Development of a multicellular co-culture model of normal and cystic fibrosis human airways in vitro

Cystic fibrosis (CF) is the most common lethal inherited disease among Caucasians and arises due to mutations in a chloride channel, called cystic fibrosis transmembrane conductance regulator. A hallmark of this disease is the chronic bacterial infection of the airways, which is usually, associated with pathogens such as Pseudomonas aeruginosa, S. aureus and recently becoming more prominent, B. cepacia. The excessive inflammatory response, which leads to irreversible lung damage, will in the long term lead to mortality of the patient at around the age of 40 years. Understanding the pathogenesis of CF currently relies on animal models, such as those employing genetically-modified mice, and on single cell culture models, which are grown either as polarised or non-polarised epithelium in vitro. Whilst these approaches partially enable the study of disease progression in CF, both types of models have inherent limitations. The overall aim of this thesis was to establish a multicellular co-culture model of normal and CF human airways in vitro, which helps to partially overcome these limitations and permits analysis of cell-to-cell communication in the airways. These models could then be used to examine the co-ordinated response of the airways to infection with relevant pathogens in order to validate this approach over animals/single cell models. Therefore epithelial cell lines of non-CF and CF background were employed in a co-culture model together with human pulmonary fibroblasts. Co-cultures were grown on collagen-coated permeable supports at air-liquid interface to promote epithelial cell differentiation. The models were characterised and essential features for investigating CF infections and inflammatory responses were investigated and analysed. A pseudostratified like epithelial cell layer was established at air liquid interface (ALI) of mono-and co-cultures and cell layer integrity was verified by tight junction (TJ) staining and transepithelial resistance measurements (TER). Mono- and co-cultures were also found to secrete the airway mucin MUC5AC. Influence of bacterial infections was found to be most challenging when intact S. aureus, B. cepacia and P. aeruginosa were used. CF mono- and co-cultures were found to mimic the hyperinflammatory state found in CF, which was confirmed by analysing IL-8 secretions of these models. These co-culture models will help to elucidate the role fibroblasts play in the inflammatory response to bacteria and will provide a useful testing platform to further investigate the dysregulated airway responses seen in CF.

Development of a co-culture model of the human lungs for toxicity testing and identification of biomarkers of inhalation toxicity

The airway epithelium is the first point of contact in the lung for inhaled material, including infectious pathogens and particulate matter, and protects against toxicity from these substances by trapping and clearance via the mucociliary escalator, presence of a protective barrier with tight junctions and initiation of a local inflammatory response. The inflammatory response involves recruitment of phagocytic cells to neutralise and remove and invading materials and is oftern modelled using rodents. However, development of valid in vitro airway epithelial models is of great importance due to the restrictions on animal studies for cosmetic compound testing implicit in the 7th amendment to the European Union Cosmetics Directive. Further, rodent innate immune responses have fundamental differences to human. Pulmonary endothelial cells and leukocytes are also involved in the innate response initiated during pulmonary inflammation. Co-culture models of the airways, in particular where epithelial cells are cultured at air liquid interface with the presence of tight junctions and differentiated mucociliary cells, offer a solution to this problem. Ideally validated models will allow for detection of early biomarkers of response to exposure and investigation into inflammatory response during exposure. This thesis describes the approaches taken towards developing an in vitro epithelial/endothelial cell model of the human airways and identification biomarkers of response to exposure to xenobiotics. The model comprised normal human primary microvascular endothelial cells and the bronchial epithelial cell line BEAS-2B or normal human bronchial epithelial cells. BEAS-2B were chosen as their characterisation at air liquid interface is limited but they are robust in culture, thereby predicted to provide a more reliable test system. Proteomics analysis was undertaken on challenged cells to investigate biomarkers of exposure. BEAS-2B morphology was characterised at air liquid interface compared with normal human bronchial epithelial cells. The results indicate that BEAS-2B cells at an air liquid interface form tight junctions as shown by expression of the tight junction protein zonula occludens-1. To this author’s knowledge this is the first time this result has been reported. The inflammatory response of BEAS-2B (measured as secretion of the inflammatory mediators interleukin-8 and -6) air liquid interface mono-cultures to Escherichia coli lipopolysaccharide or particulate matter (fine and ultrafine titanium dioxide) was comparable to published data for epithelial cells. Cells were also exposed to polymers of “commercial interest” which were in the nanoparticle range (and referred to particles hereafter). BEAS-2B mono-cultures showed an increased secretion of inflammatory mediators after challenge. Inclusion of microvascular endothelial cells resulted in protection against LPS- and particle- induced epithelial toxicity, measured as cell viability and inflammatory response, indicating the importance of co-cultures for investigations into toxicity. Two-dimensional proteomic analysis of lysates from particle-challenged cells failed to identify biomarkers of toxicity due to assay interference and experimental variability. Separately, decreased plasma concentrations of serine protease inhibitors, and the negative acute phase proteins transthyretin, histidine-rich glycoprotein and alpha2-HS glycoprotein were identified as potential biomarkers of methyl methacrylate/ethyl methacrylate/butylacrylate treatment in rats.

Creating a culture for critical and situated technology use through effective learning design

The purpose of this concise paper is to propose, with evidence gathered through a systematic evaluation of an academic development programme in the UK, that training in the use of new and emerging learning technologies should be holistically embedded in every learning and training opportunity in learning, teaching and assessment in higher education, and not only as stand-alone modules or one-off opportunities. The future of learning in higher education cannot afford to allow Universities to disregard that digital literacy is an expected professional skill for their entire staff.

Development of a neurotoxicity test-system, using human post-mitotic, astrocytic and neuronal cell lines in co-culture

Astrocytes are essential for neuronal function and survival, so both cell types were included in a human neurotoxicity test-system to assess the protective effects of astrocytes on neurons, compared with a culture of neurons alone. The human NT2.D1 cell line was differentiated to form either a co-culture of post-mitotic NT2.N neuronal (TUJ1, NF68 and NSE positive) and NT2.A astrocytic (GFAP positive) cells (∼2:1 NT2.A:NT2.N), or an NT2.N mono-culture. Cultures were exposed to human toxins, for 4 h at sub-cytotoxic concentrations, in order to compare levels of compromised cell function and thus evidence of an astrocytic protective effect. Functional endpoints examined included assays for cellular energy (ATP) and glutathione (GSH) levels, generation of hydrogen peroxide (H2O2) and caspase-3 activation. Generally, the NT2.N/A co-culture was more resistant to toxicity, maintaining superior ATP and GSH levels and sustaining smaller significant increases in H2O2 levels compared with neurons alone. However, the pure neuronal culture showed a significantly lower level of caspase activation. These data suggest that besides their support for neurons through maintenance of ATP and GSH and control of H2O2 levels, following exposure to some substances, astrocytes may promote an apoptotic mode of cell death. Thus, it appears the use of astrocytes in an in vitro predictive neurotoxicity test-system may be more relevant to human CNS structure and function than neuronal cells alone. © 2007 Elsevier Ltd. All rights reserved.

A framework of best practice of continuing professional development for the accounting profession

The International Accounting Education Standards Board (IAESB) places a strong emphasis on individual professionals taking responsibility for their Continuing Professional Development (CPD). On the other hand, the roles performed by professional accountants have evolved out of practical necessity to 'best' suit the diverse needs of business in a global economy. This diversity has meant that professional accountants are seen in highly specialised roles requiring diverse skill sets. In order to enhance the contribution of the accountant as a knowledge professional for business, it follows that CPD that leverages off an individual's experience should be designed to meet the needs of professionals across the different specialised roles within the profession. In doing so the project identifies how CPD should differ across roles and levels of organisational responsibility for accounting professionals. The study also makes a number of policy recommendations to IAESB and IFAC. © 2013 © 2013 Taylor & Francis.

A quantitative approach for understanding small-scale human mesenchymal stem cell culture - implications for large-scale bioprocess development

Human mesenchymal stem cell (hMSC) therapies have the potential to revolutionise the healthcare industry and replicate the success of the therapeutic protein industry; however, for this to be achieved there is a need to apply key bioprocessing engineering principles and adopt a quantitative approach for large-scale reproducible hMSC bioprocess development. Here we provide a quantitative analysis of the changes in concentration of glucose, lactate and ammonium with time during hMSC monolayer culture over 4 passages, under 100% and 20% dissolved oxgen (dO2), where either a 100%, 50% or 0% growth medium exchange was performed after 72h in culture. Yield coefficients, specific growth rates (h-1) and doubling times (h) were calculated for all cases. The 100% dO2 flasks outperformed the 20% dO2 flasks with respect to cumulative cell number, with the latter consuming more glucose and producing more lactate and ammonium. Furthermore, the 100% and 50% medium exchange conditions resulted in similar cumulative cell numbers, whilst the 0% conditions were significantly lower. Cell immunophenotype and multipotency were not affected by the experimental culture conditions. This study demonstrates the importance of determining optimal culture conditions for hMSC expansion and highlights a potential cost savings from only making a 50% medium exchange, which may prove significant for large-scale bioprocessing. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

An Open Virtual World for Professional Development

The paper presents a study that focuses on the issue of sup-porting educational experts to choose the right combination of educational methodology and technology tools when designing training and learning programs. It is based on research in the field of adaptive intelligent e-learning systems. The object of study is the professional growth of teachers in technology and in particular that part of their qualification which is achieved by organizing targeted training of teachers. The article presents the process of creating and testing a system to support the decision on the design of training for teachers, leading to more effective implementation of technology in education and integration in diverse educational contexts. ACM Computing Classification System (1998): H.4.2, I.2.1, I.2, I.2.4, F.4.1.

Professionalism within pharmacy – the thoughts of Professional Leaders

To ascertain the thoughts of selected professional leaders on matters relating to pharmacist professionalism. These views will help build a picture of the professional status of pharmacy. Methods - Semi-structured interviews were conducted between July and November 2013 with representatives from eight UK pharmacy leadership bodies. The bodies were selected for their roles in pharmacy policy development, regulation and professional representation. The interviews were recorded and transcribed verbatim. Analysis by constant comparison identified a number of emerging themes. Results - The following emerging themes were identified from the interview data: Influence of the Pharmacy Landscape: Participants highlighted the role that pharmacy plays within the National Health Service and wider society and how future developments may affect the professional status currently afforded to pharmacists. Vocalising Pharmacy: Communication within the profession and also with those external to the profession, including other healthcare professionals and the general public, is important to ensure a high professional standing. The Impact of Commercialism: Professionalism and commercialism were generally seen to be antithetical and a rise in commercialism may adversely impact on external perceptions of the professionalism of pharmacy. Responsibility for Professionalism: The professional image of pharmacy is maintained by the individuals operating within it regardless of their scope of practice. It is the responsibility of all those individuals to ensure that they actively demonstrate ‘professional’ behaviours. The Journey to Professionalism: Acquiring a professional ethos is a continual process but there are stages in a pharmacist’s development that are considered particularly important. These include upbringing, undergraduate education and pre-registration training. Conclusions - Pharmacy’s professional status in the UK remains open to challenge and vital to retaining that status is the public perception of pharmacists. Future research examining pharmacy’s claims to professional status should focus on exploring the attitudes of the general public in addition to the views of pharmacists.

‘Let’s talk about feedback’:an exploration of academic feedback practices in pharmacy education

Feedback is considered one of the most effective mechanisms to aid learning and achievement (Hattie and Timperley, 2007). However, in past UK National Student Surveys, perceptions of academic feedback have been consistently rated lower by final year undergraduate students than other aspects of the student experience (Williams and Kane, 2009). For pharmacy students in particular, Hall and colleagues recently reported that almost a third of students surveyed were dissatisfied with feedback and perceived feedback practice to be inconsistent (Hall et al, 2012). Aims of the Workshop: This workshop has been designed to explore current academic feedback practices in pharmacy education across a variety of settings and cultures as well as to create a toolkit for pharmacy academics to guide their approach to feedback. Learning Objectives: 1. Discuss and characterise academic feedback practices provided by pharmacy academics to pharmacy students in a variety of settings and cultures. 2. Develop academic feedback strategies for a variety of scenarios. 3. Evaluate and categorise feedback strategies with use of a feedback matrix. Description of Workshop Activities: Introduction to workshop and feedback on pre-reading exercise (5 minutes). Activity 1: A short presentation on theoretical models of academic feedback. Evidence of feedback in pharmacy education (10 minutes). Activity 2: Discussion of feedback approaches in participants’ organisations for differing educational modalities. Consideration of the following factors will be undertaken: experiential v. theoretical education, formative v. summative assessment, form of assessment and the effect of culture (20 minutes, large group discussion). Activity 3: Introduction of a feedback matrix (5 minutes). Activity 4: Development of an academic feedback toolkit for pharmacy education. Participants will be divided into 4 groups and will discuss how to provide effective feedback for 2 scenarios. Feedback strategies will be categorised with the feedback matrix. Results will be presented back to the workshop group (20 minutes, small group discussion, 20 minutes, large group presentation). Summary (10 minutes). Additional Information: Pre-reading: Participants will be provided with a list of definitions for academic feedback and will be asked to rank the definitions in order of perceived relevance to pharmacy education. References Archer, J. C. (2010). State of the science in health professional education: effective feedback. Medical education, 44(1), 101-108. Hall, M., Hanna, L. A., & Quinn, S. (2012). Pharmacy Students’ Views of Faculty Feedback on Academic Performance. American journal of pharmaceutical education, 76(1). Hattie, J., & Timperley, H. (2007). The power of feedback. Review of educational research, 77(1), 81-112. Medina, M. S. (2007). Providing feedback to enhance pharmacy students’ performance. American Journal of Health-System Pharmacy, 64(24), 2542-2545.

Vágyott kultúra és reális értékítélet - Tudásmenedzsment-rendszer kiépítésének előfeltétele egy felsőoktatási intézményben (Desired culture and real value judgement – Precondition of development a knowledge management system in a higher education institution)

A tudásmenedzsment-rendszerek kiépítése és működtetése egyre népszerűbb vállalati cél. A legnagyobb igyekezet ellenére is kudarccal végződhet egy ilyen változás megvalósítása, ha a szükséges feltételek nélkül próbálkozunk ezzel a beavatkozással a szervezet életében. Az egyik legfontosabb előfeltétel a bizalomra, közös tanulásra, fejlődésre épülő, nyitott szervezeti kultúra, mely a tanulószervezeti jellemzőkkel írható le. A szerzők kutatásukban arra voltak kíváncsiak, milyen elképzeléseik, vágyaik vannak a felsőoktatásban oktató kollégáknak az ideálisnak nevezett szervezeti kultúráról. Ezeket az elképzeléseket egy külső tanácsadó cég által végzett kérdőíves felmérésen alapuló vizsgálat segítségével tudták meg, melynek kiértékelésére a circumplex-módszer szolgált. Az eredményeket összevetették a tanulószervezeti jellemzőkkel, vizsgálva azt a hipotézist, miszerint az oktató kollégák tudat alatt is olyan ideális szervezetet képzelnek el, mely a tanulószervezeti kultúra jellemzőivel azonos. _________ To create and to operate a knowledge management system is becoming a more and more popular target of the companies. Realizing the changes above can result in a failure – in spite of the biggest will – if organizations lack certain prerequisites which are necessary in companies’ lives. One of the most important prerequisites is organizational culture which can be characterized by confidence, common learning, development and open atmosphere. This is called a learning organizational culture. In their research the authors would have liked to know what kind of dreams the colleagues have about their own organizational culture. They achieve these results from an investigation with questionnaires which were realized by an advisory team. To evaluating the results of this investigation circumplex method was used. These results were compared with the characteristics of learning organization to confirm our hypothesis. According to this idea colleagues have the same images about their successful organization as the characteristics of learning organizations.