Insulin and cellular stressors both activate p38 MAPK. Insulin protects cardiac tissue in a p38 MAPK-dependent manner. Paradoxically, inhibiting p38 MAPK is also protective. Hsp27 phosphorylation is regulated by p38 MAPK. Insulin was tested in H9c2 cardiomyocytes subjected to media exchange, 6 hours of oxygen-glucose deprivation, and reoxygenation. Insulin suppressed stress-induced phosphorylation of Hsp27 due to media-exchange or oxygen-glucose deprivation. Surprisingly, insulin increased Hsp27 phosphorylation during reoxygenation. Insulin also reduced total p38 MAPK levels. Insulin before oxygen-glucose deprivation prevented both localization of Hsp27 to the nucleus and localization of phospho-p38 MAPK to the cytoplasm. Insulin during oxygen-glucose deprivation caused the localization of phospho-p38 MAPK in the cytoplasm, but did not increase Hsp27 phosphorylation until reoxygenation. In conclusion, insulin may protect before oxygen-glucose deprivation by redirecting phospho-p38 MAPK to the nucleus away from damaging pathways in the cytoplasm and protects during oxygen-glucose deprivation by priming phospho-p38 MAPK to phosphorylate Hsp27. / Insulin was used on a model on H9c2 myotubes to determine the effect of oxygen-glucose deprivation and reoxygenation on the localization and phosphorylation of Hsp27 and p38 MAPK

Autophagy is a physiological process by which the cell eliminates damaged organelles, toxic agents, and long-lived proteins by degradation through lysosomal system. Mitophagy, the specific autophagic elimination of mitochondria, regulates mitochondrial number to match metabolic demand and is a core machinery of quality control to remove damaged mitochondria. A neuroprotective role of physiological autophagy/mitophagy has been discovered. However, recent studies suggested that highly accelerated autophagy/mitophagy might contribute to neuronal death in various pathological situations including cerebral ischemia. In this study, we aimed to investigate the activation of excessive autophagy, particularly, the more specific mitophagy, in neuronal tissues and its contribution to ischemia/hypoxia (I/H)-induced delayed neuronal death. I/H injury was induced by oxygen and glucose deprivation (OGD) followed by reperfusion (RP) on primary cortical neurons in vitro. Cerebral ischemia was induced by unilateral common carotid artery occlusion and hypoxia in neonatal mice in vivo. In order to determine the extent to which autophagy contributes to neuronal death in cerebral ischemia, we performed multiple methods and found that in both primary cortical neurons and SH-SY5Y cells exposed to OGD for 6 h and RP for 24, 48, and 72 h, respectively, an increase of autophagy was observed as determined by the increased ratio of LC3-II to LC3-I and Beclin 1 expression. Using Fluoro-Jade C and monodansylcadaverine double-staining, and electron microscopy we found the increment in autophagy after OGD/RP was accompanied by increased autophagic cell death, and this increased cell death was inhibited by the specific autophagy inhibitor, 3-methyladenine. The presence of large autolysosomes and numerous autophagosomes in cortical neurons were confirmed by electron microscopy. Autophagy activities were increased dramatically in the ischemic brains 3-7 days postinjury from a rat model of neonatal cerebral I/H as shown by increased punctate LC3 staining and Beclin-1 expression. We thus obtained the conclusion that excessive activation of autophagy contributes to neuronal death in cerebral ischemia. BNIP3 (Bcl-2/adenovirus E19 kD interacting protein 3), a member of a unique subfamily of death-inducing mitochondrial proteins, is highly associated with mitochondrial dysfunction and delayed neuronal death in stroke. It is known that BNIP3-induced neuronal death is caspase-independent and characterized by early mitochondrial damage. Recent evidence suggested that the BNIP3 family of proteins might be important regulators of mitophagy. Here, using both stroke models, we found that homodimer (60 kD) of BNIP3/NIX (BNIP3L) were highly expressed in a ‘delayed’ manner. Particularly, significant mitophagic activation was confirmed by electron microscopy. In contrast, both neonatal mitophagy and apoptosis were significantly inhibited in the BNIP3 knockout (KO) mice after I/H, which was also accompanied by a significantly increased autophagic response. In addition, the infarct volume in the BNIP3 KO mice was significantly reduced as compared to wild-type (WT) mice after 7 or 28 days recovery, showing a prominent neuroprotection of BNIP3 gene silencing. A protein-to-protein interaction of mitochondria-localized BNIP3 (60 kD) with the autophagosome marker, LC3, was confirmed by co-ip, immunocytochemistry and further quantified by ELISA, indicating BNIP3 was an effective LC3-binding target on damaged mitochondria. These data demonstrated a novel role of BNIP3 in regulating neuronal mitophagy and cell death during ischemic stroke.

Humanin (HN) is an anti-apoptotic peptide that suppresses neuronal cell death induced by Alzheimer's disease, prion protein fragments, and serum deprivation. Recently, we demonstrated that Gly14-HN (HNG), a variant of HN in which the 14th amino acid serine is replaced with glycine, can decrease apoptotic neuronal death and reduce infarct volume in a focal cerebral ischemia/reperfusion mouse model. In this study, we postulate that the mechanism of HNG's neuroprotective effect is mediated by the PI3K/Akt pathway. Oxygen-glucose deprivation (OGD) was performed in cultured mouse primary cortical neurons for 60 min. The effect of HNG and PI3K/Akt inhibitors on OGD-induced cell death was examined at 24 h after reperfusion. HNG increased cell viability after OGD in primary cortical neurons, whereas the PI3K/Akt inhibitors wortmannin and Akti-1/2 attenuated the protective effect of HNG. HNG rapidly increased Akt phosphorylation, an effect that was inhibited by wortmannin and Akti-1/2. Mouse brains were injected intraventricularly with HNG before being subjected to middle cerebral artery occlusion (MCAO). HNG treatment significantly elevated p-Akt levels after cerebral I/R injury and decreased infarct volume. The protective effect of HNG on infarct size was attenuated by wortmannin and Akti-1/2. Taken as a whole, these results suggest that PI3K/Akt activation mediates HNG's protective effect against hypoxia/ischemia reperfusion injury.

No / Objective: The aim of this study was to determine the number of OGDs (oesophago-gastro-duodenoscopies) trainees need to perform to acquire competency in terms of successful unassisted completion to the second part of the duodenum 95% of the time. Design: OGD data were retrieved from the trainee e-portfolio developed by the Joint Advisory Group on GI Endoscopy ( JAG) in the UK. All trainees were included unless they were known to have a baseline experience of >20 procedures or had submitted data for <20 procedures. The primary outcome measure was OGD completion, defined as passage of the endoscope to the second part of the duodenum without physical assistance. The number of OGDs required to achieve a 95% completion rate was calculated by the moving average method and learning curve cumulative summation (LC-Cusum) analysis. To determine which factors were independently associated with OGD completion, a mixed effects logistic regression model was constructed with OGD completion as the outcome variable. Results: Data were analysed for 1255 trainees over 288 centres, representing 243 555 OGDs. By moving average method, trainees attained a 95% completion rate at 187 procedures. By LC-Cusum analysis, after 200 procedures, >90% trainees had attained a 95% completion rate. Total number of OGDs performed, trainee age and experience in lower GI endoscopy were factors independently associated with OGD completion. Conclusions: There are limited published data on the OGD learning curve. This is the largest study to date analysing the learning curve for competency acquisition. The JAG competency requirement for 200 procedures appears appropriate

Denna studie undersöker hur myndigheter kan öppna upp data som innehåller personuppgifter genom att geografiskt aggregera datamaterial för att bevara personlig integritet. Personlig integritet definieras i denna kontext i enlighet med dataskyddsförordningen som skyddet av personuppgifter. De aggregeringar som utreddes är de arealbaserade svenska områdena SAMS, DESO och Kommuner. Även Bespoke-neighborhood-metoden k-närmaste granne utvärderades i ett spann som gick från 5 till 1280 individer. En metod skapades för att kvantifiera hur identifierbara individer är utifrån sannolikheten att uppskatta att en personuppgift tillhör en individ. Användbarheten av datamaterialet operationaliserades som hur nära rådata det var i geografisk detaljnivå. Resultatet blev att metoden ”aggregeringar av k-närmaste granne” är den mest anpassningsbara och användbara metoden för att bevara personlig integritet och användbarhet när data aggregeras.

O grande número de websites heterogêneos e sem padronização para divulgar informações sobre transparência pública é uma evidência do despreparo das instituições públicas frente ao movimento denominado Open Government Data ou Dados Abertos Governamentais. Este cenário é encontrado no mundo todo, porém com evidência nas instituições onde os princípios de dados abertos ainda são novidade, sobretudo os governos locais. Esta pesquisa objetiva definir uma arquitetura de referência para estruturação de dados a partir dos repositórios distribuídos caracterizados pelos websites de transparência. Os requisitos de qualidade da arquitetura foram identificados a partir dos problemas de disponibilização de dados em uma amostra de 561 websites mantidos por municípios brasileiros. Uma vez definida, a arquitetura passou por um processo de validação para verificar sua viabilidade e seu potencial de utilidade pela percepção dos usuários externos ao seu desenvolvimento e por meio da implementação em um protótipo de software. Com a arquitetura, e seu aspecto colaborativo envolvido, será possível implementar sistemas de software que disponibilizam ferramentas aos usuários para facilmente abrir os dados de qualquer instituição que utilize a internet para disseminar informações de transparência. / The large number of heterogeneous and non-standardized websites to disseminate information about public transparency is an evidence of the lack of preparation of public institutions in relation to the Open Government Data movement. This scenario is found all over the world, but with more frequency in institutions where open data principles are still new, especially local governments. This research aims to define a reference architecture for structuring data from distributed repositories characterized by transparency websites. The quality requirements of the architecture were identified from the problems of data availability in a sample of 561 websites maintained by Brazilian municipalities. Once defined, the architecture underwent a validation process to verify its viability and its potential of utility by the perception of external users to its development and by the implementation of a software prototype. With the architecture, and its collaborative aspect involved, it will be possible to implement software systems that provide users with tools to easily open data from any institution that uses the internet to disseminate transparency information. Keywords: Transparency. Open Government Data. OGD. Software architecture.

O grande número de websites heterogêneos e sem padronização para divulgar informações sobre transparência pública é uma evidência do despreparo das instituições públicas frente ao movimento denominado Open Government Data ou Dados Abertos Governamentais. Este cenário é encontrado no mundo todo, porém com evidência nas instituições onde os princípios de dados abertos ainda são novidade, sobretudo os governos locais. Esta pesquisa objetiva definir uma arquitetura de referência para estruturação de dados a partir dos repositórios distribuídos caracterizados pelos websites de transparência. Os requisitos de qualidade da arquitetura foram identificados a partir dos problemas de disponibilização de dados em uma amostra de 561 websites mantidos por municípios brasileiros. Uma vez definida, a arquitetura passou por um processo de validação para verificar sua viabilidade e seu potencial de utilidade pela percepção dos usuários externos ao seu desenvolvimento e por meio da implementação em um protótipo de software. Com a arquitetura, e seu aspecto colaborativo envolvido, será possível implementar sistemas de software que disponibilizam ferramentas aos usuários para facilmente abrir os dados de qualquer instituição que utilize a internet para disseminar informações de transparência. / The large number of heterogeneous and non-standardized websites to disseminate information about public transparency is an evidence of the lack of preparation of public institutions in relation to the Open Government Data movement. This scenario is found all over the world, but with more frequency in institutions where open data principles are still new, especially local governments. This research aims to define a reference architecture for structuring data from distributed repositories characterized by transparency websites. The quality requirements of the architecture were identified from the problems of data availability in a sample of 561 websites maintained by Brazilian municipalities. Once defined, the architecture underwent a validation process to verify its viability and its potential of utility by the perception of external users to its development and by the implementation of a software prototype. With the architecture, and its collaborative aspect involved, it will be possible to implement software systems that provide users with tools to easily open data from any institution that uses the internet to disseminate transparency information. Keywords: Transparency. Open Government Data. OGD. Software architecture.

No description available.

Öppna data är ett område som organisationer inom offentlig sektor förväntas arbeta med. Trotsdetta är det fortfarande ett område som visat sig vara problematiskt. En del av problematikenhärstammar i att det är svårt att veta vilket värde öppna data har, vilket är en faktor som ledertill att motivationen för kommunala verksamheter att prioritera öppna data är lågt. I den härstudien undersöker vi vilka utmaningar och framgångsfaktorer som existerar samt vilket värdesom förväntas komma fram av arbetet inom öppna data från ett kommunalt perspektiv.Resultatet av studien kommer från en flerfallstudie där fyra kommuner av olika storlek och tvårepresentanter från Sveriges Kommuner och Landsting (SKL) deltog. Genomsemistrukturerade intervjuer som tillvägagångssätt för att samla in empiriska data har mångautmaningar framkommit, främst relaterade till organisatoriska aspekter. Tekniska utmaningarhar enligt tidigare forskning varit utmanande, men upplevs inte som lika utmanande i dennastudie. Samtidigt framgår det att det är svårt att hitta konkreta framgångsfaktorer dåkommunala initiativ inom öppna data fortfarande inte kommit så långt. Det har också visat sigvara svårt att generera värde främst på grund av att det inte finns tillräckligt mångadatamängder publicerade. De data som dock finns publicerade från de olika kommunerna ärofta osammanhängande och svåra att förädla inom privat sektor. Medborgarna som i slutändanska kunna använda tjänster och produkter som baseras på öppna data har därför endast kunnatse begränsade effekter från öppna data. Avsaknaden av ett nationellt samarbete försvårararbetet med öppna data både för offentlig sektor att publicera data och för privat sektor attförädla data. Detta leder i sin tur till att inget värde kan skapas för medborgare. På grund avdetta har det inte uppvisats något konkret värdeskapande utan värdet av öppna data ärfortfarande en förväntning och inte en verklighet i termer av realiserat värde. / Open government data (OGD) is an area that organizations in the public sector expects to workwith. Despite this, OGD is an area that has been proven to be problematic. One part of theproblem can be derived from that it is currently unknown what value OGD has, which is afactor that gives OGD low priority in the public organizations. In this study, we examine thechallenges and success factors that exists for OGD initiatives in municipalities, and at the sametime we study the expected value of these initiatives. The result from the study comes from amultiple case study conducted with four municipalities and two representatives from SwedishAssociation of Local Authorities and Regions. With semi-structured interviews as an approachto gather empirical data, multiple challenges have emerged, mostly related to organizationalaspects. Technical challenges have according to earlier research been challenging, but in thisstudy, this was only perceived as a minor challenge. At the same it is difficult to find concretesuccess factors since OGD initiatives from municipalities still have much work ahead of them.We can also see clearly that it has been difficult to generate any sort of value, mostly becauseof the insufficient number of datasets published. The data that is published however, is mostlyincoherent between the different municipalities and difficult to refine from the private sector.The citizens, who ultimately is going to use services and products based on OGD have thereforeonly seen limited effects from OGD. The lack of national collaboration makes the work withOGD problematic both concerning publication of datasets from the public sector and therefinement of data from the private sector. This leads to a situation where no value can becreated for the citizens. Because of this, no concrete value creation has been shown and thevalue of OGD is still an expectation and not a reality in terms of realized value.

Due to the high metabolic demand and low capacity for energy storage of the brain, neurons are vitally reliant on a constant oxygen supply. Under chronic mild hypoxic conditions (10% oxygen), angiogenesis is induced in the brain in an attempt to restore tissue oxygen tension to normal levels. In brain hypoxia, vascular endothelial growth factor (VEGF) plays a critical role in angiogenesis; however, the role of its homolog placental growth factor (PlGF) is unknown. Using PlGF knockout (PlGF-/-) mice exposed to whole body hypoxia (10% oxygen) for 7, 14 and 21-days, we show that PlGF-/- animals exhibit a delay in the angiogenic response of the brain to hypoxia. PlGF-/- microvessels had a significant increase in fibrinogen accumulation and extravasation, which correlated with disruption of the tight-junction protein claudin-5. These vessels displayed large lumens, were surrounded by reactive astrocytes, lacked mural cell coverage and endothelial VEGF expression, and regressed after 21 days of hypoxia. The lack of PlGF, in combination with reduced VEGF expression levels observed in the brain of PlGF-/- animals during the first 5 days of hypoxia, is likely the cause of the delayed angiogenic response and the prothrombotic phenotype of these mice. In vitro studies conducted to analyze mechanisms involved in the impaired angiogenic phenotype and enhanced astrocytic reactivity to hypoxia of PlGF-/- animals indicated that: i) PlGF-/- mouse brain endothelial cells exhibit alterations in intracellular signaling pathways associated with sprouting (ERK1/2) and vessel branching morphogenesis (GSK-3β) and ii) PlGF-/- astrocytes overexpress VEGF receptor-2 (VEGFR-2) which through activation of the ERK1/2 signaling pathway leads to a more proliferative astrocytic phenotype. These astrocytes were more resistant to oxygen and glucose deprivation (OGD) than PlGF+/+ astrocytes, a characteristic that was shown to be independent of the classical antiapoptotic VEGFR-2-dependent PI3K/Akt pathway. The findings presented in this thesis demonstrated a critical role of PlGF in vascular remodeling in the hypoxic brain.