-made in Asia : transformation across Asian markets and popular culture

The thesis is an examination of how Japanese popular culture products are remade (rimeiku). Adaptation of manga, anime and television drama, from one format to another, frequently occurs within Japan. The rights to these stories and texts are traded in South Korea and Taiwan. The ‘spin-off’ products form part of the Japanese content industry. When products are distributed and remade across geographical boundaries, they have a multi-dimensional aspect and potentially contribute to an evolving cultural re-engagement between Japan and East Asia. The case studies are the television dramas Akai Giwaku and Winter Sonata and two manga, Hana yori Dango and Janguru Taitei. Except for the television drama Winter Sonata these texts originated in Japan. Each study shows how remaking occurs across geographical borders. The study argues that Japan has been slow to recognise the value of its popular culture through regional and international media trade. Japan is now taking steps to remedy this strategic shortfall to enable the long-term viability of the Japanese content industry. The study includes an examination of how remaking raises legal issues in the appropriation of media content. Unauthorised copying and piracy contributes to loss of financial value. To place the three Japanese cultural products into a historical context, the thesis includes an overview of Japanese copying culture from its early origins through to the present day. The thesis also discusses the Meiji restoration and the post-World War II restructuring that resulted in Japan becoming a regional media powerhouse. The localisation of Japanese media content in South Korea and Taiwan also brings with it significant cultural influences, which may be regarded as contributing to a better understanding of East Asian society in line with the idea of regional ‘harmony’. The study argues that the commercial success of Japanese products beyond Japan is governed by perceptions of the quality of the story and by the cultural frames of the target audience. The thesis draws on audience research to illustrate the loss or reinforcement of national identity as a consequence of cross-cultural trade. The thesis also examines the contribution to Japanese ‘soft power’ (Nye, 2004, p. x). The study concludes with recommendations for the sustainability of the Japanese media industry.

Surveillance of dengue fever virus: a review of epidemiological models and early warning systems

Dengue fever is one of the world’s most important vector-borne diseases. The transmission area of this disease continues to expand due to many factors including urban sprawl, increased travel and global warming. Current preventative techniques are primarily based on controlling mosquito vectors as other prophylactic measures, such as a tetravalent vaccine are unlikely to be available in the foreseeable future. However, the continually increasing dengue incidence suggests that this strategy alone is not sufficient. Epidemiological models attempt to predict future outbreaks using information on the risk factors of the disease. Through a systematic literature review, this paper aims at analyzing the different modeling methods and their outputs in terms of accurately predicting disease outbreaks. We found that many previous studies have not sufficiently accounted for the spatio-temporal features of the disease in the modeling process. Yet with advances in technology, the ability to incorporate such information as well as the socio-environmental aspect allowed for its use as an early warning system, albeit limited geographically to a local scale.

The causes of sinuous crustal-scale deformation patterns in hot orogens: Evidence from scaled analogue experiments and the southern Central Andes

The cause of upper-crustal segmentation into rhomb-shaped, shear zone-bound domains associated with contractional sedimentary basins in hot, wide orogens is not well understood. Here we use scaled multilayered analogue experiments to investigate the role of an orogen-parallel crustal-strength gradient on the formation of such structures. We show that the aspect ratio and size of domains, the sinuous character and abundance of transpressional shear zones vary with the integrated mechanical strength of crust. Upper-crustal deformation patterns and the degree of strain localization in the experiments are controlled by the ratio between the brittle and ductile strength in the model crust as well as gradients in tectonic and buoyancy forces. The experimental results match the first-order kinematic and structural characteristics of the southern Central Andes and provide insight on the dynamics of underlying deformation patterns in hot, wide orogens.

Do students and lecturers actively use collaboration tools in learning management systems?

In recent years there has been a large emphasis placed on the need to use Learning Management Systems (LMS) in the field of higher education, with many universities mandating their use. An important aspect of these systems is their ability to offer collaboration tools to build a community of learners. This paper reports on a study of the effectiveness of an LMS (Blackboard©) in a higher education setting and whether both lecturers and students voluntarily use collaborative tools for teaching and learning. Interviews were conducted with participants (N=67) from the faculties of Science and Technology, Business, Health and Law. Results from this study indicated that participants often use Blackboard© as an online repository of learning materials and that the collaboration tools of Blackboard© are often not utilised. The study also found that several factors have inhibited the use and uptake of the collaboration tools within Blackboard©. These have included structure and user experience, pedagogical practice, response time and a preference for other tools.

Experimental studies of the shear behavior and strength of LiteSteel beams with stiffened web openings

Abstract: LiteSteel beam (LSB) is a new cold-formed steel hollow flange channel section produced using a patented manufacturing process. It is commonly used as flexural members in residential, industrial and commercial buildings. Current practice in flooring systems is to include openings in the web element of floor joists or bearers so that building services can be located within them. Test results have shown that the shear capacity of LSBs can be reduced considerably by the inclusion of web openings. A cost effective method of eliminating the detrimental effects of a large web opening is to attach suitable stiffeners around the web openings of LSBs. A detailed experimental study consisting of 17 shear tests was therefore undertaken to investigate the shear behaviour and strength of LSBs with stiffened circular web openings. Both plate and stud stiffeners with varying sizes and thicknesses were attached to the web elements of LSBs using a number of screw-fastening arrangements in order to develop a suitable stiffening arrangement for LSBs. Simply supported test specimens of LSBs with an aspect ratio of 1.5 were loaded at mid-span until failure. This paper presents the details of this experimental study of LSBs with stiffened web openings, and the results of their shear capacities and associated behavioural characteristics. Suitable screw-fastened plate stiffener arrangements have been recommended in order to restore the original shear capacity of LSBs.

Importance of a resilient air services network to Australian remote, rural, and regional communities

Rural, regional, and remote settlements in Australia require resilient infrastructure to remain sustainable in a context characterized by frequent large-scale natural disasters, long distances between urban centers, and the pressures of economic change. A critical aspect of this infrastructure is the air services network, a system of airports, aircraft operators, and related industries that enables the high-speed movement of people, goods, and services to remote locations. A process of deregulation during the 1970s and 1980s resulted in many of these airports passing into local government and private ownership, and the rationalization of the industry saw the closure of a number of airlines and airports. This paper examines the impacts of deregulation on the resilience of air services and the contribution that they make to regional and rural communities. In particular, the robustness, redundancy, resourcefulness, and rapidity of the system are examined. The conclusion is that while the air services network has remained resilient in a situation of considerable change, the pressures of commercialization and the tendency to manage aspects of the system in isolation have contributed to a potential decrease in overall resilience.

Making sense of mental illness as a full human experience : Perspective of illness and recovery held by people with a mental illness living in the community.

Recovery is a highly contextualized concept amid divergent interpretations and unique experiences. There is substantial current interest in building evidence about recovery from mental illness in order to inform best practice founded in the ways people find to live productive and meaningful lives. This paper presents some accounts related to recovery and illness expressed by eight people through a Participatory Action Research project. The research facilitated entry to the subjective experiences of living in the community as an artist with a mental illness. The people in the research shared an integrated understanding of illness, recovery and identity. Their understanding provided insight into mental illness as an inseparable aspect of who they were. Further, specific issue was raised of recovery as a clinical term with a requirement to meet distinct conventions of recovery. This paper emphasizes that being ill and being well, for the person with a mental illness, is a dynamic and complex development not easily explained or transformed into uniform process or outcomes. Attempts to establish an integral or consensual approach to recovery has, to date, disregarded mental illness as a full human experience. This paper argues that broader frameworks for thinking and responding to the dynamic processes of mental illness and recovery are needed and require acknowledgment of competing and contradictory ideas.

Children, young people and their social and spatial citizenship rights

The use of public space by children and young people is a contentious issue in a number of developed and developing countries and a range of measures are frequently deployed to control the public space which usually deny the rights of children and young people to claim the space for their use. Child and youth curfews, oppressive camera surveillance and the unwarranted attentions of police and private security personnel as control measures in public space undermine attempts to secure greater participation by children and young people in constructing positive strategies to address concerns that impact on them and others in a local area. Evidence from research in Scotland undertaken by Article 12 (2000) suggests that young people felt strongly that they did not count in local community matters and decision making and the imposition on them of a curfew by the adult world of the local area created resentment both at the harshness of the measure and disappointment at an opportunity lost to be consulted and involved in dealing with perceived problems of the locality. This is an important cluster of linked issues as Brown (1998:116) argues that young people are ‘selectively constructed as “problem” and “other” with their concerns marginalised, their lifestyles problematised and their voices subdued’, and this flows into their use of public space as their claims to its use as an aspect of social citizenship are usually cast as inferior or rejected as they ‘stand outside the formal polity’ as ‘non persons’. This has major implications for the ways in which young people view their position in a community as many report a feeling of not being wanted, valued or tolerated. The ‘youth question’ according to Davis (1990) acts as a form of ‘screen’ on which observers and analysts project hopes and fears about the state of society, while in the view of Loader (1996:89) the ‘question of young people’ sits within a discourse comprising two elements, the one being youth, particularly young males, as the ‘harbinger of often unwelcome social change and threat’ and the other element ‘constructs young people as vulnerable’. This discourse of threat is further exemplified in the separation of children from teenagers as Valentine (1996) suggests, the treatment of younger children using public space is often dramatically different to that of older children and the most feared stage of all, 'youth'

Imaging affect : abstraction and the echo of the unknowable

As an artist my primary interest is in the abstract, that is in images of the imageless. I am curious about the emergence of pictorial significance and content from this unknowable space. To speak of the significance of an imageless image is also to speak of its affect. I aim to explore this both theoretically and practically. Theoretically I will explore affect through the late work of Lyotard and his notion of the affect-phrase. This is an under-examined aspect of Lyotard and demarcates a valuable way to look at the origins, impact and ramifications of affect for art. Practically I will apply these understandings to the development of my own creative work which includes both painting and digital work. My studio practice moves towards exploring the unfamiliar through the powerful and restless silence of affect.In this intense space each work or body of work 'leaks' into the next occasioning a sense of borderlessness, or of uncertainty. This interpenetration and co-mingling of conceptual and material terrains combines to present temporal and spatial slippages evident within the works themselves and their making, but it is also evident in bodies of work across the chronology of their making. Through a mapping of my own painting and digital arts practice and the utilisation of Lyotard’s notion of the affect -phrase I aim to describe the action of this ‘charged emptiness’ on creativity and explore and explain its significance on that we call image and its animation of what we call critical discourse.

Cultural relativity as exchange : J.W. Lindt’s 'bush character and Aboriginal' from 'The Grafton Album', circa 1872

J.W.Lindt’s Colonial man and Aborigine image from the GRAFTON ALBUM: “On chemistry and optics all does not depend, art must with these in triple union blend” (text from J.W. Lindt’s photographic backing card) In this paper, I follow an argument that Lindt held a position in his particular colonial environment where he was simultaneously both an insider and an outsider and that such a position may be considered prerequisite in stimulating exchange. A study of the transition of J.W. Lindt in Grafton, N.S.W. in the 1860s from a traveller to a migrant and subsequently to a professional photographer, as well as Lindt’s photographic career, which evolved through strategic action and technical approaches to photography, bears witness to his cultural relativity. One untitled photograph from this period of work constructs a unique commentary of Australian colonial life that illustrates a non-hegemonic position, particularly as it was included in one of the first albums of photographs of Aborigines that Lindt gifted to an illustrious person (in this case the Mayor of Grafton). As in his other studio constructions, props and backdrops were arranged and sitters were positioned with care, but this photograph is the only one in the album that includes a non-Aborigine in a relationship to an Aborigine. An analysis of the props, technical details of the album and the image suggests a reconciliatory aspect that thwarts the predominant attitudes towards Aborigines in the area at that time.

A model for enhancing assessment and teaching practice at the coalface : Insights from a Fellow-In-Residence engagement program

Australian universities are currently engaging with new governmental policies and regulations that require them to demonstrate enhanced quality and accountability in teaching and research. The development of national academic standards for learning outcomes in higher education is one such instance of this drive for excellence. These discipline-specific standards articulate the minimum, or Threshold Learning Outcomes, to be addressed by higher education institutions so that graduating students can demonstrate their achievement to their institutions, accreditation agencies, and industry recruiters. This impacts not only on the design of Engineering courses (with particular emphasis on pedagogy and assessment), but also on the preparation of academics to engage with these standards and implement them in their day-to-day teaching practice on a micro level. This imperative for enhanced quality and accountability in teaching is also significant at a meso level, for according to the Australian Bureau of Statistics, about 25 per cent of teachers in Australian universities are aged 55 and above and more than 54 per cent are aged 45 and above (ABS, 2006). A number of institutions have undertaken recruitment drives to regenerate and enrich their academic workforce by appointing capacity-building research professors and increasing the numbers of early- and mid-career academics. This nationally driven agenda for quality and accountability in teaching permeates also the micro level of engineering education, since the demand for enhanced academic standards and learning outcomes requires both a strong advocacy for a shift to an authentic, collaborative, outcomes-focused education and the mechanisms to support academics in transforming their professional thinking and practice. Outcomes-focused education means giving greater attention to the ways in which the curriculum design, pedagogy, assessment approaches and teaching activities can most effectively make a positive, verifiable difference to students’ learning. Such education is authentic when it is couched firmly in the realities of learning environments, student and academic staff characteristics, and trustworthy educational research. That education will be richer and more efficient when staff works collaboratively, contributing their knowledge, experience and skills to achieve learning outcomes based on agreed objectives. We know that the school or departmental levels of universities are the most effective loci of changes in approaches to teaching and learning practices in higher education (Knight & Trowler, 2000). Heads of Schools are being increasingly entrusted with more responsibilities - in addition to setting strategic directions and managing the operational and sometimes financial aspects of their school, they are also expected to lead the development and delivery of the teaching, research and other academic activities. Guiding and mentoring individuals and groups of academics is one critical aspect of the Head of School’s role. Yet they do not always have the resources or support to help them mentor staff, especially the more junior academics. In summary, the international trend in undergraduate engineering course accreditation towards the demonstration of attainment of graduate attributes poses new challenges in addressing academic staff development needs and the assessment of learning. This paper will give some insights into the conceptual design, implementation and empirical effectiveness to date, of a Fellow-In-Residence Engagement (FIRE) program. The program is proposed as a model for achieving better engagement of academics with contemporary issues and effectively enhancing their teaching and assessment practices. It will also report on the program’s collaborative approach to working with Heads of Schools to better support academics, especially early-career ones, by utilizing formal and informal mentoring. Further, the paper will discuss possible factors that may assist the achievement of the intended outcomes of such a model, and will examine its contributions to engendering an outcomes-focussed thinking in engineering education.

Measuring a destination’s brand equity between 2003 and 2012 using the consumer-based brand equity (CBBE) hierarchy

Given the increasing investments being made in brand development by destination marketing organisations (DMO) since the 1990s, including rebranding and repositioning, more research is needed to enhance understanding of how to effectively monitor destination brand performance over time. This paper reports the results of a study of brand performance of a competitive set of destinations, in their most important market, between 2003 and 2012. Brand performance was measured from the perspective of consumer perceptions, based on the concept of consumer-based brand equity (CBBE). A structured questionnaire was administered to different samples in 2003, 2007 and 2012. The results indicated minimal changes in perceptions of the five destinations over the 10 year period. Due to the commonality of challenges faced by DMOs worldwide, it is suggested the CBBE hierarchy provides destination marketers with a practical tool for evaluating brand performance over time; in terms of measures of effectiveness of past marketing communications, as well as indicators of future performance. In addition, and importantly, CBBE also provides transparent accountability measures for stakeholders. While the topic of destination image has been one of the most popular in the tourism literature, there has been a paucity of research published in relation to the temporal aspect of consumer perceptions. This is a rare investigation into the measurement of perceptions of destinations over a 10 year period.

The role of representative design in talent development : a comment on “Talent identification and promotion programmes of Olympic athletes”

A recent comment in the Journal of Sports Sciences (MacNamara & Collins, 2011) highlighted some major concerns with the current structure of talent identification and development (TID) programmes of Olympic athletes (e.g. Gulbin, 2008; Vaeyens, Gullich, Warr, & Philippaerts, 2009). In a cogent commentary, MacNamara and Collins (2011) provided a short review of the extant literature, which was both timely and insightful. Specifically, they criticised the ubiquitous one-dimensional ‘physically-biased’ attempts to produce world class performers, emphasising the need to consider a number of key environmental variables in a more multi-disciplinary perspective. They also lamented the wastage of talent, and alluded to the operational and opportunistic nature of current talent transfer programmes. A particularly compelling aspect of the comment was their allusion to high profile athletes who had ‘failed’ performance evaluation tests and then proceeded to succeed in that sport. This issue identifies a problem with current protocols for evaluating performance and is a line of research that is sorely needed in the area of talent development. To understand the nature of talent wastage that might be occurring in high performance programmes in sport, future empirical work should seek to follow the career paths of ‘successful’ and ‘unsuccessful’ products of TID programmes, in comparative analyses. Pertinent to the insights of MacNamara and Collins (2011), it remains clear that a number of questions have not received enough attention from sport scientists interested in talent development, including: (i) why is there so much wastage of talent in such programmes? And (ii), why are there so few reported examples of successful talent transfer programmes? These questions highlight critical areas for future investigation. The aim of this short correspondence is to discuss these and other issues researchers and practitioners might consider, and to propose how an ecological dynamics underpinning to such investigations may help the development of existing protocols...

Bubble extinction in Hele-Shaw flow with surface tension and kinetic undercooling regularisation

We perform an analytic and numerical study of an inviscid contracting bubble in a two-dimensional Hele-Shaw cell, where the effects of both surface tension and kinetic undercooling on the moving bubble boundary are not neglected. In contrast to expanding bubbles, in which both boundary effects regularise the ill-posedness arising from the viscous (Saffman-Taylor) instability, we show that in contracting bubbles the two boundary effects are in competition, with surface tension stabilising the boundary, and kinetic undercooling destabilising it. This competition leads to interesting bifurcation behaviour in the asymptotic shape of the bubble in the limit it approaches extinction. In this limit, the boundary may tend to become either circular, or approach a line or "slit" of zero thickness, depending on the initial condition and the value of a nondimensional surface tension parameter. We show that over a critical range of surface tension values, both these asymptotic shapes are stable. In this regime there exists a third, unstable branch of limiting self-similar bubble shapes, with an asymptotic aspect ratio (dependent on the surface tension) between zero and one. We support our asymptotic analysis with a numerical scheme that utilises the applicability of complex variable theory to Hele-Shaw flow.