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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
501

The agglomeration of fine iron particles in a fluidised bed cascade

Blundell, Daniel Laurence. January 2005 (has links)
Thesis (Ph.D.)--University of Wollongong, 2005. / Typescript. Includes bibliographical references: p. 198-203.
502

Efeito in vitro de vernizes fluoretados contendo NaF e TiF4 nas propriedades nanomecânicas e de superfície do esmalte dentário submetido à erosão

Medeiros, Maria Isabel Dantas de 17 February 2014 (has links)
Made available in DSpace on 2015-05-14T12:56:02Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1634368 bytes, checksum: 15995bdf9c8fa3e54e6dbaeeb501003d (MD5) Previous issue date: 2014-02-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Objectives: To analyze the effect of NaF and TiF4 varnishes on nanohardness (N) and elastic modulus (E) of enamel surface after very short exposure time to simulation of drinking a can of soft drink; and evaluate the thickness and topography of protective layer formed, by atomic force microscopy (AFM). Methods: Thirty blocks of human enamel (4x4 mm) were divided into 3 groups (n=10): control, NaF varnish and TiF4 varnish. The specimens remained in artificial saliva for 24h, and the varnishes were applied only once. After 6h, specimens were submitted to erosive challenge (10 cycles: 5s in cola drink/5s in artificial saliva). The thickness of protective layer formed and the surface topography were evaluated by optical microscope and AFM. The data were subjected to Two-Way ANOVA, Tukey and Student s-t (α=0.05). Results: The values shown for N and E (GPa) before and after erosive challenge differed statistically, only for control and TiF4 groups. After the erosive challenge, NaF group showed statistically higher values for N and E than those found for control and TiF4 groups. However, the thickness and depth of indentation values showed that N and E values found for TiF4 group were related to the protective layer formed. By AFM, both varnishes showed globular protective layer formation, and the thickness of layer was significantly higher for TiF4 than NaF group. Conclusions: NaF varnish was able to protect the nanomechanical properties of enamel after short erosive challenge, and TiF4 varnish showed a thick and homogeneous protective layer formation with nanomechanical properties. Keywords: tooth erosion; elastic modulus; fluorides; microscopy, atomic force / Objetivos: Analisar o efeito dos vernizes de NaF e TiF4 na nanodureza (N) e módulo de elasticidade (E) da superfície do esmalte após um curto tempo de exposição a bebida a base de cola, simulando a ingestão de uma lata de refrigerante, e também avaliar a espessura e topografia da camada protetora formada, por meio de microscópio de força atômica (AFM). Métodos: Trinta blocos de esmalte humano (4x4 mm) foram divididos em 3 grupos (n = 10): controle (sem verniz), verniz de NaF e verniz de TiF4. As amostras permaneceram em saliva artificial durante 24 horas, e os vernizes foram aplicados apenas uma vez. Após 6h os espécimes foram submetidos a desafio erosivo (10 ciclos: 5s em bebida a base de cola/5s em saliva artificial). A espessura da camada protetora formada e a topografia da superfície foram avaliadas por microscopia óptica e AFM. Os dados foram submetidos à ANOVA Two-Way, Tukey e T- Student (α = 0,05). Resultados: Houve diferença estatística dos valores de N e E (GPa), antes e após o desafio erosivo, apenas para os grupos controle e TiF4. Após o desafio erosivo o grupo NaF mostrou estatisticamente maiores valores de N e E que os grupos controle e TiF4. No entanto, os valores da espessura e profundidade da indentação mostraram que os valores encontrados de N e E para o grupo TiF4 estavam relacionados com a camada protetora formada e não com o esmalte erodido. Ambos os vernizes mostraram, por meio de AFM, a formação de uma camada protetora globular, e a espessura da camada formada foi significativamente maior para o grupo TiF4 que NaF. O verniz de NaF foi capaz de proteger as propriedades nanomecânicas de esmalte após curto desafio erosivo e o verniz de TiF4 apresentou formação de camada protetora espessa e homogênea com propriedades nanomecânicas . Conclusões: O verniz de NaF foi capaz de proteger as propriedades nanomecânicas de esmalte após curto desafio erosivo e o verniz de TiF4 apresentou formação de camada protetora espessa e homogênea com propriedades nanomecânicas.
503

Hétérogénéité des membranes lipidiques et propriétés mécaniques : des bicouches modèles aux membranes des globules gras du lait / Heterogeneity of biological membranes and mechanical properties : lipid bilayers model of the milk fat globules membranes

Etthakafy, Oumaima 25 October 2017 (has links)
Les globules gras du lait sont entourés d’une membrane biologique extrêmement complexe en composition et en structure, appelée MFGM (milk fat globule membrane). L’investigation de cette membrane, in situ dans le lait, par microscopie confocale nous suggère que les lipides polaires à haute température de transition de phase (Tm) forment des domaines en phase gel ou liquide ordonné, dispersés dans une phase continue fluide. Sur la base de cette observation, ce projet vise à comprendre en quoi la composition en lipides polaires laitiers et leur état de phase peuvent moduler les propriétés élastiques de la MFGM, en vue d’une meilleure maîtrise de la stabilité des globules gras en industrie laitière.L’hétérogénéité mécanique générée par la coexistence de différents types de phase a ainsi été caractérisée par spectroscopie de force AFM en utilisant des bicouches de lipides modèles de la membrane réelle, à basse (T<Tm) et haute températures (T>Tm). Pour analyser finement les déterminants de l’élasticité de la membrane, et tenir compte de la courbure, une étude approfondie des effets de l’état de phase et de la composition hétérogène en lipides polaires a été entreprise par spectroscopie de force atomique, en complément d’une analyse structurale par microscopie électronique ou diffraction des rayons X. Nous y avons montré, en particulier, que la présence de molécules de longueur de chaîne acyles et d’insaturation variables rend les membranes de sphingomyéline de lait en phase gel moins rigides qu’attendu, bien que significativement plus rigide qu’une membrane fluide. Cette approc / The milk fat globules are enveloped by a biological membrane, called MFGM, of highly complex composition and structure. Investigation of this membrane, in situ in milk, using confocal microscopy suggested that polar lipids with high transition temperature (Tm) form domains in gel or liquid-ordered phase, dispersed in a continuous fluid phase. From this observation, the aim of this project was to understand how the composition and organization of dairy polar lipids can modulate the elastic properties of the MFGM, in order to better control stability of the fat globules in the dairy industry. The mechanical heterogeneity created by the coexistence of phases was then characterized by AFM force spectroscopy using lipid bilayers models at low (T<Tm) and high temperatures (T>Tm).In order to closely analyze the factors that direct membrane elasticity, force spectroscopy measurements were undertaken on curved liposome membranes, in combination with structural characterization by TEM and SAXS. We showed, in particular, that heterogeneity in acyl chain length and unsaturation made gel-phase milk sphingomyelin membranes less rigid than expected, although more rigid than a fluid phase membrane. This approach was finally applied to native milk fat globules, where mechanical heterogeneity was visible. However, elasticity values were somewhat different from those calculated on model systems, probably because of the presence of membrane proteins.
504

A Probing System with Replaceable Tips for Three Dimensional Nano-Metrology

Mrinalini, R Sri Muthu January 2017 (has links) (PDF)
With increase in the number of three dimensional (3-D) nanometer-scale objects that are being either fabricated or studied, there is a need to accurately characterize their geometry. While the Atomic force microscope (AFM) is a versatile tool for performing nano-metrology, it suffers from issues of poor accessibility of 3-D features and inability to measure 3-D forces that limit its applicability in 3-D nano-metrology. This thesis investigates the design and development of a novel probing system based on AFM that improves accessibility and enables direct measurement of 3-D forces acting on the AFM tip. Two approaches are investigated to address the issue of poor accessibility. The first is to develop a novel system that enables in-situ replacement and reuse of specialized AFM tips that improve accessibility, and the second is to design a special AFM tip that can actively re-orient about two independent axes. In order to perform in-situ tip replacement, a liquid meniscus based micro-gripper is developed and integrated on to a conventional AFM probe. The stiffness of the gripper is analyzed and shown to be adequately high along all three axes for AFM imaging to be performed. Tip replacement and re-use are both experimentally demonstrated by employing a novel tip-exchange station. The replaced tips are employed to show artifact-free AFM imaging of a standard calibration grating in both tapping-mode and contact-mode. To actively re-orient a conventional tip, a novel magnetically-actuated micro-scale ball-and-socket joint is integrated onto an AFM probe. The quasi-static behavior of the joint is experimentally characterized, and the ability of the tip to independently re-orient about two axes is demonstrated. The achieved range is about +/- 90 degrees about both X- and Y-axes. In order to realize the potential of the proposed probes for 3-D nano-metrology, an AFM is developed in-house that possesses the capability to make direct measurement of 3-D forces. Optimization of the measurement system to achieve identical sensitivities and resolution along all three axes is studied. Subsequently, the necessary electronics for measurement, actuation and control are developed. All the subsystems are experimentally calibrated and integrated. The overall AFM is shown to have a resolution of about 0.2 nm when operated in tapping-mode. The developed AFM is employed to showcase the following applications: characterization of the coefficient of kinetic friction of Muscovite mica, force controlled nano-scribing on polymethyl methacrylate (PMMA) and tapping-mode imaging of a calibration grating with the developed re-orientable AFM probe. Finally, the unique ability of the re-orientable AFM probe to control its tip-orientation is employed to develop a nanometer-scale coordinate measurement machine (CMM). The developed nano-CMM is shown to access the vertical wall of a sample and obtain its topography.
505

Modeling Mechanisms of Water Affinity and Condensation on Si-based Surfaces via Experiments and Applications

January 2011 (has links)
abstract: Water affinity and condensation on Si-based surfaces is investigated to address the problem of fogging on silicone intraocular lenses (IOL) during cataract surgery, using Si(100), silica (SiO2) and polydimethylsiloxane (PDMS) silicone (SiOC2H6)n. Condensation is described by two step nucleation and growth where roughness controls heterogeneous nucleation of droplets followed by Ostwald ripening. Wetting on hydrophilic surfaces consists of continuous aqueous films while hydrophobic surfaces exhibit fogging with discrete droplets. Si-based surfaces with wavelength above 200 nm exhibit fogging during condensation. Below 200 nm, surfaces are found to wet during condensation. Water affinity of Si-based surfaces is quantified via the surface free energy (SFE) using Sessile drop contact angle analysis, the Young-Dupr&eacute; equation, and Van Oss theory. Topography is analyzed using tapping mode atomic force microscopy (TMAFM). Polymer adsorption and ion beam modification of materials (IBMM) can modify surface topography, composition, and SFE, and alter water affinity of the Si-based surfaces we studied. Wet adsorption of hydroxypropyl methylcellulose (HPMC) C32H60O19 with areal densities ranging from 1018 atom/cm2 to 1019 atom/cm2 characterized via Rutherford backscattering spectrometry (RBS), allows for the substrate to adopt the topography of the HPMC film and its hydrophilic properties. The HPMC surface composition maintains a bulk stoichiometric ratio as confirmed by 4.265 MeV 12C(&alpha;, &alpha;)12C and 3.045 MeV 16O(&alpha;, &alpha;)16O, and 2.8 MeV He++ elastic recoil detection (ERD) of hydrogen. Both PIXE and RBS methods give comparable areal density results of polymer films on Si(100), silica, and PDMS silicone substrates. The SFE and topography of PDMS silicone polymers used for IOLs can also be modified by IBMM. IBMM of HPMC cellulose occurs during IBA as well. Damage curves and ERD are shown to characterize surface desorption accurately during IBMM so that ion beam damage can be accounted for during analysis of polymer areal density and composition. IBMM of Si(100)-SiO2 ordered interfaces also induces changes of SFE, as ions disorder surface atoms. The SFE converges for all surfaces, hydrophobic and hydrophilic, as ions alter electrochemical properties of the surface via atomic and electronic displacements. / Dissertation/Thesis / Ph.D. Physics 2011
506

Développement d'un simulateur haptique pour la cacaractérisation et la microinjection cellulaires / Haptic simulator for cell characterization and microinjection

Ladjal, Hamid 26 May 2010 (has links)
L'objectif fondamental de cette thèse est de développer et de mettre en oeuvre un outil interactif desimulation des techniques de micromanipulation biologiques de cellules. Au moyen de cet outil, l'opérateurpourra se former, s'entraîner et améliorer sa maîtrise en développant une gestuelle proche de celle exécutéeen réalité. La conception d'un tel environnement de simulation en temps-réel nécessite de trouver uncompromis entre le réalisme des modèles de comportement biomécanique utilisés, la précision et la stabilitédes algorithmes des méthodes de résolution et de rendu haptique utilisées ainsi que la vitesse de calcul. Lamodélisation mécanique retenue repose sur l'utilisation du modèle hyperélastique de St Venant-Kirchhoff etune formulation dynamique explicite éléments-finis du type masses-tenseurs. Le bien-fondé de cettemodélisation est vérifié sur des essais de microindentation par Microscopie à Force Atomique (AFM) decellules souches embryonnaires de souris et de microinjection d'ovocytes. Nous avons développé etimplémenté des modèles d'interaction en temps-réel qui s'articulent autour de la détection et la gestionrapide des collisions entre outil/cellule.La synthèse du rendu haptique fourni à l'opérateur est également proposée par l'intermédiaire d'un couplagevirtuel. Pour chaque application, nous avons justifié nos choix méthodologiques et Algorithmiques qui sontguidés par les contraintes de "réalisme+précision" "temps-réel". Les différents modèles proposés ont étéintégrés dans le simulateur SIMIC que nous avons développé pendant cette thèse. Ce dernier est dédié à lasimulation interactive pour l'aide à l'apprentissage du geste de microinjection et de nanoindentationcellulaire. / The fundamental objective of this thesis is to develop and implementing an interactive simulation techniquesfor micromanipulation biological cells. Using this tool, the operator can form, train and improve its control bydeveloping a gesture similar to that performed in reality. The design of such a simulation environment in realtime requires a compromise between the realism of biomechanical models used the accuracy and stability ofalgorithms and solution methods used haptic rendering and computational speed. Modeling Mechanicalrestraint involves the use of hyperelastic model of St Venant-Kirchhoff formulation and explicit dynamic finiteelement-type mass tensors. The validity of this model is tested on microindentation tests by Atomic ForceMicroscopy (AFM) of mouse embryonic stem cells and microinjection of oocytes. We have developed andimplemented models of real-time interaction that revolve around the detection and management of rapidcollisions between tool / cell.The synthesis of the haptic feedback provided to the operator is also available through a virtual coupling. Foreach application, we have justified our methodological choices and Algorithms that are guided by theconstraints of realism + precision "" real time ". The various proposed models have been integrated into thesimulator SIMIC that we developed during this thesis. This is dedicated to interactive simulation to supportlearning of gesture microinjection and cell nanoindentation.
507

Estudo da viscoelasticidade de células de câncer renal por microscopia de força atômica / Viscoelasticity study of kidney cancer cells by atomic force microscopy

Alencar, Luciana Magalhães Rebêlo January 2010 (has links)
ALENCAR, Luciana Magalhães Rebêlo. Estudo da viscoelasticidade de células de câncer renal por microscopia de força atômica. 2010. 155 f. Tese (Doutorado em Física) - Programa de Pós-Graduação em Física, Departamento de Física, Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2010. / Submitted by Edvander Pires (edvanderpires@gmail.com) on 2015-05-22T18:40:21Z No. of bitstreams: 1 2010_tese_lmralencar.pdf: 15264548 bytes, checksum: e3df53eb49036e78f623d089ab7e0995 (MD5) / Approved for entry into archive by Edvander Pires(edvanderpires@gmail.com) on 2015-05-22T18:42:17Z (GMT) No. of bitstreams: 1 2010_tese_lmralencar.pdf: 15264548 bytes, checksum: e3df53eb49036e78f623d089ab7e0995 (MD5) / Made available in DSpace on 2015-05-22T18:42:17Z (GMT). No. of bitstreams: 1 2010_tese_lmralencar.pdf: 15264548 bytes, checksum: e3df53eb49036e78f623d089ab7e0995 (MD5) Previous issue date: 2010 / The mechanical properties of living cells have a crucial role in the accomplishment of their physiological functions. However, our knowledge on this subject is still limited. Is not fully understood how a cell responds structurally and mechanically to an external pressure or as the elasticity of cells is altered in diseased organisms compared to healthy ones. Recently, the biomechanics of cancer cells, in particular the elasticity or stiffness, has been identified as an important factor that is related to function, adhesion, motility, invasion and transformation of the neoplastic cells. Studies in vivo show that cancerous transformations introduce significant changes in the structure and behavior of cells. These differences can cause changes in mechanical properties, often leading to greater cell deformability. Quantifying the change of elasticity using mechanical tests in conjunction with a microscopic examination, can become a powerful method for the diagnosis of cancer, and open new routes for treatments. In this context, Atomic Force Microscopy (AFM) is presented as an ideal tool for cell research due to its high resolution capability for surface nano-manipulation, ability to work in fluids and for being a noninvasive and nondestructive technique. This study investigates the mechanical response of cancer cells (lines A-498 and ACHN), compared to normal cells (RC-124). Using an AFM and its components as a morphological tool of high resolution characterization and characterization of the cells mechanical properties using the AFM probe as a nano-indenter, and from the strength data obtained by the microscope, and appropriate theoretical models to interpret these data to obtain qualitative and quantitative values of the elastic response these cells. / As propriedades mecânicas de células vivas possuem um papel crucial no bom desempenho de suas funções fisiológicas. Porém, nosso conhecimento nesse assunto ainda é limitado. Não é totalmente compreendido como uma célula responde, estrutural e mecanicamente, a uma tensão externa ou como a elasticidade das células altera-se em organismos doentes em comparação a organismos sadios. Recentemente, a biomecânica de células do câncer (em particular, a elasticidade ou rigidez) tem sido apontada como um fator importante que está relacionado à função, adesão, motilidade, transformação e invasão da célula neoplásica. Estudos in vivo mostram que transformações cancerosas introduzem alterações significativas na estrutura e comportamento celular. Essas diferenças também podem causar alterações nas propriedades mecânicas, geralmente levando a uma maior deformabilidade da célula. A quantificação da alteração de elasticidade, utilizando ensaios mecânicos em conjunto com um exame microscópico, pode tornar-se um poderoso diagnóstico do câncer e abrir caminhos para novos tratamentos. Neste contexto, a Microscopia de Força Atômica (AFM) se apresenta como uma ferramenta ideal para a investigação de células por sua alta resolução, capacidade de nano-manipulação de superfícies, possibilidade de trabalhar em meios líquidos e por ser uma técnica não destrutiva. Neste trabalho, propõe-se a investigação da resposta mecânica de células cancerígenas (linhagens A-498 e ACHN), comparando-se com células normais (RC-124), utilizando-se um Microscópio de Força Atômica e seus componentes como ferramentas de caracterização morfológica de alta resolução e caracterização das propriedades mecânicas dessas células. Utilizando a sonda de AFM como nano-indentador e a partir dos dados de força obtidos pelo microscópio, analisados por meio de modelos teóricos adequados, temos por objetivo obter valores qualitativos e quantitativos da resposta elástica dessas células.
508

Resistência de união entre cimentos resinosos autoadesivos e cerâmicas de zircônia policristalina / Bond strengthof self-adhesive resin cements to polycrystalline zirconia ceramics

Rafael Ferrone Andreiuolo 20 February 2014 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo deste estudo foi avaliar a resistência de união a microtração de cimentos resinosos autoadesivos a cerâmicas de zircônia policristalina. Dezoito blocos cerâmicos de zircônia 3Y-TZP (9 LAVA e 9 LAVA Plus) foram jateados com partículas de 50 m de Al2O3por 20 s com pressão de 28 psi a uma distância de 10 mm. Os blocos cerâmicos foram duplicados em resina composta (Point 4, Kerr) por moldagem com silicone. Os blocos de resina composta foram cimentados à superfície jateada da zircônia usando três diferentes cimentos resinosos autoadesivos: (1) RelyX Unicem 2 (3M ESPE); (2) SmartCem 2 (Dentsply); (3) Speedcem (Ivoclar Vivadent). Após 24 h imersos em água destilada a 37oC, os blocos cimentados foram cortados em palitos para testes de microtração,com área da interface adesiva de 1 mm2 0,2 mm, e tensionados até a fratura. Os resultados foram analisados pelo teste de análise de variância de dois fatores e pelo teste de comparações múltiplas LSD (&#945;=0.05). As amostras fraturadas foram analisadas com microscopia eletrônica de varredura (MEV) e o modo de falha foi registrado. A topografia das superfícies cerâmicas antes e após o jateamento foi comparada por microscopia de força atômica (AFM). A resistência de união do cimento Speedcem à zircônia foi estatisticamente superior àquela reportada pelos cimentos RelyX Unicem 2 e SmartCem 2, independentemente da cerâmica usada (p<0,05). O fator cerâmica não teve influência estatística na resistência de união. A interação entre os dois fatores se mostrou significativa (p<0,05). O modo de fratura associado ao SmartCem 2 foi quase exclusivamente adesiva, enquanto oRelyX Unicem 2e o Speedcem exibiram um maior percentual de falhas mistas. Não foram observadas falhas coesivas. O AFM não revelou diferença no padrão de topografia de superfície entre as duas cerâmicas antes ou após o jateamento. Concluiu-se que o cimento Speedcem foi superior na adesão a cerâmicas de zircônia policristalina. / The aim of this study was to evaluate the microtensile bond strength of self-adhesive resin cements to polycrystalline zirconia. Eighteen 3Y-TZP ceramic blocks (9 LAVA and 9 LAVA Plus) were sandblasted with 50 m Al2O3 powder for 20 s with a pressure of 28 psi at a working distance of 10 mm. The ceramic blocks were duplicated in composite resin (Point 4, Kerr) using a silicon mold. Composite blocks were bonded to sandblasted zirconia using different self-adhesive resin cements: (1) RelyX Unicem 2 (3M ESPE); (2) SmartCem 2 (Dentsply); (3) Speedcem (Ivoclar Vivadent). After 24 h immersed in distilled water at 37oC the bonded specimens were cut into microtensile bond sticks of 1 mm2 0,2 mminterface areaand loaded in tension until failure. Data were analyzed by two-way ANOVA and LSDtest for multiple comparisons (&#945;=0.05). The fractured surfaces were observed using a scanning electron microscope (SEM) and the failure mode was recorded. Surface topography of the sandblasted and the as sintered zirconia materials were analyzed under an atomic force microscope (AFM).Bond strength of Speedcem cement to zirconia was significantly higher than that of RelyX Unicem 2 and SmartCem 2, irrespective of the ceramic used (p<0,05). Bond strength was not significantly influenced by the ceramic factor. Interactions were significant (p<0,05). The mode of failure of SmartCem 2 was almost exclusively adhesive, while RelyX Unicem 2 and Speedcem exhibited a good percentage of mixed fractures. No cohesive fractures were observed. AFM did not reveal differences in the topography pattern of the two ceramics before or after sandblasting. It was concluded that Speedcem presented superior adhesion to polycrystalline zirconia ceramics.
509

DNA double-strand break repair studied by atomic force microscopy

Zabolotnaya, Ekaterina January 2018 (has links)
DNA double-strand breaks (DSBs), where both strands of the DNA duplex are simultaneously fractured, are considered the most lethal type of DNA damage. The conserved Mre11-Rad50 DNA repair complex enables the catalytic activities of the Mre11 nuclease and the Rad50 ATPase to function together to coordinate the recognition and processing of DSBs prior to the recruitment of long-range end-resection machinery required to trigger the DSB repair by the homologous recombination (HR) pathway. Fast-scan atomic force microscopy (AFM) in fluid conditions was primarily used to explore the architectural arrangement, DNA binding and processing machinery of the Mre11-Rad50 complex from the thermophilic crenarchaeote Sulfolobus acidocaldarius. The structural analysis identified four distinct architectural arrangements and demonstrates the key role of the Rad50 zinc hooks in the oligomerisation of the complex. AFM imaging showed a dynamic and Velcro-like interplay between Mre11-Rad50 protein complexes and the DNA double-helix using the Rad50 coiled-coils in a novel mode of DNA binding. The complex appears to use the Rad50 zinc hook region to bind to and track along dsDNA for broken DNA-terminals. Furthermore, the present study shows that this archaeal complex can drive extensive ATP-dependent unwinding of DNA templates. It is the first time that such unwinding has been observed in a single molecule study. These observations reveal novel activities leading to the proposal of a new model for Mre11-Rad50 action during DSB repair. AFM was also used to visualise the structure and activity of the HerA-NurA protein complex, which has been predicted to combine the activity of the NurA nuclease and hexameric HerA-translocase to generate long single-stranded DNA overhangs essential for DSB repair by HR in archaea. The present data verify and clarify the presumed biological role of this complex. Overall, the present study provides new insights into the initial steps of DNA DSB repair by the HR pathway and, most importantly, the detection of the broken ends.
510

Loading of dendrimer nanoparticles into layer-by-layer assembled Poly(diallyl dimethyl ammonium) chloride-(Poly(acrylic acid))n Multilayer Films : Particle Electrokinetics, Film Structure Dynamics and Elasticity / Chargement de nanoparticules de dendrimères en films multicouches du type (poly(diallyldiméthylammonium)chlorure-poly(acide acrylique))n : électrocinétique des particules, élasticité et dynamique de la structure des films

Moussa, Mariam 04 December 2017 (has links)
Une analyse détaillée des propriétés physico-chimiques des nanoparticules (NP) anthropogéniques est nécessaire pour comprendre à un niveau mécanistique leurs interactions/toxicité potentielle avec/envers les composants biotiques des systèmes aquatiques naturels. Une telle analyse est également requise pour réaliser une évaluation complète et une optimisation de la performance des méthodes d’(ultra)filtration développées pour circonscrire le relargage des NPs dans les milieux aquatiques. Dans ce contexte, l'objectif de cette thèse de doctorat était de déchiffrer les processus physico-chimiques fondamentaux régissant la capture de nanodendrimères carboxylés (PAMAM-COOH) - utilisés fréquemment dans des applications biomédicales – par des films multicouches du type (poly(diallyldiméthylammonium)chlorure-poly(acide acrylique))n ((PDADMAC-PAA)n) assemblés par déposition séquentielle des composantes polymériques cationique et anionique. À cette fin, une étude systématique des propriétés électrohydrodynamiques des NPs PAMAM-COOH a d'abord été effectuée en fonction du pH et de la concentration en sel monovalent du milieu. Sur la base de la théorie électrocinétique de particules molles ayant une fonctionnalité zwitterionique, il est démontré que les caractéristiques électriques interfaciales des NPs considérées sont déterminées à la fois par des contributions électrostatiques de surface et volumique des nanoparticules, lesquelles dependent de l’extension intraparticulaire de la double couche électrique. L’existence de ces deux types de contributions conduit à un changement remarquable de signe de la mobilité des NPs en modifiant la concentration du sel monovalent en solution et à une dépendance prononcé du point de zéro mobilité des NPs avec la concentration de l’électrolyte. En outre, une confrontation quantitative entre résultats expérimentaux et théorie souligne comment les modifications structurales des NPs induites par des changements de pH et de salinité affectent les caractéristiques électrocinétiques des dendrimères. Dans une deuxième partie, la structure, la morphologie et les propriétés mécaniques des films PDADMAC-PAA et leur évolution temporelle dans des conditions de vieillissement naturel ou après traitement thermique ont été déterminées par microscopie à force atomique (AFM) et analyses microspectroscopie Raman. Les résultats démontrent que les films multicouches PDADMAC-PAA de type exponentiel présentent des caractéristiques mécaniques et structurelles typiques de films polyélectrolytes multicouches à croissance linéaire. En particulier, leur relaxation lente vers un état d’équilibre est accélérée après traitement thermique à 60°C et se révèle être intimement liée à l'instabilité de domaines de films riches en PDADMAC, épuisés en eau (faits confirmés par la théorie de la fonctionnelle de la densité) et marqués par la présence de structures caractéristiques en forme de ‘donuts’. Dans une dernière partie, des résultats préliminaires sont donnés pour la dépendance de l'élasticité des films multicouches PDADMAC-PAA avec la concentration en solution de nanodendrimères. Les résultats suggèrent que ces films multicouches complexes constituent une option prometteuse pour la capture et l'élimination de nanodendrimères carboxylés présents en milieux aqueux / A detailed analysis of the physicochemical properties of engineered nanoparticles (NPs) is required to understand on a mechanistic level their interactions/potential toxicity with/towards biotic components of fresh water systems. Such an analysis is further mandatory to achieve a comprehensive evaluation and optimisation of the performance of (ultra)filtration methods developed to prevent NPs release into aquatic media. Within this context, the aim of this PhD thesis was to decipher the basic physico-chemical processes governing the loading of carboxylated-poly(amidoamine) (PAMAM-COOH) nanodendrimers -commonly employed in biomedical applications- into layer-by-layer assembled (poly(diallyl dimethyl ammonium) chloride-poly(acrylic acid))n ((PDADMAC-PAA)n) multilayer films. For that purpose, a systematic investigation of the electrohydrodynamic properties of PAMAM-COOH NPs was first performed as a function of pH and monovalent salt concentration in solution. On the basis of advanced electrokinetic theory for soft particles with zwitterionic functionality, it is demonstrated that the interfacial electrostatic features of the considered NPs are determined both by surface and bulk particle contributions to an extent that depends on electrolyte concentration. This leads to a remarkable NPs mobility reversal with changing monovalent salt concentration and to a marked dependence of the point of zero NPs mobility on electrolyte content. In addition, confrontation between experiments and theory further highlights how pH- and salt-mediated modifications of the NP particle structure affect dendrimer electrokinetic features at large pH and/or low salt concentrations. In a second part, the structure, morphology and mechanical properties of PDADMAC-PAA films, and their evolution over time under natural aging conditions or after thermal treatment, were addressed from atomic force microscopy (AFM) and Raman microspectroscopy analyses. Results evidence that PDADMAC-PAA multilayer films of exponential type exhibit mechanical and structural features that are typical for polyelectrolyte multilayer films with linear growth. In particular, their slow relaxation to equilibrium is accelerated after heating treatment at 60°C and, in line with density functional theory computation, this relaxation dynamics is shown to be intimately connected to instability of film domains rich in PDADMAC, depleted in water and marked by the presence of characteristic donut-like structures. In a final part, the reported dependence of PDADMAC-PAA multilayer films elasticity on concentration of nanodendrimers in bulk solution suggests that these complex multilayer films constitute a promising option to be further investigated for the loading and removal of carboxylated nanodendrimers from aqueous environments

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