Les microorganismes occupent une place centrale dans la diversité du vivant et les processus écosystémiques, notamment dans le sol où ils sont en interaction avec les plantes. Cette thèse vise à caractériser l'influence respective des plantes et du contexte abiotique dans la distribution spatiale des microorganismes. Le travail s'appuie sur un modèle simplifié de la biologie des sols, les plantes en coussins des falaises de haute montagne. Nous avons suivi une seule espèce aux morphotypes variés, Silene acaulis, une espèce ingénieure de l'écosystème dont la croissance mène à la création d'un sol de novo. L'échantillonnage comprend le prélèvement de sol de plante et de sol extérieur comme témoin, pour des coussins distribués le long de gradients altitudinaux et géologiques. Des méthodes moléculaires ont été utilisées pour décrire la diversité microbienne et le génotypage des coussins.Les coussins structurent la beta diversité bactérienne et fongique à l'échelle régionale en agissant comme un tampon à échelle locale sur les effets de la roche mère et de l'altitude en homogénéisant le pH et par un apport de nutriments. Cet effet ingénieur est d'autant plus fort que la contrainte abiotique augmente et varie selon le génotype des coussins. La beta diversité bactérienne diffère de la beta diversité fongique. Alors que les communautés bactériennes sont sensibles au pH du sol et convergent sous les coussins, les communautés fongiques sont corrélées à la génétique des coussins, particulièrement les clades aux modes de vie biotrophes/pathogènes. Ce travail montre que les plantes sont un filtre biotique majeur de la biogéographie microbienne. / Microorganisms are key component of Hearth biodiversity and ecosystem processes, especially in soils where they interact with plants. The objectives of the PhD was to caracterize the plant and abiotic respective influence on microbial spatial distribution. The work was based on a simplified soil biology model, the alpine cushion plants. We choose one species composed of variable morphotype, Silene acaulis,can ecosystem engineer species that creates de novo soil through growth. Sampling design includes soil within cushions and outside, spanning altitudinal and geological gradients. Molecular approachs were used to describe diversity and to genotype cushions.Cushions structures bacterial and fungal regional beta diversity through a local buffering of the influence of abiotic context, homogeneizing soil pH and by nutrient supply. This engineering effect increased in stressful conditions and varied according to plant genotype. Betadiversity differed between bacteria and fungi. Bacterial communities are mainly influenced by pH and converge within cushions while fungal communities correlate to cushion genetic, especially plant-associated biotrophs fungal clades. This work shows that plants act as a major biotic filter on microbial biogeography.

Die Analyse des Standes der Technik zu Simulationsmethoden im Werkzeugentwicklungsprozess zeigt die breiten Bestrebungen ganzheitliche Modelle zur Abbildung der Maschinen- und Werkzeugeigenschaften in der Prozesssimulation zu schaffen um die Werkzeuginbetriebnahme in Zukunft in die virtuelle Welt überführen zu können. Dabei werden die Einflüsse wie die Stößelkippung unter Last, die Durchbiegung der Werkzeugaufspannplatten, die Verformung des Ziehkissens und die elastische Nachgiebigkeit der Werkzeuge separat betrachtet. Es konnte kein Simulationsmodell identifiziert werden, welche alle relevanten Eigenschaften der Werkzeuge und der Maschinen zu einem Gesamtmodell vereint. Zudem fehlten Modellierungsansätze für wesentliche Teilaspekte einer ganzheitlichen Abbildung der Wechselwirkungen von Maschine, Werkzeug und Prozess. Insbesondere ist dabei die systematische Untersuchung zur Abbildung des Pinolentragbildes sowohl örtlich als auch über dem Pressenhub und der versteifende Einfluss der Werkzeugführung zu nennen. Mit der vorliegenden Arbeit konnte ein ganzheitliches FE-Prozessmodell mit den wesentlichen elastischen Eigenschaften der Maschine und der Werkzeuge erstellt werden. Wichtig ist dabei insbesondere die systematische Abbildung relevanter mechanischer Ziehkissenstrukturen als Ersatzmodell und die Integration der tatsächlichen Ziehkissenkraft im Prozessmodell. Bekannte Modellierungsansätze wie z.B. die Abbildung der Kippsteifigkeit des Stößelsystems und die Berücksichtigung der Durchbiegung der Werkzeugaufspannplatten wurden weiterentwickelt und in das Gesamtmodell implementiert. Für jedes Ersatzmodell wurde eine Strategie zur Modellerstellung und zur zweckmäßigen Parametrierung mit dem Ziel eines minimalen Zeit- und Kostenaufwandes vorgeschlagen. Die elastische Modellierung der Werkzeuge wurde mit einer systematischen Vorgehensweise hinterlegt und zeigt bei konsequenter Anwendung signifikant reduzierte Berechnungszeiten. Die Berücksichtigung der Ziehstäbe erfolgt in dieser Arbeit aufgrund der Einschränkungen des verwendeten FE-Solvers als Strukturmodell. Zusätzlich wurde ein Modellierungsansatz vorgestellt, welcher die analytische Berücksichtigung der Ziehstäbe auf elastischen Werkzeugstrukturen ermöglicht. Damit steht jetzt ein Prozessmodell mit den relevanten Maschinen- und Werkzeugeigenschaften zur Verfügung, welches durch eine einfache Parametrierung auf die jeweilige Prozess-Maschinen-Konfiguration angepasst werden kann. Dies ermöglicht eine deutlich genauere Abbildung des Pinolentragbildes, der daraus resultierenden Niederhalterdruckverteilung und damit des Materialeinzuges in die Matrize. Dieses erweiterte Prozessmodell ist damit in der Lage eine virtuelle Inbetriebnahme der Werkzeuge zu unterstützen und in Zukunft eine weitere Verkürzung des Inbetriebnahmeprozesses zu ermöglichen.:I. Inhaltsverzeichnis I II. Formelzeichen und Abkürzungen V II.1. Formelzeichen V II.2. Abkürzungen VI 1 Einführung 2 2 Stand der Technik 4 2.1 Modellierung des Blechwerkstoffes 6 2.1.1 Strukturelle und funktionale Abbildung 6 2.1.2 Ermittlung der Parameterwerte 9 2.2 Reibungsverhältnisse in der Kontaktfuge 10 2.2.1 Strukturelle und funktionale Abbildung 11 2.2.2 Ermittlung der Parameterwerte 11 2.3 Karosserieziehwerkzeuge 12 2.3.1 Prinzip und Bedeutung für den Umformprozess 12 2.3.2 Wechselwirkungen im Gesamtsystem Presse-Werkzeug-Prozess 15 2.3.3 Strukturelle und funktionale Abbildung in der FE-Prozesssimulation 16 2.3.4 Ermittlung der Parameterwerte 18 2.4 Pressen der Blechumformung 18 2.4.1 Prinzip und Bedeutung für den Umformprozess 18 2.4.2 Wechselwirkungen im Gesamtsystem Presse-Werkzeug-Prozess 22 2.4.3 Strukturelle und funktionale Abbildung der Maschine in der Prozesssimulation 26 2.4.4 Ermittlung von Pressenparameter 30 2.5 Werkzeugentwicklung und -inbetriebnahme 33 2.5.1 Werkzeuginbetriebnahme 34 2.5.2 Simulationsmethoden zur Unterstützung der Werkzeugeinarbeitung 35 2.6 Defizite in der FE-Prozesssimulation des Karosserieziehprozesses 36 3 Zielstellung, Abgrenzung und Vorgehensweise 38 4 Modellerweiterung Werkzeug 40 4.1 Simulationsumgebung 40 4.2 Virtuelles Testwerkzeug 40 4.3 Elastische Werkzeugstruktur 43 4.4 Werkzeuganbindung an umliegende Strukturen 48 4.4.1 Matrize und Stempel 48 4.4.2 Niederhalter 48 4.4.3 Abbildung der Ziehstäbe auf elastisch modellierten Werkzeugen 50 4.5 Die Werkzeugführung 54 4.5.1 Analytische Berechnung der Wirkung der Werkzeugführung 54 4.5.2 Messtechnische Berücksichtigung 56 4.5.3 Bewertung der Modellierungsansätze 56 4.6 Zwischenfazit zur Modellerweiterung der Werkzeuge 58 5 Modellerweiterung Maschine 60 5.1 Messung der Stößel- und Tischdurchbiegung 60 5.2 Ersatzmodellierung und Parametrierung 61 5.3 Überlagerung von Stößelkippung und -durchbiegung 64 5.4 Berücksichtigung der Tischdurchbiegung 67 5.5 Modellstrukturen zur Berücksichtigung der ZK-Mechanik 67 5.6 Kippelastischer Modellierungsansatz 70 5.6.1 1-Punktziehkissen mit Führung 70 5.6.2 2- und 4-Punktziehkissen 72 5.6.3 Ziehkissen mit mehr als 4 Druckpunkten 73 5.7 Elastische Abbildung des Druckkastens 74 5.7.1 Strukturmodell des Druckkastens 74 5.7.2 Ersatzmodell des Druckkastens 76 5.8 Modellstrukturen zur Berücksichtigung der Krafterzeugung 78 5.8.1 Ziehkissenkraft als wegabhängige Größe 79 5.8.2 Ziehkissen-Istkraft als Verhaltensmodell 80 5.8.3 Integration eines reduzierten Anlagenmodells 80 5.9 Ableitung einer Standardparametrierung 84 5.10 Zwischenfazit zu Modellerweiterungen Presse 85 6 Anwendung der Modellerweiterungen auf Versuchsumgebung 88 6.1 Versuchsumgebung 88 6.1.1 Versuchsmaschine 88 6.1.2 Blechwerkstoffe 89 6.1.3 Versuchswerkzeug 91 6.1.4 Messwerterfassung 92 6.2 Modellierung des Blechwerkstoffes 95 6.3 Reibungsverhältnis zwischen Blech und Werkzeug 96 6.4 Modellerweiterungen Werkzeug 98 6.4.1 Modellerstellung 98 6.4.2 Einflussanalyse der elastischen Werkzeugeigenschaften 99 6.5 Modellerweiterung Maschine 100 6.5.1 Berücksichtigung der elastischen Eigenschaften des Stößelsystems 100 6.5.2 Berücksichtigung der Ziehkissenmechanik 105 6.5.3 Validierung der erweiterten FE-Prozessmodelle 109 6.6 Modellbewertung anhand Grenzformänderungsdiagramme 112 6.7 Modellierung der Regelabweichungen der Ziehkissenzylinderkräfte 113 6.8 Anwendung der Standardparametrierung 116 7 Anwendungsbeispiele für die Werkzeugentwicklung 118 7.1 Werkzeugkonstruktion am Beispiel der Fertigungstoleranzen 118 7.2 Werkzeuginbetriebnahme 120 7.2.1 Ermittlung der Zylindersollkräfte auf Mehrpunktziehkissen 121 7.2.2 Virtuelles Tuschieren der Werkzeughälften 123 8 Zusammenfassung 126 9 Literatur 128 III. Anhang 136

This work reports on the successful formation of supported multicomponent lipid bilayer membranes (sLBMs) from natural occurring lipids as well as synthetic lipids on a set of polymer cushions consisting of alternating maleic acid copolymers. Maleic acid copolymers provide a versatile platform to adjust the physico-chemical behaviour by the choice of the comonomer unit. The formation of sLBMs was triggered by a transient reduction of the electrostatic repulsion between the polymer cushions and the lipid vesicles by lowering the solutions pH to 4. Upon formation the stability of sLBMs was not affected by subsequent variations of the environmental pH to 7.2. Even drastic changes in the environmental pH (between pH 2 and pH 9) did not lead to delamination and proved the stability of the polymer sLBM. The degree of hydrophilicity and swelling of the anionic polymer cushions was found to determine both the kinetics of the membrane formation and the mobility of the lipid bilayer with lipid diffusion coefficients in the range from 0.26 to 2.6 µm2 s-1. An increase in cushion hydrophilicity correlated with a strong increase in the diffusion coefficient of the lipids. This trend was found to correlate with the kinetics of bilayer formation in the process of vesicle spreading. The observations strongly support the important role of the support’s polarity for the fluidity of the sLBM, which is probably related to the presence of a water layer between support and bilayer. The investigated polymer cushions are considered to open new options for the in situ modulation of lipid bilayer membranes characteristics to match the requirements for the successful integration of functional transmembrane proteins (TMPs). As each cushion exhibits different physico-chemical properties, the resulting behaviour of the sLBMs and TMPs could be exactly adjusted to the specific requirements of biological samples. This is exemplarily shown by the integration of the TMP beta amyloid precursor protein cleaving enzyme (BACE). Integrated BACE was observed to be mobile on all polymer cushions. On the contrary, no lateral mobility of BACE was found in solid sLBM. Furthermore, the activity of integrated BACE was analysed by the cleavage of an amyloid precursor protein analogue. Remarkably, the polymer cushions did not only enhance the mobility but were also found to increase the activity of BACE by a factor of 1.5 to 2.5 in comparison to solid sLBM. From the obtained results it is obvious that even small cytoplasmic domains of transmembrane proteins might not be preserved upon the integration in silica sLBM. The observed beneficial effects of the utilised polymer cushions on the mobility and activity of transmembrane proteins motivate further studies to clarify the general applicability of the polymer platform. Altogether, this polymer platform provides valuable options to form sLBM with varying characteristics to reconstitute transmembrane proteins for a wide range of possible future applications in biology. / Die vorliegende Arbeit beschreibt die Bildung von polymer unterstützten Lipiddoppelschichten zur Integration von transmembranen Proteinen. Das Polymerkissensystem besteht aus alternierenden Maleinsäurecopolymeren. Lipiddoppelschichten wurden durch die Steuerung der elektrostatischen Repulsion erzeugt: die Verringerung des pH-Wertes auf 4 wurde eine Erhöhung der adsorbierten Vesikelmenge auf den Polymeroberflächen induziert. Nach der erfolgten Bildung der Lipiddoppelschichten kann der pH-Wert beliebig variiert werden, ohne dass die Stabilität der Lipiddoppelschichten beeinflusst wird. Auch drastische Veränderungen des pH-Milieus (pH 2 - pH 9) führten zu keinen Veränderungen in der Membranintegrität. Der Grad der Hydrophilie und der Quellung der anionischen Polymerschichten beeinflusst sowohl die Bildung der Modellmembranen als auch die Mobilität der integrierten Lipidmoleküle. Dabei reichen die erzielten Lipiddiffusionskoeffizienten von 0.26 bis 2.6 µm2 s-1. Dabei ist die Mobilität direkt von der Hydrophilie des Substrates abhängig. Die beobachteten Ergebnisse zeigen deutlich die entscheidende Rolle der Polarität der verwendeten Substratoberflächen auf die Lipidmobilität, die sehr wahrscheinlich mit der Präsenz einer variablen Wasserschicht zusammenhängt. Die untersuchten Polymerkissen eröffnen neue Möglichkeiten für die insitu Modulierung der Charakteristika von Lipidschichten, um funktionale transmembrane Proteine zu integrieren. Aufgrund der unterschiedlichen physiko-chemischen Eigenschaften kann das Verhalten der Lipidschichten und der transmembranen Proteine nach den spezifischen Anforderungen des Modellsystems angepasst werden. Die funktionale Integration wurde am Beispiel des transmembranen Proteins BACE nachempfunden. Die Mobilität des integrierten BACE wurde auf allen Polymerkissen beobachtet. Im Gegensatz dazu wurde auf harten Substraten keine BACE Mobilität gefunden. Die Aktivität des integrierten BACE wurde durch die enzymatische Spaltung eines APP-Analogons nachgewiesen. Bemerkenswerteweise wurde ein Anstieg der BACE Aktivität auf den Polymerkissen um den Faktor 1,5 bis 2,5 im Vergleich zu den auf harten Substraten integrierten BACE beobachtet. Zusammenfassend, die verwendeten Polymerkissen bieten vielfältige Möglichkeiten Lipidschichten mit variierenden Eigenschaften für die Integration von transmembranen Proteinen zu erzeugen.

A Computational Fluid Dynamics (CFD) tool is developed in this thesis to solve complex fluid-structure interaction (FSI) problems. The fluid domain is based on Smoothed Particle Hydro-dynamics (SPH) and the structural domain employs large-deformation Finite Element Method (FEM). Validation tests of SPH and FEM are first performed individually. A loosely-coupled SPH-FEM model is then proposed for solving FSI problems. Validation results of two benchmark FSI problems are illustrated (Antoci et al., 2007; Souto-Iglesias et al., 2008). The first test case is flow in a sloshing tank interacting with an elastic body and the second one is dam-break flow through an elastic gate. The results obtained with the SPH-FEM model show good agreement with published results and suggest that the SPH-FEM model is a viable and effective numerical tool for FSI problems. This research is then applied to simulate a two-dimensional free-stream flow interacting with a deformable, pressurized surface, such as an ACV/SES bow seal. The dynamics of deformable surfaces such as the skirt/seal systems of the ACV/SES utilize the large-deformation FEM model. The fluid part including the air inside the chamber and water are simulated by SPH. A validation case is performed to investigate the application of SPH-FEM model in ACV/SES via comparison with experimental data (Zalek and Doctors, 2010). The thesis provides the theory of the SPH and FEM models incorporated and the derivation of the loosely-coupled SPH-FEM model. The validation results have suggested that this SPH-FEM model can be readily applied to skirt/seal dynamics of ACV/SES interacting with free-surface flow. / Ph. D.

<div>This body of work examines analytical and numerical models to simulate the response of structures in shock absorption applications. Specifically, the work examines the prediction of cushion curves of polymer foams, and a topological examination of a $\chi$ shape unit cell found in architected mechanical elastomeric metamaterials. The $\chi$ unit cell exhibits the same effective stress-strain relationship as a closed cell polymer foam. Polymer foams are commonly used in the protective packaging of fragile products. Cushion curves are used within the packaging industry to characterize a foam's impact performance. These curves are two-dimensional representations of the deceleration of an impacting mass versus static stress. The main drawback with cushion curves is that they are currently generated from an exhaustive set of experimental test data. This work examines modeling the shock response using a continuous rod approximation with a given impact velocity in order to generate cushion curves without the need of extensive testing. In examining the $\chi$ unit cell, this work focuses on the effects of topological changes on constitutive behavior and shock absorbing performance. Particular emphasis is placed on developing models to predict the onset of regions of quasi-zero-modulus (QZM), the length of the QZM region and the cushion curve produced by impacting the unit cell. The unit cell's topology is reduced to examining a characteristic angle, defining the internal geometry with the cell, and examining the effects of changing this angle.</div><div>However, the characteristic angle cannot be increased without tradeoffs; the cell's effective constitutive behavior evolves from long regions to shortened regions of quasi-zero modulus. Finally, this work shows that the basic $\chi$ unit cell can be tessellated to produce a nearly equivalent force deflection relationship in two directions. The analysis and results in this work can be viewed as new framework in analyzing programmable elastomeric metamaterials that exhibit this type of nonlinear behavior for shock absorption.</div>

Trafikinducerade vibrationer: En studie om busskuddars påverkan på vibrationsnivåer i mark och byggnader. Erika Johansson Kling I Uppsala används busskuddar som en form av medveten ojämnhet i vägbanan för att sänka hastigheten och öka trafiksäkerheten på olycksdrabbade vägsträckor. Boende i bostäder belägna nära busskuddar har dock rapporterat att de ibland kan uppleva vibrationer kopplade till då tunga fordon passerar busskuddar som störande. Denna studie syftade till att kvantifiera vibrationsnivåer i mark och byggnader nära busskuddar samt utreda centrala faktorer som kan påverka vibrationsnivån och vibrationsutbredningen. Genom vibrationsmätningar i fält på fyra platser i Uppsala samlades data in och analyserades utifrån angivna frågeställningar. Resultatet bekräftade det som litteraturstudien belyste, att det är svårt att på ett generellt sätt beskriva vibrationsutbredningen och att den tydligt är platsspecifik. Vid samtliga undersöka platser kunde dock konstateras att vibrationsnivån i marken tenderar att vara en faktor mellan 2,5–32 högre då ett tungt fordon passerar en busskudde, jämfört med då det passerar en opåverkad referenspunkt längs med samma väg. Generellt avtog dessutom vibrationsnivån i marken med ökat avstånd från källan. Vilken vibrationsled som dominerade varierade dock både mellan platserna och för olika avstånd från källan. Vid en av de fyra mätplatserna förstärktes vibrationsnivån i byggnaden, jämfört med i marken utanför. Vid samma mätplats överskreds dessutom känseltröskeln inomhus, och en skyddsvibrationsnivå på 0,5 mm/s kan behöva tillämpas vid busskudden för att förhindra detta. Mätningarna visade att både vibrationer med låga och höga frekvenser kan uppstå i marken nära busskudden då tunga fordon passerar. Dock var det enbart vibrationer med låga frekvenser som spreds vidare i marken vid ett ökat avstånd från källan. Studien visade dessutom att det kan krävas ett skyddsavstånd mellan busskudde och byggnad på cirka 100 meter för att känseltröskeln inte ska riskera att överskridas inomhus. Trots att ett sådant avstånd är platsspecifikt, belyser det vilka krav det skulle kunna ställa på samhällsbyggnaden i ett växande Uppsala. Det faktum att busskuddar kan förhöja vibrationsnivån i marken vid passage av tunga fordon ter sig något märkligt, då busskuddarna ska vara utformade på ett sätt som tillåter tunga fordon att passera relativt obehindrat. Fler studier kring busskuddarnas utformning och/eller grundläggning är därför av stor relevans. Det är vidare relevant att exempelvis utreda hur fordonens hastighet över busskuddar påverkar vibrationsnivån i marken, då litteraturstudien belyser att det främst är då tunga fordon passerar ojämnheter med hög hastighet som betydande vibrationer kan uppstå. Busskuddar anses vara en av de mest effektiva hastighetssänkande åtgärderna, men studien visar att det kan ske på bekostnad av ökade vibrationsnivåer. En grundläggande helhetsbedömning bör därför göras av busskuddarnas lämplighet vid kommande stadsplanering. Nyckelord: markvibration, vibrationsmätning, busskudde, trafik, samhällsbyggnad. Institutionen för geovetenskaper, Uppsala Universitet, Villavägen 16, SE-752 36 Uppsala. ISSN 1401–5765. / Traffic induced vibrations: A study on the effect of speed control cushions on vibration levels in surrounding land and buildings. Erika Johansson Kling In Uppsala, speed control cushions are used as a form of conscious unevenness in the road surface to reduce speed and increase road safety on accident-affected road sections. However, housing in residences located near speed control cushions have reported that they can sometimes experience vibrations linked to when heavy vehicles pass speed control cushions as disturbing. This study aimed to quantify vibration levels in land and buildings near speed control cushions and to investigate key factors that can affect the vibration level and vibration propagation. Through vibration measurements at four places in Uppsala, data was collected and analyzed based on the stated research questions. The result confirmed what the literature study highlighted, that it is difficult to describe the propagation of vibrations in a general way and that it is clearly site-specific. However, it was found at all the measurement sites that the vibration level in the ground tends to be a factor between 2.5 and 32 higher when a heavy vehicle passes a speed control cushion, compared to when it passes an unaffected reference point along the same road. Generally, the vibration level in the ground also decreased with increased distance from the source. However, the level of vibration that dominated varied between the locations and for different distances from the source. At one measurement site, the vibration level in the building was amplified compared to in the ground outside. At the same measurement site, the sensing threshold was also exceeded indoors, and a protection vibration level of 0.5 mm/s may have to be applied nearby the speed control cushions to prevent this from happening. The measurements showed that both vibrations with low and high frequencies can occur in the ground near the speed control cushions when heavy vehicles pass. However, it was only vibrations with low frequencies that propagated further in the ground at an increased distance from the source. The study also showed that a safety distance between speed control cushions and buildings of approximately 100 meters may be required in order for the sensitivity threshold not to be exceeded indoors. Although such a distance is sitespecific, it highlights what requirements it could put on the urban planning in a growing Uppsala. The fact that speed control cushions can increase the level of vibration in the ground when heavy vehicles pass seems somewhat strange, since the speed control cushions are supposed to be designed in a way that allows heavy vehicles to pass relatively unobstructed. Additional studies on the design and/or foundation of the speed control cushions are therefore of great relevance. It is furthermore relevant to investigate, for example, how the speed of the vehicles when passing over speed control cushions affects the vibration level in the ground, since the literature study highlights that it is primarily when heavy vehicles pass an unevenness in the road surface at high speed that significant vibrations can occur. Speed control cushions are considered to be one of the most effective speed reducing measures, but the study shows that they may lead to increased vibration levels. An overall 3 evaluation should therefore be made regarding the suitability of speed control cushions in future urban planning. Keywords: ground vibration, vibration measurement, speed control cushion, traffic, urban planning. Department of Earth Sciences, Uppsala University, Villavägen 16, SE-752 36 Uppsala. ISSN 1401–5765.

Artificial lipid bilayers formed on solid surface supports are widespread model systems to study physical, chemical, as well as biological aspects of cell membranes and fundamental interfacial interactions. The approach to use a thin polymer film representing a cushion for lipid bilayers prevents incorporated membrane proteins from pinning to the support and mimics the native environment of a lipid bilayer in certain aspects of the extracellular matrix and intracellular structures. A key component for cell anchorage to extracellular fibronectin is the transmembrane adhesion receptor alpha(5)beta(1) integrin. Its transport dynamics and clustering behavior plays a major role in the assembly of focal adhesions, which mediate mechanical forces and biochemical signals of cells with their surrounding. The system investigated herein is envisioned to use extrinsically controlled stimuli-responsive polymer cushions to tune the frictional drag between polymer cushion and mobile membranes with incorporated integrins to actively regulate lipid membrane characteristics. To attain this goal, a temperature- and pH-responsive polymer based on poly(N-isopropylacrylamide) copolymers containing varying amounts of carboxyl-group-terminated comonomers at different aliphatic spacer lengths (PNIPAAm-co-carboxyAAM) was surface-grafted to a poly(glycidyl methacrylate) anchorage layer. The swelling transitions were characterized using atomic force microscopy, ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D) and found to be tunable over a wide range of temperature and pH. In agreement with the behavior of the polymers in solution, longer alkyl spacers decreased the phase transition temperature T(P) and higher contents of carboxylic acid terminated comonomers increased T(P) at alkaline conditions and decreased T(P) at acidic conditions. Remarkably, the point where the degree of carboxyl group deprotonation balances the T(P)-lowering effect of the alkyl spacer was distinctive for each alkyl spacer length. These findings illustrate how the local and global balance of hydrophilic and hydrophobic interactions along the copolymer chain allows to adjust the swelling transition to temperatures below, comparable, or above those observed for PNIPAAm homopolymers. Additionally, it could be shown that surface-grafting leads to a decrease in T(P) for PNIPAAm homopolymers (7°C) and copolymers (5°C - 10°C). The main reason is the increase in local polymer concentration of the swollen film constrained by dense surface anchorage in comparison to the behavior of dilute free chains in solution. In accordance with the Flory-Huggins theory, T(P) decreases with increasing concentration up to the critical concentration. Biological functionalization of the PNIPAAm-co-carboxyAAm thin films was demonstrated for the cell adhesion ligand peptide cRGD via carbodiimide chemistry to mimic extracellular binding sites for the cell adhesion receptors integrin. The outcome of QCM-D measurements of cRGD-functionalized surfaces showed a maintained stimuli-responsiveness with slight reduction in T(P). A drying/rehydration procedure of a 9:1 lipid mixture of the cationic lipid dioleoyl-trimethylammoniumpropane (DOTAP) and the zwitterionic dioleoyl-phosphatidylcholine (DOPC) was utilized to form lipid bilayer membranes on PNIPAAm-co-carboxyAAM cushions. Fluorescence recovery after photobleaching (FRAP) revealed that lipid mobility was distinctively higher (6.3 - 9.6) µm2 s-1 in comparison to solid glass support ((3.0 - 5.9) µm2 s-1). In contradiction to the initial expectations, modulation of temperature and pH led to poor variations in lipid mobility that did not correlate with the PNIPAAm cushion swelling state. The results suggested a weak coupling of the lipid bilayer with PNIPAAm polymer cushions that can be slightly tuned by electrostatic interactions. The transmembrane adhesion receptor alpha(5)beta(1) integrin was reconstituted into liposomes consisting of DOPC/sphingomyelin/cholesterol 2:2:1 for the formation of polymer cushioned bilayers. PNIPAAm- co-carboxyAAM and maleic acid (MA) copolymers were used as cushions, both with the option for cRGD functionalization. On the MA copolymer cushions, fusion of proteoliposomes resulted in supported bilayers with mobile lipids as confirmed by FRAP. However, incorporated integrins were immobile. In an attempt to explain this observation, the medium-sized cytoplasmic integrin domain was accounted to hamper the movement by steric interactions with the underlying polymer chains in conjunction with electrostatic interactions of the cationic cytoplasmic domain with the oppositely charged MA copolymer. On the PNIPAAm-co-carboxyAAM cushion only a drying/rehydration procedure lead to bilayer formation. However, again the integrins were immobile, presumably due to the harsh treatment during preparation. Nevertheless, the results of the investigated set of PNIPAAm copolymer films suggest their application as temperature- and pH-responsive switchable layers to control interfacial phenomena in bio-systems at different physiological conditions. The PNIPAAm-co-carboxyAAm cushioned bilayer system represents a promising step towards extrinsically controlled membrane – substrate interactions.

[pt] Esta dissertação apresenta resultados de simulação física e numérica do comportamento mecânico de cavidades circulares em meios contínuos. Na simulação numérica foi possível reproduzir o comportamento tensão-deformação registrado nos ensaios. O comportamento mecânico do contínuo foi abordado de duas formas: i) Teoria clássica e ii) Modelo generalizado de Cosserat. A segunda abordagem, por dispor de um grau de liberdade extra, permite a reprodução numérica de algumas feições observadas ao redor das cavidades circulares em testes de laboratório de maneira mais realística. A teoria clássica de contínuo foi associada somente ao modelo constitutivo de Mohr-Coulomb. Já para Cosserat, foram utilizados dois modelos constitutivos: Mohr-Coulomb e Bogdanova-Lippmann Modificado. A motivação para apresentar contínuo generalizado neste trabalho é que o mesmo inclui a parcela referente ao comportamento das partículas. Em todos os testes foram utilizadas amostras do arenito Botucatu, obtidas em São Paulo e Paraná. Para caracterização mecânica deste material foram realizados ensaios uniaxiais, triaxiais e brasileiros. Já a simulação física do comportamento de cavidades circulares foi analisada segundo duas geometrias: cúbica (com aplicação de estado de tensão biaxial) e cilíndrica (TWC – Thick Walled Cylinder). O acompanhamento da ruptura das cavidades cilíndricas foi feito de forma visual (amostras cúbicas) e com monitoramento tomográfico em tempo real (amostras cilíndricas). Com base na observação experimental da ruptura das cavidades cilíndricas e nas simulações numéricas considerando o contínuo clássico e de Cosserat, foi possível verificar que, ambas as abordagens possibilitaram a reprodução das feições observadas. / [en] This work seeks to realize physical and numerical simulation of the mechanical behavior of the wellbore stability for continuum environment.The Continunm s mechanical behavior is approach by two ways: i) Classic Continuum Theory and ii) Cosserat Continuum. On the second approach, the theory allows an extra degree of freedom, which plays an important rule on instabilities and bifurcation problems; this allows a more realistic numerical simulation of the failure mechanism observed on circular cavity. The Classic Continuum Theory is associated to a Mohr-Coulomb constitutive model. On the other hand for Cosserat Theory s applied tow constitutive models: Mohr-Coulomb and Modified Bogdanova-Lippmann.The generalized continuum takes in account the microstructure of the material.It s used on all tests Botucatu s specimens, which were acquired at São Paulo and Paraná. For characterize the rock s behavior it s realized triaxial, uniaxial and brazilian tests. Then the physical simulation of the circular cavity s behavior was analyzed for two geometries: cubic samples (biaxial stress) and cylindric samples (TWC – Thick Walled Cylinder). The failure mechanism of circular cavity was followed visually (cubic samples) and with CT X-Ray in real time (cylindric samples).From the experimental observations of the failure mechanism of circular cavity and numerical simulations, with Classic Continuum and Cosserat, was possible to verify that both approaches reproduce the behavior of the rocks observed on experimental data.

Ce mémoire décrit la conception, la modélisation et la commande d’un manipulateur pneumatique,fondé sur un principe de traction aérodynamique original. De puissants jets d’air verticauxpermettent de créer un flux d’air horizontal pour manipuler des objets sans contact. Les objetssont maintenus en constante lévitation sur la surface grâce à un coussin d’air, et peuvent positionnés selon trois degrés de libert´ du plan, grâce à la combinaison adéquate et distribuéede jets d’air verticaux. Nous détaillons la conception d’un prototype original de manipulateurexploitant ce principe fluidique. Ce prototype a été intégré dans une plate-forme expérimentaleafin de valider le principe de manipulation : le système permet de déplacer des objets à unevitesse atteignant 180 mm/s. Nous avons modélisé le fonctionnement de la surface selon plusieursméthodes. Un premier modèle comportemental, fond´e sur des données expérimentales, aété établi. Il permet de simuler l’´evolution de la position d’un objet sur la surface, selon un degréde liberté . Deux modèles de connaissance, fond´es sur une étude aérodynamique fondamentale,donnent l’´evolution de la position de l’objet selon respectivement deux et trois degrés de libertédu plan. Chacun des modèles a été validé expérimentalement. Nous avons synthétisé différentscontrˆoleurs afin d’asservir la position de l’objet : un premier, de type PID, et un second, de typerobuste (méthode H1). La commande de un, puis deux degrés de liberté du système, a permisd’atteindre de bonnes performances : temps de réponse d’environ 2 s et dépassement souventinférieur à 5%. Nous avons également étudié un micro-manipulateur pneumatique permettant ded´eplacer des objets de taille millimétrique selon deux directions, grâce à des jets d’air inclinés.Ces objets peuvent atteindre des vitesses de 123 mm/s. La résolution du positionnement estinférieure à 0.4 μm. / This thesis presents the design, the modeling and the control of a pneumatic manipulatorbased on an original aerodynamic traction principle. An horizontal air flow is induced by strongvertical air jets in order to manipulate objects without contact. The objects are maintained inconstant levitation on an air cushion. Three degrees of freedom positioning of the objects canbe realized thanks to the right combination of distributed air jets. The design of an originalmanipulator using this aerodynamic principle is detailed. The device has been integrated in anexperimental setup in order to validate the manipulation principle : objects can reach velocityof 180 mm/s. Several models of the system have been established. A first model, based on experimentaldata, gives the evolution of the 1 DOF-position of an object on the device. Twoother models, based on a fundamental aerodynamic study, respectively give the evolution of the2- and 3-DOF position of the objet. The three models have been validated experimentally. Inorder to control the position of the object, different controllers have been designed : a PID oneand a robust H1 one. The control of one and two degrees of freedom of the device gives goodperformances : settling time of around 2 s and overshoot less than 5% in most of the cases. Wehave also studied a micro-manipulator that is able to position millimetric sized objects, in twodirections, thanks to inclined air jets. Objects can reach velocity of 123 mm/s, and the resolutionof the positioning is less than 0.4 μm.

Artificial lipid bilayers formed on solid surface supports are widespread model systems to study physical, chemical, as well as biological aspects of cell membranes and fundamental interfacial interactions. The approach to use a thin polymer film representing a cushion for lipid bilayers prevents incorporated membrane proteins from pinning to the support and mimics the native environment of a lipid bilayer in certain aspects of the extracellular matrix and intracellular structures. A key component for cell anchorage to extracellular fibronectin is the transmembrane adhesion receptor alpha(5)beta(1) integrin. Its transport dynamics and clustering behavior plays a major role in the assembly of focal adhesions, which mediate mechanical forces and biochemical signals of cells with their surrounding. The system investigated herein is envisioned to use extrinsically controlled stimuli-responsive polymer cushions to tune the frictional drag between polymer cushion and mobile membranes with incorporated integrins to actively regulate lipid membrane characteristics. To attain this goal, a temperature- and pH-responsive polymer based on poly(N-isopropylacrylamide) copolymers containing varying amounts of carboxyl-group-terminated comonomers at different aliphatic spacer lengths (PNIPAAm-co-carboxyAAM) was surface-grafted to a poly(glycidyl methacrylate) anchorage layer. The swelling transitions were characterized using atomic force microscopy, ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D) and found to be tunable over a wide range of temperature and pH. In agreement with the behavior of the polymers in solution, longer alkyl spacers decreased the phase transition temperature T(P) and higher contents of carboxylic acid terminated comonomers increased T(P) at alkaline conditions and decreased T(P) at acidic conditions. Remarkably, the point where the degree of carboxyl group deprotonation balances the T(P)-lowering effect of the alkyl spacer was distinctive for each alkyl spacer length. These findings illustrate how the local and global balance of hydrophilic and hydrophobic interactions along the copolymer chain allows to adjust the swelling transition to temperatures below, comparable, or above those observed for PNIPAAm homopolymers. Additionally, it could be shown that surface-grafting leads to a decrease in T(P) for PNIPAAm homopolymers (7°C) and copolymers (5°C - 10°C). The main reason is the increase in local polymer concentration of the swollen film constrained by dense surface anchorage in comparison to the behavior of dilute free chains in solution. In accordance with the Flory-Huggins theory, T(P) decreases with increasing concentration up to the critical concentration. Biological functionalization of the PNIPAAm-co-carboxyAAm thin films was demonstrated for the cell adhesion ligand peptide cRGD via carbodiimide chemistry to mimic extracellular binding sites for the cell adhesion receptors integrin. The outcome of QCM-D measurements of cRGD-functionalized surfaces showed a maintained stimuli-responsiveness with slight reduction in T(P). A drying/rehydration procedure of a 9:1 lipid mixture of the cationic lipid dioleoyl-trimethylammoniumpropane (DOTAP) and the zwitterionic dioleoyl-phosphatidylcholine (DOPC) was utilized to form lipid bilayer membranes on PNIPAAm-co-carboxyAAM cushions. Fluorescence recovery after photobleaching (FRAP) revealed that lipid mobility was distinctively higher (6.3 - 9.6) µm2 s-1 in comparison to solid glass support ((3.0 - 5.9) µm2 s-1). In contradiction to the initial expectations, modulation of temperature and pH led to poor variations in lipid mobility that did not correlate with the PNIPAAm cushion swelling state. The results suggested a weak coupling of the lipid bilayer with PNIPAAm polymer cushions that can be slightly tuned by electrostatic interactions. The transmembrane adhesion receptor alpha(5)beta(1) integrin was reconstituted into liposomes consisting of DOPC/sphingomyelin/cholesterol 2:2:1 for the formation of polymer cushioned bilayers. PNIPAAm- co-carboxyAAM and maleic acid (MA) copolymers were used as cushions, both with the option for cRGD functionalization. On the MA copolymer cushions, fusion of proteoliposomes resulted in supported bilayers with mobile lipids as confirmed by FRAP. However, incorporated integrins were immobile. In an attempt to explain this observation, the medium-sized cytoplasmic integrin domain was accounted to hamper the movement by steric interactions with the underlying polymer chains in conjunction with electrostatic interactions of the cationic cytoplasmic domain with the oppositely charged MA copolymer. On the PNIPAAm-co-carboxyAAM cushion only a drying/rehydration procedure lead to bilayer formation. However, again the integrins were immobile, presumably due to the harsh treatment during preparation. Nevertheless, the results of the investigated set of PNIPAAm copolymer films suggest their application as temperature- and pH-responsive switchable layers to control interfacial phenomena in bio-systems at different physiological conditions. The PNIPAAm-co-carboxyAAm cushioned bilayer system represents a promising step towards extrinsically controlled membrane – substrate interactions.