Influence of culture conditions on the molecular signature of mesenchymal stem cells

Cell based therapies require cells capable of self renewal and differentiation, and a prerequisite is the ability to prepare an effective dose of ex vivo expanded cells for autologous transplants. The in vivo identification of a source of physiologically relevant cell types suitable for cell therapies is therefore an integral part of tissue engineering. Bone marrow is the most easily accessible source of mesenchymal stem cells (MSCs), and harbours two distinct populations of adult stem cells; namely hematopoietic stem cells (HSCs) and bone mesenchymal stem cells (BMSCs). Unlike HSCs, there are yet no rigorous criteria for characterizing BMSCs. Changing understanding about the pluripotency of BMSCs in recent studies has expanded their potential application; however, the underlying molecular pathways which impart the features distinctive to BMSCs remain elusive. Furthermore, the sparse in vivo distribution of these cells imposes a clear limitation to their in vitro study. Also, when BMSCs are cultured in vitro there is a loss of the in vivo microenvironment which results in a progressive decline in proliferation potential and multipotentiality. This is further exacerbated with increased passage number, characterized by the onset of senescence related changes. Accordingly, establishing protocols for generating large numbers of BMSCs without affecting their differentiation potential is necessary. The principal aims of this thesis were to identify potential molecular factors for characterizing BMSCs from osteoarthritic patients, and also to attempt to establish culture protocols favourable for generating large number of BMSCs, while at the same time retaining their proliferation and differentiation potential. Previously published studies concerning clonal cells have demonstrated that BMSCs are heterogeneous populations of cells at various stages of growth. Some cells are higher in the hierarchy and represent the progenitors, while other cells occupy a lower position in the hierarchy and are therefore more committed to a particular lineage. This feature of BMSCs was made evident by the work of Mareddy et al., which involved generating clonal populations of BMSCs from bone marrow of osteoarthritic patients, by a single cell clonal culture method. Proliferation potential and differentiation capabilities were used to group cells into fast growing and slow growing clones. The study presented here is a continuation of the work of Mareddy et al. and employed immunological and array based techniques to identify the primary molecular factors involved in regulating phenotypic characteristics exhibited by contrasting clonal populations. The subtractive immunization (SI) was used to generate novel antibodies against favourably expressed proteins in the fast growing clonal cell population. The difference between the clonal populations at the transcriptional level was determined using a Stem Cell RT2 Profiler TM PCR Array which focuses on stem cell pathway gene expression. Monoclonal antibodies (mAb) generated by SI were able to effectively highlight differentially expressed antigenic determinants, as was evident by Western blot analysis and confocal microscopy. Co-immunoprecipitation, followed by mass spectroscopy analysis, identified a favourably expressed protein as the cytoskeletal protein vimentin. The stem cell gene array highlighted genes that were highly upregulated in the fast growing clonal cell population. Based on their functions these genes were grouped into growth factors, cell fate determination and maintenance of embryonic and neural stem cell renewal. Furthermore, on a closer analysis it was established that the cytoskeletal protein vimentin and nine out of ten genes identified by gene array were associated with chondrogenesis or cartilage repair, consistent with the potential role played by BMSCs in defect repair and maintaining tissue homeostasis, by modulating the gene expression pattern to compensate for degenerated cartilage in osteoarthritic tissues. The gene array also presented transcripts for embryonic lineage markers such as FOXA2 and Sox2, both of which were significantly over expressed in fast growing clonal populations. A recent groundbreaking study by Yamanaka et al imparted embryonic stem cell (ESCs) -like characteristic to somatic cells in a process termed nuclear reprogramming, by the ectopic expression of the genes Sox2, cMyc and Oct4. The expression of embryonic lineage markers in adult stem cells may be a mechanism by which the favourable behaviour of fast growing clonal cells is determined and suggests a possible active phenomenon of spontaneous reprogramming in fast growing clonal cells. The expression pattern of these critical molecular markers could be indicative of the competence of BMSCs. For this reason, the expression pattern of Sox2, Oct4 and cMyc, at various passages in heterogeneous BMSCs population and tissue derived cells (osteoblasts and chondrocytes), was investigated by a real-time PCR and immunoflourescence staining. A strong nuclear staining was observed for Sox2, Oct4 and cMyc, which gradually weakened accompanied with cytoplasmic translocation after several passage. The mRNA and protein expression of Sox2, Oct4 and cMyc peaked at the third passage for osteoblasts, chondrocytes and third passage for BMSCs, and declined with each subsequent passage, indicating towards a possible mechanism of spontaneous reprogramming. This study proposes that the progressive decline in proliferation potential and multipotentiality associated with increased passaging of BMSCs in vitro might be a consequence of loss of these propluripotency factors. We therefore hypothesise that the expression of these master genes is not an intrinsic cell function, but rather an outcome of interaction of the cells with their microenvironment; this was evident by the fact that when removed from their in vivo microenvironment, BMSCs undergo a rapid loss of stemness after only a few passages. One of the most interesting aspects of this study was the integration of factors in the culture conditions, which to some extent, mimicked the in vivo microenvironmental niche of the BMSCs. A number of studies have successfully established that the cellular niche is not an inert tissue component but is of prime importance. The total sum of stimuli from the microenvironment underpins the complex interplay of regulatory mechanisms which control multiple functions in stem cells most importantly stem cell renewal. Therefore, well characterised factors which affect BMSCs characteristics, such as fibronectin (FN) coating, and morphogens such as FGF2 and BMP4, were incorporated into the cell culture conditions. The experimental set up was designed to provide insight into the expression pattern of the stem cell related transcription factors Sox2, cMyc and Oct4, in BMSCs with respect to passaging and changes in culture conditions. Induction of these pluripotency markers in somatic cells by retroviral transfection has been shown to confer pluripotency and an ESCs like state. Our study demonstrated that all treatments could transiently induce the expression of Sox2, cMyc and Oct4, and favourably affect the proliferation potential of BMSCs. The combined effect of these treatments was able to induce and retain the endogenous nuclear expression of stem cell transcription factors in BMSCs over an extended number of in vitro passages. Our results therefore suggest that the transient induction and manipulation of endogenous expression of transcription factors critical for stemness can be achieved by modulating the culture conditions; the benefit of which is to circumvent the need for genetic manipulations. In summary, this study has explored the role of BMSCs in the diseased state of osteoarthritis, by employing transcriptional profiling along with SI. In particular this study pioneered the use of primary cells for generating novel antibodies by SI. We established that somatic cells and BMSCs have a basal level of expression of pluripotency markers. Furthermore, our study indicates that intrinsic signalling mechanisms of BMSCs are intimately linked with extrinsic cues from the microenvironment and that these signals appear to be critical for retaining the expression of genes to maintain cell stemness in long term in vitro culture. This project provides a basis for developing an “artificial niche” required for reversion of commitment and maintenance of BMSC in their uncommitted homeostatic state.

Integration of CENA practice standards for postgraduate emergency nursing assessment

The use of professional competency standards to assess postgraduate nursing student’s clinical performance has been in place since 2004, at the Queensland University of Technology, School of Nursing & Midwifery (SONAM) when the Graduate Certificate in Emergency Nursing degree commenced. Emergency nursing students were assessed in their workplace, using a Clinical Performance Appraisal Tool or CPAT which was based on the Australian College of Critical Care Nurses (ACCCN) Competency Standards. With the subsequent formation of a separate Emergency Nursing Course advisory group in 2007, there was a review of clinical assessment course component. The release of the 2008 CENA revised Practice Standards for the Emergency Nursing Specialist’s, led to the emergency nursing course advisory committee supporting the integration of the CENA practice standards for assessment of emergency nurses in preference to the less relevant ACCCN competency standards. The SONAM emergency nursing study area team commenced the phasing in and progression of the CENA practice standards across the two Graduate Certificate units, and Graduate Diploma and Master of Nursing (emergency) clinical major options in 2009. As some units undertaken in the degree are available to nurses in other disciplines a separate CPAT was devised for the clinical assessments according to speciality context. The team has had to carefully consider how the professional standards are integrated into the teaching and assessment of the unit and not just applied instead of the ACCCN competency standards. Professional standards for the emergency context has also helped tailor course content and learning outcomes to be relevant across a number of emergency nursing contexts in Australia. The assessment of the CPAT is undertaken at the workplace by QUT appointed clinical lecturers. Clinical lecturers need to apply and have suitable postgraduate qualification to undertake the position. The clinical lecturer support role is well established at QUT. The integration of the new CENA practice standards has necessitated a review of the postgraduate assessment of emergency nurses. A clinical lecturer workshop has been organised to review role, scope and how to utilise the new look CENA based CPAT, clinical assessment format.

Enterprise architecture and the integration of service-oriented architecture

In recent years, enterprise architecture (EA) has captured a growing attention as a means to systematically consolidate and interrelate diverse business and IT artefacts in order to provide holistic decision support. The recent popularity of a service-orientation has added “service “and related constructs as a new element that requires consideration within an Enterprise Architecture. Since the emergence of the Service-Oriented Architecture (SOA), many attempts have been made to incorporate SOA artefacts in existing EA frameworks. Yet, the approaches taken to achieve this goal differ substantially for the most commonly used EA frameworks to date. SOA in the context of enterprise architecture is one of the future research challenges. Several authors argue that further research is needed in order to understand how SOA impacts prior enterprise architecture frameworks. This study explores SOA integration within EA, identifies SOA integration approaches within EA and identifies factors that impact SOA integration within Enterprise Architecture.

Collaboration : developing integration in multi-purpose services in rural New South Wales, Australia

Introduction: The Multi-purpose Service (MPS) Program was introduced to rural Australia in 1991 as a solution to poor health outcomes in rural compared with metropolitan populations, difficulty in attracting healthcare staff and a lack of viability and range of health services in rural areas. The aim of this study was to describe the main concerns of participants involved in the development of multi-purpose services in rural New South Wales (NSW). This article is abstracted from a larger study and discusses the extent to which collaboration occurred within the new multi-purpose service. Methods: A constructivist grounded theory methodology was used. Participants were from 13 multi-purpose services in rural NSW and 30 in-depth interviews were conducted with 6 community members, 11 managers and 13 staff members who had been involved in the process of developing a multi-purpose service. Results: The main concern of all participants was their anticipation of risk. This anticipation of risk manifested itself in either trust or suspicion and explained their progression through a phase of collaborating. Participants who had trust in other stakeholders were more likely to embrace an integrated health service identity. Those participants, who were suspicious that they would lose status or power, maintained that the previous hospital services provided a better health service and described a coexistence of services within the multi-purpose service. Conclusions: This study provided an insight into the perceptions of community members, staff members and managers involved in the process of developing a multi-purpose service. It revealed that the anticipation of risk was intrinsic to a process of changing from a traditional hospital service to collaborating in a new model of health care provided at a multi-purpose service.

Symbolic number : the integration of magnitude and spatial representations in children aged 6 to 8 years

The process of learning symbolic Arabic digits in early childhood requires that magnitude and spatial information integrates with the concept of symbolic digits. Previous research has separately investigated the development of automatic access to magnitude and spatial information from symbolic digits. However, developmental trajectories of symbolic number knowledge cannot be fully understood when considering components in isolation. In view of this, we have synthesized the existing lines of research and tested the use of both magnitude and spatial information with the same sample of British children in Years 1, 2 and 3 (6-8 years of age). The physical judgment task of the numerical Stroop paradigm (NSP) demonstrated that automatic access to magnitude was present from Year 1 and the distance effect signaled that a refined processing of numerical information had developed. Additionally, a parity judgment task showed that the onset of the Spatial-Numerical Association of Response Codes (SNARC) effect occurs in Year 2. These findings uncover the developmental timeline of how magnitude and spatial representations integrate with symbolic number knowledge during early learning of Arabic digits and resolve inconsistencies between previous developmental and experimental research lines.

Fundamentals of process integration

Technologies and languages for integrated processes are a relatively recent innovation. Over that period many divergent waves of innovation have transformed process integration. Like sockets and distributed objects, early workflow systems ordered programming interfaces that connected the process modelling layer to any middleware. BPM systems emerged later, connecting the modelling world to middleware through components. While BPM systems increased ease of use (modelling convenience), long-standing and complex interactions involving many process instances remained di±cult to model. Enterprise Service Buses (ESBs), followed, connecting process models to heterogeneous forms of middleware. ESBs, however, generally forced modellers to choose a particular underlying middleware and to stick to it, despite their ability to connect with many forms of middleware. Furthermore ESBs encourage process integrations to be modelled on their own, logically separate from the process model. This can lead to the inability to reason about long standing conversations at the process layer. Technologies and languages for process integration generally lack formality. This has led to arbitrariness in the underlying language building blocks. Conceptual holes exist in a range of technologies and languages for process integration and this can lead to customer dissatisfaction and failure to bring integration projects to reach their potential. Standards for process integration share similar fundamental flaws to languages and technologies. Standards are also in direct competition with other standards causing a lack of clarity. Thus the area of greatest risk in a BPM project remains process integration, despite major advancements in the technology base. This research examines some fundamental aspects of communication middleware and how these fundamental building blocks of integration can be brought to the process modelling layer in a technology agnostic manner. This way process modelling can be conceptually complete without becoming stuck in a particular middleware technology. Coloured Petri nets are used to define a formal semantics for the fundamental aspects of communication middleware. They provide the means to define and model the dynamic aspects of various integration middleware. Process integration patterns are used as a tool to codify common problems to be solved. Object Role Modelling is a formal modelling technique that was used to define the syntax of a proposed process integration language. This thesis provides several contributions to the field of process integration. It proposes a framework defining the key notions of integration middleware. This framework provides a conceptual foundation upon which a process integration language could be built. The thesis defines an architecture that allows various forms of middleware to be aggregated and reasoned about at the process layer. This thesis provides a comprehensive set of process integration patterns. These constitute a benchmark for the kinds of problems a process integration language must support. The thesis proposes a process integration modelling language and a partial implementation that is able to enact the language. A process integration pilot project in a German hospital is brie°y described at the end of the thesis. The pilot is based on ideas in this thesis.

The impact of knowledge integration on enterprise system success

Despite promising benefits and advantages, there are reports of failures and low realisation of benefits in Enterprise System (ES) initiatives. Among the research on the factors that influence ES success, there is a dearth of studies on the knowledge implications of multiple end-user groups using the same ES application. An ES facilitates the work of several user groups, ranging from strategic management, management, to operational staff, all using the same system for multiple objectives. Given the fundamental characteristics of ES – integration of modules, business process views, and aspects of information transparency – it is necessary that all frequent end-users share a reasonable amount of common knowledge and integrate their knowledge to yield new knowledge. Recent literature on ES implementation highlights the importance of Knowledge Integration (KI) for implementation success. Unfortunately, the importance of KI is often overlooked and little about the role of KI in ES success is known. Many organisations do not achieve the potential benefits from their ES investment because they do not consider the need or their ability to integrate their employees’ knowledge. This study is designed to improve our understanding of the influence of KI among ES end-users on operational ES success. The three objectives of the study are: (I) to identify and validate the antecedents of KI effectiveness, (II) to investigate the impact of KI effectiveness on the goodness of individuals’ ES-knowledge base, and (III) to examine the impact of the goodness of individuals’ ES-knowledge base on the operational ES success. For this purpose, we employ the KI factors identified by Grant (1996) and an IS-impact measurement model from the work of Gable et al. (2008) to examine ES success. The study derives its findings from data gathered from six Malaysian companies in order to obtain the three-fold goal of this thesis as outlined above. The relationships between the antecedents of KI effectiveness and its consequences are tested using 188 responses to a survey representing the views of management and operational employment cohorts. Using statistical methods, we confirm three antecedents of KI effectiveness and the consequences of the antecedents on ES success are validated. The findings demonstrate a statistically positive impact of KI effectiveness of ES success, with KI effectiveness contributing to almost one-third of ES success. This research makes a number of contributions to the understanding of the influence of KI on ES success. First, based on the empirical work using a complete nomological net model, the role of KI effectiveness on ES success is evidenced. Second, the model provides a theoretical lens for a more comprehensive understanding of the impact of KI on the level of ES success. Third, restructuring the dimensions of the knowledge-based theory to fit the context of ES extends its applicability and generalisability to contemporary Information Systems. Fourth, the study develops and validates measures for the antecedents of KI effectiveness. Fifth, the study demonstrates the statistically significant positive influence of the goodness of KI on ES success. From a practical viewpoint, this study emphasises the importance of KI effectiveness as a direct antecedent of ES success. Practical lessons can be drawn from the work done in this study to empirically identify the critical factors among the antecedents of KI effectiveness that should be given attention.

An information literacy integration model and its application in higher education

Purpose - The purpose of this paper is to present a model for curricular integration of information literacy for undergraduate programs in higher education. Design/methodology/approach - Data are drawn from individual interviews at three universities in Australia and curricular integration working experience at a New Zealand university. Sociocultural theories are adopted in the research process and in the development of the model, Findings - Key characteristics of the curriculum integration of information literacy were identified and an information literacy integration model was developed. The S2J2 key behaviours for campus-wide multi-partner collaboration in information literacy integration were also identified. Research limitations/implications - The model was developed without including the employer needs. Through the process of further research, the point of view of the employer on how to provide information literacy education needs to be explored in order to strengthen the model in curricular design. Practical implications - The information literacy integration model was developed based on practical experience in higher education and has been applied in different undergraduate curricular programs. The model could be used or adapted by both librarians and academics when they integrate information literacy into an undergraduate curriculum from a lower level to a higher level. Originality/value - The information literacy integration model was developed based on recent PhD research. The model integrates curriculum, pedagogy and learning theories, information literacy theories, information literacy guidelines, people and collaborative together. The model provides a framework of how information literacy can be integrated into multiple courses across an undergraduate academic degree in higher education.

Integration of opinion into customer analysis model

Nowadays, Opinion Mining is getting more important than before especially in doing analysis and forecasting about customers’ behavior for businesses purpose. The right decision in producing new products or services based on data about customers’ characteristics means profit for organization/company. This paper proposes a new architecture for Opinion Mining, which uses a multidimensional model to integrate customers’ characteristics and their comments about products (or services). The key step to achieve this objective is to transfer comments (opinions) to a fact table that includes several dimensions, such as, customers, products, time and locations. This research presents a comprehensive way to calculate customers’ orientation for all possible products’ attributes. A use case study is also presented in this paper to show the advantages of using OLAP and data cubes to analyze costumers’ opinions.

Enhancing resilience in indigenous people: The integration of individual, family and community interventions

This paper describes a number of interventions being developed to address the emotional, social and behavioural problems experienced by Aboriginal people in Australia. These are: the We-Al-Li program to help people deal with the impact of transgenerational trauma; and the RAP Indigenous Parenting Program. It is argued that the emotional well being of indigenous people will be enhanced through the integration of interventions targeting the individual, family and community.

An example of cognitive obstacles in advanced integration: the case of scalar line integrals

Cognitive obstacles that arise in the teaching and learning of scalar line integrals, derived from cognitive aids provided to students when first learning about integration of single variable functions are described. A discussion of how and why the obstacles cause students problems is presented and possible strategies to overcome the obstacles are outlined.

Multisensory integration weightings underlying haptic feedback laparoscopic surgery

"This letter aims to highlight the multisensory integration weighting mechanisms that may account for the results in studies investigating haptic feedback in laparoscopic surgery. The current lack of multisensory theoretical knowledge in laparoscopy is evident, and “a much better understanding of how multimodal displays in virtual environments influence human performance is required” ...publisher website

Organisational knowledge integration: towards a conceptual framework

This study analyses organisational knowledge integration processes from a multi-level and systemic perspective, with particular reference to the case of Fujitsu. A conceptual framework for knowledge integration is suggested focusing on team-building capability, capturing and utilising individual tacit knowledge, and communication networks for integrating dispersed specialist knowledge required in the development of new products and services. The research highlights that knowledge integration occurring in the innovation process is a result of knowledge exposure, its distribution and embodiment and finally its transfer, which leads to innovation capability and competitive advantage in firm.