<p dir="ltr">Membrane-dependent enzymes play crucial roles in cellular signaling by transducing extracellular signals into intracellular responses. Phospholipase Cepsilon (PLCe) and diacylglycerol kinase alpha (DGKa) are membrane-associated enzymes regulated by G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs), controlling signaling pathways essential for numerous cellular processes. PLCe catalyzes the hydrolysis of phosphatidylinositol phosphates into inositol phosphates (IPX) and diacylglycerol (DAG), triggering calcium release from intracellular stores and activating protein kinase C (PKC)-dependent pathways. While PLCe is crucial for normal cardiovascular function, hyperactivation or sustained activation can lead to hypertrophy. Due to structural heterogeneity, previous studies focused on isolated regulatory domains or the catalytic core. In this work, I present the first cryo-EM reconstruction of the largest PLCe fragment to date in complex with an antigen-binding fragment (Fab). This structure reveals the domain architecture of the N-terminal regions of the lipase and defines an extended membrane-binding surface critical for maximal basal and G protein-dependent activity. These findings lay the groundwork for high-resolution structures of the full-length enzyme and its complexes with the small GTPase Rap1A. Additionally, I explored the role of mAKAP in the Rap1A–PLCe pathway, alongside the guanine nucleotide exchange factor (GEF) function of PLCe toward Rap1A. In parallel, cryo-EM studies of DGKa bound to a covalent inhibitor were initiated. DGKa reduces DAG, thereby limiting PKC activity, and its inhibition is emerging as a promising cancer immunotherapy target. We have established a protocol for structural studies of full-length DGKa, which will elucidate its structures in basal and inhibited states.</p>

Le diabète de type 2 (DT2) est une maladie métabolique complexe causée par des facteurs génétiques mais aussi environnementaux, tels la sédentarité et le surpoids. La dysfonction de la cellule β pancréatique est maintenant reconnue comme l’élément déterminant dans le développement du DT2. Notre laboratoire s’intéresse à la sécrétion d’insuline par la cellule β en réponse aux nutriments calorigéniques et aux mécanismes qui la contrôle. Alors que la connaissance des mécanismes responsables de l’induction de la sécrétion d’insuline en réponse aux glucose et acides gras est assez avancée, les procédés d’inhibition de la sécrétion dans des contextes normaux ou pathologiques sont moins bien compris. L’objectif de la présente thèse était d’identifier quelques-uns de ces mécanismes de régulation négative de la sécrétion d’insuline dans la cellule β pancréatique, et ce en situation normale ou pathologique en lien avec le DT2. La première hypothèse testée était que l’enzyme mitochondriale hydroxyacyl-CoA déshydrogénase spécifique pour les molécules à chaîne courte (short-chain hydroxyacyl-CoA dehydrogenase, SCHAD) régule la sécrétion d’insuline induite par le glucose (SIIG) par la modulation des concentrations d’acides gras ou leur dérivés tels les acyl-CoA ou acyl-carnitine dans la cellule β. Pour ce faire, nous avons utilisé la technologie des ARN interférants (ARNi) afin de diminuer l’expression de SCHAD dans la lignée cellulaire β pancréatique INS832/13. Nous avons par la suite vérifié chez la souris DIO (diet-induced obesity) si une exposition prolongée à une diète riche en gras activerait certaines voies métaboliques et signalétiques assurant une régulation négative de la sécrétion d’insuline et contribuerait au développement du DT2. Pour ce faire, nous avons mesuré la SIIG, le métabolisme intracellulaire des lipides, la fonction mitochondriale et l’activation de certaines voies signalétiques dans les îlots de Langerhans isolés des souris normales (ND, normal diet) ou nourries à la dière riche en gras (DIO) Nos résultats suggèrent que l’enzyme SCHAD est importante dans l’atténuation de la sécrétion d’insuline induite par le glucose et les acides aminés. En effet, l’oxydation des acides gras par la protéine SCHAD préviendrait l’accumulation d’acyl-CoA ou de leurs dérivés carnitine à chaîne courtes potentialisatrices de la sécrétion d’insuline. De plus, SCHAD régule le métabolisme du glutamate par l’inhibition allostérique de l’enzyme glutamate déshydrogénase (GDH), prévenant ainsi une hyperinsulinémie causée par une sur-activité de GDH. L’étude de la dysfonction de la cellule β dans le modèle de souris DIO a démontré qu’il existe une grande hétérogénéité dans l’obésité et l’hyperglycémie développées suite à la diète riche en gras. L’orginialité de notre étude réside dans la stratification des souris DIO en deux groupes : les faibles et forts répondants à la diète (low diet responders (LDR) et high diet responder (HDR)) sur la base de leur gain de poids corporel. Nous avons mis en lumières divers mécanismes liés au métabolisme des acides gras impliqués dans la diminution de la SIIG. Une diminution du flux à travers le cycle TG/FFA accompagnée d’une augmentation de l’oxydation des acides gras et d’une accumulation intracellulaire de cholestérol contribuent à la diminution de la SIIG chez les souris DIO-HDR. De plus, l’altération de la signalisation par les voies AMPK (AMP-activated protein kinase) et PKC epsilon (protéine kinase C epsilon) pourrait expliquer certaines de ces modifications du métabolisme des îlots DIO et causer le défaut de sécrétion d’insuline. En résumé, nous avons mis en lumière des mécanismes importants pour la régulation négative de la sécrétion d’insuline dans la cellule β pancréatique saine ou en situation pathologique. Ces mécanismes pourraient permettre d’une part de limiter l’amplitude ou la durée de la sécrétion d’insuline suite à un repas chez la cellule saine, et d’autre part de préserver la fonction de la cellule β en retardant l’épuisement de celle-ci en situation pathologique. Certaines de ces voies peuvent expliquer l’altération de la sécrétion d’insuline dans le cadre du DT2 lié à l’obésité. À la lumière de nos recherches, le développement de thérapies ayant pour cible les mécanismes de régulation négative de la sécrétion d’insuline pourrait être bénéfique pour le traitement de patients diabétiques. / Type 2 diabetes (T2D) is a complex metabolic disease caused by genetic as well as environmental factors, such as sedentarity and obesity. Pancreatic β cell dysfunction is now recognized as the key factor in T2D development. Our laboratory is studying the mechanisms of regulation of insulin secretion by the pancreatic β cell in response to nutrients. While the knowledge of the mechanisms responsible for initiation of insulin secretion in response to glucose and fatty acids is quite advanced, the inhibitory processes of insulin secretion in normal or pathological situations are still poorly understood. This doctoral thesis has focused on the identification of some of the mechanisms responsible for negative regulation of insulin secretion in pancreatic β cell. We have addressed this issue under normal situation or pathological conditions related to T2D. We first tested the hypothesis by which a mitochondrial enzyme, short-chain hydroxyacyl-CoA dehydrogenase (SCHAD), negatively regulates glucose-induced insulin secretion (GIIS) by limiting the concentrations of some fatty acids and their derivatives such as acyl-CoA or acyl-carnitine molecules in the β cell. For this purpose, the downregulation of SCHAD by RNA interference (RNAi) was used in the pancreatic β cell line INS832/13. Then, we tested wether a prolonged administration of high-fat diet to mice (diet-induced obesity mouse model, DIO) would modulate intracellular metabolic and molecular pathways responsible for inhibition of insulin secretion. C57BL/6 mice were therefore fed a high-fat diet for 8 weeks followed by insulin secretion, intracellular lipid metabolism, mitochondrial function and intracellular signaling measurements on isolated pancreatic islets of Langerhans of those mice. Our results suggest that SCHAD negatively regulates GIIS and amino acid-induced insulin secretion. We propose that fatty acid oxidation by SCHAD would prevent the accumulation of short-chain acyl-CoAs or acyl-carnitines capable of potentiating insulin secretion. In addition, SCHAD regulates glutamate metabolism by the allosteric inhibition of glutamate dehydrogenase (GDH) preventing the hyperinsulinemia caused by excessive GDH activity. The study of β cell dysfunction in the DIO mouse model stratified LDR and HDR highlighted various fatty acid metabolism pathways involved in the reduction of GIIS. A decrease in the triglycerides/free fatty acid (TG/FFA) cycling associated with an increase in fatty acid oxidation and intracellular accumulation of cholesterol was shown to contribute to the decreased GIIS in DIO-HDR mice. Furthermore, alteration of AMP-activated kinase (AMPK) and protein kinase C epsilon (PKC epsilon) signaling pathways would be responsible for those alterations in metabolic pathways observed in DIO islets and cause decreased insulin secretion. In summary, we have shed light on important pathways negatively regulating insulin secretion in pancreatic β cell. These pathways could either limit the amplitude or duration of insulin secretion after a meal, or help to preserve β-cell function by delaying exhaustion. Some of those signaling pathways could explain the altered insulin secretion observed in T2D obese patients. In light of our research, the development of therapies targeting pathways that negatively regulate insulin secretion may be beneficial for treating diabetic patients.

<p>This is a thesis work proposed by Sweco System in order to carry out a study related to the heating system of a circular fuel oil storage tank or cistern. The study tank is a 23m diameter and 18m height with a storage capacity of around 7500m3 of Eo5 heavy fuel oil. The content ought to be at a minimum storage temperature of 50ºC so that the fuel oil is fluid enough and operation labors can be adequately performed. In fact, these types of heavy fuel oils have fairly high viscosities at lower temperatures and the heating and pumping system can be compromised at temperatures below the pour point. For this purpose a heating system is installed to maintain the fluid warm. So far the system was operated by an oil burner but there are plans to its replacement by a District Heating-heat exchanger combo. Thereby, tank heating needs, flow and thermal patterns and heat transfer within it are principally studied.</p><p> </p><p>Tank boundaries are studied and their thermal resistances are calculated in order to dimension heat supply capacity. The study implies Finite Elements (Comsol Multiphysics) and Finite Volume (Fluent) analysis to work out some stationary heat transfer by conduction cases on some parts and thermal bridges present on these boundaries. Afterwards both cooling and heating processes of the fuel oil are studied using several strategies: basic models and Computational Fluid Dynamics (CFD). CFD work with Fluent is focused on optimizing inlet and outlet topologies. Understanding the cooling process is sought as well; Fluent CFD transient models are simulated in this way as well. Additionally the effect of filling levels is taken into account leading to a multiphase (fuel oil and air) flow cases where especially heating coupling of both phases is analyzed.</p><p> </p><p>Results show that maximum heat supply needs are around 80kW when the tank temperature is around 60ºC and 70kW when it is around 50ºC. Expectedly the main characteristic of the flow turns out to be the buoyancy driven convective pattern. K-ε turbulence viscous models are applied to both heating and cooling processes showing thermal stratification, especially at the bottom of the tank. Hotter fluid above follows very complex flow patterns. During the heating processes models used predict fairly well mixed and homogenous temperature distribution regardless small stratification at the bottom of the tank. In this way no concrete inlet-outlet configuration shows clear advantages over the rest. Due to the insulation of the tank, low thermal conductivity of the fluid and vast amount of mass present in the tank, the cooling process is slow (fluid average temperature drops around 5.7 ºC from 60ºC in 15 days when the tank is full and ambient temperature is considered to be at -20ºC) and lies somewhere in the middle between the solid rigid and perfect mixture cooling processes. However, due to stratification some parts of the fluid reach minimum admissible temperatures much faster than average temperature does. On the other hand, as expected, air phase acts as an additional thermal resistance; anyhow the cooling process is still faster for lower filling levels than the full one.</p> / <p>El presente proyecto fue propuesto por Sweco Systems para llevar a cabo un estudio relacionado con el sistema de calefacción de una cisterna o tanque de almacenamiento de fuel oil circular. Dicho tanque tiene 23 m de diámetro  y 18 m de altura con una capacidad de almacenamiento de alrededor de 7500 m³ de Eo5 fuel oil pesado. El contenido mantenerse a una temperatura mínima de 50 ºC de manera que el fuel oil es suficientemente fluido para que las labores de operación puedan ser ejecutadas adecuadamente. De hecho, estos tipos de fuel oil pesado tienen altas viscosidades a bajas temperaturas y, por tanto, tanto los sistemas de calefacción y como el de bombeo pueden verse comprometidosr a temperaturas por debajo del pour point. Con este fin un sistema de calefacción es instalado para mantener el fluido suficientemente caliente. Hasta el momento, el sistema era operado por un quemador de fuel, sin embargo, hay planes que éste sea sustituido por un combo intercambiador de calor-District Heating. Por lo tanto, principalmente son estudiadas las necesidades de calefacción así como los flujos térmicos y fluidos.</p><p>Se estudian las fronteras del tanque, y sus respectivas resistencias térmicas son calculadas con el fin de dimensionar la capacidad necesaria de suministro de calor. El estudio implica Elementos Finitos (Comsol Multiphysics) y Volúmenes Finitos (Fluent) para elaborar análisis estacionarios de transferencia de calor por conducción en algunos casos. Existen puentes térmicos en las paredes y su importancia es también anallizada. Posteriormente se estudian tanto los procesos de calentamiento y enfriamiento del fuel oil utilizando diversas estrategias: modelos básicos y Dinámica de Fluidos Computacional (CFD). El trabajo con CFD se centra en la optimización de topologías de entradas y salidas del sistema. También es solicitado entender el proceso de enfriamiento; En este sentido, se simulan modelos CFD transitorios de Fluent. Además, el efecto de los niveles de llenado se tiene en cuenta dando lugar a estudios de flujo multifase (fuel oil y aire), haciendo hincapié en el análisis de acoplamiento de transferencia de calor entre las dos fases.</p><p>Los resultados muestran que las necesidades de calefacción máximas son de alrededor de 80kW cuando la temperatura del tanque es de alrededor de 60 º C y 70kW cuando está alrededor de 50 ºC. Como era de esperar, la principal característica de este tipo de flujos es la convección natural resultante de las fuerzas de flotabilidad. Se aplican modelos turbulentos k-ε a los procesos de calentamiento y enfriamiento, mostrando estratificación térmica, sobre todo en la parte inferior de la cisterna. El líquido más caliente que se sitúa encima muestra complejos patrones de flujo. Durante los procesos de calentamiento, los modelos utilizados predicen un buen mezclado y distribución homogénea de la temperatura independientemente de esta pequeña estratificación en la parte inferior de la cisterna. De esta manera, ninguna concreta configuración de entradas-salidas simuladas muestra claras ventajas sobre el resto. Debido al aislamiento de la cisterna, la baja conductividad térmica del fluido y la gran cantidad de masa presente en el tanque el proceso de enfriamiento es lento (la temperatura media del fluido desciende 5.7 º C desde 60 º C en 15 días cuando el tanque está lleno y la temperatura ambiente es de -20 º C) y se encuentra en algún lugar en medio de los procesos de enfriamiento del sólido rígido y perfecta mezcla. Sin embargo, debido a la estratificación, algunas partes el líquido alcanzan la temperatura mínima admisible mucho más rápido que la media de temperatura. Por otra parte, como se esperaba, la fase de aire actúa como una resistencia térmica adicional, de todos modos, el proceso de enfriamiento es aún más rápido para niveles de llenado más bajos que el lleno.</p>

This is a thesis work proposed by Sweco System in order to carry out a study related to the heating system of a circular fuel oil storage tank or cistern. The study tank is a 23m diameter and 18m height with a storage capacity of around 7500m3 of Eo5 heavy fuel oil. The content ought to be at a minimum storage temperature of 50ºC so that the fuel oil is fluid enough and operation labors can be adequately performed. In fact, these types of heavy fuel oils have fairly high viscosities at lower temperatures and the heating and pumping system can be compromised at temperatures below the pour point. For this purpose a heating system is installed to maintain the fluid warm. So far the system was operated by an oil burner but there are plans to its replacement by a District Heating-heat exchanger combo. Thereby, tank heating needs, flow and thermal patterns and heat transfer within it are principally studied.   Tank boundaries are studied and their thermal resistances are calculated in order to dimension heat supply capacity. The study implies Finite Elements (Comsol Multiphysics) and Finite Volume (Fluent) analysis to work out some stationary heat transfer by conduction cases on some parts and thermal bridges present on these boundaries. Afterwards both cooling and heating processes of the fuel oil are studied using several strategies: basic models and Computational Fluid Dynamics (CFD). CFD work with Fluent is focused on optimizing inlet and outlet topologies. Understanding the cooling process is sought as well; Fluent CFD transient models are simulated in this way as well. Additionally the effect of filling levels is taken into account leading to a multiphase (fuel oil and air) flow cases where especially heating coupling of both phases is analyzed.   Results show that maximum heat supply needs are around 80kW when the tank temperature is around 60ºC and 70kW when it is around 50ºC. Expectedly the main characteristic of the flow turns out to be the buoyancy driven convective pattern. K-ε turbulence viscous models are applied to both heating and cooling processes showing thermal stratification, especially at the bottom of the tank. Hotter fluid above follows very complex flow patterns. During the heating processes models used predict fairly well mixed and homogenous temperature distribution regardless small stratification at the bottom of the tank. In this way no concrete inlet-outlet configuration shows clear advantages over the rest. Due to the insulation of the tank, low thermal conductivity of the fluid and vast amount of mass present in the tank, the cooling process is slow (fluid average temperature drops around 5.7 ºC from 60ºC in 15 days when the tank is full and ambient temperature is considered to be at -20ºC) and lies somewhere in the middle between the solid rigid and perfect mixture cooling processes. However, due to stratification some parts of the fluid reach minimum admissible temperatures much faster than average temperature does. On the other hand, as expected, air phase acts as an additional thermal resistance; anyhow the cooling process is still faster for lower filling levels than the full one. / El presente proyecto fue propuesto por Sweco Systems para llevar a cabo un estudio relacionado con el sistema de calefacción de una cisterna o tanque de almacenamiento de fuel oil circular. Dicho tanque tiene 23 m de diámetro  y 18 m de altura con una capacidad de almacenamiento de alrededor de 7500 m3 de Eo5 fuel oil pesado. El contenido mantenerse a una temperatura mínima de 50 ºC de manera que el fuel oil es suficientemente fluido para que las labores de operación puedan ser ejecutadas adecuadamente. De hecho, estos tipos de fuel oil pesado tienen altas viscosidades a bajas temperaturas y, por tanto, tanto los sistemas de calefacción y como el de bombeo pueden verse comprometidosr a temperaturas por debajo del pour point. Con este fin un sistema de calefacción es instalado para mantener el fluido suficientemente caliente. Hasta el momento, el sistema era operado por un quemador de fuel, sin embargo, hay planes que éste sea sustituido por un combo intercambiador de calor-District Heating. Por lo tanto, principalmente son estudiadas las necesidades de calefacción así como los flujos térmicos y fluidos. Se estudian las fronteras del tanque, y sus respectivas resistencias térmicas son calculadas con el fin de dimensionar la capacidad necesaria de suministro de calor. El estudio implica Elementos Finitos (Comsol Multiphysics) y Volúmenes Finitos (Fluent) para elaborar análisis estacionarios de transferencia de calor por conducción en algunos casos. Existen puentes térmicos en las paredes y su importancia es también anallizada. Posteriormente se estudian tanto los procesos de calentamiento y enfriamiento del fuel oil utilizando diversas estrategias: modelos básicos y Dinámica de Fluidos Computacional (CFD). El trabajo con CFD se centra en la optimización de topologías de entradas y salidas del sistema. También es solicitado entender el proceso de enfriamiento; En este sentido, se simulan modelos CFD transitorios de Fluent. Además, el efecto de los niveles de llenado se tiene en cuenta dando lugar a estudios de flujo multifase (fuel oil y aire), haciendo hincapié en el análisis de acoplamiento de transferencia de calor entre las dos fases. Los resultados muestran que las necesidades de calefacción máximas son de alrededor de 80kW cuando la temperatura del tanque es de alrededor de 60 º C y 70kW cuando está alrededor de 50 ºC. Como era de esperar, la principal característica de este tipo de flujos es la convección natural resultante de las fuerzas de flotabilidad. Se aplican modelos turbulentos k-ε a los procesos de calentamiento y enfriamiento, mostrando estratificación térmica, sobre todo en la parte inferior de la cisterna. El líquido más caliente que se sitúa encima muestra complejos patrones de flujo. Durante los procesos de calentamiento, los modelos utilizados predicen un buen mezclado y distribución homogénea de la temperatura independientemente de esta pequeña estratificación en la parte inferior de la cisterna. De esta manera, ninguna concreta configuración de entradas-salidas simuladas muestra claras ventajas sobre el resto. Debido al aislamiento de la cisterna, la baja conductividad térmica del fluido y la gran cantidad de masa presente en el tanque el proceso de enfriamiento es lento (la temperatura media del fluido desciende 5.7 º C desde 60 º C en 15 días cuando el tanque está lleno y la temperatura ambiente es de -20 º C) y se encuentra en algún lugar en medio de los procesos de enfriamiento del sólido rígido y perfecta mezcla. Sin embargo, debido a la estratificación, algunas partes el líquido alcanzan la temperatura mínima admisible mucho más rápido que la media de temperatura. Por otra parte, como se esperaba, la fase de aire actúa como una resistencia térmica adicional, de todos modos, el proceso de enfriamiento es aún más rápido para niveles de llenado más bajos que el lleno.

Le diabète de type 2 (DT2) est une maladie métabolique complexe causée par des facteurs génétiques mais aussi environnementaux, tels la sédentarité et le surpoids. La dysfonction de la cellule β pancréatique est maintenant reconnue comme l’élément déterminant dans le développement du DT2. Notre laboratoire s’intéresse à la sécrétion d’insuline par la cellule β en réponse aux nutriments calorigéniques et aux mécanismes qui la contrôle. Alors que la connaissance des mécanismes responsables de l’induction de la sécrétion d’insuline en réponse aux glucose et acides gras est assez avancée, les procédés d’inhibition de la sécrétion dans des contextes normaux ou pathologiques sont moins bien compris. L’objectif de la présente thèse était d’identifier quelques-uns de ces mécanismes de régulation négative de la sécrétion d’insuline dans la cellule β pancréatique, et ce en situation normale ou pathologique en lien avec le DT2. La première hypothèse testée était que l’enzyme mitochondriale hydroxyacyl-CoA déshydrogénase spécifique pour les molécules à chaîne courte (short-chain hydroxyacyl-CoA dehydrogenase, SCHAD) régule la sécrétion d’insuline induite par le glucose (SIIG) par la modulation des concentrations d’acides gras ou leur dérivés tels les acyl-CoA ou acyl-carnitine dans la cellule β. Pour ce faire, nous avons utilisé la technologie des ARN interférants (ARNi) afin de diminuer l’expression de SCHAD dans la lignée cellulaire β pancréatique INS832/13. Nous avons par la suite vérifié chez la souris DIO (diet-induced obesity) si une exposition prolongée à une diète riche en gras activerait certaines voies métaboliques et signalétiques assurant une régulation négative de la sécrétion d’insuline et contribuerait au développement du DT2. Pour ce faire, nous avons mesuré la SIIG, le métabolisme intracellulaire des lipides, la fonction mitochondriale et l’activation de certaines voies signalétiques dans les îlots de Langerhans isolés des souris normales (ND, normal diet) ou nourries à la dière riche en gras (DIO) Nos résultats suggèrent que l’enzyme SCHAD est importante dans l’atténuation de la sécrétion d’insuline induite par le glucose et les acides aminés. En effet, l’oxydation des acides gras par la protéine SCHAD préviendrait l’accumulation d’acyl-CoA ou de leurs dérivés carnitine à chaîne courtes potentialisatrices de la sécrétion d’insuline. De plus, SCHAD régule le métabolisme du glutamate par l’inhibition allostérique de l’enzyme glutamate déshydrogénase (GDH), prévenant ainsi une hyperinsulinémie causée par une sur-activité de GDH. L’étude de la dysfonction de la cellule β dans le modèle de souris DIO a démontré qu’il existe une grande hétérogénéité dans l’obésité et l’hyperglycémie développées suite à la diète riche en gras. L’orginialité de notre étude réside dans la stratification des souris DIO en deux groupes : les faibles et forts répondants à la diète (low diet responders (LDR) et high diet responder (HDR)) sur la base de leur gain de poids corporel. Nous avons mis en lumières divers mécanismes liés au métabolisme des acides gras impliqués dans la diminution de la SIIG. Une diminution du flux à travers le cycle TG/FFA accompagnée d’une augmentation de l’oxydation des acides gras et d’une accumulation intracellulaire de cholestérol contribuent à la diminution de la SIIG chez les souris DIO-HDR. De plus, l’altération de la signalisation par les voies AMPK (AMP-activated protein kinase) et PKC epsilon (protéine kinase C epsilon) pourrait expliquer certaines de ces modifications du métabolisme des îlots DIO et causer le défaut de sécrétion d’insuline. En résumé, nous avons mis en lumière des mécanismes importants pour la régulation négative de la sécrétion d’insuline dans la cellule β pancréatique saine ou en situation pathologique. Ces mécanismes pourraient permettre d’une part de limiter l’amplitude ou la durée de la sécrétion d’insuline suite à un repas chez la cellule saine, et d’autre part de préserver la fonction de la cellule β en retardant l’épuisement de celle-ci en situation pathologique. Certaines de ces voies peuvent expliquer l’altération de la sécrétion d’insuline dans le cadre du DT2 lié à l’obésité. À la lumière de nos recherches, le développement de thérapies ayant pour cible les mécanismes de régulation négative de la sécrétion d’insuline pourrait être bénéfique pour le traitement de patients diabétiques. / Type 2 diabetes (T2D) is a complex metabolic disease caused by genetic as well as environmental factors, such as sedentarity and obesity. Pancreatic β cell dysfunction is now recognized as the key factor in T2D development. Our laboratory is studying the mechanisms of regulation of insulin secretion by the pancreatic β cell in response to nutrients. While the knowledge of the mechanisms responsible for initiation of insulin secretion in response to glucose and fatty acids is quite advanced, the inhibitory processes of insulin secretion in normal or pathological situations are still poorly understood. This doctoral thesis has focused on the identification of some of the mechanisms responsible for negative regulation of insulin secretion in pancreatic β cell. We have addressed this issue under normal situation or pathological conditions related to T2D. We first tested the hypothesis by which a mitochondrial enzyme, short-chain hydroxyacyl-CoA dehydrogenase (SCHAD), negatively regulates glucose-induced insulin secretion (GIIS) by limiting the concentrations of some fatty acids and their derivatives such as acyl-CoA or acyl-carnitine molecules in the β cell. For this purpose, the downregulation of SCHAD by RNA interference (RNAi) was used in the pancreatic β cell line INS832/13. Then, we tested wether a prolonged administration of high-fat diet to mice (diet-induced obesity mouse model, DIO) would modulate intracellular metabolic and molecular pathways responsible for inhibition of insulin secretion. C57BL/6 mice were therefore fed a high-fat diet for 8 weeks followed by insulin secretion, intracellular lipid metabolism, mitochondrial function and intracellular signaling measurements on isolated pancreatic islets of Langerhans of those mice. Our results suggest that SCHAD negatively regulates GIIS and amino acid-induced insulin secretion. We propose that fatty acid oxidation by SCHAD would prevent the accumulation of short-chain acyl-CoAs or acyl-carnitines capable of potentiating insulin secretion. In addition, SCHAD regulates glutamate metabolism by the allosteric inhibition of glutamate dehydrogenase (GDH) preventing the hyperinsulinemia caused by excessive GDH activity. The study of β cell dysfunction in the DIO mouse model stratified LDR and HDR highlighted various fatty acid metabolism pathways involved in the reduction of GIIS. A decrease in the triglycerides/free fatty acid (TG/FFA) cycling associated with an increase in fatty acid oxidation and intracellular accumulation of cholesterol was shown to contribute to the decreased GIIS in DIO-HDR mice. Furthermore, alteration of AMP-activated kinase (AMPK) and protein kinase C epsilon (PKC epsilon) signaling pathways would be responsible for those alterations in metabolic pathways observed in DIO islets and cause decreased insulin secretion. In summary, we have shed light on important pathways negatively regulating insulin secretion in pancreatic β cell. These pathways could either limit the amplitude or duration of insulin secretion after a meal, or help to preserve β-cell function by delaying exhaustion. Some of those signaling pathways could explain the altered insulin secretion observed in T2D obese patients. In light of our research, the development of therapies targeting pathways that negatively regulate insulin secretion may be beneficial for treating diabetic patients.

<p dir="ltr">The generalization error of a classifier is related to the complexity of the set of functions among which the classifier is chosen. We study a family of low-complexity classifiers consisting of thresholding a random one-dimensional feature. The feature is obtained by projecting the data on a random line after embedding it into a higher-dimensional space parametrized by monomials of order up to k. More specifically, the extended data is projected n-times and the best classifier among those n, based on its performance on training data, is chosen. </p><p dir="ltr">We show that this type of classifier is extremely flexible, as it is likely to approximate, to an arbitrary precision, any continuous function on a compact set as well as any Boolean function on a compact set that splits the support into measurable subsets. In particular, given full knowledge of the class conditional densities, the error of these low-complexity classifiers would converge to the optimal (Bayes) error as k and n go to infinity. On the other hand, if only a training dataset is given, we show that the classifiers will perfectly classify all the training points as k and n go to infinity. </p><p dir="ltr">We also bound the generalization error of our random classifiers. In general, our bounds are better than those for any classifier with VC dimension greater than O(ln(n)). In particular, our bounds imply that, unless the number of projections n is extremely large, there is a significant advantageous gap between the generalization error of the random projection approach and that of a linear classifier in the extended space. Asymptotically, as the number of samples approaches infinity, the gap persists for any such n. Thus, there is a potentially large gain in generalization properties by selecting parameters at random, rather than optimization. </p><p dir="ltr">Given a classification problem and a family of classifiers, the Rashomon ratio measures the proportion of classifiers that yield less than a given loss. Previous work has explored the advantage of a large Rashomon ratio in the case of a finite family of classifiers. Here we consider the more general case of an infinite family. We show that a large Rashomon ratio guarantees that choosing the classifier with the best empirical accuracy among a random subset of the family, which is likely to improve generalizability, will not increase the empirical loss too much. </p><p dir="ltr">We quantify the Rashomon ratio in two examples involving infinite classifier families in order to illustrate situations in which it is large. In the first example, we estimate the Rashomon ratio of the classification of normally distributed classes using an affine classifier. In the second, we obtain a lower bound for the Rashomon ratio of a classification problem with a modified Gram matrix when the classifier family consists of two-layer ReLU neural networks. In general, we show that the Rashomon ratio can be estimated using a training dataset along with random samples from the classifier family and we provide guarantees that such an estimation is close to the true value of the Rashomon ratio.</p>

Spark ignited engines are still important for conventional as well as for hybrid power trains and are thus objective to optimization. Today a lot of functionalities arise from software solutions, which have to be calibrated. Modern engine technologies provide an extensive variability considering their valve train, fuel injection and load control. Thus, calibration efforts are really high and shall be reduced by introduction of virtual methods. In this work a physical 0D combustion model is set up, which can cope with a new generation of spark ignition engines. Therefore, at first cylinder thermodynamics are modeled and validated in the whole engine map with the help of a real-time capable approach. Afterwards an up to date turbulence model is introduced, which is based on a quasi-dimensional k-epsilon-approach and can cope with turbulence production from large scale shearing. A simplified model for ignition delay is implemented which emphasizes the transfer from laminar to turbulent flame propagation after ignition. The modeling is completed with the calculation of overall heat release rates in a 0D entrainment approach with the help of turbulent flame velocities. After validation of all sub-models, the 0D combustion prediction is used in combination with a 1D gas exchange analysis to virtually calibrate the modern engine torque structure and the ECU function for exhaust gas temperature with extensive simulations. / Moderne Ottomotoren spielen heute sowohl in konventionellen als auch hybriden Fahrzeugantrieben eine große Rolle. Aktuelle Konzepte sind hochvariabel bezüglich Ventilsteuerung, Kraftstoffeinspritzung und Laststeuerung und ihre Optimierungspotentiale erwachsen zumeist aus neuen Softwarefunktionen. Deren Applikation ist zeit- und kostenintensiv und soll durch virtuelle Methoden unterstützt werden. In der vorliegenden Arbeit wird ein physikalisches 0D Verbrennungsmodell für Ottomotoren aufgebaut und bis zur praktischen Anwendung geführt. Dafür wurde zuerst die Thermodynamik echtzeitfähig modelliert und im gesamten Motorenkennfeld abgeglichen. Der Aufbau eines neuen Turbulenzmodells auf Basis der quasidimensionalen k-epsilon-Gleichung ermöglicht anschließend, die veränderlichen Einflüsse globaler Ladungsbewegung auf die Turbulenz abzubilden. Für den Brennverzug wurde ein vereinfachtes Modell abgeleitet, welches den Übergang von laminarer zu turbulenter Flammenausbreitung nach der Zündung in den Vordergrund stellt. Der restliche Brennverlauf wird durch die physikalische Ermittlung der turbulenten Brenngeschwindigkeit in einem 0D Entrainment-Ansatz dargestellt. Nach Validierung aller Teilmodelle erfolgt die virtuelle Bedatung der Momentenstruktur und der Abgastemperaturfunktion für das Motorsteuergerät.

Spark ignited engines are still important for conventional as well as for hybrid power trains and are thus objective to optimization. Today a lot of functionalities arise from software solutions, which have to be calibrated. Modern engine technologies provide an extensive variability considering their valve train, fuel injection and load control. Thus, calibration efforts are really high and shall be reduced by introduction of virtual methods. In this work a physical 0D combustion model is set up, which can cope with a new generation of spark ignition engines. Therefore, at first cylinder thermodynamics are modeled and validated in the whole engine map with the help of a real-time capable approach. Afterwards an up to date turbulence model is introduced, which is based on a quasi-dimensional k-epsilon-approach and can cope with turbulence production from large scale shearing. A simplified model for ignition delay is implemented which emphasizes the transfer from laminar to turbulent flame propagation after ignition. The modeling is completed with the calculation of overall heat release rates in a 0D entrainment approach with the help of turbulent flame velocities. After validation of all sub-models, the 0D combustion prediction is used in combination with a 1D gas exchange analysis to virtually calibrate the modern engine torque structure and the ECU function for exhaust gas temperature with extensive simulations.:Contents 1 Introduction. 2 Thermodynamic modeling with real-time capability. 3 Quasi-dimensional modeling of turbulence and global charge motion. 4 Physical modeling of ignition delay. 5 Combustion modeling based on a 0D entrainment approach. 6 Virtual engine calibration with a quasi-dimensional combustion model. 7 Summary and outlook. / Moderne Ottomotoren spielen heute sowohl in konventionellen als auch hybriden Fahrzeugantrieben eine große Rolle. Aktuelle Konzepte sind hochvariabel bezüglich Ventilsteuerung, Kraftstoffeinspritzung und Laststeuerung und ihre Optimierungspotentiale erwachsen zumeist aus neuen Softwarefunktionen. Deren Applikation ist zeit- und kostenintensiv und soll durch virtuelle Methoden unterstützt werden. In der vorliegenden Arbeit wird ein physikalisches 0D Verbrennungsmodell für Ottomotoren aufgebaut und bis zur praktischen Anwendung geführt. Dafür wurde zuerst die Thermodynamik echtzeitfähig modelliert und im gesamten Motorenkennfeld abgeglichen. Der Aufbau eines neuen Turbulenzmodells auf Basis der quasidimensionalen k-epsilon-Gleichung ermöglicht anschließend, die veränderlichen Einflüsse globaler Ladungsbewegung auf die Turbulenz abzubilden. Für den Brennverzug wurde ein vereinfachtes Modell abgeleitet, welches den Übergang von laminarer zu turbulenter Flammenausbreitung nach der Zündung in den Vordergrund stellt. Der restliche Brennverlauf wird durch die physikalische Ermittlung der turbulenten Brenngeschwindigkeit in einem 0D Entrainment-Ansatz dargestellt. Nach Validierung aller Teilmodelle erfolgt die virtuelle Bedatung der Momentenstruktur und der Abgastemperaturfunktion für das Motorsteuergerät.:Contents 1 Introduction. 2 Thermodynamic modeling with real-time capability. 3 Quasi-dimensional modeling of turbulence and global charge motion. 4 Physical modeling of ignition delay. 5 Combustion modeling based on a 0D entrainment approach. 6 Virtual engine calibration with a quasi-dimensional combustion model. 7 Summary and outlook.

Apesar de as EDPS que modelam leis de conservação e problemas em dinâmica dos fluídos serem bem estabelecidas, suas soluções numéricas continuam ainda desafiadoras. Em particular, há dois desafios associados à computação e ao entendimento desses problemas: um deles é a formação de descontinuidades (choques) e o outro é o fenômeno turbulência. Ambos os desafios podem ser atribuídos ao tratamento dos termos advectivos não lineares nessas equações de transporte. Dentro deste canário, esta tese apresenta o estudo do desenvolvimento de um novo esquema \"upwind\" de alta resolução e sua associação com modelagem da turbulência. O desempenho do esquema é investigado nas soluções da equação de advecção 1D com dados iniciais descontínuos e de problemas de Riemann 1D para as equações de Burgers, Euler e águas rasas. Além disso, são apresentados resultados numéricos de escoamentos incompressíveis 2D e 3D no regime laminar a altos números de Reynolds. O novo esquema é então associado à modelagem \'capa\' - \'epsilon\' da turbulência para a simulação numérica de escoamentos incompressíveis turbulentos 2D e 3D com superfícies livres móveis. Aplicação, verificação e validação dos métodos numéricos são também fornecidas / Althought the PDEs that model conservation laws and fluid dynamics problems are well established, their numerical solutions have presented a continuing challenge. In particular, there are two challenges associated with the computation and the understanding of these problems, namely, formation of shocks and turbulence. Both challenges can be attributed to the nonlinear advection terms of these transport equations. In this scenario, this thesis presents the study of the development of a new high-resolution upwind scheme and its association with turbulence modelling. The performance of the scheme is investigated by solving the 1D advection equation with discontinuous initial data 1D Riemann problems for Burgers, Euler and shallow water equations. Besides, numerical results for 2D and 3D incompressible laminar flows at high Reynolds number are presented. The new scheme is then associated with the \'capa - \' epsilon\' turbulence model for the simulation of 2D and 3D incompressible turbulent flows with moving free surfaces. Application, verification and validation of the numerical methods are also provided

Apesar de as EDPS que modelam leis de conservação e problemas em dinâmica dos fluídos serem bem estabelecidas, suas soluções numéricas continuam ainda desafiadoras. Em particular, há dois desafios associados à computação e ao entendimento desses problemas: um deles é a formação de descontinuidades (choques) e o outro é o fenômeno turbulência. Ambos os desafios podem ser atribuídos ao tratamento dos termos advectivos não lineares nessas equações de transporte. Dentro deste canário, esta tese apresenta o estudo do desenvolvimento de um novo esquema \"upwind\" de alta resolução e sua associação com modelagem da turbulência. O desempenho do esquema é investigado nas soluções da equação de advecção 1D com dados iniciais descontínuos e de problemas de Riemann 1D para as equações de Burgers, Euler e águas rasas. Além disso, são apresentados resultados numéricos de escoamentos incompressíveis 2D e 3D no regime laminar a altos números de Reynolds. O novo esquema é então associado à modelagem \'capa\' - \'epsilon\' da turbulência para a simulação numérica de escoamentos incompressíveis turbulentos 2D e 3D com superfícies livres móveis. Aplicação, verificação e validação dos métodos numéricos são também fornecidas / Althought the PDEs that model conservation laws and fluid dynamics problems are well established, their numerical solutions have presented a continuing challenge. In particular, there are two challenges associated with the computation and the understanding of these problems, namely, formation of shocks and turbulence. Both challenges can be attributed to the nonlinear advection terms of these transport equations. In this scenario, this thesis presents the study of the development of a new high-resolution upwind scheme and its association with turbulence modelling. The performance of the scheme is investigated by solving the 1D advection equation with discontinuous initial data 1D Riemann problems for Burgers, Euler and shallow water equations. Besides, numerical results for 2D and 3D incompressible laminar flows at high Reynolds number are presented. The new scheme is then associated with the \'capa - \' epsilon\' turbulence model for the simulation of 2D and 3D incompressible turbulent flows with moving free surfaces. Application, verification and validation of the numerical methods are also provided