Sensitivity of edge detection methods for quantifying cell migration assays

Quantitative imaging methods to analyze cell migration assays are not standardized. Here we present a suite of two–dimensional barrier assays describing the collective spreading of an initially–confined population of 3T3 fibroblast cells. To quantify the motility rate we apply two different automatic image detection methods to locate the position of the leading edge of the spreading population after 24, 48 and 72 hours. These results are compared with a manual edge detection method where we systematically vary the detection threshold. Our results indicate that the observed spreading rates are very sensitive to the choice of image analysis tools and we show that a standard measure of cell migration can vary by as much as 25% for the same experimental images depending on the details of the image analysis tools. Our results imply that it is very difficult, if not impossible, to meaningfully compare previously published measures of cell migration since previous results have been obtained using different image analysis techniques and the details of these techniques are not always reported. Using a mathematical model, we provide a physical interpretation of our edge detection results. The physical interpretation is important since edge detection algorithms alone do not specify any physical measure, or physical definition, of the leading edge of the spreading population. Our modeling indicates that variations in the image threshold parameter correspond to a consistent variation in the local cell density. This means that varying the threshold parameter is equivalent to varying the location of the leading edge in the range of approximately 1–5% of the maximum cell density.

Background to the development of a curriculum for the history of “cyber” and “communications” security

For any discipline to be regarded as a professional undertaking by which its members may be treated as true “professionals” in a specific area, practitioners must clearly understand that discipline’s history as well as the place and significance of that history in current practice as well as its relevance to available technologies and artefacts at the time. This is common for many professional disciplines such as medicine, pharmacy, engineering, law and so on but not yet, this paper submits, in information technology. Based on twenty five elapsed years of experience in developing and delivering Cybersecurity courses at undergraduate and postgraduate levels, this paper proposes a rationale and set of differing perspectives for the planning and development of curricula relevant to the delivery of appropriate courses in the history of cybersecurity or information assurance to information and communications technology (ICT) students and thus to potential information technology professionals.

The importance of individual characteristics in experimental economic research

This thesis advances the knowledge of behavioural economics on the importance of individual characteristics – such as gender, personality or culture – for choices relevant to labour and insurance markets. It does so using economic experiments, survey tools and physiological data, collected in economic laboratories and in the field. More specifically, the thesis includes 5 experimental economic studies investigating individual-specific characteristics (gender, age, personality, cultural background) in decisions influenced by risk attitudes and social preferences. One of these characteristics is the physiological state of decision-makers, measured by heart rate variability. The results show that individual-specific characteristics play an important role for choices affected by social preferences, a finding to a lesser degree observable for risk preferences. This finding is confirmed under revealed incentivised choices and when studying (latent) physiological responses of decision-makers.

Cell migration and proliferation on homogeneous and non-homogeneous domains : modelling on the scale of individuals and populations

Cell migration is a behaviour critical to many key biological effects, including wound healing, cancerous cell invasion and morphogenesis, the development of an organism from an embryo. However, given that each of these situations is distinctly different and cells are extremely complicated biological objects, interest lies in more basic experiments which seek to remove conflating factors and present a less complex environment within which cell migration can be experimentally examined. These include in vitro studies like the scratch assay or circle migration assay, and ex vivo studies like the colonisation of the hindgut by neural crest cells. The reduced complexity of these experiments also makes them much more enticing as problems to mathematically model, like done here. The primary goal of the mathematical models used in this thesis is to shed light on which cellular behaviours work to generate the travelling waves of invasion observed in these experiments, and to explore how variations in these behaviours can potentially predict differences in this invasive pattern which are experimentally observed when cell types or chemical environment are changed. Relevant literature has already identified the difficulty of distinguishing between these behaviours when using traditional mathematical biology techniques operating on a macroscopic scale, and so here a sophisticated individual-cell-level model, an extension of the Cellular Potts Model (CPM), is been constructed and used to model a scratch assay experiment. This model includes a novel mechanism for dealing with cell proliferations that allowed for the differing properties of quiescent and proliferative cells to be implemented into their behaviour. This model is considered both for its predictive power and used to make comparisons with the travelling waves which result in more traditional macroscopic simulations. These comparisons demonstrate a surprising amount of agreement between the two modelling frameworks, and suggest further novel modifications to the CPM that would allow it to better model cell migration. Considerations of the model’s behaviour are used to argue that the dominant effect governing cell migration (random motility or signal-driven taxis) likely depends on the sort of invasion demonstrated by cells, as easily seen by microscopic photography. Additionally, a scratch assay simulated on a non-homogeneous domain consisting of a ’fast’ and ’slow’ region is also used to further differentiate between these different potential cell motility behaviours. A heterogeneous domain is a novel situation which has not been considered mathematically in this context, nor has it been constructed experimentally to the best of the candidate’s knowledge. Thus this problem serves as a thought experiment used to test the conclusions arising from the simulations on homogeneous domains, and to suggest what might be observed should this non-homogeneous assay situation be experimentally realised. Non-intuitive cell invasion patterns are predicted for diffusely-invading cells which respond to a cell-consumed signal or nutrient, contrasted with rather expected behaviour in the case of random-motility-driven invasion. The potential experimental observation of these behaviours is demonstrated by the individual-cell-level model used in this thesis, which does agree with the PDE model in predicting these unexpected invasion patterns. In the interest of examining such a case of a non-homogeneous domain experimentally, some brief suggestion is made as to how this could be achieved.

Fractional models for the migration of biological cells in complex spatial domains

Travelling wave phenomena are observed in many biological applications. Mathematical theory of standard reaction-diffusion problems shows that simple partial differential equations exhibit travelling wave solutions with constant wavespeed and such models are used to describe, for example, waves of chemical concentrations, electrical signals, cell migration, waves of epidemics and population dynamics. However, as in the study of cell motion in complex spatial geometries, experimental data are often not consistent with constant wavespeed. Non-local spatial models have successfully been used to model anomalous diffusion and spatial heterogeneity in different physical contexts. In this paper, we develop a fractional model based on the Fisher-Kolmogoroff equation and analyse it for its wavespeed properties, attempting to relate the numerical results obtained from our simulations to experimental data describing enteric neural crest-derived cells migrating along the intact gut of mouse embryos. The model proposed essentially combines fractional and standard diffusion in different regions of the spatial domain and qualitatively reproduces the behaviour of neural crest-derived cells observed in the caecum and the hindgut of mouse embryos during in vivo experiments.

Vietnamese in Australia

Vietnamese-Australians live in Australia, a large island continent. The physical contrast between Vietnam and Australia is remarked upon by many Vietnamese in their migration stories. Whereas Vietnam is remembered as an interlinked sensual and social world, Australia is often viewed as a harsh, spacious, empty, dry continent. Australia is located in a regional Asian context, but this location has always been culturally and politically problematic, as it historically attempted to define itself as a "white" European nation in the Southern Hemisphere(Ang, 2000, p. xiii; McNamara & Coughlan, 1997, p. 1). During the Gold Rush period in the late 1800s, when there was widespread opposition to Chinese labor, Australia implemented a "White Australia" policy, although there were historically a significant number of Australians of Asian background. This exclusionary immigration policy was effectively overturned in the 1970s with the acceptance of a large number of refugees from Vietnam, Cambodia, and Laos in 1975. Vietnamese-Australians live predominantly in urban areas with over three quarters living in Sydney and Melbourne, the two largest cities. Within these two cities they are also highly concentrated in ethnically diverse suburbs, most living in areas with more than 1,000 residents born in Vietnam (Viviani, 1996, p. 49). However, Jupp (Jupp et al., 1990; Jupp, 1993) has argued that these areas are also zones of transition, with much movement in and out...

Environments for Healthy Living (EFHL) Griffith Birth Cohort Study : background and methods

The health of an individual is determined by the interaction of genetic and individual factors with wider social and environmental elements. Public health approaches to improving the health of disadvantaged populations will be most effective if they optimise influences at each of these levels, particularly in the early part of the life course. In order to better ascertain the relative contribution of these multi-level determinants there is a need for robust studies, longitudinal and prospective in nature, that examine individual, familial, social and environmental exposures. This paper describes the study background and methods, as it has been implemented in an Australian birth cohort study, Environments for Healthy Living (EFHL): The Griffith Study of Population Health. EFHL is a prospective, multi-level, multi-year longitudinal birth cohort study, designed to collect information from before birth through to adulthood across a spectrum of eco-epidemiological factors, including genetic material from cord-blood samples at birth, individual and familial factors, to spatial data on the living environment. EFHL commenced the pilot phase of recruitment in 2006 and open recruitment in 2007, with a target sample size of 4000 mother/infant dyads. Detailed information on each participant is obtained at birth, 12-months, 3-years, 5-years and subsequent three to five yearly intervals. The findings of this research will provide detailed evidence on the relative contribution of multi-level determinants of health, which can be used to inform social policy and intervention strategies that will facilitate healthy behaviours and choices across sub-populations.

PEGylation of lysine residues reduces the pro-migratory activity of IGF-I

Background: The insulin-like growth factor (IGF) system is composed of ligands and receptors which regulate cell proliferation, survival, differentiation and migration. Some functions are regulated via intracellular signaling cascades, others by involvement of the extracellular milieu, including binding proteins and other extracellular matrix proteins. However, understanding of their functions and the exact nature of these interactions remains incomplete. Methods: IGF-I was PEGylated at its lysine sites - K27, K65 and K68. Binding of PEG-IGF-I to the IGFBPs was analyzed using BIAcore and its ability to activate the IGF-IR was assessed using IGF-IR phosphorylation assay. Furthermore, functional consequences of PEGylating the lysine residues of IGF-I was investigated using cell viability and cell migration assays. In addition, particular downstream signaling pathways regularly implicated in these mechanisms were also dissected using phospho-AKT and phospho-ERK1/2 assays. Results: In this study, IGF-I specifically PEGylated at lysine 27 (PEG-K27), 65 (PEG-K65) or 68 (PEG-K68) were employed. Receptor phosphorylation was only reduced by 2-fold with PEG-K65 and PEG-K68 over all the time points tested, and as observed in two cell types, 3T3 fibroblasts and MCF-7 breast cancer cells. PEGylation at K27 resulted in a much larger effect, with more than 10-fold lower activation for 3T3 fibroblasts and a ~3 fold reduced IGF-IR activation for MCF-7 breast cancer cells over 15 minutes. In addition, all PEG-IGF-I variants demonstrated a ten-fold reduction in the association rate to IGF binding proteins (IGFBPs). Functionally, all PEG variants completely lost their ability to induce cell migration in the presence of IGFBP-3/vitronectin (VN) complexes as compared to IGF-I; in contrast, cell viability was fully preserved. Further investigations into the downstream signaling pathways revealed that the PI3-K/AKT pathway was preferentially affected upon treatment with the PEG-IGF-I variants compared to the MAPK/ERK pathway. Conclusion: PEGylation of IGF-I has an impact on cell migration but not cell viability. General significance: PEG-IGF-I may differentially modulate IGF-I mediated functions that are dependent on its interaction with its receptor as well as key extracellular proteins such as VN and IGFBPs.

Re-orientations : East Asian popular cultures in contemporary Vietnam

Vietnamese-Australians live in Australia, a large island continent. The physical contrast between Vietnam and Australia is remarked upon by many Vietnamese in their migration stories. Whereas Vietnam is remembered as an interlinked sensual and social world, Australia is often viewed as a harsh, spacious, empty, dry continent. Australia is located in a regional Asian context, but this location has always been culturally and politically problematic, as it historically attempted to define itself as a "white" European nation in the Southern Hemisphere (Ang, 2000, p. xiii; McNamara & Coughlan, 1997, p. 1). During the Gold Rush period in the late 1800s, when there was widespread opposition to Chinese labor, Australia implemented a "White Australia" policy, although there were historically a significant number of Australians of Asian background. This exclusionary immigration policy was effectively overturned in the 1970s with the acceptance of a large number of refugees from Vietnam, Cambodia, and Laos in 1975. Vietnamese-Australians live predominantly in urban areas with over three quarters living in Sydney and Melbourne, the two largest cities. Within these two cities they are also highly concentrated in ethnically diverse suburbs, most living in areas with more than 1,000 residents born in Vietnam (Viviani, 1996, p. 49). However, Jupp (Jupp et al., 1990; Jupp, 1993) has argued that these areas are also zones of transition, with much movement in and out.

Introduction : the anthropology of migration

This introduction to the volume places the study of migration within the wider processes of social change and the complex actions of class, gender and ethnicity that are integrated into the experience of displacement. It argues that the articles in this volume contribute important new ethnographic and theoretical analyses to our knowledge and understanding of the constraints and the opportunities provided by cultural diversity in several societies by elaborating upon recent advances made in conceptualisations of migrancy. It places particular emphasis on developing phenomenological approaches to migration studies which incorporate the lived experience of migrants into any analysis. It also suggests that redressing the neglect of this area of study in Australia has the possibility of generating informed interruptions into the current political shift towards divisive public debates on immigration and cultural pluralism.

Quantifying alterations in cell migration : tracking fluorescently-tagged migrating cells by FACs and live-imaging

Cell migration is fundamental to many different physiological processes including embryonic development, inflammation and wound healing. Given the range and importance cell migration plays a number of assays have been developed to measure different aspects of cell migration. Here we describe two different methods to analyze cell migration. The first method analyzes the migration of fluorescently tagged cells using Boyden chambers and FACs and the second looks at migration properties using time-lapse microscopy.

Multiple types of data are required to identify the mechanisms influencing the spatial expansion of melanoma cell colonies

Background The expansion of cell colonies is driven by a delicate balance of several mechanisms including cell motility, cell-to-cell adhesion and cell proliferation. New approaches that can be used to independently identify and quantify the role of each mechanism will help us understand how each mechanism contributes to the expansion process. Standard mathematical modelling approaches to describe such cell colony expansion typically neglect cell-to-cell adhesion, despite the fact that cell-to-cell adhesion is thought to play an important role. Results We use a combined experimental and mathematical modelling approach to determine the cell diffusivity, D, cell-to-cell adhesion strength, q, and cell proliferation rate, ?, in an expanding colony of MM127 melanoma cells. Using a circular barrier assay, we extract several types of experimental data and use a mathematical model to independently estimate D, q and ?. In our first set of experiments, we suppress cell proliferation and analyse three different types of data to estimate D and q. We find that standard types of data, such as the area enclosed by the leading edge of the expanding colony and more detailed cell density profiles throughout the expanding colony, does not provide sufficient information to uniquely identify D and q. We find that additional data relating to the degree of cell-to-cell clustering is required to provide independent estimates of q, and in turn D. In our second set of experiments, where proliferation is not suppressed, we use data describing temporal changes in cell density to determine the cell proliferation rate. In summary, we find that our experiments are best described using the range D = 161 - 243 ?m2 hour-1, q = 0.3 - 0.5 (low to moderate strength) and ? = 0.0305 - 0.0398 hour-1, and with these parameters we can accurately predict the temporal variations in the spatial extent and cell density profile throughout the expanding melanoma cell colony. Conclusions Our systematic approach to identify the cell diffusivity, cell-to-cell adhesion strength and cell proliferation rate highlights the importance of integrating multiple types of data to accurately quantify the factors influencing the spatial expansion of melanoma cell colonies.

Beyond Siam : the Tjideng Apron

This article follows a previous (2008) article titled 'Siam' in the same publication and it examines the role of serendipity and chance in art, research, through and the migrant story. This is against the background of Women POW's in WWII; post-war migration to regional Australia and artefacts which reflect global history through a very personal story.