No description available.

As células iPS surgiram com a promessa de contornar as limitações das células-tronco embrionárias, como questões éticas, segurança, compatibilidade e disponibilidade. Essas células podem ser obtidas a partir de células somáticas de indivíduos normais ou de pacientes com doenças genéticas, fazendo destas uma importante ferramenta para o screening de drogas, modelos de doenças e testes toxicológicos. Grandes avanços ocorreram na reprogramação de células diferenciadas pela expressão forçada de fatores de transcrição (FT), principalmente, através de vetores lentivirais (VL), que proporcionam uma reprogramação eficiente. Entretanto, a inserção lentiviral no genoma humano e sua influência na reprogramação é pouco conhecida. Neste trabalho, avaliamos o perfil de inserção dos VL utilizados na geração de iPS. As iPS foram geradas e caracterizadas por nosso grupo a partir de fibroblastos humanos transduzidos com VL contendo 3 FT [SOX2, TCL-1A e C-MYC (célula TSM)], e de células mesenquimais derivadas de tecido adiposo com um vetor lentiviral policistrônico contendo 4 FT [OCT4, SOX2, KLF4 e C-MYC (iPS 4FT)]. Cinco colônias isoladas de cada iPS foram mapeadas e analisadas quanto aos sítios de inserção pela técnica de LM-PCR. O DNA genômico digerido foi amplificado com um primer específico para o LTR viral e outro para um linker sintético. Os produtos foram clonados, sequenciados, e analisados em bancos de dados para identificar similaridades com o genoma humano, entre outras análises. Na célula TSM, 176 sequências, obtidas com a técnica de LM-PCR, apresentaram identidade com o genoma humano, sendo que cerca de 50% ocorreram em regiões gênicas com 94% destas em introns. Já nas iPS 4FT, 251 sequências apresentaram identidade, com cerca de 45% atingindo genes, 92% destas em introns. As inserções distribuíram-se por todos os cromossomos, com preferência pelos cromossomos 16, 17 e 20 para a TSM e pelos cromossomos 11, 15 e 17 para a iPS 4FT. Analisamos a distância da inserção ao sítio de início de transcrição (TSS), e inserções próximas a ilhas CpG, que em geral correspondem a regiões regulatórias. A maior proporção de inserção ocorreu a partir de ±30Kb de distância desses sítios. Os sítios frágeis e as regiões repetitivas do genoma foram atingidas, mas com uma frequência baixa. Os resultados mostraram uma preferência de inserção lentiviral por regiões gênicas nas iPS, indicando a possível participação de proteínas como LEDGF/p75 na integração nas células estudadas. Este trabalho mostrou que o local da integração pode contribuir para a reprogramação e, apesar de possíveis efeitos negativos das integrações, estas as células iPS ainda são uma ferramenta importante para estudos in vitro. E identificar fatores que influenciem a seleção do sítio de inserção é importante para determinar regiões cromossômicas \"seguras\" para a integração, aumentando a segurança no uso clínico. / The induced pluripotent stem (iPS) cells came with the promise of circumvent some of the limitations in the use of embryonic stem cells, like ethical issues, biological safety, immune compatibility and availability. This cells can be generated from somatic cells of normal individuals or from patients with some genetic disease, making then an important tool for drug screening, construction of disease models and toxicological trials. Great advances have happened in reprogramming differentiated cells through the forced exogenous expression of transcription factors (TF), mostly by lentiviral vectors (LV), which provide an efficient reprogramming. However, the lentiviral insertion in the human genome and its influence in reprogramming is not well known. In this work, we evaluate the insertion profile of LV used to generate human iPS cells. The iPS cells were generated, by our group, from human fibroblasts transduced by LV containing 3 TF [SOX2, TCL-1A and C-MYC (TSM reprogrammed cell)], and from mesenchymal cells derived from human adipose tissue transduced by a polycistronic LV containing 4 TF [OCT4, SOX2, KLF4 and C-MYC (iPS 4TF)]. Five isolated colonies of each iPS cell were mapped and analyzed for the insertion sites through LM-PCR technique. The digested genomic DNA was amplified with a primer for the viral LTR e another for a synthetic linker. The products were cloned, sequenced and analyzed in database to identify similarities with the human genome, among other analyzes. In TSM cell, 176 sequences, derived from the LM-PCR technique, presented identity with the human genome, and about 50% of those occurred in genic regions with 94% in introns. In iPS 4TF, 251 sequences showed identity, with about 45% reaching genes, 92% of these in introns. The insertions were distributed on all chromosomes, with preference for the 16, 17 and 20 for the TSM cell, and for the 11, 15 and 17 for the iPS 4TF. We analyzed the distance of the insertion from de transcription start site, and insertions near CpG islands, which, overall, correspond to regulatory regions. The highest proportion of insertion occurred starting ±30Kb distance from these sites. The fragile sites and the repetitive regions of the genome were also reached, but with low frequency. The results showed a preference of lentiviral insertion for genic regions in iPS, indicating the potential participation of proteins like LEDGF/p75 in integration in the cells of this work. This work shows that the integration site may contribute to the reprogramming, and, despite possible negative effects of integration, these iPS cells are still an important tool for in vitro studies. Identify factors that influence the selection of insertion site is important for determination of \"safe\" chromosomal regions for the integration, increasing the safe in clinical use.

This master thesis deals with automation process of a data mining in the LISp-Miner program. The aim of this thesis is to create an automated process that analyzes analytical questions in the data about traffic accidents in the Czech Republic using a LMCL scripting language and LM Exec module. Theoretical part of thesis describes the process of knowledge discovery in databases and most widely used methodology. It also describes the relevant topics for the work with LISp-Miner. The practical part is focused on description of traffic accidents in the Czech Republic, a description of the used data, creation and evaluation of analytical questions and especially a description of created scripts. The output of the thesis is a group of scripts and manual how to use them again, so they can be reused for analysis of actual data on traffic accidents not only in the Czech Republic, if they have the same data structure.

In Hicks’s IS-LM model, where it is assumed that production is determined in the goods marketand the interest rate is determined in the money market, when the marginal propensity to spend is greater than one, the IS has a positive slope. Modigliani (1944), Varian (1977) and Sargent (1987) determined that in this special case the IS-LM model is stable when the LM slope isgreater than the IS.In line with Samuelson (1941), this article shows that in this case the model is stable when the IS slope is greater than the LM slope. However, in this stable case the model does not have a useful economic meaning.One solution to this theoretical problem is to abandon the Keynesian adjustment mechanism and replace it with the Classical mechanism where the interest rate is determined in the goods market and production is determined in the money market. In this case, the IS-LM model is stable when the LM is steeper than the IS. / En el modelo IS-LM de Hicks, en el que se asume que la producción se determina en el mercado de bienes y la tasa de interés en el mercado de dinero, cuando la propensión marginal a gastar es mayor que uno, la IS tiene pendiente positiva. Modigliani (1944), Varian (1977) y Sargent (1987), determinaron que en este caso especial el modelo IS-LM es estable cuando la pendiente de la LM es mayor que la de la IS.En línea con Samuelson (1941), este artículo muestra que en este caso especial el modelo es estable cuando la pendiente de la IS es mayor que la de la LM. Sin embargo, en este caso estable, el modelo no tiene un significado económico útil.Una solución a este problema teórico es abandonar el mecanismo de ajuste keynesiano y reemplazarlo por el mecanismo clásico, donde la tasa de interés se determina en el mercado de bienes y la producción en el mercado de dinero. En este caso el modelo IS-LM es estable cuando la LM es más empinada que la IS.

As células iPS surgiram com a promessa de contornar as limitações das células-tronco embrionárias, como questões éticas, segurança, compatibilidade e disponibilidade. Essas células podem ser obtidas a partir de células somáticas de indivíduos normais ou de pacientes com doenças genéticas, fazendo destas uma importante ferramenta para o screening de drogas, modelos de doenças e testes toxicológicos. Grandes avanços ocorreram na reprogramação de células diferenciadas pela expressão forçada de fatores de transcrição (FT), principalmente, através de vetores lentivirais (VL), que proporcionam uma reprogramação eficiente. Entretanto, a inserção lentiviral no genoma humano e sua influência na reprogramação é pouco conhecida. Neste trabalho, avaliamos o perfil de inserção dos VL utilizados na geração de iPS. As iPS foram geradas e caracterizadas por nosso grupo a partir de fibroblastos humanos transduzidos com VL contendo 3 FT [SOX2, TCL-1A e C-MYC (célula TSM)], e de células mesenquimais derivadas de tecido adiposo com um vetor lentiviral policistrônico contendo 4 FT [OCT4, SOX2, KLF4 e C-MYC (iPS 4FT)]. Cinco colônias isoladas de cada iPS foram mapeadas e analisadas quanto aos sítios de inserção pela técnica de LM-PCR. O DNA genômico digerido foi amplificado com um primer específico para o LTR viral e outro para um linker sintético. Os produtos foram clonados, sequenciados, e analisados em bancos de dados para identificar similaridades com o genoma humano, entre outras análises. Na célula TSM, 176 sequências, obtidas com a técnica de LM-PCR, apresentaram identidade com o genoma humano, sendo que cerca de 50% ocorreram em regiões gênicas com 94% destas em introns. Já nas iPS 4FT, 251 sequências apresentaram identidade, com cerca de 45% atingindo genes, 92% destas em introns. As inserções distribuíram-se por todos os cromossomos, com preferência pelos cromossomos 16, 17 e 20 para a TSM e pelos cromossomos 11, 15 e 17 para a iPS 4FT. Analisamos a distância da inserção ao sítio de início de transcrição (TSS), e inserções próximas a ilhas CpG, que em geral correspondem a regiões regulatórias. A maior proporção de inserção ocorreu a partir de ±30Kb de distância desses sítios. Os sítios frágeis e as regiões repetitivas do genoma foram atingidas, mas com uma frequência baixa. Os resultados mostraram uma preferência de inserção lentiviral por regiões gênicas nas iPS, indicando a possível participação de proteínas como LEDGF/p75 na integração nas células estudadas. Este trabalho mostrou que o local da integração pode contribuir para a reprogramação e, apesar de possíveis efeitos negativos das integrações, estas as células iPS ainda são uma ferramenta importante para estudos in vitro. E identificar fatores que influenciem a seleção do sítio de inserção é importante para determinar regiões cromossômicas \"seguras\" para a integração, aumentando a segurança no uso clínico. / The induced pluripotent stem (iPS) cells came with the promise of circumvent some of the limitations in the use of embryonic stem cells, like ethical issues, biological safety, immune compatibility and availability. This cells can be generated from somatic cells of normal individuals or from patients with some genetic disease, making then an important tool for drug screening, construction of disease models and toxicological trials. Great advances have happened in reprogramming differentiated cells through the forced exogenous expression of transcription factors (TF), mostly by lentiviral vectors (LV), which provide an efficient reprogramming. However, the lentiviral insertion in the human genome and its influence in reprogramming is not well known. In this work, we evaluate the insertion profile of LV used to generate human iPS cells. The iPS cells were generated, by our group, from human fibroblasts transduced by LV containing 3 TF [SOX2, TCL-1A and C-MYC (TSM reprogrammed cell)], and from mesenchymal cells derived from human adipose tissue transduced by a polycistronic LV containing 4 TF [OCT4, SOX2, KLF4 and C-MYC (iPS 4TF)]. Five isolated colonies of each iPS cell were mapped and analyzed for the insertion sites through LM-PCR technique. The digested genomic DNA was amplified with a primer for the viral LTR e another for a synthetic linker. The products were cloned, sequenced and analyzed in database to identify similarities with the human genome, among other analyzes. In TSM cell, 176 sequences, derived from the LM-PCR technique, presented identity with the human genome, and about 50% of those occurred in genic regions with 94% in introns. In iPS 4TF, 251 sequences showed identity, with about 45% reaching genes, 92% of these in introns. The insertions were distributed on all chromosomes, with preference for the 16, 17 and 20 for the TSM cell, and for the 11, 15 and 17 for the iPS 4TF. We analyzed the distance of the insertion from de transcription start site, and insertions near CpG islands, which, overall, correspond to regulatory regions. The highest proportion of insertion occurred starting ±30Kb distance from these sites. The fragile sites and the repetitive regions of the genome were also reached, but with low frequency. The results showed a preference of lentiviral insertion for genic regions in iPS, indicating the potential participation of proteins like LEDGF/p75 in integration in the cells of this work. This work shows that the integration site may contribute to the reprogramming, and, despite possible negative effects of integration, these iPS cells are still an important tool for in vitro studies. Identify factors that influence the selection of insertion site is important for determination of \"safe\" chromosomal regions for the integration, increasing the safe in clinical use.

Im Hippokampus gibt es verschiedene Netzwerkoszillationen mit unterschiedlichen Frequenzen. Ein Typ dieser Oszillationen sind die ”Ripple” mit einer Frequenz von etwa 200 Hz, welche in Komplexen mit einer Aktivitätswelle, der ”Sharp wave” auftreten. Sharp wave-ripple Komplexe (SWR) werden mit der Konsolidierung von Gedächtnis in Zusammenhang gebracht. Das Netzwerk, das den SWR unterliegt, hat bestimmte Mechanismen, von denen einige in der vorliegenden Arbeit näher untersucht werden. Im ersten Teil wird untersucht, wie ein hemmendes Interneuron in der hippokampalen Region CA1, das ”oriens-lacunosum moleculare” (O-LM) Interneuron, während der SWR in das Netzwerk eingebunden ist. Wir konnten zeigen, dass O-LM Zellen während der SWR starke synaptische Exzitation erhalten. Die Exzitation tritt spät während des Ripples im lokalen Feldpotential (LFP) auf und zeigt eine Phasenankopplung an die Ripple. In etwa der Hälfte der O-LM Zellen konnten wir Aktionspotentiale während der SWR zeigen, die an die Ripple-Phase im LFP gebunden sind und nach dem Ripple-Maximum auftreten. Der zweite Teil der Arbeit bezieht sich auf die hippokampale Region CA1 und vergleicht während SWR den synaptischen Eingang in zwei Untertypen von Pyramidenzellen, die tiefen und die oberflächlichen Pyramidenzellen. Beide Untertypen bekommen synaptische Eingänge während der SWR. Diese Eingänge sind eine Mischung aus exzitatorischen und inhibitorischen Eingängen, die in den Untertypen in ihrer Stärke vergleichbar sind. Im dritten Teil untersuchen wir die SWR in der Region CA2 des Hippokampus und zeigen, dass Pyramidenzellen in CA2 in das Netzwerk während SWR eingebunden sind. Wir können sowohl exzitatorische als auch inhibitorische synaptische Eingänge in den Pyramidenzellen darstellen und konnten eine Phasenkopplung der synaptischen Eingänge an die SWR im LFP zeigen. Aufgrund der Phasenverschiebung bei verschiedenen Haltepotentialen vermuten wir einen Oszillator für die Exzitation und einen für die Hemmung. / In the hippocampus there are different patterns of activity also known as network oscillations. These oscillations express different frequencies, and one oscillation is the ripple oscillation at around 200 Hz. It is associated with an activity wave called sharp wave and form a so-called sharp wave-ripple complex (SWR). SWRs are implicated in memory consolidation. In this thesis we investigate mechanisms underlying sharp wave-ripple complexes. In the first part of this thesis I examine one type of inhibitory neurons in the region CA1 of the hippocampus during SWR. Oriens-lacunosum moleculare (O-LM) interneurons receive strong excitatory synaptic input during ripples. This input arrives after the ripple maximum and is phase locked with the ripple cycles. Around half of the probed O-LM cells fire during the SWR and thereby show an active participation during SWR. The magnitude of excitation in O-LM cells and the ratio between excitation and inhibition determine if an O-LM cell is active during the SWR. Action potentials in these cells occur late during the SWR and are phase locked. In the second part the synaptic input onto excitatory pyramidal cells were investigated during ripple oscillations. Previous work has identified two different types of pyramidal cells in area CA1. We recorded from deep and superficial pyramidal cells. For both types of pyramidal cells the inhibitory and excitatory synaptic inputs temporally associated with ripples express comparable strength. In the last and third part, I recorded SWR in the CA2 region of the hippocampus and showed incidence, frequency and amplitude of ripples and SWR. Pyramidal cells in the CA2 region are integrated into the network during SWR. They receive SWR associated synaptic input during SWR. The excitatory and inhibitory synaptic inputs in CA2 pyramidal cells were investigated in detail. Phase analysis show phase locking of local field potential ripples and synaptic inputs to the ascending phase of the ripple cycle.

No description available.

Présentation détaillée de la place de ce travail dans le cadre du projet PANDORE, de la définition du langage LM pour la description de taches de manipulation et d'assemblage, d'une analyse fonctionnelle de ce langage, du matériel mis en œuvre, de l'implantation actuelle de LM sur ce matériel et de l'expérimentation faite du matériel à travers le logiciel implanté.

IS-LM methodology was not developed by Hicks alone. Hicks, together with Harrod and Meade jointly contributed to the idea of general equilibrium analysis of products and money markets which were separated treated by Keynes in his General Theory. In order to honor the contribution of the other two economists, we suggested that Hicksian IS-LM framework be renamed Harrod-Hicks-Meade IS-LM model. In IS-LM graphical analysis, the slopes of IS-LM curves and the effectiveness of fiscal and monetary policy are closed related. This thesis has surveyed domestic¡]in Chinese¡^and international¡]in English¡^economics textbooks thoroughly on this matter and discovered that the mistakes are often made or not even mentioned at all. The slope of LM¡]IS¡^curve is determined by interest rate factor , Li¡]Ii¡^and income factor, Ly¡]Cy¡^. If the interest rate factor causes the slopes of two LM¡]IS¡^curves to differ, the expansionary monetary¡]fiscal¡^policy will make LM¡]IS¡^curves horizontal shift to the right. As a result, monetary¡]fiscal¡^policy will be more effective if the LM¡]IS¡^curve is steeper. On the contrary, if the income factor is the cause of the different slopes, the same policy will make LM¡]IS¡^curves shift right vertically. Monetary¡]fiscal¡^policy will be more effective if the LM¡]IS¡^curve is flatter . Once Pigou Effect is present, the aggregate demand curve will become flatter. As a result, monetary¡]fiscal¡^ policy is more¡]less¡^effective than without. Hence, monetary policy is relatively more effective than fiscal policy. In general, a policy-induced shift of aggregate demand curve can¡¦t adopt the horizontal moving method, specifically, for monetary policy, we have to adopt the vertical moving method. As for fiscal policy, the vertical moving method should be adopted , if and only if the income factor causes the different slopes of aggregate demand curves. Neither horizontal nor vertical shift in aggregate demand can be taken for the case when interest rate is the reason leading to the different slopes of aggregate demand curves.

No description available.