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 alpha-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, alpha-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.

The influence of sensitivity to reward on reactivity to alcohol-related cues

Aims. To investigate the role of sensitivity to reward in mediating social drinkers' reactivity to alcohol cues. Design. A standard cue-reactivity paradigm was employed. Two groups of social drinkers (heavy and ight) were assessed after exposure to the sight, smell and taste of a neutral cue (water) and then an alcohol cue (glass of beer). Setting. Sessions were conducted in a laboratory based environment. Participants. Twenty heavy (12 males, eight females) and 18 light social drinkers (seven males, 11 females) were recruited; mean age was 23.6 years. Measurements. The Card Arranging Reward Responsivity Objective Test (CARROT), assessing behavioural responsiveness to a monetary incentive; urge to drink; positive affect; and the BAS scales, assessing sensitivity to reward. Findings. Heavy drinkers displayed a significant increase in responsivity to rewards (i.e. CARROT) and self-reported urge to drink, bur not positive affect, after exposure to alcohol. For the heavy drinkers, heightened sensitivity to reward (i.e. BAS scales) was significantly related to cue-elicited urge to drink and positive affect. Conclusion. The results are consistent with a conditioned appetitive motivational model of alcohol use and suggest that Gray's theory of personality may be of some benefit in explaining variation in reactivity responses.

Environmental benchmarking of Hong Kong buildings

A method for assessing the environmental performance of Hong Kong’s buildings has been developed, known as the Hong Kong Building Environmental Assessment Method or HK-BEAM. The assessment is a type of expert-based survey, either of the design, in the case of proposed buildings, or an evaluation of building performance for newly built or existing buildings. The assessment essentially provides a benchmark of environmental performance against a series of qualitative and quantitative measures that earn “credits”. Buildings can be rated as “excellent”, “very good”, “good” or “fair”. The assessment covers global, local and indoor issues. The original assessment has been in use since 1996 and allowed appraisal of new and existing air-conditioned offices. A new version has been recently produced for residential buildings. The latest version has addressed some of the criticisms of the earlier versions and covers a wider range of issues, taking a life-cycle approach. Reviews the latest new residential version, making comparisons with the earlier new offices scheme.

Truth is God harm minimization through the practice of ecologically sustainable design and buddhism

The emergence of the global ecological crisis is presenting unique opportunities for the coordination of ethical thinking across cultural boundaries. Harm minimization as an ethical imperative operates as the ‘modus operandi’ behind both Ecologically Sustainable Design (ESD) and Buddhist practice. The architectural response to ESD is founded upon the ‘Declaration of Interdependence for a Sustainable Future’ adopted in 1993 by the International Union of Architects, of which the RAIA is a member.

Buddhism is a response to existential concerns universal to humanity. It developed as a set of principles for personal transformation known as the Four Noble Truths elucidated two and a half thousand years ago. Buddhist meditation practise ‘interrupts automatic patterns of conditioned behaviour’ recognised as the major obstacle to be overcome in any programme for change. Unsustainable egocentric behaviour is considered fundamental to our global ecological crisis and calls for radical behavioural change are increasingly being heard at the professional as well as the personal level. Emerging synergies between the Western cognitive sciences and Buddhist study of the mind increasingly validate the Tibetan Buddhist mind development phenomenon. Buddhists argue that their programme for enhancing ethical behaviour through mind development is a step-by step process of observation and analysis built upon empirical observation – a fundamental pre-requisite of any ‘scientific’ enquiry. Collaborative research programmes currently underway are an attempt to re-interpret Buddhist meditation techniques within a framework acceptable to Western scientific understanding. A truly holistic approach to harm minimization requires its consideration.

Introducing new building technologies : lessons from a case study

Hypocaust systems, where conditioned or non-conditioned air is passed through ducts within the concrete floor or ceiling of a building prior to its delivery to the rooms, are starting to appear in new buildings in Australia.

This paper describes the lessons that can be learned from the early experiences with a hypocaust system, installed in a new building in Melbourne. It concludes that a more cooperative process between all those involved in introducing and using a new 'technology' is essential if the problems described are to be avoided or at least minimized.

Emotional turmoil at the Bauhaus : problems of internal and external legitimacy : an historical case study

Walter Gropius established the Bauhaus in Germany in 1919. The organization established one of the most important design movements of the twentieth century. The organization had a very brief existence and was fraught with disruptions and emotional turmoil. Despite the difficulties, Gropius managed to keep the organization alive long enough for its extraordinary creativity to be harnessed and developed. The organization closed in 1933, but by that time its legitimacy as a source of design and pedagogy was assured. Organizational survival is often dependent on government subsidies, support through sales, donations or sponsorships. A factor in attracting this support is the perceived legitimacy of the organisation. Legitimacy is defined as a degree of consensus that the meanings and behavior of an organisation are valid and desirable by society in
general. Legitimacy remains an undeveloped concept. This paper reviews relevant theories of legitimacy, considers the role of emotions in shaping legitimacy and the emotions evoked as legitimacy is negotiated by internal and external stakeholders. A historical case study of the Bauhaus provides the backdrop for portraying the focal role emotions can play in institutionalization. The paper concludes with a discussion of the lessons of legitimacy available to contemporary cultural organisations.

An exploratory investigation of attitudes toward cause-related marketing

This paper reports on a study concerning consumer perceptions of cause-related marketing in Australia, and seeks to identify a direct link between consumer attitudes toward donating to charity (pro-social behaviour) and corporate philanthropy, and attitudes toward cause-related marketing. After a discussion on the related literature, there follows discussion on the research
model employed, and the fmdings of he study. Among other fmdings, statistically significant relationships between attitudes toward charitable giving and attitudes toward cause-related marketing are in evidence. In this study, respondents express a preference for local causes, as opposed to national and international causes, and also demonstrate a preference for cause types similar to those that already receive the highest level of charitable donations. The paper concludes with a discussion of the limitations of the study and presents suggestions for future research.

α2β1 integrin affects metastatic potential of ovarian carcinoma spheroids by supporting disaggregation and proteolysis

Background: Ovarian cancer is characterized by a wide-spread intra-abdominal metastases which represents a major clinical hurdle in the prognosis and management of the disease. A significant proportion of ovarian cancer cells in peritoneal ascites exist as multicellular aggregates or spheroids. We hypothesize that these cellular aggregates or spheroids are invasive with the capacity to survive and implant on the peritoneal surface. This study was designed to elucidate early inherent mechanism(s) of spheroid survival, growth and disaggregation required for peritoneal metastases.

Methods: In this study, we determined the growth pattern and adhesive capacity of ovarian cancer cell lines (HEY and OVHS1) grown as spheroids, using the well established liquid overlay technique, and compared them to a normal ovarian cell line (IOSE29) and cancer cells grown as a monolayer. The proteolytic capacity of these spheroids was compared with cells grown as a monolayer using a gelatin zymography assay to analyze secreted MMP-2/9 in conditioned serum-free medium. The disaggregation of cancer cell line spheroids was determined on extracellular matrices (ECM) such as laminin (LM), fibronectin (FN) and collagen (CI) and the expression of α2, α3, αv, α6 and β1 interin was determined by flow cytometric analysis. Neutralizing antibodies against α2, β1 subunits and α2β1 integrin was used to inhibit disaggregation as well as activation of MMPs in spheroids.

Results: We demonstrate that ovarian cancer cell lines grown as spheroids can sustain growth for 10 days while the normal ovarian cell line failed to grow beyond 2 days. Compared to cells grown as a monolayer, cancer cells grown as spheroids demonstrated no change in adhesion for up to 4 days, while IOSE29 cells had a 2–4-fold loss of adhesion within 2 days. Cancer cell spheroids disaggregated on extracellular matrices (ECM) and demonstrated enhanced expression of secreted pro-MMP2 as well as activated MMP2/MMP9 with no such activation of MMP's observed in monolayer cells. Flow cytometric analysis demonstrated enhanced expression of α2 and diminution of α6 integrin subunits in spheroids
versus monolayer cells. No change in the expression of α3, αv and β1 subunits was evident. Conversely, except for αv integrin, a 1.5–7.5-fold decrease in α2, α3, α6 and β1 integrin subunit expression was observed in IOSE29 cells within 2 days. Neutralizing antibodies against α2, β1 subunits and α2β1 integrin inhibited disaggregation as well as activation of
MMPs in spheroids.

Conclusion: Our results suggest that enhanced expression of α2β1 integrin may influence spheroid disaggregation and
proteolysis responsible for the peritoneal dissemination of ovarian carcinoma. This may indicate a new therapeutic target
for the suppression of the peritoneal metastasis associated with advanced ovarian carcinomas.

Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors

Background
Breast carcinoma is accompanied by changes in the acellular and cellular components of the microenvironment, the latter typified by a switch from fibroblasts to myofibroblasts.


Methods
We utilised conditioned media cultures, Western blot analysis and immunocytochemistry to investigate the differential effects of normal mammary fibroblasts (NMFs) and mammary cancer-associated fibroblasts (CAFs) on the phenotype and behaviour of PMC42-LA breast cancer cells. NMFs were obtained from a mammary gland at reduction mammoplasty, and CAFs from a mammary carcinoma after resection.


Results
We found greater expression of myofibroblastic markers in CAFs than in NMFs. Medium from both CAFs and NMFs induced novel expression of α-smooth muscle actin and cytokeratin-14 in PMC42-LA organoids. However, although conditioned media from NMFs resulted in distribution of vimentin-positive cells to the periphery of PMC42-LA organoids, this was not seen with CAF-conditioned medium. Upregulation of vimentin was accompanied by a mis-localization of E-cadherin, suggesting a loss of adhesive function. This was confirmed by visualizing the change in active β-catenin, localized to the cell junctions in control cells/cells in NMF-conditioned medium, to inactive β-catenin, localized to nuclei and cytoplasm in cells in CAF-conditioned medium.


Conclusion
We found no significant difference between the influences of NMFs and CAFs on PMC42-LA cell proliferation, viability, or apoptosis; significantly, we demonstrated a role for CAFs, but not for NMFs, in increasing the migratory ability of PMC42-LA cells. By concentrating NMF-conditioned media, we demonstrated the presence of factor(s) that induce epithelial-mesenchymal transition in NMF-conditioned media that are present at higher levels in CAF-conditioned media. Our in vitro results are consistent with observations in vivo showing that alterations in stroma influence the phenotype and behaviour of surrounding cells and provide evidence for a role for CAFs in stimulating cancer progression via an epithelial-mesenchymal transition. These findings have implications for our understanding of the roles of signalling between epithelial and stromal cells in the development and progression of mammary carcinoma.

What determines Australia's response to emergency and natural disasters

This article examines the determinants of Australia's response to emergencies and natural disasters. It examines the response from the Australian public by examining contributions made to the appeals of the country's largest Non-Governmental Organisation: World Vision of Australia. It also examines the response of the Australian Government. The data include 43 emergencies and natural disasters since 1998. Results suggest that the responses from both the public and government are positively associated with the number of people affected, media coverage, and the level of political and civil freedom in the country where the event occurred. The type and location of the emergency or disaster are important for the public's response. Differences between public and government donations exist: support from the Australian Government is positively associated with smaller countries and there is some evidence that the public donates more to events occurring in larger and poorer countries.

Blueprinting perspective to managing giving

This paper examines the potential of using service blueprinting to assist non-profit organizations in better managing their donation efficiency by identifying the complexities of the donation process from a customer perspective, which in turn should allow them to better meet the needs of those they seek to assist. A disregard of donors’ expectations in relation to the overall experience could potentially result in “dissatisfaction” and thus reductions in future donations to a given organization.

Service blueprinting : a potential tool for improving cause-donor exchanges

This paper examines the potential for using service blueprinting to assist non-profit organizations in better managing donor exchanges. In many cases organizations are unclear of donors' expectations in relation to the overall giving experience. Failure to deliver on expectations could potentially result in donor 'dissatisfaction', which in turn may result in reductions of future donations to these organizations. This article begins with a review of blueprinting and the benefits of using such as tool as to assist in understanding the donation exchange. Zeithaml and Bitner's (2000) steps in building a blueprint are explained in the context of non-profit-donor exchanges. An example of a hypothetical blueprint provided to highlight the complexity and divergence encountered in such exchanges and used to identify potential problems associated with the various aspects of the donation experience. Finally, implications for managing giving and suggestions for future research are offered.

Prominent messages in education and applied linguistic abstracts : how do authors appeal to their prospective readers?

This study investigates the rhetorical structure of abstracts of papers published in Applied Linguistics and Education. It examines how abstract authors in these two fields emphasise the significance of their research, and how they appeal to their prospective readership. Although abstracts in both disciplinary groups are found to display a coordinate textual development they exhibit a utilization of different relational schemata to indicate the functional prominence of textual propositions. In particular, different relational patterns are seen to be employed to fulfil the two primary objectives of an abstract: to provide a synopsis of the accompanying article, and to promote it to relevant research and professional communities. The way authors demonstrate the value of their research and their professional credibility appears to be conditioned by disciplinary writing conventions. It is proposed that relational choices, which result in differences in the accentuation of communicative messages in Applied Linguistics and Education abstracts, depend on the perceived relationship between the author and the discourse community in terms of expectations of prior knowledge.

Characterization of proADAMTS5 processing by proprotein convertases.

ADAMTS5 (aggrecanase-2), a key metalloprotease mediating cartilage destruction in arthritis, is synthesized as a zymogen, proADAMTS5. We report a detailed characterization of the propeptide excision mechanism and demonstrate that it is a major regulatory step with unusual characteristics. Using furin-deficient cells and a furin inhibitor, we found that proADAMTS5 was processed by proprotein convertases, specifically furin and PC7, but not PC6B. Mutagenesis of three sites containing basic residues within the ADAMTS5 propeptide (RRR⁴⁶, RRR⁶⁹ and RRRRR²⁶¹) suggested that proADAMTS5 processing occurs after ^Arg261. That furin processing was essential for ADAMTS5 activity was illustrated using the known ADAMTS5 substrate aggrecan, as well as a new substrate, versican, an important regulatory proteoglycan during mammalian development. When compared to other ADAMTS proteases, proADAMTS5 processing has several distinct features. In contrast to ADAMTS1, whose furin processing products were clearly present intracellularly, cleaved ADAMTS5 propeptide and mature ADAMTS5 were found exclusively in the conditioned medium. Despite attempts to enhance detection of intracellular proADAMTS5 processing, such as by immunoprecipitation of total ADAMTS5, overexpression of furin, and secretion blockade by monensin, neither processed ADAMTS5 propeptide nor the mature enzyme were found intracellularly, which was strongly suggestive of extracellular processing. Extracellular ADAMTS5 processing was further supported by activation of proADAMTS5 added exogenously to HEK293 cells stably expressing furin. Unlike proADAMTS9, which is processed by furin at the cell-surface, to which it is bound, ADAMTS5 does not bind the cell-surface. Thus, the propeptide processing mechanism of ADAMTS5 has several points of distinction from those of other ADAMTS proteases, which may have considerable significance in the context of osteoarthritis.

Sport sponsorship

OBJECTIVES
By the end of this chapter, readers will be able to:
• describe the nature and role of sponsorship as part of a company's marketing mix;
• be familiar with terminology relating to sponsorship and integrated sponsorship marketing;
• discuss the background and growth of sponsorship as a marketing discipline globally and in New Zealand;
• understand corporate methodology for measuring sponsorship effectiveness;
• identify models for sponsorship selection, planning, management and evaluation;
• understand the role of research in sponsorship;
• describe techniques for developing a sponsorship proposal;
• be familiar with global and local trends in sponsorship;
• understand legal implications, including contracts/agreements, ambush marketing and cause-related sponsorship.