Interactions between Mycoplasma bovis and bovine lymphocytes: characterization of a lympho-inhibitory peptide produced by Mycoplasma bovis

The effects of Mycoplasma bovis on bovine lympho-proliferation and viability were studied. Incubation of bovine peripheral blood mononuclear cells (PBMC) in vitro with M. bovis induced lymphocyte apoptosis as monitored by Annexin V binding, propidium iodide incorporation, and DNA fragmentation. The induction of lymphocyte death was abrogated by chloramphenicol, indicating that lymphocyte death was dependent upon prokaryotic protein production.;In attempts to better define the lymphotoxic factors associated with M. bovis, the task of isolating the M. bovis lymphocyte inhibitory or cytotoxic factor was undertaken. Using size exclusion chromatography a fraction able to suppress bovine lympho-proliferatioe responses to ConA was isolated. MALDI-TOF analysis of the suppressive fraction revealed one dominant peak (M.W. of 2,970 daltons) unique to the suppressive fraction. N-terminal sequencing of the suppressive fraction revealed a 19 amino acid sequence homologous to the C-terminus of the M. bovis theoretical gene encoding Vsp-L (variable surface protein-L) as determined by a BLAST search against the NCBI national protein database. The sequence data was used to construct primers for PCR amplification of the vspL gene domain corresponding to the last 26 amino acids of the VspL\u27s protein C-terminus. This PCR product was cloned and inserted into an expression plasmid as a GST-fusion protein engineered with a thrombin recognition cleavage site between GST and the recombinant peptide (to facilitate purification of the peptide). Upon purification, the peptide Mb-LIP (M. bovis lymphocyte inhibitory peptide) was tested for lympho-proliferatioe inhibition. The recombinant peptide inhibited bovine lymphocyte proliferation responses to ConA and CD3 stimulation in vitro. Further study indicated that incubation of PBMCs with the recombinant peptide decreased the number of viable lymphocytes in culture. The recombinant peptide Mb-LIP is therefore considered a lymphotoxic peptide. This marks the first described characterization of a mycoplasmal product able to inhibit naive lymphocyte function

Sports coach as transformative leader: Arresting school disengagement through community sport based initiatives.

Reducing social exclusion through interventions designed to sustain school engagement is a key aim of the education and social policy of any government. This paper is a response to the call for there to be more focused empirical sports coaching research through examining the transformative potential of community based sports coaches to support schools in arresting school disengagement. By embracing an understanding that challenges the definitional core of sports coaching as simply improving the sporting performance of an individual or team, and, drawing theoretically on the work of Carlisle et al. (2006) and Shields (2010), the role of ‘coach as transformative leader’ is articulated. Analysis of data collected by means of semi-structured interviews with a group of community based sports coaches (n=8), revealed three factors salient to our understanding of re-engaging young people with formal education through sport. These were the impact of the community sport programme, the relationship between schools and community sports groups and, the implementation of transformative leadership qualities by sport coaching practitioners. Importantly, this paper explicates the pivotal function that coaching practice which embraces transformative leadership principles can have on re-orienting young people from disadvantaged backgrounds towards more optimistic futures and educational objectives

Chaordic learning systems: reconceptualising pedagogy for the digital age

This article focuses on an explorative and experimental project seeking to implement Chaordic Learning Systems (CLS) as a pedagogic approach in Higher Education. We outline a project that embraced technologies of Web 2.0 to show how both physical and virtual spaces can be used to support and develop a strong and dynamic learning community in which staff and students work alongside each other to co-produce learning resources. Drawing on theories of Communities of Practice and Situated Learning a new teaching framework was introduced to a Level 5 undergraduate module (7.5 ECTS credits) that had not, until this project, used both face-to-face and online learning tools to engage students in the critical and discursive debates pertaining to sport and physical culture. We undertook this project with the belief that Higher Education should be concerned with answering the calls of an increasingly digital society for whom learning is not restricted by the physical boundaries of the university or the political landscape within which learning finds itself

Expression of Epstein–Barr Virus–Encoded Small RNA (by the EBER-1 Gene) in Liver Specimens from Transplant Recipients with Post-Transplantation Lymphoproliferative Disease

Epstein-Barr virus (EBV)—associated post-transplantation lymphoproliferative disease (PTLD) develops in 1 to 10 percent of transplant recipients, in whom it can be treated by a reduction in the level of immunosuppression. We postulated that the tissue expression of the small RNA transcribed by the EBER-1 gene during latent EBV infection would identify patients at risk for PTLD. We studied EBER-1 gene expression in liver specimens obtained from 24 patients 2 days to 22 months before the development of PTLD, using in situ hybridization with an oligonucleotide probe. Control specimens were obtained from 20 recipients of allografts with signs of injury due to organ retrieval, acute graft rejection, or viral hepatitis in whom PTLD had not developed 9 to 71 months after the biopsy. Of the 24 patients with PTLD, 17 (71 percent) had specimens in which 1 to 40 percent of mononuclear cells were positive for the EBER-1 gene. In addition, 10 of these 17 patients (59 percent) had specimens with histopathological changes suggestive of EBV hepatitis. In every case, EBER-1—positive cells were found within the lymphoproliferative lesions identified at autopsy. Only 2 of the 20 controls (10 percent) had specimens with EBER-1—positive cells (P<0.001), and such cells were rare. EBER-1 gene expression in liver tissue precedes the occurrence of clinical and histologic PTLD. The possibility of identifying patients at risk by the method we describe here and preventing the occurrence of PTLD by a timely reduction of immunosuppression needs to be addressed by future prospective studies. (N Engl J Med 1992;327:1710–4.), POST-TRANSPLANTATION lymphoproliferative disease (PTLD), either polyclonal or monoclonal, complicates the clinical course of 1 to 10 percent of organ-transplant recipients.123 Immunohistochemical studies have demonstrated that the lymphoid cells within the lesions of PTLD almost invariably contain Epstein–Barr virus (EBV), primarily in a state of latent infection.4,5 The EBER-1 gene is expressed early during latent EBV infection and codes for a small messenger RNA (mRNA) expressed at up to 107 copies per cell.6 We and others have previously demonstrated the value of the detection of EBER-1 RNA for identifying EBV-infected cells in formalin-fixed paraffin-embedded tissues.7,8 In the current investigation, we used… © 1992, Massachusetts Medical Society. All rights reserved

Chaordic learning systems: reconceptualising pedagogy for the digital age

This article focuses on an explorative and experimental project seeking to implement Chaordic Learning Systems (CLS) as a pedagogic approach in Higher Education. We outline a project that embraced technologies of Web 2.0 to show how both physical and virtual spaces can be used to support and develop a strong and dynamic learning community in which staff and students work alongside each other to co-produce learning resources. Drawing on theories of Communities of Practice and Situated Learning a new teaching framework was introduced to a Level 5 undergraduate module (7.5 ECTS credits) that had not, until this project, used both face-to-face and online learning tools to engage students in the critical and discursive debates pertaining to sport and physical culture. We undertook this project with the belief that Higher Education should be concerned with answering the calls of an increasingly digital society for whom learning is not restricted by the physical boundaries of the university or the political landscape within which learning finds itself

Combined exome and whole-genome sequencing identifies mutations in ARMC4 as a cause of primary ciliary dyskinesia with defects in the outer dynein arm

Primary ciliary dyskinesia (PCD) is a rare, genetically heterogeneous ciliopathy disorder affecting cilia and sperm motility. A range of ultrastructural defects of the axoneme underlie the disease, which is characterised by chronic respiratory symptoms and obstructive lung disease, infertility and body axis laterality defects. We applied a next-generation sequencing approach to identify the gene responsible for this phenotype in two consanguineous families

Promotional Tone in Reviews of Menopausal Hormone Therapy After the Women's Health Initiative: An Analysis of Published Articles

Adriane Fugh-Berman and colleagues analyzed a selection of published opinion pieces on hormone therapy and show that there may be a connection between receiving industry funding for speaking, consulting, or research and the tone of such opinion pieces