Hypoxia-induced DNA hypermethylation in human pulmonary fibroblasts is associated with Thy-1 promoter methylation and the development of a pro-fibrotic phenotype

BACKGROUND: Pulmonary fibrosis is a debilitating and lethal disease with no effective treatment options. Understanding the pathological processes at play will direct the application of novel therapeutic avenues. Hypoxia has been implicated in the pathogenesis of pulmonary fibrosis yet the precise mechanism by which it contributes to disease progression remains to be fully elucidated. It has been shown that chronic hypoxia can alter DNA methylation patterns in tumour-derived cell lines. This epigenetic alteration can induce changes in cellular phenotype with promoter methylation being associated with gene silencing. Of particular relevance to idiopathic pulmonary fibrosis (IPF) is the observation that Thy-1 promoter methylation is associated with a myofibroblast phenotype where loss of Thy-1 occurs alongside increased alpha smooth muscle actin (α-SMA) expression. The initial aim of this study was to determine whether hypoxia regulates DNA methylation in normal human lung fibroblasts (CCD19Lu). As it has been reported that hypoxia suppresses Thy-1 expression during lung development we also studied the effect of hypoxia on Thy-1 promoter methylation and gene expression.

METHODS: CCD19Lu were grown for up to 8 days in hypoxia and assessed for global changes in DNA methylation using flow cytometry. Real-time PCR was used to quantify expression of Thy-1, α-SMA, collagen I and III. Genomic DNA was bisulphite treated and methylation specific PCR (MSPCR) was used to examine the methylation status of the Thy-1 promoter.

RESULTS: Significant global hypermethylation was detected in hypoxic fibroblasts relative to normoxic controls and was accompanied by increased expression of myofibroblast markers. Thy-1 mRNA expression was suppressed in hypoxic cells, which was restored with the demethylating agent 5-aza-2'-deoxycytidine. MSPCR revealed that Thy-1 became methylated following fibroblast exposure to 1% O2.

CONCLUSION: These data suggest that global and gene-specific changes in DNA methylation may play an important role in fibroblast function in hypoxia.

Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli

Calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) has been implicated in a range of conditions and pathologies from prostate to hepatic cancer. Here, we describe the expression in Escherichia coli and the purification protocol for the following constructs: full-length CaMKK2 in complex with CaM, CaMKK2 'apo', CaMKK2 (165-501) in complex with CaM, and the CaMKK2 F267G mutant. The protocols described have been optimized for maximum yield and purity with minimal purification steps required and the proteins subsequently used to develop a fluorescence-based assay for drug binding to the kinase, "Using the fluorescent properties of STO-609 as a tool to assist structure-function analyses of recombinant CaMKK2"

CASIMIRO: a robot head for human-computer interaction

The physical appearance and behavior of a robot is an important asset in terms of Human-Computer Interaction. Multimodality is also fundamental, as we humans usually expect to interact in a natural way with voice, gestures, etc. People approach complex interaction devices with stances similar to those used in their interaction with other people. In this paper we describe a robot head, currently under development, that aims to be a multimodal (vision, voice, gestures,...) perceptual user interface.

Handmade Education : Interactive Food for Children`s Learning

Back to the early age of human history, Human thought of food as any substance that provides nutrition for the body. Man needed to hunt animals for food. Nowadays, Food is consumed not just to stop hunger of the body, but it also feeds the souls. Food has already been used for many other objectives apart from eating such as for artistic creations and creative marketing campaigns. Likewise, this research will also show a new perspective of food – Food as an educational material for children.

Safety and Efficacy of Oral Fexofenadine in Children with Seasonal Allergic Rhinitis - a Pooled Analysis of Three Studies

Allergic rhinitis is one of the most common clinical conditions in children; however, data regarding the safety of antihistamines in children with seasonal allergic rhinitis are limiting. To evaluate the safety and efficacy of fexofenadine in children with seasonal allergic rhinitis, data were pooled from three, double-blind, randomized, placebo-controlled, parallel-group, 2-week trials in children (6-11 year) with seasonal allergic rhinitis. All studies assessed fexofenadine HCl 30 mg b.i.d.; two studies included fexofenadine HCl at 15 and 60 mg b.i.d. Patients (and investigators) reported any adverse events during the trial. Physical examinations, including measurements of vital signs and laboratory tests, were performed. Efficacy assessments (total symptom score and individual symptom scores) were evaluated. Exposure to fexofenadine HCl 30 mg b.i.d. and to any fexofenadine dose exceeded 10,000 and 17,000 patient days, respectively. Incidences of adverse events, and discontinuations because of adverse events, were low and similar across treatment groups. In the placebo group, 24.4% of subjects reported adverse events compared with 24.1% for fexofenadine HCl 30 mg b.i.d., and 28.4% for all fexofenadine-treated groups. The most common adverse event overall was headache (4.3% placebo; 5.8% fexofenadine HCl 30 mg b.i.d.; and 7.2% any fexofenadine doses). Treatment-related adverse events were similar across treatment groups with no sedative effects. Fexofenadine HCl 30 mg b.i.d. was significantly superior to placebo in reducing the total symptom score and all individual seasonal allergic rhinitis symptoms, including nasal congestion (p < 0.05). Fexofenadine, at doses of up to 60 mg b.i.d., is safe and non-sedating, and fexofenadine HCl 30 mg b.i.d. effectively reduces all seasonal allergic rhinitis symptoms in children aged 6-11 years.

The Use of a Computer-Based Simulation Game (CSG) for Learning and Teaching

Introduction A computer-based simulation game (CSG) was used for the first time in a final-year undergraduate module. A change management simulation game was used in the seminar classes as a formative exercise that was linked to parts of the students’ summative assessment. The module evaluation suggests that most students learned from using the CSG.

Professional Practice in Learning and Development: How to Design and Deliver Plans for the Workplace

Introduction The world is changing! It is volatile, uncertain, complex and ambiguous. As cliché as it may sound the evidence of such dynamism in the external environment is growing. Business-as-usual is more of the exception than the norm. Organizational change is the rule; be it to accommodate and adapt to change, or instigate and lead change. A constantly changing environment is a situation that all organizations have to live with. What makes some organizations however, able to thrive better than others? Many scholars and practitioners believe that this is due to the ability to learn. Therefore, this book on developing Learning and Development (L&D) professionals is timely as it explores and discusses trends and practices that impact organizations, the workforce and L&D professionals. Being able to learn and develop effectively is the cornerstone of motivation as it helps to address people’s need to be competent and to be autonomous (Deci & Ryan, 2002; Loon & Casimir, 2008; Ryan & Deci, 2000). L&D stimulates and empowers people to perform. Organizations that are better at learning at all levels; the individual, group and organizational level, will always have a better chance of surviving and performing. Given the new reality of a dynamic external environment and constant change, L&D professionals now play an even more important role in their organizations than ever before. However, L&D professionals themselves are not immune to the turbulent changes as their practices are also impacted. Therefore, the challenges that L&D professionals face are two-pronged. Firstly, in relation to helping and supporting their organization and its workforce in adapting to the change, whilst, secondly developing themselves effectively and efficiently so that they are able to be one-step ahead of the workforce that they are meant to help develop. These challenges are recognised by the CIPD, as they recently launched their new L&D qualification that has served as an inspiration for this book. L&D plays a crucial role at both strategic (e.g. organizational capability) and operational (e.g. delivery of training) levels. L&D professionals have moved from being reactive (e.g. following up action after performance appraisals) to being more proactive (e.g. shaping capability). L&D is increasingly viewed as a driver for organizational performance. The CIPD (2014) suggest that L&D is increasingly expected to not only take more responsibility but also accountability for building both individual and organizational knowledge and capability, and to nurture an organizational culture that prizes learning and development. This book is for L&D professionals. Nonetheless, it is also suited for those studying Human Resource Development HRD at intermediate level. The term ‘Human Resource Development’ (HRD) is more common in academia, and is largely synonymous with L&D (Stewart & Sambrook, 2012) Stewart (1998) defined HRD as ‘the practice of HRD is constituted by the deliberate, purposive and active interventions in the natural learning process. Such interventions can take many forms, most capable of categorising as education or training or development’ (p. 9). In fact, many parts of this book (e.g. Chapters 5 and 7) are appropriate for anyone who is involved in training and development. This may include a variety of individuals within the L&D community, such as line managers, professional trainers, training solutions vendors, instructional designers, external consultants and mentors (Mayo, 2004). The CIPD (2014) goes further as they argue that the role of L&D is broad and plays a significant role in Organizational Development (OD) and Talent Management (TM), as well as in Human Resource Management (HRM) in general. OD, TM, HRM and L&D are symbiotic in enabling the ‘people management function’ to provide organizations with the capabilities that they need.