Hepatocyte growth factor stimulates invasion across reconstituted basement membranes by a new human small intestinal cell line

Hepatocyte growth factor/scatter factor (HGF/SF) is a protein growth factor whose pleiotropic effects on epithelial cells include the stimulation of motility, mitosis and tubulogenesis. These responses are mediated by the cell surface tyrosine kinase receptor c-met. Because both the cytokine and receptor are found in the gastrointestinal tract, we have studied the effects of HGF/SF on transformed gut epithelial cells which express c-met. Here we describe the response of a new transformed human jejunal epithelioid cell line (HIE-7) to HGF/SF. Morphologically HIE-7 cells are immature. Their epithelial lineage was confirmed by reactivity with the epithelial specific antibodies AE1/AE3, Cam 5.2, Ber-EP4 and anti-EMA and is consistent with their expression of c-met mRNA and protein. In addition, electron microscopic analysis revealed the presence of primitive junctions and rudimentary microvilli, but features of polarization were absent. When grown on reconstituted basement membranes, HIE-7 cells formed closely associated multicellular cord-like structures adjacent to acellular spaces. However, the cells did not mature structurally, form lumen-like structures or express disaccharidase mRNA, even in the presence of recombinant HGF (rHGF). On the other hand, rHGF induced HIE-7 cells to scatter and stimulated their rapid migration in a modified wound assay. To determine whether the motogenic effect caused by rHGF is associated with HIE-7 cell invasiveness across reconstituted basement membranes, a Boyden chamber chemoinvasion assay was performed. rHGF stimulated a 10-fold increase in the number of HIE-7 cells that crossed the basement membrane barrier, while only stimulating a small increase in chemotaxis across a collagen IV matrix, suggesting that the cytokine activates matrix penetration by these cells. rHGF also stimulated the invasion of basement membranes by an undifferentiated rat intestinal cell line (IEC-6) and by two human colon cancer cell lines which are poorly differentiated (DLD-1 and SW 948). In contrast, two moderately well differentiated colon cancer cell lines (Caco-2 and HT-29) did not manifest an invasive response when exposed to rHGF. These results suggest that HGF/SF may play a significant role in the invasive behavior of anaplastic and poorly differentiated gut epithelial tumors.

A cisgender girl in a transgender world : The methodological challenges of researching in the transgender community

The available research literature on intimate partner violence is often centred around a heteronormative understanding of gender, relationships and violence. When it comes to intimate partner violence in the transgender community, the research is limited or nonexistent due in part to the methodological issues of visibility and access by those outside this community. Drawing from Renzetti (1992, 1995), McClennen (2003), and the feminist participatory research model, this paper examines the techniques for overcoming the methodological barriers as a cisgender or 'normatively gendered' woman in a transgender community. Throughout the research with the transgender community, five strategies for overcoming methodological barriers were developed: Cultural Immersion, Commitment and Visibility, Sensitivity and Acceptance, Honesty, and Communication. This paper explores how utilising these strategies enabled access to the transgender community in order to conduct effective research.

Hunting and gathering : new imperatives in mapping and collecting student learning data to assure quality outcomes

Assurance of learning (AOL) is a quality enhancement and quality assurance process used in higher education. It involves a process of determining programme learning outcomes and standards, and systematically gathering evidence to measure students' performance on these. The systematic assessment of whole-of-programme outcomes provides a basis for curriculum development and management, continuous improvement, and accreditation. To better understand how AOL processes operate, a national study of university practices across one discipline area, business and management, was undertaken. To solicit data on AOL practice, interviews were undertaken with a sample of business school representatives (n = 25). Two key processes emerged: (1) mapping of graduate attributes and (2) collection of assurance data. External drivers such as professional accreditation and government legislation were the primary reasons for undertaking AOL outcomes but intrinsic motivators in relation to continuous improvement were also evident. The facilitation of academic commitment was achieved through an embedded approach to AOL by the majority of universities in the study. A sustainable and inclusive process of AOL was seen to support wider stakeholder engagement in the development of higher education learning outcomes.

Myoepithelial molecular markers in human breast carcinoma PMC42-LA cells are induced by extracellular matrix and stromal cells

The microenvironment plays a key role in the cellular differentiation of the two main cell lineages of the human breast, luminal epithelial, and myoepithelial. It is not clear, however, how the components of the microenvironment control the development of these cell lineages. To investigate how lineage development is regulated by 3-D culture and microenvironment components, we used the PMC42-LA human breast carcinoma cell line, which possesses stem cell characteristics. When cultured on a two-dimensional glass substrate, PMC42-LA cells formed a monolayer and expressed predominantly luminal epithelial markers, including cytokeratins 8, 18, and 19; E-cadherin; and sialomucin. The key myoepithelial-specific proteins α-smooth muscle actin and cytokeratin 14 were not expressed. When cultured within Engelbreth-Holm- Swarm sarcoma-derived basement membrane matrix (EHS matrix), PMC42-LA cells formed organoids in which the expression of luminal markers was reduced and the expression of other myoepithelial-specific markers (cytokeratin 17 and P-cadherin) was promoted. The presence of primary human mammary gland fibroblasts within the EHS matrix induced expression of the key myoepithelial-specific markers, α-smooth muscle actin and cytokeratin 14. Immortalized human skin fibroblasts were less effective in inducing expression of these key myoepithelial-specific markers. Confocal dual-labeling showed that individual cells expressed luminal or myoepithelial proteins, but not both. Conditioned medium from the mammary fibroblasts was equally effective in inducing myoepithelial marker expression. The results indicate that the myoepithelial lineage is promoted by the extracellular matrix, in conjunction with products secreted by breast-specific fibroblasts. Our results demonstrate a key role for the breast microenvironment in the regulation of breast lineage development.

eHealth-as-a-Service (eHaaS) : a data-driven decision making approach in Australian context

A commitment in 2010 by the Australian Federal Government to spend $466.7 million dollars on the implementation of personally controlled electronic health records (PCEHR) heralded a shift to a more effective and safer patient centric eHealth system. However, deployment of the PCEHR has met with much criticism, emphasised by poor adoption rates over the first 12 months of operation. An indifferent response by the public and healthcare providers largely sceptical of its utility and safety speaks to the complex sociotechnical drivers and obstacles inherent in the embedding of large (national) scale eHealth projects. With government efforts to inflate consumer and practitioner engagement numbers giving rise to further consumer disillusionment, broader utilitarian opportunities available with the PCEHR are at risk. This paper discusses the implications of establishing the PCEHR as the cornerstone of a holistic eHealth strategy for the aggregation of longitudinal patient information. A viewpoint is offered that the real value in patient data lies not just in the collection of data but in the integration of this information into clinical processes within the framework of a commoditised data-driven approach. Consideration is given to the eHealth-as-a-Service (eHaaS) construct as a disruptive next step for co-ordinated individualised healthcare in the Australian context.

Pleistocene climate fluctuations influence phylogeographical patterns in Melomys cervinipes across the mesic forests of eastern Australia

Aim Our aim was to clarify the lineage-level relationships for Melomys cervinipes and its close relatives and investigate whether the patterns of divergence observed for these wet-forest-restricted mammals may be associated with recognized biogeographical barriers. Location Mesic closed forest along the east coast of Australia, from north Queensland to mid-eastern New South Wales. Methods To enable rigorous phylogenetic reconstruction, divergence-date estimation and phylogeographical inference, we analysed DNA sequence and microsatellite data from 307 specimens across the complete distribution of M. cervinipes (45 localities). Results Three divergent genetic lineages were found within M. cervinipes, corresponding to geographically delineated northern, central and southern clades. Additionally, a fourth lineage, comprising M. rubicola and M. capensis, was identified and was most closely related to the northern M. cervinipes lineage. Secondary contact of the northern and central lineages was identified at one locality to the north of the Burdekin Gap. Main conclusions Contemporary processes of repeated habitat fragmentation and contraction, local extinction events and subsequent re-expansion across both small and large areas, coupled with the historical influence of the Brisbane Valley Barrier, the St Lawrence Gap and the Burdekin Gap, have contributed to the present phylogeographical structure within M. cervinipes. Our study highlights the need to sample close to the periphery of putative biogeographical barriers or risk missing vital phylogeographical information that may significantly alter the interpretation of biogeographical hypotheses.

The responsibility to protect in the Asia-Pacific region

In 2005, governments around the world unanimously agreed to the principle of the responsibility to protect (R2P), which holds that all states have a responsibility to protect their populations from genocide and mass atrocities, that the international community should assist them to fulfil this duty, and that the international community should take timely and decisive measures to protect populations from such crimes when their host state fails to do so. Progressing R2P from words to deeds requires international consensus about the principle’s meaning and scope. To achieve a global consensus on this, we need to better understand the position of governments around the world, including in the Asia-Pacific region, which has long been associated with an enduring commitment to a traditional concept of sovereignty. The present article contributes to such an endeavour through its three sections. The first part charts the nature of the international consensus on R2P and examines the UN secretary-general’s approach. The second looks in detail at the positions of the Asia-Pacific region’s governments on the R2P principle. The final part explores the way forward for progressing the R2P principle in the Asia-Pacific region.

A preliminary framework for DNA barcoding, incorporating the multispecies coalescent

The capacity to identify an unknown organism using the DNA sequence from a single gene has many applications. These include the development of biodiversity inventories (Janzen et al. 2005), forensics (Meiklejohn et al. 2011), biosecurity (Armstrong and Ball 2005), and the identification of cryptic species (Smith et al. 2006). The popularity and widespread use (Teletchea 2010) of the DNA barcoding approach (Hebert et al. 2003), despite broad misgivings (e.g., Smith 2005; Will et al. 2005; Rubinoff et al. 2006), attest to this. However, one major shortcoming to the standard barcoding approach is that it assumes that gene trees and species trees are synonymous, an assumption that is known not to hold in many cases (Pamilo and Nei 1988; Funk and Omland 2003). Biological processes that violate this assumption include incomplete lineage sorting and interspecific hybridization (Funk and Omland 2003). Indeed, simulation studies indicate that the concatenation approach (in which these two processes are ignored) can lead to statistically inconsistent estimation of the species tree (Kubatko and Degnan 2007)...

How to sequence and annotate insect mitochondrial genomes for systematic and comparative genomics research

Over the past decade the mitochondrial (mt) genome has become the most widely used genomic resource available for systematic entomology. While the availability of other types of ‘–omics’ data – in particular transcriptomes – is increasing rapidly, mt genomes are still vastly cheaper to sequence and are far less demanding of high quality templates. Furthermore, almost all other ‘–omics’ approaches also sequence the mt genome, and so it can form a bridge between legacy and contemporary datasets. Mitochondrial genomes have now been sequenced for all insect orders, and in many instances representatives of each major lineage within orders (suborders, series or superfamilies depending on the group). They have also been applied to systematic questions at all taxonomic scales from resolving interordinal relationships (e.g. Cameron et al., 2009; Wan et al., 2012; Wang et al., 2012), through many intraordinal (e.g. Dowton et al., 2009; Timmermans et al., 2010; Zhao et al. 2013a) and family-level studies (e.g. Nelson et al., 2012; Zhao et al., 2013b) to population/biogeographic studies (e.g. Ma et al., 2012). Methodological issues around the use of mt genomes in insect phylogenetic analyses and the empirical results found to date have recently been reviewed by Cameron (2014), yet the technical aspects of sequencing and annotating mt genomes were not covered. Most papers which generate new mt genome report their methods in a simplified form which can be difficult to replicate without specific knowledge of the field. Published studies utilize a sufficiently wide range of approaches, usually without justification for the one chosen, that confusion about commonly used jargon such as ‘long PCR’ and ‘primer walking’ could be a serious barrier to entry. Furthermore, sequenced mt genomes have been annotated (gene locations defined) to wildly varying standards and improving data quality through consistent annotation procedures will benefit all downstream users of these datasets. The aims of this review are therefore to: 1. Describe in detail the various sequencing methods used on insect mt genomes; 2. Explore the strengths/weakness of different approaches; 3. Outline the procedures and software used for insect mt genome annotation, and; 4. Highlight quality control steps used for new annotations, and to improve the re-annotation of previously sequenced mt genomes used in systematic or comparative research.

An efficient eAuction potocol

A secure protocol for electronic, sealed-bid, single item auctions is presented. The protocol caters to both first and second price (Vickrey) auctions and provides full price flexibility. Both computational and communication cost are linear with the number of bidders and utilize only standard cryptographic primitives. The protocol strictly divides knowledge of the bidder's identity and their actual bids between, respectively, a registration authority and an auctioneer, who are assumed not to collude but may be separately corrupt. This assures strong bidder-anonymity, though only weak bid privacy. The protocol is structured in two phases, each involving only off-line communication. Registration, requiring the use of the public key infrastructure, is simultaneous with hash-sealed bid-commitment and generates a receipt to the bidder containing a pseudonym. This phase is followed by encrypted bid-submission. Both phases involve the registration authority acting as a communication conduit but the actual message size is quite small. It is argued that this structure guarantees non-repudiation by both the winner and the auctioneer. Second price correctness is enforced either by observing the absence of registration of the claimed second-price bid or, where registered but lower than the actual second price, is subject to cooperation by the second price bidder - presumably motivated through self-interest. The use of the registration authority in other contexts is also considered with a view to developing an architecture for efficient secure multiparty transactions

Selective neuronal differentiation of neural stem cells induced by nanosecond microplasma agitation

An essential step for therapeutic and research applications of stem cells is their ability to differentiate into specific cell types. Neuronal cells are of great interest for medical treatment of neurodegenerative diseases and traumatic injuries of central nervous system (CNS), but efforts to produce these cells have been met with only modest success. In an attempt of finding new approaches, atmospheric-pressure room-temperature microplasma jets (MPJs) are shown to effectively direct in vitro differentiation of neural stem cells (NSCs) predominantly into neuronal lineage. Murine neural stem cells (C17.2-NSCs) treated with MPJs exhibit rapid proliferation and differentiation with longer neurites and cell bodies eventually forming neuronal networks. MPJs regulate ~. 75% of NSCs to differentiate into neurons, which is a higher efficiency compared to common protein- and growth factors-based differentiation. NSCs exposure to quantized and transient (~. 150. ns) micro-plasma bullets up-regulates expression of different cell lineage markers as β-Tubulin III (for neurons) and O4 (for oligodendrocytes), while the expression of GFAP (for astrocytes) remains unchanged, as evidenced by quantitative PCR, immunofluorescence microscopy and Western Blot assay. It is shown that the plasma-increased nitric oxide (NO) production is a factor in the fate choice and differentiation of NSCs followed by axonal growth. The differentiated NSC cells matured and produced mostly cholinergic and motor neuronal progeny. It is also demonstrated that exposure of primary rat NSCs to the microplasma leads to quite similar differentiation effects. This suggests that the observed effect may potentially be generic and applicable to other types of neural progenitor cells. The application of this new in vitro strategy to selectively differentiate NSCs into neurons represents a step towards reproducible and efficient production of the desired NSC derivatives. © 2013.

Balancing organizational and professional commitments in professional service firms : the HR practices that matter

Due to their potential to positively influence sales quality and performance and reduce employee turnover in service organizations, HR practices targeting employee commitment have received considerable attention in the HRM literature in recent years. Parallel to this, there has been increasing focus on the nature of commitment, and in particular the existence of multiple commitment foci. In this paper, we examine how HR practices influence professionals' commitment to their organization, to their profession or to both organization and profession, in a qualitative study of three Danish financial investment firms. Our findings suggest that in professional service firms, HR practices encourage high levels of organizational commitment primarily and most often through their influence on professional commitment and that HR practices related to flexible work design are essential in creating balance between an employee's commitment to organization and commitment to their profession. Further, the findings suggest that these same HR practices may foster such high levels of professional commitment that labor turnover will increase when opportunities for pursuing professional goals afforded by work design are restricted.

Coming into an inheritance : family support and Chinese heritage language learning

The critical role that family plays in Chinese Heritage Language learning has gained increasing attention from psychological, political and sociological scholarship. Guided by Bourdieu’s notion of ‘habitus’, our mixed methods sociological study firstly addresses the need for quantitative evidence on the relationship between family support and Chinese Heritage Language proficiency through a survey of 230 young Chinese Australians; and then explores the dynamics of family support of Chinese Heritage Language learning through multiple interviews with five participants. The interview data demonstrate ongoing intergenerational reproduction of Chinese Heritage Language through various forms of family inculcation. Learners’ transition from resistance to commitment is a focus of the analysis. Extant research struggles to theorise the reasons behind this transition. We offer a Bourdieusian explanation that construes the transition as ‘habitus realisation’. Our study has implications for Chinese Heritage Language researchers, Chinese immigrant parents and Chinese teachers.

Course quality assurance at a glance

From 2014, QUT will be adopting a life-cycle approach to Course Quality Assurance informed by a wider and richer range of historic, ‘live’ and ‘predictive’ course data. Key data elements continue to be grouped according to the three broad categories – Viability, Quality of Learning Environment and Outcomes – and are further supported with analytic data presented within tables and charts. Course Quality Assurance and this Consolidated Courses Performance Report illuminate aspects of courses from a data evidence base highlighting the strengths and weaknesses of our courses. It provides the framework and tools to achieve QUT's commitment to excellent graduate outcomes by drawing attention and focus to the quality of our courses and providing a structured approach for bringing about change. Our portfolio of courses forms a vital part of QUT, generating almost $600 million in 2013 alone. Real world courses are fundamental to the strength of the Institution; they are what our many thousands of current and future students are drawn to and invest their time and aspirations in. As we move through a period of some regulatory and deregulatory uncertainty, there is a greater need for QUT to monitor and respond to the needs and expectations of our students. The life-cycle approach, with its rich and predicative data, provides the best source of evidence we have had, to date, to assure the quality of our courses and their relevance in a rapidly changing higher education context.

Influence and increased funding in Canadian public libraries : the case of Alberta in fiscal year 2009–10

This exploratory case study examined the role of social influence in the decision-making process to increase public library funding in the Canadian province of Alberta in the 2009–10 fiscal year. Using Robert Cialdini’s theory of factors of influence (i.e., commitment and consistency, authority, liking, social proof, scarcity, and reciprocity) as a framework for analysis, findings show that consistency and commitment and authority were relevant and that liking was also important. These findings are consistent with Cialdini’s theory, which suggests that the quality of relationships is one factor that can most strongly influence a decision maker. This study gives insight into the factors motivating those involved in public library funding allocation decisions. No prior studies have examined the construct of influence in decision making about funding for public libraries at any level of government.