Etude clinique et analyse de liaison au locus 3q28 de deux familles suisses avec atrophie optique dominante de Kjer (OPA1) [Clinical study and genetic 3q28 locus linkage in 2 Swiss families with Kjer dominant optic atrophy (OPA1)]

METHODS: We examined 20 patients from 2 unrelated Swiss families to describe their clinical phenotype. In addition, a linkage analysis was performed in an attempt to confirm the reported genetic homogeneity of this condition as well as to refine its genomic localization. RESULTS: Two point analysis provided a cumulative LOD-score of 3.03 with marker D3S 2305. The absence of recombination precluded further refinement of the disease interval. CONCLUSIONS: Our data confirm the genetic homogeneity and the extreme variability of expression, occasionally mimicking low tension glaucoma.

Somatostatin Subtype‑2 Receptor-Targeted Metal-Based Anticancer Complexes

Conjugates of a dicarba analogue of octreotide, a potent somatostatin agonist whose receptors are overexpressed on tumor cells, with [PtCl2(dap)] (dap = 1-(carboxylic acid)-1,2-diaminoethane) (3), [(η6-bip)Os(4-CO2-pico)Cl] (bip = biphenyl, pico = picolinate) (4), [(η6-p-cym)RuCl(dap)]+ (p-cym = p-cymene) (5), and [(η6-p-cym)RuCl(imidazole-CO2H)(PPh3)]+ (6), were synthesized by using a solid-phase approach. Conjugates 35 readily underwent hydrolysis and DNA binding, whereas conjugate 6 was inert to ligand substitution. NMR spectroscopy and molecular dynamics calculations showed that conjugate formation does not perturb the overall peptide structure. Only 6 exhibited antiproliferative activity in human tumor cells (IC50 = 63 ± 2 μM in MCF-7 cells and IC50 = 26 ± 3 μM in DU-145 cells) with active participation of somatostatin receptors in cellular uptake. Similar cytotoxic activity was found in a normal cell line (IC50 = 45 ± 2.6 μM in CHO cells), which can be attributed to a similar level of expression of somatostatin subtype-2 receptor. These studies provide new insights into the effect of receptor-binding peptide conjugation on the activity of metal-based anticancer drugs, and demonstrate the potential of such hybrid compounds to target tumor cells specifically.

El Land-Art a l'educació 0-3: l'expressió lliure i la comprensió del medi a través de l'art i la natura

Actualment el Land – Art és un moviment poc conegut i difós en la vessant educativa. Aquest treball pretén treure a la llum i transmetre l’essència d’aquest moviment enriquidor cap als infants. Tant és així, que el Land – Art esdevé una eina d’aprenentatge innovadora en el primer cicle d’Educació Infantil a través d’una metodologia constructivista que involucra a tota la comunitat educativa: professionals, família i alumnes. L’objectiu del present projecte és l’ús d’aquest moviment com instrument d’aprenentatge per tal d’observar i analitzar uns resultats que s’han anat construint a partir d’una hipòtesi: “El Land – Art a l’Educació 0-3 fomenta l’expressió lliure i la comprensió del medi”. En aquest treball trobarem, en primer lloc, el marc teòric del projecte; en segon lloc, el seu mètode d’investigació; en tercer lloc, la vessant pràctica del projecte; i, en quart i darrer lloc, les conclusions que reflecteixen la part comparativa entre el marc teòric i la part pràctica, on es finalitzarà amb les reflexions finals.

Monocarboxylate transporters: new players in body weight regulation.

Over the last two decades, several genes have been identified that appear to play a role in the regulation of energy homeostasis and body weight. For a small subset of them, a reduction or an absence of expression confers a resistance to the development of obesity. Recently, a knockin mouse for a member of the monocarboxylate transporter (MCT) family, MCT1, was demonstrated to exhibit a typical phenotype of resistance to diet-induced obesity and a protection from its associated metabolic perturbations. Such findings point out at MCTs as putatively new therapeutic targets in the context of obesity. Here, we will review what is known about MCTs and their possible metabolic roles in different organs and tissues. Based on the description of the phenotype of the MCT1 knockin mouse, we will also provide some insights about their putative roles in weight gain regulation.

Nanoparticles for gene delivery to retinal pigment epithelial cells.

PURPOSE: To evaluate the safety and potential use of poly(lactic) acid (PLA) and poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) as vectors for gene transfer to RPE cells. METHODS: Experiments were conducted with primary bovine RPE cells and with the ARPE-19 human RPE cell line. Rhodamine loaded NPs were used to study factors influencing the internalization process by the various RPE cells: concentrations of NPs, duration of contact time, stage of cell culture and ambient temperature. The extent of NPs internalization was evaluated by fluorescence and phase microscopy. Potential NP toxicity was measured by the trypan blue exclusion dye test and the MTT method. Green fluorescent protein (GFP) plasmid or red nuclear fluorescent protein (RNFP) plasmid were sequestered in NPs. The ability ot these "loaded" NPs to generate gene transfection and protein expression in RPE cells was assessed both in vivo and in vitro by fluorescence and confocal microscopy. RESULTS: The extent of NP internalization in cultured cells increases with their concentration reaching a plateau at 1 mg/ml and a contact time of up to 6 h. Temperature and culture stage did not influence the in vitro internalization process. No toxic effects on RPE cells could be detected when these were incubated with up to 4 mg/ml of NPs. In human and bovine RPE cells incubated with GFP loaded NPs, cytoplasmic green fluorescence was observed in 14+/-1.65% of the cultured cells. Incubation with RNFP loaded NPs yielded a nuclear red fluorescence in 18.9+/-1.6% of the cells. These percentage levels of expression initially detected after 48 h of incubation remained unchanged during the following 8 additional days in culture. No significant differences in the extent of cytoplasm or nuclear fluorescence expression were observed between bovine or human RPE cultured cells. In vivo, a preferential RNFP expression within the RPE cell layer was detected after intra vitreous injection of RNFP plasmid loaded NPs. CONCLUSIONS: The ability of PLGA NPs to sequester plasmids, their nontoxic characteristics, and rapid internalization enables gene transfer and expression in RPE cells. These findings may be of potential use when designing future gene therapy strategies for ocular diseases of the posterior segment.

Capsid modified replicating adenovirus to selectively target colon cancer

Résumé: Pratiquement tous les cancers du colon contiennent des mutations dans la voie de signalisation de Wnt qui active constitutivement cette voie. Cette activation mène à la stabilisation de la β-catenine. La β-catenin est transportée dans le noyau ou elle active des gènes cible en interagissant avec le facteur de transcription de TCF/LEF. Des adénovirus qui peuvent sélectivement se répliquer dans les cellules tumorales sont les agents qui peuvent permettre la déstruction de la tumeur mais pas le tissu normal. In vitro, les adénovirus avec des sites d'attachement du facteur de transcription TCF dans les promoteurs de l'adénovirus montrent une sélectivité et une activité dans une large sélection de lignées cellulaires de cancer du colon. Au contraire, in vivo, quand les adénovirus modifiés sont injectés dans la circulation, ils sont moins efficaces à cause de leur fixation par le foie et à cause de l'absence d'expression du récepteur du Coxsackie-Adénovirus (CAR). Le but de ma thèse était de modifier la protéine principale de capside de l'adénovirus, fibre, pour augmenter l'infection des tumeurs du cancer du colon. La fibre de l'adénovirus est responsable de l'attachement aux cellules et de l'entrée virale. J'ai inséré un peptide RGD dans la boucle HI de la fibre qui dirige sélectivement le virus aux récepteurs des integrines. Les integrines sont surexprimées par les cellules du cancer du colon et l'endothélium des vesseaux de la tumeur. Le virus re-ciblé, vKH6, a montré une activité accrue dans toutes les lignées cellulaires de cancer du colon, tandis que la sélectivité était maintenue. In vivo, vKH6 était supérieur au virus avec une capside de type sauvage en retardant la croissance de la tumeur. Le virus s'est répliqué plus vite et dispersé graduellement dans la tumeur. Cet effet a été montré par hybridation in situ et par PCR quantitative. Cependant, la monothérapie avec le virus n'a pu retarder la croissance des cellules tumorales SW620 greffées que de 2 semaines, mais à cause des régions non infectées la tumeur n'a pas pu être éliminée. Bien que la combinaison avec les chimiothérapies conventionnelles soit d'intérêt potentiel, presque toutes interfèrent avec la réplication virale. Les drogues antiangiogéniques sont des agents anti-tumoraux efficaces et prometteurs. Ces drogues n'interfèrent pas avec le cycle de vie de l'adénovirus. RAD001 est un dérivé de la rapamycine et il inhibe mTOR, une protéine kinase de la voie de PI3K. RAD001 empêche la croissance des cellules et il a aussi des effets anti-angiogénique et immunosuppressifs. RAD001 in vitro n'affecte pas l'expression des gènes viraux et la production virale. La combinaison de VKH6 et RAD001 in vivo a un effet additif en retardant la croissance de la tumeur. Des nouveaux peptides plus efficaces dans le ciblage de l'adénovirus sont nécessaires pour augmenter l'infection des tumeurs. J'ai créé un système de recombinaison qui permettra la sélection de nouveaux peptides dans le contexte du génome de l'adénovirus. Summary Virtually all colon cancers have mutations in the Wnt signalling pathway which result in the constitutive activation of the pathway. This activation leads to stabilization of β-catenin. β-catenin enters the nucleus and activates its target genes through interaction with the TCF transcription factor. Selectively replicating adenoviruses are promising novel agents that can destroy the tumour but not the surrounding normal tissue. In vitro, adenoviruses with TCF binding sites in the early viral promoters show selectivity and activity in a broad panel of viruses but in vivo they are less effective due to the lack of expression of the Coxsackie-Adenovirus receptor (CAR). The aim of my thesis was to modify the major capsid protein of the adenovirus, fibre, to increase the infection of colon tumours. Fibre of adenovirus is responsible for the binding to cells and for the viral uptake. I inserted an RGD binding peptide into the HI loop of fibre that selectively targets the virus to integrins that are overexpressed on tumour cells and on tumour endothelium. The retargeted virus, vKH6, showed increased activity in all colon cancer cell lines while selectivity was maintained. In vivo, vKH6 is superior to a matched virus with a wild type capsid in delaying tumour growth. vKH6 replicates and gradually spreads within the tumour as shown by in situ hybridization and Q-PCR. The virus alone can delay the growth of SW620 xenografts by 2 weeks but due to uninfected tumour regions the tumour cannot be cured. Although combination with conventional chemotherapeutics is of potential interest, almost all of them interfere with the viral replication. Growing evidence supports that anti-angiogenic drugs are effective and promising anti-tumour agents. These drugs interfere less with the viral life cycle. RAD001 is a rapamycin derivative and it blocks mTOR, a protein kinase in the PI3K pathway. RAD001 inhibits cell growth and has strong anti-angiogenic and immunosuppressive effects. RAD001 in vitro does not affect viral gene expression and viral burst size. In vivo vKH6 and RAD001 have an additive effect in delaying tumour growth, but tumour growth is still not completely inhibited. To further increase tumour infection new tumour specific targeting peptides are needed. I created an adenovirus display library that will allow the selection of targeting peptides. This system may also facilitate the production of fibre modified viruses.

Vaarinpidosta virtuaaliaikaan : Sata vuotta suomalaista tilintarkastusta

Tämän tutkimuksen aiheena on tilintarkastuksen historiallinen kehittyminen Suomessa runsaan sadan vuoden aikana. Tutkimuksen tavoitteena on analysoida osakeyhtiön tilintarkastuksen kehitystä ja yhdistää vuosisadan kehityspiirteet tilintarkastuksen kokonaiskuvaksi. Tutkittava periodi alkaa 1800-luvun lopulta ja päättyy 2000-luvun taitteeseen. Tutkimuksessa tarkastellaan suomalaista tilintarkastusinstituutiota, joka jaetaan kolmeen osaan: tilintarkastusta säätelevään normistoon (normit), tilintarkastajajärjestelmään (toimijat) ja tilintarkastuksen sisältöön (tehtävät). Tutkimuksessa tavoitellaan vastauksia kysymyksiin: mitä tarkastettiin, milloin tarkastettiin, kuka tarkasti ja miten tarkastettiin eri aikakausina? Tutkimus perustuu historialliseen lähdeaineistoon, jonka muodostavat tutkimusajanjakson lainsäädäntö, lainvalmisteluasiakirjat, viranomaisten ohjeet ja päätökset, alan järjestöjen suositukset, ammattilehtien artikkelit sekä laskentatoimen ja tilintarkastuksen ammattikirjallisuus. Metodologisesti tutkimus on teoreettinen, kvalitatiivinen historiantutkimus, jossa lähdeaineistoa käsitellään lähdekriittisesti ja osittain sisältöanalyysin keinoin. Tilintarkastusta säätelevässä normistossa keskeisiä lakeja ovat olleet osakeyhtiölaki, kirjanpitolaki ja tilintarkastuslaki. Lakisääteinen tilintarkastus alkoi vuoden 1895 osakeyhtiölaista, joka uudistui vuonna 1978 ja jälleen vuonna 1997. Kirjanpitolainsäädäntö on uudistunut viidesti: 1925 ja 1928, 1945, 1973, 1993 sekä 1997. Vuoden 1994 tilintarkastuslakiin koottiin tilintarkastuksen säädökset useista laeista. Muita normistoja ovat olleet EY:n direktiivit, Kilan ohjeet, KHT-yhdistyksen suositukset, Keskuskauppakamarin säännökset ja viimeisimpinä IAS- ja ISA-standardit. Ammattimainen tilintarkastajajärjestelmä saatiin maahamme kauppiaskokousten ansiosta. Ammattimaisena tilintarkastuksen toimijana aloitti Suomen Tilintarkastajainyhdistys vuonna 1911, ja sen toimintaa jatkoi KHT-yhdistys vuodesta 1925 alkaen. Tilintarkastajien auktorisointi siirtyi Keskuskauppakamarille vuonna 1924. HTM-tilintarkastajat ovat olleet alalla vuodesta 1950 lähtien. Kauppakamarijärjestö on toiminut hyväksyttyjen tilintarkastajien valvojana koko ammattimaisen tilintarkastustoiminnan ajan. Valtion valvontaa suorittaa VALA (Valtion tilintarkastuslautakunta). Koko tutkittavan periodin ajan auktorisoitujen tilintarkastajien rinnalla osakeyhtiöiden tarkastajina ovat toimineet myös maallikot.Tilintarkastuksen tehtäviin kuului vuoden 1895 osakeyhtiölain mukaan hallinnon ja tilien tarkastus. Myöhemmin sisältö täsmentyi tilinpäätöksen, kirjanpidon ja hallinnon tarkastukseksi. Tutkimusajanjakson alussa tilintarkastus oli manuaalista kaikkien tositteiden prikkausta ja virheiden etsimistä. Myöhemmin tarkastus muuttui pistokokeiksi. Kertatarkastuksesta siirryttiin jatkuvaan valvontatarkastukseen 1900-luvun alkupuolella. Dokumentoinnista ja työpapereista alkaa olla havaintoja 1930-luvulta lähtien. Atk-tarkastus yleistyi 1970- ja 1980-luvuilla, jolloin myös riskianalyyseihin alettiin kiinnittää huomiota. Hallinnon tarkastuksen merkitys on kasvanut kaiken aikaa. Tilintarkastuskertomukset olivat tutkimusajanjakson alussa vapaamuotoisia ja sisällöltään ilmaisurikkaita ja kuvailevia. Kertomus muuttui julkiseksi vuoden 1978 osakeyhtiölain myötä. Myöhemmin KHT-yhdistyksen vakiokertomusmallit yhdenmukaistivat ja pelkistivät raportointia. Tutkimuksen perusteella tilintarkastuksen historia voidaan jakaa kolmeen kauteen, jotka ovat tilintarkastusinstituution rakentumisen kausi (1895 - 1950), vakiintumisen kausi (1951 - 1985) ja kansainvälistymisen ja julkisuuden kausi (1986 alkaen). Tutkimusajanjakson jokaisella vuosikymmenellä keskusteltiin jatkuvasti tilintarkastajien riittävyydestä, alalle pääsyn ja tutkintojen vaikeudesta, tilintarkastajien ammattitaidon tasosta,hallinnon tarkastuksen sisällöstä, tilintarkastuskertomuksesta sekä maallikkotarkastajien asemasta. 1990-luvun keskeisimmät keskusteluaiheet olivat konsultointi, riippumattomuus, odotuskuilu sekä tilintarkastuksen taso ja laadunvalvonta. Analysoitaessa tilintarkastuksen muutoksia runsaan sadan vuoden ajalta voidaan todeta, että tilintarkastuksen ydintehtävät eivät juurikaan ole muuttuneet vuosikymmenien kuluessa. Osakeyhtiön tilintarkastus on edelleenkin laillisuustarkastusta. Sen tarkoituksena on yhä kirjanpidon, tilinpäätöksen ja hallinnon tarkastus. Tilintarkastajat valvovat osakkeenomistajien etua ja raportoivat heille tarkastuksen tuloksista. Tilintarkastuksen ulkoinen maailma sen sijaan on muuttunut vuosikymmenten saatossa. Kansainvälistyminen on lisännyt säännösten määrää, odotuksia ja vaatimuksia on nykyisin enemmän, uusi tekniikka mahdollistaa nopean tiedonkulun ja valvonta on lisääntynyt nykypäivää kohti tultaessa. Tilintarkastajan pätevyys perustuu nykyään tietotekniikan, tietojärjestelmien ja yrityksen toimialantuntemukseen. Runsaan sadan vuoden takaisen lain vaarinpitovaatimuksesta on tultu virtuaaliaikaiseen maailmaan!

Genome-wide transcriptional responses to shade: Linking shade avoidance and auxin

Plants have the ability to use the composition of incident light as a cue to adapt development and growth to their environment. Arabidopsis thaliana as well as many crops are best adapted to sunny habitats. When subjected to shade, these plants exhibit a variety of physiological responses collectively called shade avoidance syndrome (SAS). It includes increased growth of hypocotyl and petioles, decreased growth rate of cotyledons and reduced branching and crop yield. These responses are mainly mediated by phytochrome photoreceptors, which exist either in an active, far-red light (FR) absorbing or an inactive, red light (R) absorbing isoform. In direct sunlight, the R to FR light (R/FR) ratio is high and converts the phytochromes into their physiologically active state. The phytochromes interact with downstream transcription factors such as PHYTOCHROME INTERACTING FACTOR (PIF), which are subsequently degraded. Light filtered through a canopy is strongly depleted in R, which result in a low R/FR ratio and renders the phytochromes inactive. Protein levels of downstream transcription factors are stabilized, which initiates the expression of shade-induced genes such as HFR1, PIL1 or ATHB-2. In my thesis, I investigated transcriptional responses mediated by the SAS in whole Arabidopsis seedlings. Using microarray and chromatin immunoprecipitation data, we identified genome-wide PIF4 and PIF5 dependent shade regulated gene as well as putative direct target genes of PIF5. This revealed evidence for a direct regulatory link between phytochrome signaling and the growth promoting phytohormone auxin (IAA) at the level of biosynthesis, transport and signaling. Subsequently, it was shown, that free-IAA levels are upregulated in response to shade. It is assumed that shade-induced auxin production takes predominantly place in cotyledons of seedlings. This implies, that IAA is subsequently transported basipetally to the hypocotyl and enhances elongation growth. The importance of auxin transport for growth responses has been established by chemical and genetic approaches. To gain a better understanding of spatio-temporal transcriptional regulation of shade-induce auxin, I generated in a second project, an organ specific high throughput data focusing on cotyledon and hypocotyl of young Arabidopsis seedlings. Interestingly, both organs show an opposite growth regulation by shade. I first investigated the spatio-transcriptional regulation of auxin re- sponsive gene, in order to determine how broad gene expression pattern can be explained by the hypothesized movement of auxin from cotyledons to hypocotyls in shade. The analysis suggests, that several genes are indeed regulated according to our prediction and others are regulated in a more complex manner. In addition, analysis of gene families of auxin biosynthetic and transport components, lead to the identification of essential family members for shade-induced growth re- sponses, which were subsequently experimentally confirmed. Finally, the analysis of expression pattern identified several candidate genes, which possibly explain aspects of the opposite growth response of the different organs.

What to compare and how: Comparative transcriptomics for Evo-Devo.

Evolutionary developmental biology has grown historically from the capacity to relate patterns of evolution in anatomy to patterns of evolution of expression of specific genes, whether between very distantly related species, or very closely related species or populations. Scaling up such studies by taking advantage of modern transcriptomics brings promising improvements, allowing us to estimate the overall impact and molecular mechanisms of convergence, constraint or innovation in anatomy and development. But it also presents major challenges, including the computational definitions of anatomical homology and of organ function, the criteria for the comparison of developmental stages, the annotation of transcriptomics data to proper anatomical and developmental terms, and the statistical methods to compare transcriptomic data between species to highlight significant conservation or changes. In this article, we review these challenges, and the ongoing efforts to address them, which are emerging from bioinformatics work on ontologies, evolutionary statistics, and data curation, with a focus on their implementation in the context of the development of our database Bgee (http://bgee.org). J. Exp. Zool. (Mol. Dev. Evol.) 324B: 372-382, 2015. © 2015 Wiley Periodicals, Inc.

Micrornas involvement in cortical plasticity in adult mouse barrel cortex

In rodents, sensory experience alters the whisker representation in layer IV of the barrel cortex (Woolsey and Van der Loos, 1970). Excitatory and inhibitory interneurons, together with the astrocytic network, modify the functional representation in an integrated manner. Our group showed that continuous whisker stimulation induces structural and functional changes in the corresponding barrel. These modifications include the depression of neuronal responses and an insertion of new inhibitory synapses on dendritic spines (Knott et al., 2002; Genoud et al., 2006; Quairiaux et al., 2007). This form of cortical plasticity is controlled by several gene regulatory mechanisms including the activation of genetic programs controlling the expression of microRNAs (miRNAs). The transitory and localized expression of miRNAs in dendrites and their capacity to respond in an activity-dependent manner make them ideal candidates for the fine tuning of gene expression associated with neural plasticity. In a previous study of our group (Johnston- Wenger, 2010) using microarray analysis on laser-dissected barrels in order to compare the gene expression levels in stimulated and non-stimulated barrels after whisker stimulation, 261 genes were found significantly regulated, among these genes there were two miRNAs (miR- 132 and miR-137). In this study I tested the initial observation on the up-regulation of miR-132 and miR-137 after whisker stimulation and the possible involvement of two other miRNAs (miR-138 and miR-125b) that are known play a role in other form of synaptic plasticity. I used in situ hybridization (ISH) after unilateral stimulation of three whiskers (Cl-3) in the adult mouse. We found that sensory stimulation increases the expression, of miR-132 after 3hours of stimulation (p<0.01) and miR-137 (pO.Ol; 24 hrs of stim.), whereas it reduces the level of miR-125b (pO.Ol; 9 hrs of stim.). No significant difference was detected for miR-138. We further determined a correlation between the level of expression of the four selected miRNAs in the cortical barrels (measured by ISH) and in blood plasma (measured by qPCR). In addition to this quantitative comparison, we combined miRNAs ISH and immunolabeling for various neuronal markers that were chosen for the localization in both excitatory and inhibitory circuits as well as in astrocytes. Analysis of three-dimensional confocal acquisitions showed that stimulation alters significantly the degree of co-localization in the stimulated barrel of miR-132 with GAD65/67 and VGLUT2; miR-125b with GAD65/67 and parvalbumin; miR-138 with parvalbumin, VGLUT1 and PSD95; and miR-137 with VGLUT1 and astrocytic markers (GS; GFAP and SlOOß). To conclude, using increased neuronal activity in the whisker-to-barrel pathway; our results suggest that miRNAs can be regulated in an activity-dependent manner and they may regulate local mRNA translation to shape neuronal responses. These findings motivate further investigation of the different modes in which miRNAs may regulate cortical plasticity. -- Chez les rongeurs, l'expérience sensorielle modifie la représentation des vibrisses au niveau du cortex somatosensoriel primaire (Woolsey and Van der Loos, 1970). Les interneurones excitateurs et inhibiteurs, en collaboration avec le réseau astrocytaire, modifient la représentation fonctionnelle d'une manière intégrée. Notre groupe a montré que la stimulation continue des vibrisses induit des changements structuraux et fonctionnels dans le tonneau correspondant. Ces modifications incluent la dépression des réponses neuronales et une insertion de nouvelles synapses inhibitrices sur les épines dendritiques (Knott et al., 2002 ; Genoud et al., 2006 ; Quairiaux et al., 2007). Cette forme de plasticité corticale est contrôlée par plusieurs mécanismes de régulation génique dont l'activation des programmes géniques contrôlant l'expression des microARNs (miARNs). Par leur expression transitoire et localisée dans les dendrites et leur capacité à réagir d'une manière dépendante de l'activité, les miARNs sont des candidats idéaux pour le réglage fin de l'expression des gènes associée à la plasticité neuronale. Afin de comparer le niveau d'expression des gènes dans les tonneaux stimulés et non-stimulés après stimulation des vibrisses, une étude antérieure dans notre groupe (Johnston-Wenger, 2010), utilisant l'analyse par microarray sur des tonneaux disséqués par laser, a montré l'altération significative de 261 gènes. Parmi ces gènes, il y avait deux miARNs (miR-132 et miR-137). Dans la présente étude, j'ai testé l'observation initiale sur la régulation de miR-132 et miR-137 après stimulation des vibrisses et la possible implication de deux autres miARNs (miR-138 et miR-125b) connus avoir jouer un rôle important dans d'autres formes de plasticité synaptique. J'ai utilisé l'hybridation in situ (ISH) après stimulation unilatérale de trois vibrisses (Cl-3) chez la souris adulte. J'ai trouvé que la stimulation sensorielle augmente l'expression, de miR-132 après 3 heures de stimulation (p < 0.01) et miR-137 (p < 0.01 ; 24 hrs de stim.), alors qu'elle réduit le niveau de miR-125b (p < 0.01; 9 hrs de stim.). Aucune différence significative n'a été détectée pour miR-138. J'ai aussi déterminé une corrélation entre le niveau d'expression des quatre miARNs sélectionnés dans les tonneaux (mesurés par ISH) et dans le plasma sanguin (mesuré par qPCR). En plus de cette comparaison quantitative, j'ai combiné le miR-ISH et l'immunomarquage pour divers marqueurs neuronaux qui ont été choisis pour étudier la localisation dans les circuits excitateurs et inhibiteurs, ainsi que dans les astrocytes. Les acquisitions tridimensionnelles montrent que la stimulation modifie considérablement le degré de co-localisation dans le tonneau stimulé de miR-132 avec GAD65/67 et VGLUT2; miR-125b avec GAD65/67 et parvalbumine; miR-138 avec parvalbumine, VGLUT1 et PSD95; et miR-137 avec VGLUT1 et les marqueurs astrocytaires (GS ; GFAP et SlOOß). En conclusion, à l'aide de l'activité neuronale accrue dans la voie de vibrisses-au-baril; les résultats suggèrent que les miARNs peuvent être régulé d'une manière dépendante de l'activité et peuvent résulter la stabilité des ARNm et la traduction pour façonner les réponses neuronales ultérieures. Ces résultats incitent d'investiguer davantage les voies importantes par lesquels les miARNs peuvent réguler la plasticité corticale.

Net Neutrality: Measuring the Problem, Assessing the Legal Risks

Network neutrality is a growing policy controversy. Traffic management techniques affect not only high-speed, high-money content, but by extension all other content too. Internet regulators and users may tolerate much more discrimination in the interests of innovation. For instance, in the absence of regulatory oversight, ISPs could use Deep Packet Inspection (DPI) to block some content altogether, if they decide it is not to the benefit of ISPs, copyright holders, parents or the government. ISP blocking is currently widespread in controlling spam email, and in some countries in blocking sexually graphic illegal images. In 1999 this led to scrutiny of foreclosure of Instant Messaging and video and cable-telephony horizontal merger. Fourteen years later, there were in 2013 net neutrality laws implemented in Slovenia, the Netherlands, Chile and Finland, regulation in the United States and Canada , co-regulation in Norway, and self-regulation in Japan, the United Kingdom and many other European countries . Both Germany and France in mid-2013 debated new net neutrality legislation, and the European Commission announced on 11 September 2013 that it would aim to introduce legislation in early 2014. This paper analyses these legal developments, and in particular the difficulty in assessing reasonable traffic management and ‘specialized’ (i.e. unregulated) faster services in both EU and US law. It also assesses net neutrality law against the international legal norms for user privacy and freedom of expression

Deleuze, lector de Spinoza. Del problema de la expresión a la filosofía práctica

[spa]El objetivo de este artículo es presentar la interpretación deleuziana de la filosofía de Spinoza como una vía posible para la crítica del primer proyecto moderno. Para ello partimos de la premisa de que la expresión es un problema insoluble para el cartesianismo, y que el spinozismo intentó explorar este asunto para mostrar la naturaleza paradójica de este concepto. El estudio se centra en dos obras de Deleuze: Spinoza y el problema de la expresión y Spinoza: filosofía práctica.

Análisis del proceso de adquisición del código audiovisual y de la intervención de los agentes que forman a los ciudadanos en el conocimiento de los medios

The research group Gre‐TICE (Grupo de investigación en tecnologías de la Información y la Comunicación en Educación) has the acquisition of the multimedia language and their use as a form of expression as one of their lines of research. During the academic year 2002‐ 2003, following previous work in the use of ICT in Education, commenced upon the project: “The acquisition of visual and sound codes and the processes related to the visual media”. The intention of this project is to study how formal or non‐formal education context can help young adults and children to acquire visual and sound codes to become ‘critical consumers’ with the media and to use the tools in a creative way. To achieve this objective, the project team has developed a partner group which includes professional from different European regions; including teachers and managers from across the age spectrum, government institutions and cultural organisations. Whilst the project will call upon qualitative analysis of the previous projects / research, it will seek to develop ‘Good Practice’ guides and other resources/ materials to be disseminated to project partners (and others) to build innovative actions throughout the European region

Produção, propriedades e aplicação de celulases na hidrólise de resíduos agroindustriais

Cellulases have been intensively studied in the past few years, due to the interests in biofuels production from lignocellulosic materials, since they permit maintaining mild conditions during the conversion process. These enzymes can be produced by a broad variety of naturally occurring microorganisms, such as from genera Aspergillus, Trichoderma, Penicillium and Humicola. Targeting the increasing of expression levels, molecular biology tools have been used for heterologous genes insertion in host cells, e. g., Pichia pastoris and Escherichia coli. Enzymes from fungal cellulolytic complex usually act best at pH between 4 and 5 under temperatures from 40 to 60 °C and can be used for either sequential (SHF) or simultaneous (SSF) hydrolysis together alcoholic fermentation. In this review, the main raw materials for production of cellulases are identified, as well as the state of art of enzymes' properties, production and main applications.

Salivary gland cancer in Finland: Incidence, histological distribution, outcome and prognostic factors

Salivary gland cancer (SGC) is a rare cancer. The histological classification of SGC is complex and its biological behavior highly variable: it may vary from a low-grade tumor to a high-grade and often fatal malignancy. These circumstances make this cancer a diagnostic and therapeutic challenge. Older age and exposure to ionizing radiation are known risk factors. The mainstay of treatment is surgery combined with adjuvant radiation therapy, when appropriate. In addition to the histological type, the only well known prognostic factor is the TNM classification, which describes the tumor size and the amount of metastases. This study was performed using a full population-based nationwide cohort of SGC patients and tumors diagnosed in Finland in 1991-1996. The annual incidence of SGC in the entire population was, on average, 47.7 per year. By histological re-evaluation of 237 specimens the most frequent histological types were the adenoid cystic carcinoma (n=65; 27%), the mucoepidermoid carcinoma (n=45; 19%) and the acinic cell carcinoma (n=41; 17%). The highest 10-year disease-specific survival rate occurred among patients with acinic cell carcinoma (90%), followed by mucoepidermoid carcinoma (81%) and adenoid cystic carcinoma (60%). A high volume-corrected index (VCI) of Ki-67 correlated with worse survival of patients with SGC. Computer-assisted morphometric analyses of CD34-positive vessels indicated an unfavorable prognosis for patients with mucoepidermoid carcinoma and an association with poor survival among patients with acinic cell carcinoma. A high level of expression of matrix metalloproteinase-9 (MMP-9) showed a trend for a poorer prognosis in salivary duct carcinoma, and a high level of MMP-13 and a low level of MMP-1 had a trend for a poorer prognosis of patients with SGC. A low level of MMP-7 was associated with a poor prognosis of patients with acinic cell and mucoepidermoid carcinoma.