Mannheim, Univ., Diplomarbeit, 2007.

Regensburg, Univ., Diss., 2008.

Universiẗat, Diss., 1994--Konstanz.

Techn. Universiẗat, Diss., 2002--Berlin.

Techn. Universiẗat, Diss., 2002--Braunschweig.

The eukaryotic cell cycle orchestrates the precise duplication and distribution of the genetic material, cytoplasm and membranes to daughter cells. In multicellular eukaryotes, cell cycle regulation also governs various organisatorial processes ranging from gametogenesis over multicellular development to tissue formation and repair. Consequently, defects in cell cycle regulation provoke a variety of human cancers. A global view of genes and pathways governing the human cell cycle would advance many research areas and may also deliver novel cancer targets. Therefore this work aimed on the genome-wide identification and systematic characterisation of genes required for cell cycle progression in human cells. I developed a highly specific and efficient RNA interference (RNAi) technology to realize the potential of RNAi for genome-wide screening of the genes essential for cell cycle progression in human tissue culture cells. This approach is based on the large-scale enzymatic digestion of long dsRNAs for the rapid and cost-efficient generation of libraries of highly complex pools of endoribonuclease-prepared siRNAs (esiRNAs). The analysis of the silencing efficiency and specificity of esiRNAs and siRNAs revealed that esiRNAs are as efficient for mRNA degradation as chemically synthesized siRNA designed with state-of-the-art design algorithms, while exhibiting a markedly reduced number of off-target effects. After demonstrating the effectiveness of this approach in a proof-of-concept study, I screened a genome-wide esiRNA library and used three assays to generate a quantitative and reproducible multi-parameter profile for the 1389 identified genes. The resulting phenotypic signatures were used to assign novel cell cycle functions to genes by combining hierarchical clustering, bioinformatics and proteomic data mining. This global perspective on gene functions in the human cell cycle presents a framework for the systematic documentation necessary for the understanding of cell cycle progression and its misregulation in diseases. The identification of novel genes with a role in human cell cycle progression is a starting point for an in-depth analysis of their specific functions, which requires the validation of the observed RNAi phenotype by genetic rescue, the study of the subcellular localisation and the identification of interaction partners of the expressed protein. One strategy to achieve these experimental goals is the expression of RNAi resistant and/or tagged transgenes. A major obstacle for transgenesis in mammalian tissue culture cells is the lack of efficient homologous recombination limiting the use of cultured mammalian cells as a real genetic system like yeast. I developed a technology circumventing this problem by expressing an orthologous gene from a closely related species including its regulatory sequences carried on a bacterial artificial chromosome (BAC). This technology allows physiological expression of the transgene, which cannot be achieved with conventional cDNA expression constructs. The use of the orthologous gene from a closely related species confers RNAi resistance to the transgene allowing the depletion of the endogenous gene by RNAi. Thus, this technology mimics homologous recombination by replacing an endogenous gene with a transgene while maintaining normal gene expression. In combination with recombineering strategies this technology is useful for RNAi rescue experiments, protein localisation and the identification of protein interaction partners in mammalian tissue culture cells. In summary, this thesis presents a major technical advance for large-scale functional genomic studies in mammalian tissue culture cells and provides novel insights into various aspects of cell cycle progression. (Die Druckexemplare enthalten jeweils eine CD-ROM als Anlagenteil: 217 MB: Movies, Rohdaten - Nutzung: Referat Informationsvermittlung der SLUB)

This dissertation aims at designing a three-phase voltage source inverter based on the SiC devices and mainly the SiC-MOSFET. The designed inverter offers a possibility to drive the power inverter with a very high efficiency, which can reach up to 99% for 16 kW rated power. The design is dedicated to the electric vehicle application, and it aims at • Providing a comparative study on some of the current discrete SiC devices in terms of the total losses and the thermal conductivity. In addition, a behavioral study of the effective channel mobility with temperature variation in the SiC MOSFET will be investigated. • Designing a gate driver which fits with the driving requirements of the SiC-MOSFET and provides a trade-off between the switching losses and the EMI behavior. • Designing a three-phase voltage source inverter with 16 kW rated power; the design includes minimizing the inverter losses and extracts the EMI model of the power inverter by considering the effects of the parasitic parameters; moreover a short guideline for selecting the heat-sink based on the static network is introduced. • Proposing a new and simplified carried-based PWM, this will reduce the harmonics in the output waveforms and enhance the utilization of the DC-link voltage. • Proposing a new strategy for compensating the dead-time effect in carrier based-PWM and to find out the proper dead-time level in VSI based on SiC –MOSFET. • Designing faults diagnosis and protection circuits in order to protect the power inverter from the common faults; overcurrent, short-circuit, overvoltage, and overtemperature faults.

The eukaryotic cell cycle orchestrates the precise duplication and distribution of the genetic material, cytoplasm and membranes to daughter cells. In multicellular eukaryotes, cell cycle regulation also governs various organisatorial processes ranging from gametogenesis over multicellular development to tissue formation and repair. Consequently, defects in cell cycle regulation provoke a variety of human cancers. A global view of genes and pathways governing the human cell cycle would advance many research areas and may also deliver novel cancer targets. Therefore this work aimed on the genome-wide identification and systematic characterisation of genes required for cell cycle progression in human cells. I developed a highly specific and efficient RNA interference (RNAi) technology to realize the potential of RNAi for genome-wide screening of the genes essential for cell cycle progression in human tissue culture cells. This approach is based on the large-scale enzymatic digestion of long dsRNAs for the rapid and cost-efficient generation of libraries of highly complex pools of endoribonuclease-prepared siRNAs (esiRNAs). The analysis of the silencing efficiency and specificity of esiRNAs and siRNAs revealed that esiRNAs are as efficient for mRNA degradation as chemically synthesized siRNA designed with state-of-the-art design algorithms, while exhibiting a markedly reduced number of off-target effects. After demonstrating the effectiveness of this approach in a proof-of-concept study, I screened a genome-wide esiRNA library and used three assays to generate a quantitative and reproducible multi-parameter profile for the 1389 identified genes. The resulting phenotypic signatures were used to assign novel cell cycle functions to genes by combining hierarchical clustering, bioinformatics and proteomic data mining. This global perspective on gene functions in the human cell cycle presents a framework for the systematic documentation necessary for the understanding of cell cycle progression and its misregulation in diseases. The identification of novel genes with a role in human cell cycle progression is a starting point for an in-depth analysis of their specific functions, which requires the validation of the observed RNAi phenotype by genetic rescue, the study of the subcellular localisation and the identification of interaction partners of the expressed protein. One strategy to achieve these experimental goals is the expression of RNAi resistant and/or tagged transgenes. A major obstacle for transgenesis in mammalian tissue culture cells is the lack of efficient homologous recombination limiting the use of cultured mammalian cells as a real genetic system like yeast. I developed a technology circumventing this problem by expressing an orthologous gene from a closely related species including its regulatory sequences carried on a bacterial artificial chromosome (BAC). This technology allows physiological expression of the transgene, which cannot be achieved with conventional cDNA expression constructs. The use of the orthologous gene from a closely related species confers RNAi resistance to the transgene allowing the depletion of the endogenous gene by RNAi. Thus, this technology mimics homologous recombination by replacing an endogenous gene with a transgene while maintaining normal gene expression. In combination with recombineering strategies this technology is useful for RNAi rescue experiments, protein localisation and the identification of protein interaction partners in mammalian tissue culture cells. In summary, this thesis presents a major technical advance for large-scale functional genomic studies in mammalian tissue culture cells and provides novel insights into various aspects of cell cycle progression. (Die Druckexemplare enthalten jeweils eine CD-ROM als Anlagenteil: 217 MB: Movies, Rohdaten - Nutzung: Referat Informationsvermittlung der SLUB)

Vor dem Hintergrund der PSI-Theorie werden Fragen der Vorhersage von Leistung entwickelt. Neben einer kurzen Darstellung der Theorie werden zunächst zur Methodenexploration in 4 Experimenten im Stroop-Paradigma Fragen der Grundlagenforschung zum Stroop-Interferenz-Reduktions-Effekt (SIRE) durch nonverbale emotionale Primes beantwortet. In 3 weiteren Studien konnte gezeigt werden, dass Leistung mit der Interaktion aus Selbststeuerung insbesondere der Komponente Zielumsetzung und dem nichtreaktiven Maß der Stroop-Interferenz nach Leistungsprimes vorhergesagt werden kann. Es scheint Hinweise auf eine gewisse Gesetzmäßigkeit dieser Interaktion zu geben. Bei der Leistungsvorhersage wird eine invers sinusförmig verlaufende Charakteristik der Interaktion vermutet, die in 3 Studentenstichproben zum Problemlösen, zur Klausurleistung sowie zum Fortschritt bei Leistungszielen während eines Semesters untersucht werden konnte. Die wesentlichen Erkenntnisse der Vorhersage aus reaktiven Maßen (Selbststeuerung, insbesondere Willensbahnung) und nichtreaktiven Maßen (Intentionsgedächtnisnutzung) werden dargestellt. Implikationen für die Anwendung gehen in Richtung größerer Studien zur Erforschung dieser prädiktiven Interaktion. Dieses Muster ist relevant für die Anwendungswissenschaften Klinische Psychologie als auch die Arbeitspsychologie i.S. von persönlichkeitsfördernder Gestaltung der Arbeit.

This dissertation aims at designing a three-phase voltage source inverter based on the SiC devices and mainly the SiC-MOSFET. The designed inverter offers a possibility to drive the power inverter with a very high efficiency, which can reach up to 99% for 16 kW rated power. The design is dedicated to the electric vehicle application, and it aims at • Providing a comparative study on some of the current discrete SiC devices in terms of the total losses and the thermal conductivity. In addition, a behavioral study of the effective channel mobility with temperature variation in the SiC MOSFET will be investigated. • Designing a gate driver which fits with the driving requirements of the SiC-MOSFET and provides a trade-off between the switching losses and the EMI behavior. • Designing a three-phase voltage source inverter with 16 kW rated power; the design includes minimizing the inverter losses and extracts the EMI model of the power inverter by considering the effects of the parasitic parameters; moreover a short guideline for selecting the heat-sink based on the static network is introduced. • Proposing a new and simplified carried-based PWM, this will reduce the harmonics in the output waveforms and enhance the utilization of the DC-link voltage. • Proposing a new strategy for compensating the dead-time effect in carrier based-PWM and to find out the proper dead-time level in VSI based on SiC –MOSFET. • Designing faults diagnosis and protection circuits in order to protect the power inverter from the common faults; overcurrent, short-circuit, overvoltage, and overtemperature faults.