Recombinant adeno-associated virus serotype 6 (rAAV2/6)-mediated gene transfer to nociceptive neurons through different routes of delivery.

BACKGROUND: Gene transfer to nociceptive neurons of the dorsal root ganglia (DRG) is a promising approach to dissect mechanisms of pain in rodents and is a potential therapeutic strategy for the treatment of persistent pain disorders such as neuropathic pain. A number of studies have demonstrated transduction of DRG neurons using herpes simplex virus, adenovirus and more recently, adeno-associated virus (AAV). Recombinant AAV are currently the gene transfer vehicles of choice for the nervous system and have several advantages over other vectors, including stable and safe gene expression. We have explored the capacity of recombinant AAV serotype 6 (rAAV2/6) to deliver genes to DRG neurons and characterized the transduction of nociceptors through five different routes of administration in mice. RESULTS: Direct injection of rAAV2/6 expressing green fluorescent protein (eGFP) into the sciatic nerve resulted in transduction of up to 30% eGFP-positive cells of L4 DRG neurons in a dose dependent manner. More than 90% of transduced cells were small and medium sized neurons (< 700 microm 2), predominantly colocalized with markers of nociceptive neurons, and had eGFP-positive central terminal fibers in the superficial lamina of the spinal cord dorsal horn. The efficiency and profile of transduction was independent of mouse genetic background. Intrathecal administration of rAAV2/6 gave the highest level of transduction (approximately 60%) and had a similar size profile and colocalization with nociceptive neurons. Intrathecal administration also transduced DRG neurons at cervical and thoracic levels and resulted in comparable levels of transduction in a mouse model for neuropathic pain. Subcutaneous and intramuscular delivery resulted in low levels of transduction in the L4 DRG. Likewise, delivery via tail vein injection resulted in relatively few eGFP-positive cells within the DRG, however, this transduction was observed at all vertebral levels and corresponded to large non-nociceptive cell types. CONCLUSION: We have found that rAAV2/6 is an efficient vector to deliver transgenes to nociceptive neurons in mice. Furthermore, the characterization of the transduction profile may facilitate gene transfer studies to dissect mechanisms behind neuropathic pain.

Proposition d'un concept de contrôle de gestion pour la division "infrastructure routière" de l'Office fédéral des routes (OFROU)

La présente étude propose une méthodologie aboutissant à la conception d'un système de contrôle de gestion pour la division « infrastructure routière » ou « I » de l'Office fédéral des routes (OFROU). La première partie de ce travail situe l'OFROU dans son contexte de réformes et identifie les raisons qui justifient l'implantation d'un système de contrôle de gestion à l'OFROU. La deuxième partie pose les jalons permettant d'établir un contrôle de gestion dans sa division « I ». Elle cherche d'abord à donner une définition du contrôle de gestion dans le secteur public, puis présente un état des lieux du système de gestion existant. Finalement, les systèmes de contrôle de gestion de deux autres offices GMEB, MétéoSuisse et armasuisse immobilier, sont analysés afin d'en tirer des éléments pertinents pouvant servir à la conception du contrôle de gestion à l'OFROU. La troisième propose un contrôle de gestion pour la division « I ». Le travail montre que la réalisation d'un concept de contrôle de gestion pour la division « I » n'est pas évidente. Cette dernière, qui est en train de se mettre en place, manque encore d'expérience pour identifier clairement les éléments clé à insérer dans le système. On observe un grand embarras entre le stade du savoir et celui du savoir faire concret. La méthodologie et l'analyse présentées dans ce travail pourraient contribuer à développer un savoir faire interne aboutissant rapidement à la mise sur pied d'un système de contrôle de gestion pour la division « I ». Die vorliegende Studie schlägt eine Methodik zur Erarbeitung eines Management-Kontrollsystems für die Abteilung « Strasseninfrastruktur » oder « I » des Bundesamts für Strassen (ASTRA) vor. Der erste Teil dieser Arbeit stellt das ASTRA im Spannungsfeld dieser Reformen. Ihre Konsequenzen lassen die Einführung eines Management-Kontrollsystems im ASTRA folgerichtig erscheinen. Im zweiten Teil geht es darum, Wegmarken zu setzen, die die Einführung einer Managementkontrolle in der Abteilung « I » in die richtigen Bahnen lenken. Es wird versucht, eine Begriffsbestimmung der Managementkontrolle im öffentlichen Sektor sowie eine Bestandesaufnahme der bestehenden Systeme vorzunehmen. Schliesslich werden die Management-Kontrollsysteme zweier anderer FLAG-Ämter, MeteoSchweiz und armasuisse immobilier, auf mögliche Nutzen für den Aufbau der Managementkontrolle des ASTRA analysiert. Der dritte Teil beinhaltet einen Vorschlag, wie die Management-Kontrolle für die Abteilung « I » aussehen könnte. Die Arbeit zeigt, dass die Erarbeitung eines Management-Kontrollkonzepts für die Abteilung « I » keine einfache Aufgabe ist. Es fehlt an Erfahrung, um die Schlüsselbausteine des Systems erkennen zu können. Zwischen dem Stadium des Wissens und dem des konkreten Know-hows klafft ein empfindlicher Graben. Methodik und Analyse, wie sie in der vorliegenden Arbeit beschrieben werden, könnten zum Aufbau eines internen Know-hows beitragen, das die rasche Verwirklichung eines Management-Kontrollsystems für die Abteilung « I » erlauben könnte.

Routes of administration of cannabis used for nonmedical purposes and associations with patterns of drug use.

PURPOSE: Little is known regarding cannabis administration routes for nonmedical use-that is, its delivery methods (e.g., joints, water pipe, food). Therefore, we examined the prevalence rates of different cannabis delivery methods and assessed the relationship of the distinct administration routes with problematic drug use. Subgroups of cannabis users were also investigated (i.e., "pure" cannabis users, previously described as employing a harmless route of administration, and water pipe users, previously described as using a harmful route of administration). METHODS: As part of the Cohort Study on Substance Use Risk Factors, 1,763 cannabis users answered questions concerning their drug use (i.e., routes of administration, problematic cannabis use, other illicit drug use). Descriptive statistics, latent class analysis, correlations and t-tests were assessed. RESULTS: The main administration route was "joints with tobacco"; other routes of administration had prevalence rates from 23.99% to 38.23%. In addition, increasing the number of administration routes was associated with more problematic cannabis use, as well as heavier illicit drug use. Water pipes without tobacco were especially linked to heavy drug use patterns, whereas "pure" cannabis use seemed less harmful. CONCLUSIONS: Our findings highlighted that diversification in routes of administration can be associated with heavier illicit drug use. This was especially true for water pipe users, whereas "pure" cannabis users, who did not mix cannabis with tobacco, were an exception. Indeed, these results may be useful for future preventive programs, which may need to focus on those who have diversified routes of administration for cannabis.

Effects of rat anti-VEGF antibody in a rat model of corneal graft rejection by topical and subconjunctival routes.

PURPOSE: To compare the effect of a rat anti-VEGF antibody, administered either by topical or subconjunctival (SC) routes, on a rat model of corneal transplant rejection.METHODS: Twenty-four rats underwent corneal transplantation and were randomized into four treatment groups (n=6 in each group). G1 and G2 received six SC injections (0.02 ml 10 µg/ml) of denatured (G1) or active (G2) anti-VEGF from Day 0 to Day 21 every third day. G3 and G4 were instilled three times a day with denatured (G3) or active (G4) anti-VEGF drops (10 µg/ml) from Day 0 to Day 21. Corneal mean clinical scores (MCSs) of edema (E), transparency (T), and neovessels (nv) were recorded at Days 3, 9, 15, and 21. Quantification of neovessels was performed after lectin staining of vessels on flat mounted corneas.RESULTS: Twenty-one days after surgery, MCSs differed significantly between G1 and G2, but not between G3 and G4, and the rejection rate was significantly reduced in rats receiving active antibodies regardless of the route of administration (G2=50%, G4=66.65% versus G1 and G3=100%; p<0.05). The mean surfaces of neovessels were significantly reduced in groups treated with active anti-VEGF (G2, G4). However, anti-VEGF therapy did not completely suppress corneal neovessels.CONCLUSIONS: Specific rat anti-VEGF antibodies significantly reduced neovascularization and subsequent corneal graft rejection. The SC administration of the anti-VEGF antibody was more effective than topical instillation.

What is the influence of vaccination's routes on the regression of tumors located at mucosal sites?

Tumor-regressions following tumor-associated-antigen vaccination in animal models contrast with the limited clinical outcomes in cancer patients. Most animal studies however used subcutaneous-tumor-models and questions arise as whether these are relevant for tumors growing in mucosae; whether specific mucosal-homing instructions are required; and how this may be influenced by the tumor.

Formica pratensis (Hyménoptères: Formicidae) dans le canton de Vaud: état des peuplements et importance des talus de routes comme milieu de substitution.

Formica pratensis (Hyménoptères Formicidae) figure sur la liste rouge des espèces menacées de Suisse. Cette espèce de fourmis des bois colonise les milieux herbacés ouverts comme les prés et prairies sèches peu exploitées, les talus bien exposés. Face à la raréfaction de ces habitats liée à l'intensification de l'agriculture, elle colonise de plus en plus souvent les talus herbeux en bordure de routes. Afin de mieux comprendre la situation actuelle de l'espèce et ses besoins en milieux naturels, nous avons mené une étude à large échelle dans le cnaton de Vaud (Suisse). F.pratensis se rencontre sur l'ensemble du territoire, principalement en dessous de 800 m, mais elle ne semble plus aussi fréquente qu'autrefois. F.pratensis recherche surtout des stations en pente bien exposées au soleil et avec une végétation assez ouverte. De tels milieux se faisant de plus en plus rares dans le paysage agricole moderne, les talus de route apparaissent souvent comme une alternative (ou milieu de subsitution) pour F.pratensis. les importantes perturbations (pollution, fauche répétitive, pauvreté du milieu) ne permettent toutefois pas aux sociétés de fourmis de se développer pleinement et la survie de l'espèce passe par la sauvegarde d'autres milieux ouverts exploités extensivement. Grâce aux données collectées, un suivi comparatif des populations pourra être effectuée dans les années à venir

Dispersal routes reconstruction and the minimum cost arborescence problem.

We show that the dispersal routes reconstruction problem can be stated as an instance of a graph theoretical problem known as the minimum cost arborescence problem, for which there exist efficient algorithms. Furthermore, we derive some theoretical results, in a simplified setting, on the possible optimal values that can be obtained for this problem. With this, we place the dispersal routes reconstruction problem on solid theoretical grounds, establishing it as a tractable problem that also lends itself to formal mathematical and computational analysis. Finally, we present an insightful example of how this framework can be applied to real data. We propose that our computational method can be used to define the most parsimonious dispersal (or invasion) scenarios, which can then be tested using complementary methods such as genetic analysis.

Comparative phylogeography and postglacial colonization routes in Europe.

The Quaternary cold periods in Europe are thought to have heavily influenced the amount and distribution of intraspecific genetic variation in both animals and plants. The phylogeographies of 10 taxa, including mammals (Ursus arctos, Sorex spp., Crocidura suaveolens, Arvicola spp.), amphibians (Triturus spp.), arthropods (Chorthippus parallelus), and plants (Abies alba, Picea abies, Fagus sylvatica, Quercus spp.), were analysed to elucidate general trends across Europe. Only a small degree of congruence was found amongst the phylogeographies of the 10 taxa, but the likely postglacial colonization routes exhibit some similarities. A Brooks parsimony analysis produced an unrooted area phylogram, showing that: (i) the northern regions were colonized generally from the Iberic and Balkanic refugia; and (ii) the Italian lineages were often isolated due to the presence of the Alpine barrier. The comparison of colonization routes highlighted four main suture-zones where lineages from the different refugia meet. Some of the intraspecific genetic distances among lineages indicated a prequaternary divergence that cannot be connected to any particular cold period, but are probably related mainly to the date of arrival of each taxon in the European continent. As a consequence, molecular genetics so far appears to be of limited use in dating Quaternary events.