The SRC gene encodes pp60c-Src, a 60 kDa non-receptor tyrosine kinase that is frequently activated and/or overexpressed in many cancers including colon cancer. In a subset of colon cancer cell lines, it has been shown, that the overexpression of c-Src can be explained, in part, by the transcriptional activation of the SRC gene. As a result, the general goal of this thesis was to further characterize how SRC is transcriptionally regulated in human cancer cell lines. Two highly dissimilar promoters, the housekeeping-like SRC1A promoter, as well as the HIF-1Ñ regulated tissue-specific SRC1Ñ promoter, regulate SRC expression. hnRNP K and the Sp family of factors regulate the SRC1A promoter; however, the true impact of Sp3 on SRC1A activity was not understood. In this thesis, a comprehensive analysis of the effect of Sp3 on SRC1A activity was performed. Physiologically, Sp3 exists as four translational isoforms that, in part, dictate the activation potential of Sp3. In general, the longer forms of Sp3 were modest transcriptional activators of the SRC1A promoter whereas the shorter forms were unable to activate the SRC1A promoter. An analysis of all Sp3 isoforms identified that the shorter Sp3 isoforms could be converted into transcriptional activators of SRC1A if the SUMOylation of a critical lysine residue within the inhibitory domain was prevented. Conversely, SUMOylation of the same isoform had little effect on the activation potential of the longer Sp3 isoforms at the SRC1A promoter. These results suggest that transcriptional activation by Sp3 is promoter context-, isoform- and modification-dependent.<p>SRC is transcriptionally repressed by histone deacetylase inhibitors (HDIs) and despite unsuccessful studies attempting to identify HDI-responsive elements within the SRC promoter regions none could be identified. This finding also suggests that histone deacetylases (HDACs) may be required for SRC expression. Historically, it was believed that HDIs act at the histone level to alter chromatin dynamics through the inactivation of HDACs to result in histone hyperacetylation and increased transcriptional activation. As such, a systematic investigation of the changes in histone H3 and H4 acetylation status at the transcriptionally repressed SRC promoter regions and the transcriptionally activated p21WAF1 promoter region was performed. The p21WAF1 promoter was used as control in this study as p21WAF1 is a classical example of a gene transcriptionally activated by HDIs. Interestingly, similar changes in histone acetylation at the p21WAF1 promoter and both SRC promoter regions were observed. Upon closer examination of acetylation changes at discreet histone residues, it was observed that in the rare case that a particular residue was differentially acetylated upon treatment at the promoter regions analyzed, the SRC1Ñ and p21WAF1 promoter regions demonstrated more similar changes in acetylation as compared to SRC1A. Taken together, these results suggest that histone acetylation status is not an accurate indicator of transcriptional activity following HDI treatment. To further investigate HDI-mediated SRC repression, RNA Pol. II occupancy at the promoter and regions downstream of the promoter were assessed. Despite the continued occupancy of RNA Pol. II at the promoter regions, RNA Pol. II was lost from the 3¡¦ UTR upon treatment with HDIs. These findings suggest that RNA Pol. II . may be sequestered at the promoter regions upon treatment with HDIs possibly as a result of impeded transcription initiation and/or elongation. Further analysis of the phosphorylation status of RNA Pol. II identified that transcriptional initiation was indeed occurring despite HDI treatment; however, productive transcriptional elongation could not be confirmed thus suggesting a role for abrogated elongation in HDI mediated SRC repression. Complimentary analysis of the effects of HDACs on SRC expression suggested that while class I HDACs abrogated SRC expression, class II HDACs were required for the maintenance of SRC transcript levels in a promoter-independent fashion. Together, these results provide the basis for a model whereby HDIs repress SRC transcriptional expression through the inhibition of class II HDAC activity to eventually result in curtailed SRC transcriptional elongation.

The work describes the development of a new class of protein-inhibitors for protein-protein interactions, based on metallopeptides comprised of a dirhodium metal center. The metal incorporation in the peptide sequence leads to high increase in binding affinity of the inhibitors. The source of this strong affinity is the interaction of histidine on the protein surface with the rhodium center. In addition to this work, rhodium-based small molecule inhibitors for FK-506 binding proteins are investigated. Also, methodology for rhodium-catalyzed modification of proteins containing surface cysteine has been developed where a simple rhodium(II) complex catalyzes cysteine modification with diazo reagents. The reaction is marked by clean cysteine selectivity and mild reaction conditions. The resulting linkage is significantly more stable in human plasma serum, when compared to common maleimide reagents. Apart from this body of work in chemical-biology, the thesis contains the discussion of development of copper-catalyzed remote chlorination of alkyl hydroperoxides. The atom transfer chlorination utilizes simple ammonium chloride salts as the chlorine source and the internal redox process requires no external redox reagents.

The SRC gene encodes pp60c-Src, a 60 kDa non-receptor tyrosine kinase that is frequently activated and/or overexpressed in many cancers including colon cancer. In a subset of colon cancer cell lines, it has been shown, that the overexpression of c-Src can be explained, in part, by the transcriptional activation of the SRC gene. As a result, the general goal of this thesis was to further characterize how SRC is transcriptionally regulated in human cancer cell lines. Two highly dissimilar promoters, the housekeeping-like SRC1A promoter, as well as the HIF-1Ñ regulated tissue-specific SRC1Ñ promoter, regulate SRC expression. hnRNP K and the Sp family of factors regulate the SRC1A promoter; however, the true impact of Sp3 on SRC1A activity was not understood. In this thesis, a comprehensive analysis of the effect of Sp3 on SRC1A activity was performed. Physiologically, Sp3 exists as four translational isoforms that, in part, dictate the activation potential of Sp3. In general, the longer forms of Sp3 were modest transcriptional activators of the SRC1A promoter whereas the shorter forms were unable to activate the SRC1A promoter. An analysis of all Sp3 isoforms identified that the shorter Sp3 isoforms could be converted into transcriptional activators of SRC1A if the SUMOylation of a critical lysine residue within the inhibitory domain was prevented. Conversely, SUMOylation of the same isoform had little effect on the activation potential of the longer Sp3 isoforms at the SRC1A promoter. These results suggest that transcriptional activation by Sp3 is promoter context-, isoform- and modification-dependent.<p>SRC is transcriptionally repressed by histone deacetylase inhibitors (HDIs) and despite unsuccessful studies attempting to identify HDI-responsive elements within the SRC promoter regions none could be identified. This finding also suggests that histone deacetylases (HDACs) may be required for SRC expression. Historically, it was believed that HDIs act at the histone level to alter chromatin dynamics through the inactivation of HDACs to result in histone hyperacetylation and increased transcriptional activation. As such, a systematic investigation of the changes in histone H3 and H4 acetylation status at the transcriptionally repressed SRC promoter regions and the transcriptionally activated p21WAF1 promoter region was performed. The p21WAF1 promoter was used as control in this study as p21WAF1 is a classical example of a gene transcriptionally activated by HDIs. Interestingly, similar changes in histone acetylation at the p21WAF1 promoter and both SRC promoter regions were observed. Upon closer examination of acetylation changes at discreet histone residues, it was observed that in the rare case that a particular residue was differentially acetylated upon treatment at the promoter regions analyzed, the SRC1Ñ and p21WAF1 promoter regions demonstrated more similar changes in acetylation as compared to SRC1A. Taken together, these results suggest that histone acetylation status is not an accurate indicator of transcriptional activity following HDI treatment. To further investigate HDI-mediated SRC repression, RNA Pol. II occupancy at the promoter and regions downstream of the promoter were assessed. Despite the continued occupancy of RNA Pol. II at the promoter regions, RNA Pol. II was lost from the 3¡¦ UTR upon treatment with HDIs. These findings suggest that RNA Pol. II . may be sequestered at the promoter regions upon treatment with HDIs possibly as a result of impeded transcription initiation and/or elongation. Further analysis of the phosphorylation status of RNA Pol. II identified that transcriptional initiation was indeed occurring despite HDI treatment; however, productive transcriptional elongation could not be confirmed thus suggesting a role for abrogated elongation in HDI mediated SRC repression. Complimentary analysis of the effects of HDACs on SRC expression suggested that while class I HDACs abrogated SRC expression, class II HDACs were required for the maintenance of SRC transcript levels in a promoter-independent fashion. Together, these results provide the basis for a model whereby HDIs repress SRC transcriptional expression through the inhibition of class II HDAC activity to eventually result in curtailed SRC transcriptional elongation.

Es wurde eine SP3-Transportmethode entwickelt, die neutronenkinetische Rechnungen für die Kerne von Leichtwasserreaktoren mit höherer Genauigkeit als die gegenwärtig in der Kernauslegung angewandten Standardmethoden auf Basis der Zweigruppendiffusionsnäherung er-laubt. Eine Verbesserung der Genauigkeit von Abbrandrechnungen und der Berechnung von Tran-sienten ist für heterogene Kerne notwendig, in denen neben UO2-Brennelementen auch Mischoxyd – Brennelemente eingesetzt werden. In einem ersten Schritt wird die in dem Rechenprogramm DYN3D verwendete Zweigruppendiffusi-onsmethode auf viele Energiegruppen erweitert. Auf der Basis von Untersuchungen zu einer optima-len Gruppenstruktur wird die Verwendung von 8-10 Energiegruppen der Neutronen als optimal erach-tet. Das Verfahren wurde anhand von stationären und transienten Rechnungen für das OECD/NEA und US NRC PWR MOX/UO2 Core Transient Benchmark verifiziert. In den nächsten Schritten erfolgte die Entwicklung und Implementierung einer SP3-Näherung in DYN3D. Dabei besteht die Möglichkeit, ein feineres Gitter im BE zu benutzen. Das Verfahren wurde zunächst durch pinweise Berechnung stationärer Zustände des obigen Benchmarks verifiziert. Untersuchungen für das Benchmarkproblem zeigen, dass das Verhältniss des 2-ten Momentes zum 0-ten Moment des Flusses klein ist. Die beiden SP3-Gleichungen können deshalb separat in iterativer Weise gelöst werden. Dies reduziert den benötigten Speicherplatz und erfordert weniger CPU-Zeit. Dieses vereinfachte Verfahren wurde deshalb ebenfalls in das Programm implementiert. Es wird ge-zeigt, dass mit diesem Verfahren eine vergleichbare Genauigkeit erreicht wird. Stabweise Rechnun-gen mit 4, 8 und 16 Energiegrupppen wurden für einen stationären Zustand des Benchmarks durch-geführt. Eine 3-dimensionale Aufgabe des Benchmarks mit Rückkopplung und Vollleistung wurde mit dem optimierten SP3-Verfahren gerechnet. A SP3 transport approximation was developed for neutron kinetic calculations of cores of light water reactors with a higher accuracy than the present standard methods of core design based on the two group diffusion approximation. An improvement of accuracy for burnup and transient calculations is required for cores loaded with UO2 and MOX fuel assemblies. In the first step, the two group diffusion method applied in the computer code DYN3D was extended to an arbitrary number of groups. Investigations for an optimal group structure have shown that a number of 8 to 10 energy groups of neutrons seems to be reasonable. The multi-group technique was verified for steady states and transients of the OECD/NEA und US NRC PWR MOX/UO2 Core Tran-sient Benchmark. In the next steps, a SP3-approximation was developed and implemented into DYN3D. The possibility of using finer meshes inside the fuel assemblies is involved in this method. The technique was veri-fied by pinwise calculations for steady states of the above mentioned benchmark. The investigations to the benchmark problem have shown that ratio of the 2nd moment of flux to the 0th moment is small. Therefore the two coupled SP3 equations can be solved separately in an iterative way. The required computer memory and the CPU time can be reduced by this technique. This sim-pler method was also implemented in the code. It is shown that the reached accuracy is comparable to accuracy of the original technique. Pinwise calculations with 4, 8 and 16 energy groups were per-formed for a steady state of this benchmark. A three-dimensional problem of the benchmark at full power and with feedback was calculated with the optimized SP3 technique. The optimized method was used for the time integration of the transient SP3 equations. The pinwise calculation of a control rod ejection was tested for a simple system and the results were compared with the diffusion solution.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

Afin de faciliter la synthèse totale des principes actifs utilisés en industrie pharmaceutique ou agrochimique, les chimistes ont pour objectif de mettre au point de nouvelles méthodes générales, faciles à mettre en œuvre et éco-compatibles. D’après une étude réalisée en 2014, la pipéridine serait l’hétérocycle azoté le plus fréquent dans les médicaments approuvés par l’Agence américaine des produits alimentaires et médicamenteux (FDA).Dans ce contexte, trois méthodes distinctes ont été développées au cours de cette thèse pour synthétiser des pipéridines fonctionnalisées. Une bibliothèque de pipéridines substituées par des groupes aromatiques et vinyliques a d’abord été efficacement obtenue par couplage croisé catalysé par un complexe de cobalt entre des 4- et 3 halogénopipéridines et des réactifs de Grignard. L’utilisation d’un sel de cobalt, moins cher que les complexes de palladium et moins toxique que les complexes de nickel, permet de limiter les réactions parasites de déshalogénation ou de β-H élimination. Une variété de 2 diénylpipéridines, motifs présents dans plusieurs alcaloïdes, a ensuite été préparée par cyclisation catalysée par un sel de fer, peu cher et peu toxique, à partir d’amino-alcools diallyliques. Pour finir, la mise au point de conditions permettant la monoarylation de pipéridines par activation de liaisons C(sp3)‒H catalysée par un complexe de ruthénium a été envisagée. Plus particulièrement, l’influence sur la réaction d’arylation des propriétés électroniques et stériques du groupement directeur présent sur l’azote de la pipéridine a été étudiée. Ces méthodes ont également été élargies à la synthèse d’autres cycles azotés. / In order to facilitate the total synthesis of active molecules used in pharmaceutical or agrochemical industries, chemists try constantly to develop new, general, practical and sustainable methods. In 2014, a study revealed that piperidine was the most frequently present aza-heterocycle in medicines approved by the Food and Drug Administration (FDA). In this context, three different methods were developed during this Ph.D in order to synthesize functionalized piperidines. A wide variety of substituted piperidines was first efficiently obtained by a cobalt catalyzed cross-coupling reaction between 4- and 3-halogenopiperidines and Grignard reagents. Cobalt has appeared as a good alternative to the expensive palladium salts or the toxic nickel salts. Moreover, it can prevent side reactions such as dehydrohalogenation or β-H elimination. Next, 2-dienylpiperidines, present in a myriad of alkaloids, were prepared by iron catalyzed cyclization from diallylic amino-alcools. Finally, new conditions for the ruthenium catalyzed C(sp3)‒H monoarylation of piperidines were developed. The influence of the electronic and steric properties of the directing group attached to the nitrogen of the piperidine was fully studied. These methods were then applied to the synthesis of other azacycles.

Es wurde eine SP3-Transportmethode entwickelt, die neutronenkinetische Rechnungen für die Kerne von Leichtwasserreaktoren mit höherer Genauigkeit als die gegenwärtig in der Kernauslegung angewandten Standardmethoden auf Basis der Zweigruppendiffusionsnäherung er-laubt. Eine Verbesserung der Genauigkeit von Abbrandrechnungen und der Berechnung von Tran-sienten ist für heterogene Kerne notwendig, in denen neben UO2-Brennelementen auch Mischoxyd – Brennelemente eingesetzt werden. In einem ersten Schritt wird die in dem Rechenprogramm DYN3D verwendete Zweigruppendiffusi-onsmethode auf viele Energiegruppen erweitert. Auf der Basis von Untersuchungen zu einer optima-len Gruppenstruktur wird die Verwendung von 8-10 Energiegruppen der Neutronen als optimal erach-tet. Das Verfahren wurde anhand von stationären und transienten Rechnungen für das OECD/NEA und US NRC PWR MOX/UO2 Core Transient Benchmark verifiziert. In den nächsten Schritten erfolgte die Entwicklung und Implementierung einer SP3-Näherung in DYN3D. Dabei besteht die Möglichkeit, ein feineres Gitter im BE zu benutzen. Das Verfahren wurde zunächst durch pinweise Berechnung stationärer Zustände des obigen Benchmarks verifiziert. Untersuchungen für das Benchmarkproblem zeigen, dass das Verhältniss des 2-ten Momentes zum 0-ten Moment des Flusses klein ist. Die beiden SP3-Gleichungen können deshalb separat in iterativer Weise gelöst werden. Dies reduziert den benötigten Speicherplatz und erfordert weniger CPU-Zeit. Dieses vereinfachte Verfahren wurde deshalb ebenfalls in das Programm implementiert. Es wird ge-zeigt, dass mit diesem Verfahren eine vergleichbare Genauigkeit erreicht wird. Stabweise Rechnun-gen mit 4, 8 und 16 Energiegrupppen wurden für einen stationären Zustand des Benchmarks durch-geführt. Eine 3-dimensionale Aufgabe des Benchmarks mit Rückkopplung und Vollleistung wurde mit dem optimierten SP3-Verfahren gerechnet. A SP3 transport approximation was developed for neutron kinetic calculations of cores of light water reactors with a higher accuracy than the present standard methods of core design based on the two group diffusion approximation. An improvement of accuracy for burnup and transient calculations is required for cores loaded with UO2 and MOX fuel assemblies. In the first step, the two group diffusion method applied in the computer code DYN3D was extended to an arbitrary number of groups. Investigations for an optimal group structure have shown that a number of 8 to 10 energy groups of neutrons seems to be reasonable. The multi-group technique was verified for steady states and transients of the OECD/NEA und US NRC PWR MOX/UO2 Core Tran-sient Benchmark. In the next steps, a SP3-approximation was developed and implemented into DYN3D. The possibility of using finer meshes inside the fuel assemblies is involved in this method. The technique was veri-fied by pinwise calculations for steady states of the above mentioned benchmark. The investigations to the benchmark problem have shown that ratio of the 2nd moment of flux to the 0th moment is small. Therefore the two coupled SP3 equations can be solved separately in an iterative way. The required computer memory and the CPU time can be reduced by this technique. This sim-pler method was also implemented in the code. It is shown that the reached accuracy is comparable to accuracy of the original technique. Pinwise calculations with 4, 8 and 16 energy groups were per-formed for a steady state of this benchmark. A three-dimensional problem of the benchmark at full power and with feedback was calculated with the optimized SP3 technique. The optimized method was used for the time integration of the transient SP3 equations. The pinwise calculation of a control rod ejection was tested for a simple system and the results were compared with the diffusion solution.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第17780号 / 医博第3806号 / 新制||医||999(附属図書館) / 30587 / 京都大学大学院医学研究科医学専攻 / (主査)教授 妻木 範行, 教授 武藤 学, 教授 松田 道行 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第20411号 / 工博第4348号 / 新制||工||1674(附属図書館) / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 杉野目 道紀, 教授 村上 正浩, 教授 中尾 佳亮 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

京都大学 / 新制・課程博士 / 博士(工学) / 甲第24639号 / 工博第5145号 / 新制||工||1983(附属図書館) / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 杉野目 道紀, 教授 中尾 佳亮, 教授 藤原 哲晶 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM