Les travaux entrepris au cours de cette thèse ont eu pour but d'étudier la modulation de la transmission synaptique inhibitrice au niveau des réseaux moteurs spinaux, à la fois au cours du développement et après lésion de la moelle épinière. Le nouveau-né présente des activités motrices spontanées qui jouent un rôle important dans la maturation des muscles et des réseaux de neurones de la moelle épinière. Dans une première étude, nous avons identifié l'un des mécanismes impliqués dans la genèse de ces activités chez le rat nouveau-né in vitro. Nous avons démontré que l'activation des récepteurs GABAᴀ au niveau des terminales d'afférences primaires joue un rôle majeur dans le déclenchement et la propagation de ces activités spontanées. Dans une deuxième étude, nous avons testé la robustesse des dépolarisations de nature GABAergique enregistrées in vitro, c'est-à-dire leur dépendance vis-à-vis des paramètres du milieu de perfusion. Nous avons démontré que l'action dépolarisante des neurotransmetteurs GABA/glycine au niveau des motoneurones et celle du GABA au niveau des terminales d'afférences primaires ne sont pas dues à une fourniture énergétique insuffisante. La dernière étude a été consacrée à la modulation de la transmission synaptique inhibitrice après lésion de la moelle épinière. Nous avons montré que l'activation des récepteurs 5-HT2 (R5-HT2), particulièrement celle de l'isoforme 5-HT2ᴀ, renforce le poids synaptique inhibiteur via une hyperpolarisation du potentiel d'équilibre des ions chlorure (ECl) et une augmentation d'expression de KCC2 au niveau de la membrane des motoneurones. / The aim of this thesis was to explore the modulation of the inhibitory synaptic transmission within the spinal motor networks, both during development and after SCI. Spontaneous movements are an ubiquitous feature of fetal and infant behavior. They provide signals that are important for the development of muscles and the assembly of neuronal networks in the spinal cord. In a first study, we characterized one of the mechanisms underlying spontaneous motor behaviors in the in vitro spinal cord preparation isolated from neonatal rats. We demonstrated that the GABA is playing a key role in promoting spontaneous activity through primary afferent depolarizations which reach firing threshold. In the second part of my thesis, we tested the robustness of the in vitro GABAergic depolarizations and their dependence on the aCSF parameters. We demonstrated that during development the depolarizing actions of GABA/glycine on motoneurons and GABA on primary afferent terminals are not due to inadequate energy supply. In the last part of my thesis, we focused on the modulation of the inhibitory synaptic transmission following SCI. We demonstrated that activation of the 5-HT2 receptors, particularly the 5-HT2ᴀ subtype, strengthens inhibitory synaptic transmission to spinal motoneurons by hyperpolarizing the reversal potential of Cl- ions (ECl) and by increasing the cell-membrane expression of KCC2. This phenomenon reduces spasticity after SCI in rats. Upregulation of KCC2 function by targeting 5-HT2ᴀ receptors therefore opens new therapeutic strategies for the treatment of spasticity following SCI.

- 4 - ABSTRACT During the revision of epiphytic species of the Lecanora subfusca group in the Czech Republic, nine taxa have been recorded. L. cinereofusca and L. exspersa are reported for the first time from the country. L. circumborealis has been excluded from the list of Czech lichens. L. cinereofusca and L. horiza could be considered as extinct. L. rugosella and L. subrugosa are regarded as extreme morpholocial forms from L. chlarotera and L. argentata. These morphotypes corelate with ecological conditions: substrate (nutrient enriched bark) and habitat (eutrophisation effect). In case of L. subrugosa, this speculation was confirmed by molecular data (ITS rDNA sequences). Several new secondary metabolites have been discovered during the detailed research of chemical lichen compounds. These substances belonging to terpenoids are taxonomically important and very helpful for distinguishing single species. The abundance of L. allophana and L. chlarotera has decreased during last decades. Contrarily, L. pulicaris expanded slightly. The main reasons of changes in distribution are air pollution and acidification of substrates as the impact of acid rains. The rate of substrate acidification has been shown on example of L. pulicaris. In the past, this lichen predominated on acid-barked porophytes. Nowadays, it...

Numa primeira etapa, estudamos as conexões da VTA para o complexo VTA-substância negra (SN) utilizando a leucoaglutinina do Phaseolus vulgaris (PHA-L). Estas conexões são substanciais, topograficamente organizadas, com destaque para o pólo caudal da VTA que inerva bilateralmente toda a extensão deste complexo. Numa segunda etapa, estudamos as projeções da VTA para o hipotálamo. A VTA se projeta principalmente para a área pré-óptica lateral e área hipotalâmica lateral, a região subfornical posterior e o núcleo dorsomedial. Foram vistas poucas aposições entre varicosidades PHA-L+ e neurônios imunorreativos para orexina ou para hormônio concentrador de melanina. Por fim, estudamos a colocalização do substrato do receptor de insulina (IRS-1), IRS-1 fosforilado e fosfatidilinositol-3 quinase (PI3K) com tirosina hidroxilase (TH) ou com a subunidade B da toxina colérica (CTb) injetada no estriado. A maioria dos neurônios TH+ da VTA-SN expressa IRS-1; injeções de CTb no estriado resultaram em células duplamente marcadas para CTb/IRS-1, CTb/PI3K e CTb/IRS-1 fosforilado. / In a first step, we studied the connections of the VTA to the complex VTA-substantia nigra (SN) using the Phaseolus vulgaris leucoagglutinin (PHA-L). These connections are substantial, topographically organized, especially the caudal pole of the VTA, which innervates bilaterally throughout the length of this complex. In a second step, we studied the projections of the VTA to the hypothalamus. The VTA projected mainly to the lateral preoptic area, lateral hypothalamic area, posterior subfornical region and dorsomedial nucleus. Were observed few appositions between PHA-L+ varicosities and neurons immunoreactive for orexin or melanin-concentrating hormone. Finally, we studied the co-localization of the insulin receptor substrate-1 (IRS-1), IRS-1-phosphorylated and phosphatidylinositol-3 kinase (PI3K) with tyrosine hydroxylase (TH) or cholera toxin B subunit (CTb) injected into the striatum. Most TH+ neurons of the VTA-SN expressed IRS-1; CTb injections in the striatum resulted in cells double-labeled for CTb/IRS-1, CTb/PI3K and CTb/IRS-1 phosphorylated.

A palha de cana-de-açúcar está se tornando uma biomassa lignocelulósica disponível a partir da progressiva introdução da colheita mecanizada da cana-deaçúcar no Brasil, situação que possibilita a utilização de uma parte desta como matéria-prima em processos de conversão termoquímica ou bioquímica. Além de pesquisas de uso da palha de cana para produção de bioenergia, a conversão bioquímica dos açúcares constituintes de sua fração hemicelulósica, particularmente a xilose, é uma rota potencial para seu aproveitamento na obtenção de produtos de alto valor agregado, como o xilitol. A importância deste poliol se deve às suas peculiares propriedades que permitem sua aplicação nas indústrias alimentícia, odontológica e farmacêutica, aliado ao fato do continuo e rápido crescimento de seu mercado mundial. No presente trabalho foi estudado o aproveitamento da fração hemicelulósica da palha de cana como matéria-prima na produção biotecnológica de xilitol, visando a valorização e incorporação desta biomassa em uma biorrefinaria de cana-de-açúcar. O elevado conteúdo de hemicelulose da palha de cana (27%), similar ao encontrado em outras biomassas lignocelulósicas avaliadas para produção de xilitol, e a maior proporção de xilose no hidrolisado hemicelulósico (71%) em relação aos outros açúcares constituintes, tornam esta biomassa potencial matéria-prima para este bioprocesso. A utilização do hidrolisado hemicelulósico de palha de cana concentrado e destoxificado como meio de fermentação para a bioconversão de xilose em xilitol por Candida guilliermondii FTI 20037 foi avaliada em diferentes fases da pesquisa. Na primeira, foi estudada a necessidade de suplementação nutricional do hidrolisado e a disponibilidade inicial de oxigênio, sendo realizadas fermentações em batelada em frascos Erlenmeyer de 125mL com 25mL ou 50mL de meio, 30oC, 200rpm e 48h. Foi demonstrado que a suplementação do hidrolisado com extrato de farelo de arroz, (NH4)2SO4 e CaCl2·2H2O resultou em aumento do valor da produtividade volumétrica de xilitol, enquanto que a menor disponibilidade inicial de oxigênio favoreceu a eficiência de bioconversão. A avaliação do efeito dos co-substratos maltose, sacarose, celobiose e glicerol sobre este bioprocesso revelou que o maior favorecimento foi obtido com sacarose (10gL-1), já que resultou nos máximos valores de concentração final de xilitol (41,36 ± 1,69 gL-1), eficiência de bioconversão (75,70 ± 0,73%) e produtividade volumétrica (0,61 ± 0,02 gL-1h-1), correspondentes a incrementos de 9,04%, 5,01% e 6,56%, respectivamente, em relação à condição ausente de cosubstratos. A adição ao hidrolisado hemicelulósico de palha de cana de Dimetilsulfóxido (DMSO), composto com capacidade de permeabilizante de membrana celular, não resultou no incremento da produção de xilitol, a qual, pelo contrário, foi reduzida em razão da diminuição no consumo de xilose e crescimento celular de C. guilliermondii FTI 20037. Os resultados obtidos no presente estudo indicam que a produção biotecnológica de xilitol a partir de hidrolisado hemicelulósico de palha de cana suplementado com sacarose pode ser considerada uma rota de conversão bioquímica promissora para a valorização e integração desta biomassa em uma biorrefinaria de cana-de-açúcar. / Sugarcane straw is becoming an available lignocellulosic biomass from the progressive introduction of non-burning harvest in Brazil, situation that enables the utilization of a portion of this material as feedstock in thermochemical and biochemical conversion processes. Besides the use of sugarcane straw for bioenergy production, biochemical conversion of the constituent sugars of its hemicellulosic fraction, particularly xylose, is a potential route for the use of this biomass to obtain high added value products, such as xylitol. The importance of this product is due to its particular properties that enable its application in food, dental and pharmaceutical industries, coupled with the fact of the continuous and rapid growth of its market. In the present work it was studied the utilization of sugarcane straw hemicellulosic fraction as feedstock for biotechnological production of xylitol, aiming at the valorization and integration of this biomass in a sugarcane biorefinery. The high hemicellulosic content of sugarcane straw (27%), similar to that found in other lignocellulosic biomasses evaluated for xylitol production, and the higher proportion of xylose in the hemicellulosic hydrolysate (71%) in relation to the other constituent sugars, make this biomass potential feedstock for this bioprocess. The utilization of the concentrated and detoxified sugarcane straw hemicellulosic hydrolysate as fermentation medium for xylose-toxylitol bioconversion by Candida guilliermondii FTI 20037 was evaluated in different stages. In the first one, it was studied the necessity of nutritional supplementation of the hydrolysate and initial oxygen availability, being carried out batch fermentations in 125mL Erlenmeyer flasks with 25mL or 50mL of medium, 30oC, 200rpm and 48h. It was demonstrated that the supplementation of the hydrolysate with rice bran extract, (NH4)2SO4 and CaCl2·2H2O resulted on the increment of the value of xylitol volumetric productivity, whereas the higher initial oxygen availability favored the bioconversion efficiency. The evaluation of the effect of the co-substrates maltose, sucrose, cellobiose and glycerol on this bioprocess revealed that the higher improvement was obtained with sucrose (10gL-1), since it resulted in the maximum values of final concentration of xylitol (41.36 ± 1.69 gL-1), bioconversion efficiency (75.70 ± 0.73%) and volumetric productivity (0.61 ± 0.02 gL-1h-1), corresponding to increments of 9.04%, 5.01% and 6.56%, respectively, in relation to the condition absent of co-substrates. The addition to the sugarcane straw hemicellulosic hydrolysate of Dimethyl-sulfoxide, a cell membrane permeabilizer, did not resulted on the increasing of the xylitol production, which, in fact, was reduced due to the diminution on xylose consumption and cell growth of C. guilliermondii FTI 20037. The results obtained in this study indicate that biotechnological production of xylitol from sugarcane straw hemicellulosic hydrolysate supplemented with sucrose can be considered a promissory biochemical conversion route for valorization and integration of this biomass in a sugarcane biorefinery.

A palha de cana-de-açúcar está se tornando uma biomassa lignocelulósica disponível a partir da progressiva introdução da colheita mecanizada da cana-deaçúcar no Brasil, situação que possibilita a utilização de uma parte desta como matéria-prima em processos de conversão termoquímica ou bioquímica. Além de pesquisas de uso da palha de cana para produção de bioenergia, a conversão bioquímica dos açúcares constituintes de sua fração hemicelulósica, particularmente a xilose, é uma rota potencial para seu aproveitamento na obtenção de produtos de alto valor agregado, como o xilitol. A importância deste poliol se deve às suas peculiares propriedades que permitem sua aplicação nas indústrias alimentícia, odontológica e farmacêutica, aliado ao fato do continuo e rápido crescimento de seu mercado mundial. No presente trabalho foi estudado o aproveitamento da fração hemicelulósica da palha de cana como matéria-prima na produção biotecnológica de xilitol, visando a valorização e incorporação desta biomassa em uma biorrefinaria de cana-de-açúcar. O elevado conteúdo de hemicelulose da palha de cana (27%), similar ao encontrado em outras biomassas lignocelulósicas avaliadas para produção de xilitol, e a maior proporção de xilose no hidrolisado hemicelulósico (71%) em relação aos outros açúcares constituintes, tornam esta biomassa potencial matéria-prima para este bioprocesso. A utilização do hidrolisado hemicelulósico de palha de cana concentrado e destoxificado como meio de fermentação para a bioconversão de xilose em xilitol por Candida guilliermondii FTI 20037 foi avaliada em diferentes fases da pesquisa. Na primeira, foi estudada a necessidade de suplementação nutricional do hidrolisado e a disponibilidade inicial de oxigênio, sendo realizadas fermentações em batelada em frascos Erlenmeyer de 125mL com 25mL ou 50mL de meio, 30oC, 200rpm e 48h. Foi demonstrado que a suplementação do hidrolisado com extrato de farelo de arroz, (NH4)2SO4 e CaCl2·2H2O resultou em aumento do valor da produtividade volumétrica de xilitol, enquanto que a menor disponibilidade inicial de oxigênio favoreceu a eficiência de bioconversão. A avaliação do efeito dos co-substratos maltose, sacarose, celobiose e glicerol sobre este bioprocesso revelou que o maior favorecimento foi obtido com sacarose (10gL-1), já que resultou nos máximos valores de concentração final de xilitol (41,36 ± 1,69 gL-1), eficiência de bioconversão (75,70 ± 0,73%) e produtividade volumétrica (0,61 ± 0,02 gL-1h-1), correspondentes a incrementos de 9,04%, 5,01% e 6,56%, respectivamente, em relação à condição ausente de cosubstratos. A adição ao hidrolisado hemicelulósico de palha de cana de Dimetilsulfóxido (DMSO), composto com capacidade de permeabilizante de membrana celular, não resultou no incremento da produção de xilitol, a qual, pelo contrário, foi reduzida em razão da diminuição no consumo de xilose e crescimento celular de C. guilliermondii FTI 20037. Os resultados obtidos no presente estudo indicam que a produção biotecnológica de xilitol a partir de hidrolisado hemicelulósico de palha de cana suplementado com sacarose pode ser considerada uma rota de conversão bioquímica promissora para a valorização e integração desta biomassa em uma biorrefinaria de cana-de-açúcar. / Sugarcane straw is becoming an available lignocellulosic biomass from the progressive introduction of non-burning harvest in Brazil, situation that enables the utilization of a portion of this material as feedstock in thermochemical and biochemical conversion processes. Besides the use of sugarcane straw for bioenergy production, biochemical conversion of the constituent sugars of its hemicellulosic fraction, particularly xylose, is a potential route for the use of this biomass to obtain high added value products, such as xylitol. The importance of this product is due to its particular properties that enable its application in food, dental and pharmaceutical industries, coupled with the fact of the continuous and rapid growth of its market. In the present work it was studied the utilization of sugarcane straw hemicellulosic fraction as feedstock for biotechnological production of xylitol, aiming at the valorization and integration of this biomass in a sugarcane biorefinery. The high hemicellulosic content of sugarcane straw (27%), similar to that found in other lignocellulosic biomasses evaluated for xylitol production, and the higher proportion of xylose in the hemicellulosic hydrolysate (71%) in relation to the other constituent sugars, make this biomass potential feedstock for this bioprocess. The utilization of the concentrated and detoxified sugarcane straw hemicellulosic hydrolysate as fermentation medium for xylose-toxylitol bioconversion by Candida guilliermondii FTI 20037 was evaluated in different stages. In the first one, it was studied the necessity of nutritional supplementation of the hydrolysate and initial oxygen availability, being carried out batch fermentations in 125mL Erlenmeyer flasks with 25mL or 50mL of medium, 30oC, 200rpm and 48h. It was demonstrated that the supplementation of the hydrolysate with rice bran extract, (NH4)2SO4 and CaCl2·2H2O resulted on the increment of the value of xylitol volumetric productivity, whereas the higher initial oxygen availability favored the bioconversion efficiency. The evaluation of the effect of the co-substrates maltose, sucrose, cellobiose and glycerol on this bioprocess revealed that the higher improvement was obtained with sucrose (10gL-1), since it resulted in the maximum values of final concentration of xylitol (41.36 ± 1.69 gL-1), bioconversion efficiency (75.70 ± 0.73%) and volumetric productivity (0.61 ± 0.02 gL-1h-1), corresponding to increments of 9.04%, 5.01% and 6.56%, respectively, in relation to the condition absent of co-substrates. The addition to the sugarcane straw hemicellulosic hydrolysate of Dimethyl-sulfoxide, a cell membrane permeabilizer, did not resulted on the increasing of the xylitol production, which, in fact, was reduced due to the diminution on xylose consumption and cell growth of C. guilliermondii FTI 20037. The results obtained in this study indicate that biotechnological production of xylitol from sugarcane straw hemicellulosic hydrolysate supplemented with sucrose can be considered a promissory biochemical conversion route for valorization and integration of this biomass in a sugarcane biorefinery.

<p>Thin films play an important role in science and technology today. By combining different materials, properties for specific applications can be optimised. In this thesis growth of copper, copper(I) oxide and copper(I) nitride on two different substrates, amorphous SiO₂ and single crystalline α-Al₂O₃ by the so called Atomic Layer Deposition (ALD) techniques has been studied. This technique allows precise control of the growth process at monolayer level on solid substrates. Other characteristic features of ALD are that it produces films with excellent step coverage and good uniformity even as extremely thin films on complicated shaped substrates.</p><p>Alternative deposition schemes were developed for the materials of interest. It was demonstrated that use of intermediate water pulses affected the deposition pathways considerably. By adding water, the films are thought to grow via formation of an oxide over-layer instead of through a direct reaction between the precursors as in the case without water.</p><p>For growth of copper(I) nitride from Cu(hfac)₂ and ammonia no film growth occurred without adding water to the growth process. The Cu₃N films could be transformed into conducting copper films by post annealing. In copper growth from CuCl and H₂ the water affected film growth on the alumina substrates considerably more than on the fused silica substrates. The existence of surface -OH and/or -NH_x groups was often found to play an important role, according to both theoretical calculations and experimental results.</p>

In the present work, we discuss two subjects related to the nonequilibrium dynamics of polymers or biological filaments adsorbed to two-dimensional substrates. <br><br> The first part is dedicated to thermally activated dynamics of polymers on structured substrates in the presence or absence of a driving force. The structured substrate is represented by double-well or periodic potentials. We consider both homogeneous and point driving forces. Point-like driving forces can be realized in single molecule manipulation by atomic force microscopy tips. Uniform driving forces can be generated by hydrodynamic flow or by electric fields for charged polymers. <br><br> In the second part, we consider collective filament motion in motility assays for motor proteins, where filaments glide over a motor-coated substrate. The model for the simulation of the filament dynamics contains interactive deformable filaments that move under the influence of forces from molecular motors and thermal noise. Motor tails are attached to the substrate and modeled as flexible polymers (entropic springs), motor heads perform a directed walk with a given force-velocity relation. We study the collective filament dynamics and pattern formation as a function of the motor and filament density, the force-velocity characteristics, the detachment rate of motor proteins and the filament interaction. In particular, the formation and statistics of filament patterns such as nematic ordering due to motor activity or clusters due to blocking effects are investigated. Our results are experimentally accessible and possible experimental realizations are discussed. / In der vorliegenden Arbeit behandeln wir zwei Probleme aus dem Gebiet der Nichtgleichgewichtsdynamik von Polymeren oder biologischen Filamenten, die an zweidimensionale Substrate adsorbieren. <br><br> Der erste Teil befasst sich mit der thermisch aktivierten Dynamik von Polymeren auf strukturierten Substraten in An- oder Abwesenheit einer treibenden Kraft. Das strukturierte Substrat wird durch Doppelmulden- oder periodische Potentiale dargestellt. Wir betrachten sowohl homogene treibende Kräfte als auch Punktkräfte. Punktkräfte können bei der Manipulation einzelner Moleküle mit die Spitze eines Rasterkraftmikroskops realisiert werden. Homogene Kräfte können durch einen hydrodynamischen Fluss oder ein elektrisches Feld im Falle geladener Polymere erzeugt werden. <br><br> Im zweiten Teil betrachten wir die kollektive Bewegung von Filamenten in Motility-Assays, in denen Filamente über ein mit molekularen Motoren überzogenes Substrat gleiten. Das Modell zur Simulation der Filamentdynamik beinhaltet wechselwirkende, deformierbare Filamente, die sich unter dem Einfluss von Kräften, die durch molekulare Motoren erzeugt werden, sowie thermischem Rauschen bewegen. Die Schaftdomänen der Motoren sind am Substrat angeheftet und werden als flexible Polymere (entropische Federn) modelliert. Die Kopfregionen der Motoren vollführen eine gerichtete Schrittbewegung mit einer gegebenen Kraft-Geschwindigkeitsbeziehung. Wir untersuchen die kollektive Filamentdynamik und die Ausbildung von Mustern als Funktion der Motor- und der Filamentdichte, der Kraft-Geschwindigkeitscharakteristik, der Ablöserate der Motorproteine und der Filamentwechselwirkung. Insbesondere wird die Bildung und die Statistik der Filamentmuster, wie etwa die nematische Anordnung aufgrund der Motoraktivität oder die Clusterbildung aufgrund von Blockadeeffekten, untersucht. Unsere Ergebnisse sind experimentell zugänglich und mögliche experimentelle Realisierungen werden diskutiert.

Das Verhalten adhärenter Zellen hängt stark von den chemischen, topographischen und mechanischen Eigenschaften ihrer Umgebung ab. Experimentelle Untersuchungen der letzten Jahre haben gezeigt, dass adhärente Zellen aktiv die elastischen Eigenschaften ihrer Umgebung erkunden, indem sie an dieser ziehen. Der resultierende Kraftaufbau hängt von den elastischen Eigenschaften der Umgebung ab und wird an den Adhäsionskontakten in entsprechende biochemische Signale umgewandelt, die zelluläre Programme wie Wachstum, Differenzierung, programmierten Zelltod und Zellbewegung mitbestimmen. Im Allgemeinen sind Kräfte wichtige Einflussgrößen in biologischen Systemen. Weitere Beispiele dafür sind Hör- und Tastsinn, Wundheilung sowie die rollende Adhäsion von weißen Blutkörperchen auf den Wänden der Blutgefäße. In der Habilitationsschrift von Ulrich Schwarz werden mehrere theoretische Projekte vorgestellt, die die Rolle von Kräften und Elastizität in der Zelladhäsion untersuchen.<br /> (1) Es wurde eine neue Methode entwickelt, um die Kräfte auszurechnen, die Zellen an den Kontaktpunkten auf mikro-strukturierte elastische Substrate ausüben. Das Hauptergebnis ist, dass Zell-Matrix-Kontakte als Mechanosensoren funktionieren, an denen interne Kräfte in Proteinaggregation umgewandelt werden.<br /> (2) Eine Ein-Schritt-Master-Gleichung, die die stochastische Dynamik von Adhäsionsclustern als Funktion von Clustergröße, Rückbindungsrate und Kraft beschreibt, wurde sowohl analytisch als auch numerisch gelöst. Zudem wurde dieses Modell auf Zell-Matrix-Kontakte, dynamische Kraftspektroskopie sowie die rollende Adhäsion angewandt.<br /> (3) Im Rahmen der linearen Elastizitätstheorie und mit Hilfe des Konzepts der Kraftdipole wurde ein Modell formuliert und gelöst, das die Positionierung und Orientierung von Zellen in weicher Umgebung vorhersagt. Diese Vorhersagen sind in guter Übereinstimmung mit zahlreichen experimentellen Beobachtungen für Fibroblasten auf elastischen Substraten und in Kollagen-Gelen. / The behaviour of an adhering cell is strongly influenced by the chemical, topographical and mechanical properties of the surface it attaches to. During recent years, it has been found experimentally that adhering cells actively sense the elastic properties of their environment by pulling on it through numerous sites of adhesion. The resulting build-up of force at sites of adhesion depends on the elastic properties of the environment and is converted into corresponding biochemical signals, which can trigger cellular programmes like growth, differentiation, apoptosis, and migration. In general, force is an important regulator of biological systems, for example in hearing and touch, in wound healing, and in rolling adhesion of leukocytes on vessel walls. In the habilitation thesis by Ulrich Schwarz, several theoretical projects are presented which address the role of forces and elasticity in cell adhesion.<br /> (1) A new method has been developed for calculating cellular forces exerted at sites of focal adhesion on micro-patterned elastic substrates. The main result is that cell-matrix contacts function as mechanosensors, converting internal force into protein aggregation.<br /> (2) A one-step master equation for the stochastic dynamics of adhesion clusters as a function of cluster size, rebinding rate and force has been solved both analytically and numerically. Moreover this model has been applied to the regulation of cell-matrix contacts, to dynamic force spectroscopy, and to rolling adhesion.<br /> (3) Using linear elasticity theory and the concept of force dipoles, a model has been introduced and solved which predicts the positioning and orientation of mechanically active cells in soft material, in good agreement with experimental observations for fibroblasts on elastic substrates and in collagen gels.

Thin films play an important role in science and technology today. By combining different materials, properties for specific applications can be optimised. In this thesis growth of copper, copper(I) oxide and copper(I) nitride on two different substrates, amorphous SiO2 and single crystalline α-Al2O3 by the so called Atomic Layer Deposition (ALD) techniques has been studied. This technique allows precise control of the growth process at monolayer level on solid substrates. Other characteristic features of ALD are that it produces films with excellent step coverage and good uniformity even as extremely thin films on complicated shaped substrates. Alternative deposition schemes were developed for the materials of interest. It was demonstrated that use of intermediate water pulses affected the deposition pathways considerably. By adding water, the films are thought to grow via formation of an oxide over-layer instead of through a direct reaction between the precursors as in the case without water. For growth of copper(I) nitride from Cu(hfac)2 and ammonia no film growth occurred without adding water to the growth process. The Cu3N films could be transformed into conducting copper films by post annealing. In copper growth from CuCl and H2 the water affected film growth on the alumina substrates considerably more than on the fused silica substrates. The existence of surface -OH and/or -NHx groups was often found to play an important role, according to both theoretical calculations and experimental results.

<p>Enzymes are useful tools as specific analytical reagents. Two different analysis methods were developed for use in the separate fields of protein science and organic synthesis. Both methods rely on the substrate specificity of enzymes. Enzyme catalysis and substrate specificity is described and put in context with each of the two developed methods.</p><p>In <strong>paper I </strong>a method for C-terminal peptide sequencing was developed based on conventional Carboxypeptidase Y digestion combined with matrix assisted laser desorption/ionization mass spectrometry. An alternative nucleophile was used to obtain a stable peptide ladder and improve sequence coverage.</p><p>In paper<strong> II </strong>and <strong>III</strong>, three different enzymes were used for rapid analysis of enantiomeric excess and conversion of O-acylated cyanohydrins synthesized by a defined protocol. Horse liver alcohol dehydrogenase,<em> Candida antarctica</em> lipase<strong> </strong>B<strong> </strong>and pig liver esterase were sequentially added to a solution containing the O-acylated cyanohydrin. Each enzyme caused a drop in absorbance from oxidation of NADH to NAD⁺. The conversion and enantiomeric excess of the sample could be calculated from the relative differences in absorbance.</p>