Up-regulation of NADPH oxidase components and increased production of interferon-gamma by leukocytes from sickle cell disease patients

We have previously demonstrated that mononuclear leukocytes from patients with sickle cell disease (SCD) release higher amounts of superoxide compared with normal controls. The aim of this study was to further study the NADPH oxidase system in these patients by investigating gene expression of NADPH oxidase components, phosphorylation of p47(phox) component, and the release of cytokines related to NADPH oxidase activation in mononuclear leukocytes from patients with SCD. gp91(phox) gene expression was significantly higher in monocytes from SCD patients compared with normal controls (P = 0.036). Monocytes from SCD patients showed higher levels of p47 phox phosphorylation compared with normal controls. INF-gamma release by lymphocytes from SCD patients was significantly higher compared with normal controls, after 48 h culture with phytohemagglutinin (P = 0.02). The release of TNF-alpha by monocytes from SCD patients and normal controls was similar after 24 and 48 h culture with lipopolysaccharide (P > 0.05). We conclude that monocytes from SCD patients show higher levels of gp91(phox) gene expression and p47(phox) phosphorylation, along with increased IFN-gamma release by SCD lymphocytes. These findings help to explain our previous observation showing the increased respiratory burst activity of mononuclear leukocytes from SCD patients and may contribute to inflammation and tissue damage in these patients.

Diapocynin versus apocynin as pretranscriptional inhibitors of NADPH oxidase and cytokine production by peripheral blood mononuclear cells

Apocynin has been extensively used as an inhibitor of NADPH oxidase (NOX) in many experimental models using phagocytic and non-phagocytic cells. Currently, there is some controversy about the efficacy of apocynin in non-phagocytic cells, but in phagocytes the reported results are consistent, which could be due to the presence of myeloperoxidase in these cells. This enzyme has been proposed as responsible for activating apocynin by generating its dimer, diapocynin, which is supposed to be the active compound that prevents NADPH oxidase complex assembly and activation. Here, we synthesized diapocynin and studied its effect on inhibition of gp91(phox) RNA expression. We found that diapocynin strongly inhibited the expression of gp91(phox)mRNA in peripheral blood mononuclear cells (PBMC). Only at a higher concentration, apocynin was able to exert the same effect. We also compared the apocynin and diapocynin efficacy as inhibitors of tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) production in response to lipopolysaccharide (LPS)-activated PBMC. Although apocynin did inhibit TNF-alpha production, diapocynin had a much more pronounced effect, on both TNF-alpha and IL-10 production. In conclusion, these findings suggest that the bioconversion of apocynin to diapocynin is an important issue not limited to enzymatic activity inhibition, but also for other biological effects as gp91(phox) mRNA expression and cytokine production. Hence, as diapocynin can be easily prepared from apocynin, a one-step synthesis, we recommend its use in studies where the biological effects of apocynin are searched. (C) 2010 Elsevier Inc. All rights reserved.

Crucial cytokine interactions in nitric oxide production induced by Mycoplasma arthritidis superantigen

Mycoplasma arthritidis causes autoimmune arthritis in rodents. It produces a superantigen (MAM) that simultaneously activates antigen presenting cells and T cells inducing nitric oxide and cytokine release. Nitric oxide is a key inducer and regulator of the immune system activation. Here, we investigated nitric oxide and cytokine production and interactions of these molecules in MAM-stimulated co-cultures of macrophages (J774A.1 cell line) with spleen lymphocytes. We found that: a) MAM-induced nitric oxide, interferon-gamma, membrane-associated tumor necrosis factor and interleukin-2 production in co-cultures of macrophages with lymphocytes from BALB/c and C3H/HePas but not from C57B1/6 mice; b) production of nitric oxide was dependent on interferon-gamma whereas that of interferon-gamma was dependent on interleukin-2 and membrane-associated tumor necrosis factor; c) these cytokines up regulated MAM-induced nitric oxide production. Unraveling the mechanisms of cell activation induced by MAM might be helpful to design strategies to prevent immune system activation by superantigens and therefore in seeking amelioration of associated immunopathologies. (C) 2008 Elsevier Masson SAS. All rights reserved.

Use of L-Proline and ATP Production by Trypanosoma cruzi Metacyclic Forms as Requirements for Host Cell Invasion

The process of host cell invasion by Trypanosoma cruzi depends on parasite energy. What source of energy is used for that event is not known. To address this and other questions related to T. cruzi energy requirements and cell invasion, we analyzed metacyclic trypomastigote forms of the phylogenetically distant CL and G strains. For both strains, the nutritional stress experienced by cells starved for 24, 36, or 48 h in phosphate-buffered saline reduced the ATP content and the ability of the parasite to invade HeLa cells proportionally to the starvation time. Inhibition of ATP production by treating parasites with rotenone plus antimycin A also diminished the infectivity. Nutrient depletion did not alter the expression of gp82, the surface molecule that mediates CL strain internalization, but increased the expression of gp90, the negative regulator of cell invasion, in the G strain. When L-proline was given to metacyclic forms starved for 36 h, the ATP levels were restored to those of nonstarved controls for both strains. Glucose had no such effect, although this carbohydrate and L-proline were transported in similar fashions. Recovery of infectivity promoted by L-proline treatment of starved parasites was restricted to the CL strain. The profile of restoration of ATP content and gp82-mediated invasion capacity by L-proline treatment of starved Y-strain parasites was similar to that of the CL strain, whereas the Dm28 and Dm30 strains, whose infectivity is downregulated by gp90, behaved like the G strain. L-Proline was also found to increase the ability of the CL strain to traverse a gastric mucin layer, a property important for the establishment of T. cruzi infection by the oral route. Efficient translocation of parasites through gastric mucin toward the target epithelial cells in the stomach mucosa is an essential requirement for subsequent cell invasion. By relying on these closely associated ATP-driven processes, the metacyclic trypomastigotes effectively accomplish their internalization.

Interactive Lightning Session: Game-based Learning in Higher Education

Games are known for leveraging enthusiasm, engagement, energy, knowledge, and passion on gamers; areas that are fundamentally important in higher education. Our panelists will share their perspectives on how Higher Education can take advantage of the potential of game based learning to create a more engaging student learning experien

Learning as movements in interspace

My research issues (in rearch project for PhD-degree, named Drama in School) concern how learning takes place in drama education in compulsory school. One part is to explore and problematize approaches to learning (in and through drama). In this paper will the concept learning be discussed by using a thought derived from Deleuze and Guattari’s nomad philosophy. They describe learning as a movement in the interspace. Focus is on process and inquiry, not on achievement of predetermined skills and competences. 

Literacy Practices in and out of School in Karagwe : The case of primary school literacy education in rural Tanzania

This study has investigated the question of relation between literacy practices in and out of school in rural Tanzania. By using the perspective of linguistic anthropology, literacy practices in five villages in Karagwe district in the northwest of Tanzania have been analysed. The outcome may be used as a basis for educational planning and literacy programs. The analysis has revealed an intimate relation between language, literacy and power. In Karagwe, traditional élites have drawn on literacy to construct and reconstruct their authority, while new élites, such as individual women and some young people have been able to use literacy as one tool to get access to power. The study has also revealed a high level of bilingualism and a high emphasis on education in the area, which prove a potential for future education in the area. At the same time discontinuity in language use, mainly caused by stigmatisation of what is perceived as local and traditional, such as the mother-tongue of the majority of the children, and the high status accrued to all that is perceived as Western, has turned out to constitute a great obstacle for pupils’ learning. The use of ethnographic perspectives has enabled comparisons between interactional patterns in schools and outside school. This has revealed communicative patterns in school that hinder pupils’ learning, while the same patterns in other discourses reinforce learning. By using ethnography, relations between explicit and implicit language ideologies and their impact in educational contexts may be revealed. This knowledge may then be used to make educational plans and literacy programmes more relevant and efficient, not only in poor post-colonial settings such as Tanzania, but also elsewhere, such as in Western settings.

E-learning opens the door to the global community. Novice users experiences of e-learning in a Somali University

E-learning has become one of the primary ways of delivering education around the globe. In Somalia, which is a country torn within and from the global community by a prolonged civil war, University of Hargeisa has in collaboration with Dalarna University in Sweden adopted, for the first time, e-learning. This study explores barriers and facilitators to e-learning usage, experienced by students in Somalia’s higher education, using the University of Hargeisa as case study. Interviews were conducted with students to explore how University of Hargeisa’s novice users perceived elearning, and what factors positively and negatively affected their e-learning experiences. The Unified Theory of Acceptance and Use of Technology (UTAUT) model was used as a framework for interpreting the results. The findings show that, in general, the students have a very positive attitude towards e-learning, and they perceived that e-learning enhanced their educational experience. The communication aspect was found to be especially important for Somali students, as it facilitated a feeling of belonging to the global community of students and scholars and alleviated the war-torn country’s isolation. However, some socio-cultural aspects of students’ communities negatively affected their e-learning experience. This study ends with recommendations based on the empirical findings to promote the use and enhance the experience of e-learning in post conflict Somali educational institutions

Facilitation of learning in specialist nursing training in the PICU : the supervisors´ concerns in the learning situation

With the aim to unfold nurses’ concerns of the supervision of the student in the clinical caring situation of the vulnerable child, clinical nurses situated supervision of postgraduate nursing students in the Pediatric Intensive Care Unit (PICU) are explored. A qualitative approach, interpretive phenomenology, with participant observations and narrative interviews, was used. Two qualitative variations of patterns of meaning for the nurses’ clinical facilitation were disclosed in this study. Learning by doing theme supports the students learning by doing through performing skills and embracing routines. The reflecting theme supports thinking and awareness of the situation. As the supervisor often serves as a role model for the student this might have an immediate impact on how the student applies nursing care in the beginning of his or her career. If the clinical supervisor narrows the perspective and hinders room for learning the student will bring less knowledge from the clinical education than expected, which might result in reduced nursing quality.