Antisense-mediated suppression of hyaluronan synthase 2 inhibits the tumorigenesis and progression of breast cancer

The progression of several cancers is correlated with the increased synthesis of the glycosaminoglycan, hyaluronan. Hyaluronan is synthesized at the plasma membrane by various isoforms of hyaluronan synthases (HAS). The importance of HAS2 expression in highly invasive breast cancer was characterized by the antisense inhibition of HAS2 (ASHAS2). The effect of HAS2 inhibition on cell proliferation, migration, hyaluronan metabolism, and receptor status was characterized in vitro, whereas the effect on tumorigenicity and metastasis was established in vivo. HAS2 inhibition resulted in a 24-hour lag in proliferation that was concomitant to transient arrest of 79% of the cell population in G 0-G1. Inhibition of HAS2 did not alter the expression of the other HAS isoforms, whereas hyaluronidase (HYAL2) and the hyaluronan receptor, CD44, were significantly down-regulated. ASHAS2 cells accumulated greater amounts of high molecular weight hyaluronan (>10,000 kDa) in the culture medium, whereas mock and parental cells liberated less hyaluronan of three distinct molecular weights (100, 400, and 3,000 kDa). The inhibition of HAS2 in the highly invasive MDA-MB-231 breast cancer cell line inhibited the initiation and progression of primary and secondary tumor formation following s.c. and intracardiac inoculation into nude mice, whereas controls readily established both primary and secondary tumors. The lack of primary and secondary tumor formation was manifested by increased survival times where ASHAS2 animals survived 172% longer than the control animals. Collectively, these unique results strongly implicate the central role of HAS2 in the initiation and progression of breast cancer, potentially highlighting the codependency between HAS2, CD44, and HYAL2 expression. ©2005 American Association for Cancer Research.

Contribution of fibroblast and mast cell (afferent) and tumor (efferent) IL-6 effects within the tumor microenvironment

Hyperactive inflammatory responses following cancer initiation have led to cancer being described as a 'wound that never heals'. These inflammatory responses elicit signals via NFκB leading to IL-6 production, and IL-6 in turn has been shown to induce epithelial to mesenchymal transition in breast cancer cells in vitro, implicating a role for this cytokine in cancer cell invasion. We previously have shown that conditioned medium derived from cancer-associated fibroblasts induced an Epithelial to Mesenchymal transition (EMT) in PMC42-LA breast cancer cells and we have now identify IL-6 as present in this medium. We further show that IL-6 is expressed approximately 100 fold higher in a cancer-associated fibroblast line compared to normal fibroblasts. Comparison of mouse-specific (stroma) and human-specific (tumor) IL-6 mRNA expression from MCF-7, MDA MB 468 and MDA MB 231 xenografts also indicated the stroma rather than tumor as a significantly higher source of IL-6 expression. Mast cells (MCs) feature in inflammatory cancer-associated stroma, and activated MCs secrete IL-6. We observed a higher MC index (average number of mast cells per xenograft section/average tumor size) in MDA MB 468 compared to MDA MB 231 xenografts, where all MC were observed to be active (degranulating). This higher MC index correlated with greater mouse-specific IL-6 expression in the MDA MB 468 xenografts, implicating MC as an important source of stromal IL-6. Furthermore, immunohistochemistry on these xenografts for pSTAT3, which lies downstream of the IL-6 receptor indicated frequent correlations between pSTAT3 and mast cell positive cells. Analysis of publically available databases for IL-6 expression in patient tissue revealed higher IL-6 in laser capture microdissected stroma compared to adjacent tissue epithelium from patients with inflammatory breast cancer (IBC) and invasive non-inflammatory breast cancer (non-IBC) and we show that IL-6 expression was significantly higher in Basal versus Luminal molecular/phenotypic groupings of breast cancer cell lines. Finally, we discuss how afferent and efferent IL-6 pathways may participate in a positive feedback cycle to dictate tumor progression.

Hip and knee kinematics display complex and time-varying sagittal kinematics during repetitive stepping: Implications for design of a functional fatigue model of the knee extensors and flexors

The validity of fatigue protocols involving multi-joint movements, such as stepping, has yet to be clearly defined. Although surface electromyography can monitor the fatigue state of individual muscles, the effects of joint angle and velocity variation on signal parameters are well established. Therefore, the aims of this study were to i) describe sagittal hip and knee kinematics during repetitive stepping ii) identify periods of high inter-trial variability and iii) determine within-test reliability of hip and knee kinematic profiles. A group of healthy men (N = 15) ascended and descended from a knee-high platform wearing a weighted vest (10%BW) for 50 consecutive trials. The hip and knee underwent rapid flexion and extension during step ascent and descent. Variability of hip and knee velocity peaked between 20-40% of the ascent phase and 80-100% of the descent. Significant (p<0.05) reductions in joint range of motion and peak velocity during step ascent were observed, while peak flexion velocity increased during descent. Healthy individuals use complex hip and knee motion to negotiate a knee-high step with kinematic patterns varying across multiple repetitions. These findings have important implications for future studies intending to use repetitive stepping as a fatigue model for the knee extensors and flexors.

Translations of industry-based student learning and academic performances

We hypothesized that Industry based learning and teaching, especially through industry assigned student projects or training programs, is an integral part of science, technology, engineering and mathematics (STEM) education. In this paper we show that industry-based student training and experience increases students’ academic performances independent to the organizational parameters and contexts. The literature on industry-based student training focuses on employability and the industry dimension, and neglects in many ways the academic dimension. We observed that the association factors between academic attributes and contributions of industry-based student training are central and vital to the technological learning experiences. We explore international initiatives and statistics collected of student projects in two categories: Industry based learning performances and on campus performances. The data collected were correlated to five (5) universities in different industrialized countries, e.g., Australia N=545, Norway N=279, Germany N=74, France N=107 and Spain N=802 respectively. We analyzed industry-based student training along with company assigned student projects compared with in comparisons to campus performance. The data that suggests a strong correlation between industry-based student training per se and improved performance profiles or increasing motivation shows that industry-based student training increases student academic performance independent of organizational parameters and contexts. The programs we augmented were orthogonal to each other however, the trend of the students’ academic performances are identical. An isolated cohort for the reported countries that opposed our hypothesis warrants further investigation.

Transformation in educational practices through STEM

The need for strong science, technology and innovation linkages between Higher Education Institutions (HEIs) and industries is a pivotal point for middle-income countries in their endeavor to enhance human capital in socioeconomic development. Currently, the University-Industry partnerships are at an infant stage in Sri Lankan higher education context. Technological maturity and effective communication skills are contributing factors for an efficient graduate profile. Also, expanding internship programs in particular for STEM disciplines provide work experience to students that would strengthen the relevance of higher education programs. This study reports historical overviews and current trends in STEM education in Sri Lanka. Emphasis will be drawn to recent technological and higher education curricular reforms. Data from the last 10 years were extracted from the higher education sector and Ministry of Higher Education Policy portfolios. Associations and trend analysis of the sector growth were compared with STEM existence, merger and predicted augmentations. Results were depicted and summarised based on STEM streams and disciplines. It was observed that the trend of STEM augmentation in the Sri Lankan Higher Education context is growing at a slow but steady pace. Further analysis with other sectors in particular, Industry information, would be useful and a worthwhile exercise.

STEM education for sustainable development : a sociotechnical analysis

It is debated that for sustainable STEM education and knowledge investment, human centered learning design approach is critical and important. Sustainability in this context is enduring maintenance of technological trajectories for productive economical and social interactions by demonstrating life critical scenarios through life critical system development and life experiences. Technology influences way of life and the learning and teaching process. Social software application development is more than learning of how to program a software application and extracting information from the Internet. Hence, our research challenge is, how do we attract learners to STEM social software application development? Our realisation processes begin with comparing Science and Technology education in developed (e.g., Australia) and developing (e.g., Sri Lanka) countries with distinction on final year undergraduates’ industry ready training programmes. Principal components analysis was performed to separate patterns of important factors. To measure behavioural intention of perceived usefulness and attitudes of the training, the measurement model was analysed to test its validity and reliability using partial least square (PLS) analysis of structural equation modelling (SEM). Our observation is that the relationship is more complex than we argue for. Our initial conclusions were that life critical system development and life experience trajectories as determinant factors while technological influences were unavoidable. A further investigation should involve correlations between human centered learning design approach and economical development in the long run.

Does usability engineering matters for STEM education?

Technological maturity and the exponential growth of digital applications are contributing to lifestyle changes worldwide. Consequently, learning and teaching is demanding more effective sociotechnical interactions involving emerging technologies, as opposed to traditional, conventional face-to-face learning and teaching approaches. In this context, usability engineering is making significant contributions for improving computer and distance-based learning, both for learners and instructors, which have often been ignored when designing online learning and teaching applications. Usability testing is a central part of the human centered learning approach for developing sustainable STEM education from the socio-technological perspective. Our experiences with usability engineering and the impact of teaching low-cost rapid usability testing methods on knowledge translation from undergraduate to graduate courses to real-world practice (i.e. getting the methods out there in real use) are diverse and multi-modal. Our sample space has been hundreds of trained students who have learned how to do effective usability engineering in real-world situations at higher levels of realism (i.e. fidelity) and at a much lower cost than using traditional fixed usability labs. Furthermore, this low-cost rapid approach to usability engineering has been adopted by many of our graduates who are now managers, CIOs etc and who are using the methods routinely in their organizations in real world applications and scenarios. This knowledge has been used to improve design and implementation of a wide range of applications, including applications designed for teaching and learning.

Contribution of industry-based student learning performance for the STEM education

We hypothesized that Industry based learning and teaching, especially through company assigned student projects or training programs, is an integral part of science, technology, engineering and mathematics (STEM) education. In this paper we show that industry-based student training and experience increases students’ academic performances independent to the organizational parameters and contexts. The literature on industry-based student training focuses on employability and the industry dimension, and neglects in many ways the academic dimension. We observed that the association factors between academic attributes and contributions of industry-based student training are central and vital to the technological learning experiences. We explore international initiatives and statistics collected of student projects in two categories: Industry based learning performances and on campus performances. The data collected were correlated to five (5) universities in different industrialized countries, e.g., Australia N=545 projects, Norway N=279, Germany N=74, France N=107 and Spain N=802. We analyzed industry-based student training along with company assigned student projects compared with in comparisons to campus performance. The data that suggests a strong correlation between industry-based student training per se and improved performance profiles or increasing motivation shows that industry-based student training increases student academic performance independent of organizational parameters and contexts. The programs we augmented were orthogonal to each other however, the trend of the students’ academic performances are identical. An isolated cohort for the reported countries that opposed our hypothesis warrants further investigation.

Upregulation of matrix metalloproteinases (MMPs) in breast cancer xenografts : a major induction of stromal MMP-13

In human breast cancer (HBC), as with many carcinoma systems, most matrix metalloproteinases (MMPs) are largely expressed by the stromal cells, whereas the tumour cells are relatively silent in MMP expression. To determine the tissue source of the most relevant MMPs, we xenografted HBC cell lines and HBC tissues into the mammary fat pad (MFP) or bone of immunocompromised mice and measured the expression of human and mouse MMP-2, -9, -11, -13, membrane-type-1 MMP (MT1-MMP), MT2-MMP and MT3-MMP by species-specific real-time quantitative RT-PCR. Our data confirm a stromal origin for most tumour-associated MMPs and indicate marked and consistent upregulation of stromal (mouse) MMP-13 and MT1-MMP in all xenografts studied, irrespective of implantation in the MFP or bone environments. In addition, we show increased expression of both human MMP-13 and human MT1-MMP by the MDA-MB-231 tumour cells grown in the MFP compared to in vitro production. MMP protein and activity data confirm the upregulation of MMP mRNA production and indicate an increase in the activated MMP-2 species as a result of tumour implantation. These data directly demonstrate tumour induction of MMP production by stromal cells in both the MFP and bone environments. These xenografts are a valuable means for examining in vivo production of MMPs and suggest that MMP-13 and MT1-MMP will be relevant targets for inhibiting breast cancer progression.

Epithelial mesenchymal transition traits in human breast cancer cell lines parallel the CD44HI/CD24lO/-stem cell phenotype in human breast cancer

We review here the recently emerging relationship between epithelial-mesenchymal transition (EMT) and breast cancer stem cells (BCSC), and provide analyses of published data on human breast cancer cell lines, supporting their utility as a model for the EMT/BCSC state. Genome-wide transcriptional profiling of these cell lines has confirmed the existence of a subgroup with mesenchymal tendencies and enhanced invasive properties ('Basal B'/Mesenchymal), distinct from subgroups with either predominantly luminal ('Luminal') or mixed basal/luminal ('Basal A') features (Neve et al. Cancer Cell, 2006). A literature-derived EMT gene signature has shown specific enrichment within the Basal B subgroup of cell lines, consistent with their over-expression of various EMT transcriptional drivers. Basal B cell lines are found to resemble BCSC, being CD44highCD24low. Moreover, gene products that distinguish Basal B from Basal A and Luminal cell lines (Basal B Discriminators) showed close concordance with those that define BCSC isolated from clinical material, as reported by Shipitsin et al. (Cancer Cell, 2007). CD24 mRNA levels varied across Basal B cell lines, correlating with other Basal B Discriminators. Many gene products correlating with CD24 status in Basal B cell lines were also differentially expressed in isolated BCSC. These findings confirm and extend the importance of the cellular product of the EMT with Basal B cell lines, and illustrate the value of analysing these cell lines for new leads that may improve breast cancer outcomes. Gene products specific to Basal B cell lines may serve as tools for the detection, quantification, and analysis of BCSC/EMT attributes.

The type I collagen induction of MT1-MMP-mediated MMP-2 activation is repressed by αVβ3 integrin in human breast cancer cells

The influence of αVβ3 integrin on MT1-MMP functionality was studied in human breast cancer cells of differing β3 integrin status. Overexpression of β3 integrin caused increased cell surface expression of αV integrin and increased cellular adhesion to extracellular matrix (ECM) substrates in BT-549, MDA-MB-231 and MCF-7 cells. β3 integrin expression also enhanced the migration of breast cancer cells on ECM substrates and enhanced collagen gel contraction. In vivo, αVβ3 cooperated with MT1-MMP to increase the growth of MCF-7 cells after orthotopic inoculation in immunocompromised mice, but had no influence on in vitro proliferation. Despite these stimulatory effects, overexpression of β3 integrin suppressed the type I collagen (Col I) induced MMP-2 activation in all breast cancer cell lines analyzed. This was also evident in extracts from the MCF-7 tumors in vivo, where MMP-2 activation was stimulated by MT1-MMP transfection, but attenuated with β3 integrin expression. Although our studies confirm important biological effects of αVβ3 integrin on enhancing cell adhesion and migration, ECM remodeling and tumor growth, β3 integrin caused reduced MMP-2 activation in response to Col I in vitro, which appears to be physiologically relevant, as it was also seen in tumor xenografts in vivo. The reduction of MMP-2 activation (and thus MT1-MMP activity) by αVβ3 in response to Col I may be important in scenarios where cells which are activated for matrix degradation need to preserve some pericellular collagen, perhaps as a substrate for cell adhesion and migration, thus maintaining a balanced level of proteolysis required for efficient tumor growth.

eHealth-as-a-Service (eHaaS) : towards universal stakeholder engagement

With the introduction of the Personally Controlled Health Record (PCEHR), the Australian public is being asked to accept greater responsibility for their healthcare by taking an active role in the management of personal health information. Although well designed, constructed and intentioned, policy and privacy concerns have resulted in an eHealth model that may impact future health sharing requirements. Hence, as a case study for a consumer eHealth initative in the Australian context, eHealth-as-a-Service (eHaaS) serves as a disruptive step in in the aggregation and transformation of health information for use as real-world knowledge. The strategic value of extending the community Health Record Bank (HRB) model lies in the ability to automatically draw on a multitude of relevant data repositories and sources to create a single source of the truth and to engage market forces to create financial sustainability. The opportunity to transform the beleaguered Australian PCEHR into a realisable and sustainable technology consumption model for patient safety is explored. Moreover, the current clerical focus of healthcare practitioners acting in the role of de facto record keepers is renegotiated to establish a shared knowledge creation landscape of action for safer patient interventions. To achieve this potential however requires a platform that will facilitate efficient and trusted unification of all health information available in real-time across the continuum of care. eHaaS provides a sustainable environment and encouragement to realise this potential.

Consumer acceptance of Accountable-eHealth systems

In this paper, we present the results of a survey conducted to measure the attitudes of the consumers of eHealth towards Accountable-eHealth systems which are designed for information privacy management. A research model is developed that can identify the factors contributing to system acceptance and is validated using quantitative data from 187 completed survey responses from university students studying non-health related courses at a university in Queensland, Australia. The research model is validated using structural equation modelling and can be used to identify how specific characteristics of Accountable-eHealth systems would affect their overall acceptance by future eHealth consumers.

Barriers to implementing eHealth : a multi-dimensional perspective

This paper strives to identify barriers that hamper eHealth implementation from different perspectives. The benefits offered by eHealth and the need for eHealth preparedness is first discussed. This is followed by a discussion on the integral components of a robust eHealth infrastructure. Then, the barriers to eHealth such as technical interoperability issues, lack of holistic approach and technology disconnect are explained in detail. Finally, solutions to promote better adoption of eHealth through government policies, standardisation and training are also discussed.

The role of perceived usefulness and attitude on electronic health record acceptance

Information and communications technologies are a significant component of the healthcare domain, and electronic health records play a major role in it. Therefore, it is important that they are accepted en masse by healthcare professionals. How healthcare professionals perceive the usefulness of electronic health records and their attitudes towards them have been shown to have significant effects on the overall acceptance in many healthcare systems around the world. This paper investigates the role of perceived usefulness and attitude on the intention to use electronic health records by future healthcare professionals using polynomial regression with response surface analysis. Results show that the relationships between these variables are more complex than predicted in prior research. The paper concludes that the properties of the above determinants must be further investigated to clearly understand: (i) their role in predicting the intention to use electronic health records; and (ii) in designing systems that are better adopted by healthcare professionals of the future.