Single-stranded oligonucleotide-mediated in vivo gene repair in the rd1 retina.

PURPOSE: The aim of this study was to test whether oligonucleotide-targeted gene repair can correct the point mutation in genomic DNA of PDE6b(rd1) (rd1) mouse retinas in vivo. METHODS: Oligonucleotides (ODNs) of 25 nucleotide length and complementary to genomic sequence subsuming the rd1 point mutation in the gene encoding the beta-subunit of rod photoreceptor cGMP-phosphodiesterase (beta-PDE), were synthesized with a wild type nucleotide base at the rd1 point mutation position. Control ODNs contained the same nucleotide bases as the wild type ODNs but with varying degrees of sequence mismatch. We previously developed a repeatable and relatively non-invasive technique to enhance ODN delivery to photoreceptor nuclei using transpalpebral iontophoresis prior to intravitreal ODN injection. Three such treatments were performed on C3H/henJ (rd1) mouse pups before postnatal day (PN) 9. Treatment outcomes were evaluated at PN28 or PN33, when retinal degeneration was nearly complete in the untreated rd1 mice. The effect of treatment on photoreceptor survival was evaluated by counting the number of nuclei of photoreceptor cells and by assessing rhodopsin immunohistochemistry on flat-mount retinas and sections. Gene repair in the retina was quantified by allele-specific real time PCR and by detection of beta-PDE-immunoreactive photoreceptors. Confirmatory experiments were conducted using independent rd1 colonies in separate laboratories. These experiments had an additional negative control ODN that contained the rd1 mutant nucleotide base at the rd1 point mutation site such that the sole difference between treatment with wild type and control ODN was the single base at the rd1 point mutation site. RESULTS: Iontophoresis enhanced the penetration of intravitreally injected ODNs in all retinal layers. Using this delivery technique, significant survival of photoreceptors was observed in retinas from eyes treated with wild type ODNs but not control ODNs as demonstrated by cell counting and rhodopsin immunoreactivity at PN28. Beta-PDE immunoreactivity was present in retinas from eyes treated with wild type ODN but not from those treated with control ODNs. Gene correction demonstrated by allele-specific real time PCR and by counts of beta-PDE-immunoreactive cells was estimated at 0.2%. Independent confirmatory experiments showed that retinas from eyes treated with wild type ODN contained many more rhodopsin immunoreactive cells compared to retinas treated with control (rd1 sequence) ODN, even when harvested at PN33. CONCLUSIONS: Short ODNs can be delivered with repeatable efficiency to mouse photoreceptor cells in vivo using a combination of intravitreal injection and iontophoresis. Delivery of therapeutic ODNs to rd1 mouse eyes resulted in genomic DNA conversion from mutant to wild type sequence, low but observable beta-PDE immunoreactivity, and preservation of rhodopsin immunopositive cells in the outer nuclear layer, suggesting that ODN-directed gene repair occurred and preserved rod photoreceptor cells. Effects were not seen in eyes treated with buffer or with ODNs having the rd1 mutant sequence, a definitive control for this therapeutic approach. Importantly, critical experiments were confirmed in two laboratories by several different researchers using independent mouse colonies and ODN preparations from separate sources. These findings suggest that targeted gene repair can be achieved in the retina following enhanced ODN delivery.

Proyecto "Da Vinci School"

Proyecto de migración/implantación de un sistema libre en un centro de educación secundaria.

Da Vinci school

Implantación de una plataforma libre en un centro de educación secundaria.

A role for T-bet-mediated tumour immune surveillance in anti-IL-17A treatment of lung cancer.

Lung cancer is the leading cause of cancer deaths worldwide. The cytokine interleukin-17A supports tumour vascularization and growth, however, its role in lung cancer is unknown. Here we show, in the lungs of patients with lung adenocarcinoma, an increase in interleukin-17A that is inversely correlated with the expression of T-bet and correlated with the T regulatory cell transcription factor Foxp3. Local targeting of interleukin-17A in experimental lung adenocarcinoma results in a reduction in tumour load, local expansion of interferon-γ-producing CD4(+) T cells and a reduction in lung CD4(+)CD25(+)Foxp3(+) regulatory T cells. T-bet((-/-)) mice have a significantly higher tumour load compared with wild-type mice. This is associated with the local upregulation of interleukin-23 and induction of interleukin-17A/interleukin-17R-expressing T cells infiltrating the tumour. Local anti-interleukin-17A antibody treatment partially improves the survival of T-bet((-/-)) mice. These results suggest that local anti-interleukin-17A antibody therapy could be considered for the treatment of lung tumours.

Disseny i implantació de l'estructura informàtica de DaVinci School

Creació d'una infraestructura informàtica d'un centre educatiu, amb xarxes separades per a aules, secretaria i seminaris. Serveis centralitzats en un servidor amb màquines virtuals.

Actualització del sistema informàtic de l'escola Da Vinci School

Projecte d'actualització del sistema informàtic d'un centre escolar.

The European School Survey Project on Alcohol and Drugs : Swiss Findings

L'enquête 'European School Survey Project on Alcohol and other Drugs' (ESPAD) est menée tous les quatre ans dans de nombreux pays européens auprès d'écolières et d'écoliers âgés de 15 ans, un âge important en matière de prévention. La dernière vague de l'enquête a été réalisée en 2003; il s'agissait de la première participation de la Suisse à l'enquête. Un peu moins de 7000 élèves des 8e, 9e et 10e années scolaires ont participé au volet suisse de l'enquête. Les résultats montrent notamment que 21.1% des adolescente- s de 15 ans fument quotidiennement, la part des fumeurs quotidiens passant de 8.5% chez les 13 ans à 23.2% chez les 16 ans. Plus de 50% des adolescent-e-s de 13 ans ont consommé de l'alcool durant les 30 jours précédant l'enquête. Chez les 15 ans, pour lesquels l'achat d'alcool est d'ailleurs aussi interdit par la loi, cette proportion passe à 75%. L'enquête 2003 montre en outre qu'environ 20% des adolescent-e-s de 13 ans ont consommé au moins une fois du cannabis dans leur vie et que près de la moitié des 16 ans sont dans le même cas. La prévalence au cours des 12 derniers mois est à peine inférieure. En matière de prévention de la consommation de substances psychotropes chez les jeunes, les mesures structurelles devraient être renforcées, tout comme la limitation de l'accessibilité des produits, en contrôlant par exemple la vente d'alcool aux plus jeunes. Des programmes éducatifs et des campagnes médiatiques peuvent également exercer une certaine influence, pour autant qu'ils s'intègrent à une politique cohérente en matière de drogues.

Obesity prevention opinions of school stakeholders: a qualitative study.

BACKGROUND: In general, schools are an important setting to implement current recommendations for obesity prevention in children because the vast majority of children attend school. This study investigated the opinions of different school stakeholders on the feasibility and acceptability of current obesity prevention strategies that could be implemented in Swiss schools. METHODS: Research methods were comprised of a qualitative study which included school directors, physical education teachers, catering staff, school nurses and health educators, parents of young adolescents, and young adolescents interviewed (N = 40) categorized into 6 focus groups. Open-ended questions were used to determine the participants' opinion regarding current obesity prevention recommendations, and healthy eating and physical activity promotion strategies. RESULTS: All participants approved the implementation of nutritional standards for food and drinks sold in schools, but thought that increasing the attractiveness of healthy options was the best strategy to improve eating habits. Enjoying participation in physical activity classes or after-school activities was stressed. Participants suggested offering classes for all students with poor physical condition, independent of weight status. Stakeholders called for governmental support and global coordination of interventions balanced with providing schools with enough autonomy to adapt programs relevant to their individual circumstances. They recommended integrating all school stakeholders in obesity prevention initiatives, with special attention to students and local authorities. CONCLUSIONS: Participants agreed that schools are a crucial setting to implement childhood obesity prevention strategies. They called on school stakeholders to join efforts aiming to encourage healthy behaviors and to support and reinforce parents' efforts by spreading consistent and coherent health messages.

Determinants of the educational use of digital learning materials: The mediating role of self-efficacy, perceived norm and attitude

Initiatives to stimulate the development and propagation of open educational resources (OER) need a sufficiently large community that can be mobilized to participate in this endeavour. Failure to achieve this could lead to underuse of OER. In the context of the Wikiwijs initiative a large scale survey was undertaken amongst primary and secondary school teachers to explore possible determinants of the educational use of digital learning materials (DLMs). Basing on the Integrative Model of Behaviour Prediction it was conjectured that self-efficacy, attitude and perceived norm would take a central role in explaining the intention to use DLMs. Several other predictors were added to the model as well whose effects were hypothesized to be mediated by the three central variables.All conjectured relationships were found using path analysis on survey data from 1484 teachers. Intention to DLMs was most strongly determined by self-efficacy, followed by attitude. ICT proficiency was in its turn the strongest predictor of self-efficacy. Perceived norm played only a limited role in the intention to use DLMs. Concluding, it seems paramount for the success of projects such as Wikiwijs to train teachers in the use of digital learning materials and ICT (e.g. the digital blackboard) and to impact on their attitude.

In Vivo Selection of Fluoroquinolone-Resistant Escherichia coli Isolates Expressing Plasmid-Mediated Quinolone Resistance and Expanded-Spectrum β-Lactamase

A ciprofloxacin-resistant Escherichia coli isolate, isolate 1B, was obtained from a urinary specimen of a Canadian patient treated with norfloxacin for infection due to a ciprofloxacin-susceptible isolate, isolate 1A. Both isolates harbored a plasmid-encoded sul1-type integron with qnrA1 and blaVEB-1 genes. Isolate 1B had amino acid substitutions in gyrase and topoisomerase.

Plasmid-mediated quinolone resistance in Australia

The aim of this study was to search for plasmid-encoded quinolone resistance determinants QnrA and QnrS in fluoroquinolone-resistant and extended-spectrum beta-lactamase (ESBL)-producing enterobacterial isolates recovered in Sydney, Australia, in 2002. Twenty-three fluoroquinolone-resistant, of which 16 were also ESBL-positive, enterobacterial and nonrelated isolates were studied. PCR with primers specific for qnrA and qnrS genes and primers specific for a series of ESBL genes were used. A qnrA gene was identified in two ESBL-positive isolates, whereas no qnrS-positive strain was found. The QnrA1 determinant was identified in an Enterobacter cloacae isolate and in a carbapenem-resistant Klebsiella pneumoniae isolate, both of which expressed the same ESBL SHV- 12. Whereas no plasmid was identified in the E. cloacae isolate, K. pneumoniae K149 possessed two conjugative plasmids, one that harbored the qnrA and bla (SHV)-12 genes whereas the other expressed the carbapenemase gene bla (IMP-4). The qnrA gene, was located in both cases downstream of the orf513 recombinase gene and upstream of the qnrA1 gene, a structure identical to that found in sul1-type integron In36 and qnrA-positive strains from Shanghai, China. However, the gene cassettes of the sul1-type integrons were different. This study identified the first plasmid-mediated quinolone resistance determinant in Enterobacteriaceae in Australia.