The role of immunoglobulins and their transporters in urogenital Chlamydial infections

Chlamydia trachomatis infections of the male and female reproductive tracts are the world's leading sexually transmitted bacterial disease, and can lead to damaging pathology, scarring and infertility. The resolution of chlamydial infection requires the development of adaptive immune responses to infection, and includes cell-mediated and humoral immunity. Whilst cluster of differentiation (CD)4+ T cells are known to be essential in clearance of infection [1], they are also associated with immune cell infiltration, autoimmunity and infertility in the testes [2-3]. Conversely, antibodies are less associated with inflammation, are readily transported into the reproductive tracts, and can offer lumenal neutralization of chlamydiae prior to infection. Antibodies, or immunoglobulins (Ig), play a supportive role in the resolution of chlamydial infections, and this thesis sought to define the function of IgA and IgG, against a variety of chlamydial antigens expressed during the intracellular and extracellular stages of the chlamydial developmental cycle. Transport of IgA and IgG into the mucosal lumen is facilitated by receptor-mediated transcytosis yet the expression profile (under normal conditions and during urogenital chlamydial infection) of the polymeric immunoglobulin receptor (pIgR) and the neonatal Fc receptor (FcRn) remains unknown. The expression profile of pIgR and FcRn in the murine male reproductive tract was found to be polarized to the lower and upper reproductive tract tissues respectively. This demonstrates that the two receptors have a tissue tropism, which must be considered when targeting pathogens that colonize different sites. In contrast, the expression of pIgR and FcRn in the female mouse was found to be distributed in both the upper and lower reproductive tracts. When urogenitally infected with Chlamydia muridarum, both male and female reproductive tracts up-regulated expression of pIgR and down-regulated expression of FcRn. Unsurprisingly, the up-regulation of pIgR increased the concentration of IgA in the lumen. However, down-regulation of FcRn, prevented IgG uptake and led to an increase or pooling of IgG in lumenal secretions. As previous studies have identified the importance of pIgR-mediated delivery of IgA, as well as the potential of IgA to bind and neutralize intracellular pathogens, IgA against a variety of chlamydial antigens was investigated. The protection afforded by IgA against the extracellular antigen major outer membrane protein (MOMP), was found to be dependent on pIgR expression in vitro and in vivo. It was also found that in the absence of pIgR, no protection was afforded to mice previously immunized with MOMP. The protection afforded from polyclonal IgA against the intracellular chlamydial antigens; inclusion membrane protein A (IncA), inclusion membrane proteins (IncMem) and secreted chlamydial protease-like activity factor (CPAF) were produced and investigated in vitro. Antigen-specific intracellular IgA was found to bind to the respective antigen within the infected cell, but did not significantly reduce inclusion formation (p > 0.05). This suggests that whilst IgA specific for the selected antigens was transported by pIgR to the chlamydial inclusion, it was unable to prevent growth. Similarly, immunization of male mice with intracellular chlamydial antigens (IncA or IncMem), followed by depletion CD4+ T cells, and subsequent urogenital C. muridarum challenge, provided minimal pIgR-mediated protection. Wild type male mice immunized with IncA showed a 57 % reduction (p < 0.05), and mice deficient in pIgR showed a 35 % reduction (p < 0.05) in reproductive tract chlamydial burden compared to control antigen, and in the absence of CD4+ T cells. This suggests that pIgR and secretory IgA (SIgA) were playing a protective role (21 % pIgR-mediated) in unison with another antigen-specific immune mechanism (36 %). Interestingly, IgA generated during a primary respiratory C. muridarum infection did not provide a significant amount of protection to secondary urogenital C. muridarum challenge. Together, these data suggest that IgA specific for an extracellular antigen (MOMP) can play a strong protective role in chlamydial infections, and that IgA targeting intracellular antigens is also effective but dependent on pIgR expression in tissues. However, whilst not investigated here, IgA targeting and blocking other intracellular chlamydial antigens, that are more essential for replication or type III secretion, may be more efficacious in subunit vaccines. Recently, studies have demonstrated that IgG can neutralize influenza virus by trafficking IgG-bound virus to lysosomes [4]. We sought to determine if this process could also traffic chlamydial antigens for degradation by lysosomes, despite Chlamydia spp. actively inhibiting fusion with the host endocytic pathway. As observed in pIgR-mediated delivery of anti-IncA IgA, FcRn similarly transported IgG specific for IncA which bound the inclusion membrane. Interestingly, FcRn-mediated delivery of anti-IncA IgG significantly decreased inclusion formation by 36 % (p < 0.01), and induced aberrant inclusion morphology. This suggests that unlike IgA, IgG can facilitate additional host cellular responses which affect the intracellular niche of chlamydial growth. Fluorescence microscopy revealed that IgG also bound the inclusion, but unlike influenza studies, did not induce the recruitment of lysosomes. Notably, anti-IncA IgG recruited sequestosomes to the inclusion membrane, markers of the ubiquitin/proteasome pathway and major histocompatibility complex (MHC) class I loading. To determine if the protection against C. muridarum infection afforded by IncA IgG in vitro translated in vivo, wild type mice and mice deficient in functional FcRn and MHC-I, were immunized, depleted of CD4+, and urogenitally infected with C. muridarum. Unlike in pIgR-deficient mice, the protection afforded from IncA immunization was completely abrogated in mice lacking functional FcRn and MHC-I/CD8+. Thus, both anti-IncA IgA and IgG can bind the inclusion in a pIgR and FcRn-mediated manner, respectively. However, only IgG mediates a higher reduction in chlamydial infection in vitro and in vivo suggesting more than steric blocking of IncA had occurred. Unlike anti-MOMP IgA, which reduced chlamydial infection of epithelial cells and male mouse tissues, IgG was found to enhance infectivity in vitro, and in vivo. Opsonization of EBs with MOMP-IgG enhanced inclusion formation of epithelial cells in a MOMP-IgG dose-dependent and FcRn-dependent manner. When MOMP-IgG opsonized EBs were inoculated into the vagina of female mice, a small but non-significant (p > 0.05) enhancement of cervicovaginal C. muridarum shedding was observed three days post infection in mice with functional FcRn. Interestingly, infection with opsonized EBs reduced the intensity of the peak of infection (day six) but protracted the duration of infection by 60 % in wild type mice only. Infection with EBs opsonized in IgG also significantly increased (p < 0.05) hydrosalpinx formation in the oviducts and induced lymphocyte infiltration uterine horns. As MOMP is an immunodominant antigen, and is widely used in vaccines, the ability of IgG specific to extracellular chlamydial antigens to enhance infection and induce pathology needs to be considered. Together, these data suggest that immunoglobulins play a dichotomous role in chlamydial infections, and are dependent on antigen specificity, FcRn and pIgR expression. FcRn was found to be highly expressed in upper male reproductive tract, whilst pIgR was dominantly expressed in the lower reproductive tract. Conversely, female mice expressed FcRn and pIgR in both the lower and upper reproductive tracts. In response to a normal chlamydial infection, pIgR is up-regulated increasing secretory IgA release, but FcRn is down-regulated preventing IgG uptake. Similarly to other studies [5-6], we demonstrate that IgA and IgG generated during primary chlamydial infections plays a minor role in recall immunity, and that antigen-specific subunit vaccines can offer more protection. We also show that both IgA and IgG can be used to target intracellular chlamydial antigens, but that IgG is more effective. Finally, IgA against the extracellular antigen MOMP can afford protection, whist IgG plays a deleterious role by increasing infectivity and inducing damaging immunopathology. Further investigations with additional antigens or combination subunit vaccines will enhance our understanding the protection afforded by antibodies against intracellular and extracellular pathogenic antigens, and help improve the development of an efficacious chlamydial vaccine.

Molecular approaches to diagnosis and management of ovarian cancer

The recent advances in the understanding of the pathogenesis of ovarian cancer have been helpful in addressing issues in diagnosis, prognosis and management. The study of ovarian tumours by novel techniques such as immunohistochemistry, fluorescent in situ hybridisation, comparative genomic hybridisation, polymerase chain reaction and new tumour markers have aided the evaluation and application of new concepts into clinical practice. The correlation of novel surrogate tumour specific features with response to treatment and outcome in patients has defined prognostic factors which may allow the future design of tailored therapy based on a molecular profile of the tumour. These have also been used to design new approaches to therapy such as antibody targeting and gene therapy. The delineation of roles of c-erbB2, c-fms and other novel receptor kinases in the pathogenesis of ovarian cancer has led initially to the development of anti-c-erbB2 monoclonal antibody therapy. The discovery of BRCA1 and BRCA2 genes will have an impact in the diagnosis and the prevention of familial ovarian cancer. The important role played by recessive genes such as p53 in cancer has raised the possibility of restoration of gene function by gene therapy. Although the pathological diagnosis of ovarian cancer is still confirmed principally on morphological features, addition of newer investigations will increasingly be useful in addressing difficult diagnostic problems. The increasingly rapid pace of discovery of genes important in disease, makes it imperative that the evaluation of their contribution in the pathogenesis of ovarian cancer is undertaken swiftly, thus improving the overall management of patients and their outcome.

A Satellite Selection Algorithm for Achieving High Reliability of Ambiguity Resolution with GPS and Beidou Constellations

Reliability of carrier phase ambiguity resolution (AR) of an integer least-squares (ILS) problem depends on ambiguity success rate (ASR), which in practice can be well approximated by the success probability of integer bootstrapping solutions. With the current GPS constellation, sufficiently high ASR of geometry-based model can only be achievable at certain percentage of time. As a result, high reliability of AR cannot be assured by the single constellation. In the event of dual constellations system (DCS), for example, GPS and Beidou, which provide more satellites in view, users can expect significant performance benefits such as AR reliability and high precision positioning solutions. Simply using all the satellites in view for AR and positioning is a straightforward solution, but does not necessarily lead to high reliability as it is hoped. The paper presents an alternative approach that selects a subset of the visible satellites to achieve a higher reliability performance of the AR solutions in a multi-GNSS environment, instead of using all the satellites. Traditionally, satellite selection algorithms are mostly based on the position dilution of precision (PDOP) in order to meet accuracy requirements. In this contribution, some reliability criteria are introduced for GNSS satellite selection, and a novel satellite selection algorithm for reliable ambiguity resolution (SARA) is developed. The SARA algorithm allows receivers to select a subset of satellites for achieving high ASR such as above 0.99. Numerical results from a simulated dual constellation cases show that with the SARA procedure, the percentages of ASR values in excess of 0.99 and the percentages of ratio-test values passing the threshold 3 are both higher than those directly using all satellites in view, particularly in the case of dual-constellation, the percentages of ASRs (>0.99) and ratio-test values (>3) could be as high as 98.0 and 98.5 % respectively, compared to 18.1 and 25.0 % without satellite selection process. It is also worth noting that the implementation of SARA is simple and the computation time is low, which can be applied in most real-time data processing applications.

Development of an inter-vehicle communications & positioning platform for transport safety applications

This project is a breakthrough in developing new scientific approaches for the design, development and evaluation of inter-vehicle communications, networking and positioning systems as part of Cooperative Intelligent Transportation Systems ensuring the safety of both roads and rail networks. This research focused on the elicitation, specification, analysis and validation of requirements for Vehicle-to-Vehicle communications and networking, and Vehicle-to-Vehicle positioning, which are accomplished with the research platform developed for this study. A number of mathematical models for communications, networking and positioning were developed from which simulations and field experiments were conducted to evaluate the overall performance of the platform. The outcomes of this research significantly contribute to improving the performance of the communications and positioning components of Cooperative Intelligent Transportation Systems.

Radial glia phagocytose axonal debris from degenerating overextending axons in the developing olfactory bulb

Axon targeting during the development of the olfactory system is not always accurate, and numerous axons overextend past the target layer into the deeper layers of the olfactory bulb. To date, the fate of the mis-targeted axons has not been determined. We hypothesized that following overextension, the axons degenerate, and cells within the deeper layers of the olfactory bulb phagocytose the axonal debris. We utilized a line of transgenic mice that expresses ZsGreen fluorescent protein in primary olfactory axons. We found that overextending axons closely followed the filaments of radial glia present in the olfactory bulb during embryonic development. Following overextension into deeper layers of the olfactory bulb, axons degenerated and radial glia responded by phagocytosing the resulting debris. We used in vitro analysis to confirm that the radial glia had phagocytosed debris from olfactory axons. We also investigated whether the fate of overextending axons was altered when the development of the olfactory bulb was perturbed. In mice that lacked Sox10, a transcription factor essential for normal olfactory bulb development, we observed a disruption to the morphology and positioning of radial glia and an accumulation of olfactory axon debris within the bulb. Our results demonstrate that during early development of the olfactory system, radial glia play an important role in removing overextended axons from the deeper layers of the olfactory bulb.

Barriers and supports for development of a clinician scientist role in cerebrovascular nursing: A position paper

This position paper will: 1. Provide an update on relevant current developments in the education, training and positioning of clinician nurse scientists; 2. Provide and promote a rational argument for the development of the clinician nurse scientist role, and; 3. Discuss issues related to capacity building in clinical research in neuroscience nursing, with specific reference to and support for the cerebrovascular (stroke) specialty nursing area.

Beyond social justice agendas: Indigenous knowledges in pre-service teacher education and practice in Australia

The space and positioning of Indigenous knowledges (IK) within Australian curricula and pedagogy are often contentious, informed by the broader Australian socio-cultural, political and economic landscape. Against changing educational policy, historically based on the myth of terra nullius, we discuss the shifting priorities for embedding Indigenous knowledges in educational practice in university and school curricula and pedagogy. In this chapter, we argue that personal and professional commitment to social justice is an important starting point for embedding Indigenous knowledges in the Australian school curricula and pedagogy. Developing teacher knowledge around embedding IK is required to enable teachers’ preparedness to navigate a contested historical/colonising space in curriculum decision-making, teaching and learning. We draw one mpirical data from a recent research project on supporting pre-service teachers as future curriculum leaders; the project was funded by the Office of Learning and Teaching (OLT). This project aimed to support future curriculum leaders to develop their knowledge of embedding IK at one Australian university. We propose supporting the embedding of IK in situ with pre-service teachers and their supervising teachers on practicum in real, sustained and affirming ways that shifts the recognition of IK from personal commitment to social justice in education, to one that values Indigenous knowledges as content to educate (Connell, 1993). We argue that sustained engagement with and appreciation of IKhas the potential to decolonise Australian curricula, shift policy directions and enhance race relations between Indigenous and non-Indigenous Australians .

Study of reproducibility of human arterial plaque reconstruction and its effects on stress analysis based on multispectral in vivo magnetic resonance imaging

Purpose: To quantify the uncertainties of carotid plaque morphology reconstruction based on patient-specific multispectral in vivo magnetic resonance imaging (MRI) and their impacts on the plaque stress analysis. Materials and Methods: In this study, three independent investigators were invited to reconstruct the carotid bifurcation with plaque based on MR images from two subjects to study the geometry reconstruction reproducibility. Finite element stress analyses were performed on the carotid bifurcations, as well as the models with artificially modified plaque geometries to mimic the image segmentation uncertainties, to study the impacts of the uncertainties to the stress prediction. Results: Plaque reconstruction reproducibility was generally high in the study. The uncertainties among interobservers are around one or the subpixel level. It also shows that the predicted stress is relatively less sensitive to the arterial wall segmentation uncertainties, and more affected by the accuracy of lipid region definition. For a model with lipid core region artificially increased by adding one pixel on the lipid region boundary, it will significantly increase the maximum Von Mises Stress in fibrous cap (>100%) compared with the baseline model for all subjects. Conclusion: The current in vivo MRI in the carotid plaque could provide useful and reliable information for plaque morphology. The accuracy of stress analysis based on plaque geometry is subject to MRI quality. The improved resolution/quality in plaque imaging with newly developed MRI protocols would generate more realistic stress predictions.

Effects of fish density distribution and effort distribution on catchability

The effects of fish density distribution and effort distribution on the overall catchability coefficient are examined. Emphasis is also on how aggregation and effort distribution interact to affect overall catch rate [catch per unit effort (cpue)]. In particular, it is proposed to evaluate three indices, the catchability index, the knowledge parameter, and the aggregation index, to describe the effectiveness of targeting and the effects on overall catchability in the stock area. Analytical expressions are provided so that these indices can easily be calculated. The average of the cpue calculated from small units where fishing is random is a better index for measuring the stock abundance. The overall cpue, the ratio of lumped catch and effort, together with the average cpue, can be used to assess the effectiveness of targeting. The proposed methods are applied to the commercial catch and effort data from the Australian northern prawn fishery. The indices are obtained assuming a power law for the effort distribution as an approximation of targeting during the fishing operation. Targeting increased catchability in some areas by 10%, which may have important implications on management advice.

Combining propensity and influence models for product adoption prediction

This paper studies the problem of selecting users in an online social network for targeted advertising so as to maximize the adoption of a given product. In previous work, two families of models have been considered to address this problem: direct targeting and network-based targeting. The former approach targets users with the highest propensity to adopt the product, while the latter approach targets users with the highest influence potential – that is users whose adoption is most likely to be followed by subsequent adoptions by peers. This paper proposes a hybrid approach that combines a notion of propensity and a notion of influence into a single utility function. We show that targeting a fixed number of high-utility users results in more adoptions than targeting either highly influential users or users with high propensity.

Computer vision onboard UAVs for civilian tasks

Computer vision is much more than a technique to sense and recover environmental information from an UAV. It should play a main role regarding UAVs’ functionality because of the big amount of information that can be extracted, its possible uses and applications, and its natural connection to human driven tasks, taking into account that vision is our main interface to world understanding. Our current research’s focus lays on the development of techniques that allow UAVs to maneuver in spaces using visual information as their main input source. This task involves the creation of techniques that allow an UAV to maneuver towards features of interest whenever a GPS signal is not reliable or sufficient, e.g. when signal dropouts occur (which usually happens in urban areas, when flying through terrestrial urban canyons or when operating on remote planetary bodies), or when tracking or inspecting visual targets—including moving ones—without knowing their exact UMT coordinates. This paper also investigates visual serving control techniques that use velocity and position of suitable image features to compute the references for flight control. This paper aims to give a global view of the main aspects related to the research field of computer vision for UAVs, clustered in four main active research lines: visual serving and control, stereo-based visual navigation, image processing algorithms for detection and tracking, and visual SLAM. Finally, the results of applying these techniques in several applications are presented and discussed: this study will encompass power line inspection, mobile target tracking, stereo distance estimation, mapping and positioning.

Health protection

Our understanding of how the environment can impact human health has evolved and expanded over the centuries, with concern and interest dating back to ancient times. For example, over 4000 years ago, a civilisation in northern India tried to protect the health of its citizens by constructing and positioning buildings according to strict building laws, by having bathrooms and drains, and by having paved streets with a sewerage system (Rosen 1993). In more recent times, the ‘industrial revolution’ played a dominant role in shaping the modern world, and with it the modern public health system. This era was signified by rapid progress in technology, the growth of transportation and the expansion of the market economy, which lead to the organisation of industry into a factory system. This meant that labour had to be brought to the factories and by the 1820s, poverty and social distress (including overcrowding and infrequent sewage and garbage disposal) was more widespread than ever. These circumstances, therefore, lead to the rise of the ‘sanitary revolution’ and the birth of modern public health (Rosen 1993). The sanitary revolution has also been described as constituting the beginning of the first wave of environmental concern, which continued until after World War 2 when major advances in engineering and chemistry substantially changed the face of industry, particularly the chemical sector. The second wave of environmental concern came in the mid to late 20th century and was dominated by the environmental or ecology movement. A landmark in this era was the 1962 publication of the book Silent Spring by Rachel Carson. This identified for the first time the dramatic effects on the ecosystem of the widespread use of the organochlorine pesticide, DDT. The third wave of environmental concern commenced in the 1980s and continues today. The accelerated rate of economic development, the substantial increase in the world population and the globalisation of trade have dramatically changed the production methods and demand for goods in both developed and developing countries. This has lead to the rise of ‘sustainable development’ as a key driver in environmental planning and economic development (Yassi et al 2001). The protection of health has, therefore, been a hallmark of human history and is the cornerstone of public health practice. This chapter introduces environmental health and how it is managed in Australia, including a discussion of the key generic management tools. A number of significant environmental health issues and how they are specifically managed are then discussed, and the chapter concludes by discussing sustainable development and its links with environmental health.

Workplace design in the knowledge economy : a case of the NetWorkPlace™©

This thesis explores a way to inform the architectural design process for contemporary workplace environments. It reports on both theoretical and practical outcomes through an exclusively Australian case study of a network enterprise comprised of collaborative, yet independent business entities. The internet revolution, substantial economic and cultural shifts, and an increased emphasis on lifestyle considerations have prompted a radical re-ordering of organisational relationships and the associated structures, processes, and places of doing business. The social milieu of the information age and the knowledge economy is characterised by an almost instantaneous flow of information and capital. This has culminated in a phenomenon termed by Manuel Castells as the network society, where physical locations are joined together by continuous communication and virtual connectivity. A new spatial logic encompassing redefined concepts of space and distance, and requiring a comprehensive shift in the approach to designing workplace environments for today’s adaptive, collaborative organisations in a dynamic business world, provides the backdrop for this research. Within the duality of space and an augmentation of the traditional notions of place, organisational and institutional structures pose new challenges for the design professions. The literature revealed that there has always been a mono-organisational focus in relation to workplace design strategies. The phenomenon of inter-organisational collaboration has enabled the identification of a gap in the knowledge relative to workplace design. This new context generated the formulation of a unique research construct, the NetWorkPlace™©, which captures the complexity of contemporary employment structures embracing both physical and virtual work environments and practices, and provided the basis for investigating the factors that are shaping and defining interactions within and across networked organisational settings. The methodological orientation and the methods employed follow a qualitative approach and an abductively driven strategy comprising two distinct components, a cross-sectional study of the whole of the network and a longitudinal study, focusing on a single discrete workplace site. The complexity of the context encountered dictated that a multi-dimensional investigative framework was required to be devised. The adoption of a pluralist ontology and the reconfiguration of approaches from traditional paradigms into a collaborative, trans-disciplinary, multi-method epistemology provided an explicit and replicatable method of investigation. The identification and introduction of the NetWorkPlace™© phenomenon, by necessity, spans a number of traditional disciplinary boundaries. Results confirm that in this context, architectural research, and by extension architectural practice, must engage with what other disciplines have to offer. The research concludes that no single disciplinary approach to either research or practice in this area of design can suffice. Pierre Bourdieau’s philosophy of ‘practice’ provides a framework within which the governance and technology structures, together with the mechanisms enabling the production of social order in this context, can be understood. This is achieved by applying the concepts of position and positioning to the corporate power dynamics, and integrating the conflict found to exist between enterprise standard and ferally conceived technology systems. By extending existing theory and conceptions of ‘place’ and the ‘person-environment relationship’, relevant understandings of the tensions created between Castells’ notions of the space of place and the space of flows are established. The trans-disciplinary approach adopted, and underpinned by a robust academic and practical framework, illustrates the potential for expanding the range and richness of understanding applicable to design in this context. The outcome informs workplace design by extending theoretical horizons, and by the development of a comprehensive investigative process comprising a suite of models and techniques for both architectural and interior design research and practice, collectively entitled the NetWorkPlace™© Application Framework. This work contributes to the body of knowledge within the design disciplines in substantive, theoretical, and methodological terms, whilst potentially also influencing future organisational network theories, management practices, and information and communication technology applications. The NetWorkPlace™© as reported in this thesis, constitutes a multi-dimensional concept having the capacity to deal with the fluidity and ambiguity characteristic of the network context, as both a topic of research and the way of going about it.

The emergent male shopper : an identification of male supermarket shopper types

Grocery shopping is a routine activity widely considered the responsibility of the female spouse, yet modern social and demographic shifts are causing men to engage in this task. This study develops a retail shopping typology of male grocery shoppers, employing a cluster analysis technique. Five distinct cohorts emerge from the data of eight constructs, measured by seventy one items. One new shopper type emerges from this research. This shopper presented as a younger man, at the commencement of their family lifecycle, attracted by a strong value offer, focusing on price and promotional discounts. Our research offers a contribution to the marketing, consumer behaviour and supermarket retailing disciplines in three ways. By examining and identifying male shopping behaviour in the context of grocery shopping, the development of a retail shopping typology of male grocery shoppers and the extension and employment of a cluster analysis in identifying distinct groups. This research has implications for gender, segmentation studies and consumer behaviour disciplines in regard to grocery shopping. The identification of specific groups of male grocery shoppers will enable grocery retailers to effectively implement important, targeted marketing strategies.

From supply to demand-driven opportunities : unravelling the role of the customer in opportunity identification processes

Successful identification and exploitation of opportunities has been an area of interest to many entrepreneurship researchers. Since Shane and Venkataraman’s seminal work (e.g. Shane and Venkataraman, 2000; Shane, 2000), several scholars have theorised on how firms identify, nurture and develop opportunities. The majority of this literature has been devoted to understanding how entrepreneurs search for new applications of their technological base or discover opportunities based on prior knowledge (Zahra, 2008; Sarasvathy et al., 2003). In particular, knowledge about potential customer needs and problems that may present opportunities is vital (Webb et al., 2010). Whereas the role of prior knowledge of customer problems (Shane, 2003; Shepherd and DeTienne, 2005) and positioning oneself in a so-called knowledge corridor (Fiet, 1996) has been researched, the role of opportunity characteristics and their interaction with customer-related mechanisms that facilitate and hinder opportunity identification has received scant attention.