Bacterial adhesion efficiency on implant abutments: a comparative study

The attachment of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 28213 onto six different materials used to manufacture dental implant abutments was quantitatively determined after 2 and 24 h of contact between the materials and the bacterial cultures. The materials were topographically characterized and their wettability determined, with both parameters subsequently related to bacterial adhesion. Atomic force microscopy, interferometry, and contact angle measurement were used to characterize the materials" surfaces. The results showed that neither roughness nor nano-roughness greatly influenced bacterial attachment whereas wettability strongly correlated with adhesion. After 2 h the degree of E. coli attachment markedly differed depending on the material whereas similar differences were not observed for S. aureus, which yielded consistently higher counts of adhered cells. Nevertheless, after 24 h the adhesion of the two species to the different test materials no longer significantly differed, although on all surfaces the numbers of finally adhered E. coli were higher than those of S. aureus

Interferometría SAR en las islas Shetlend del Sur: modelo numérico de elevaciones de la Isla Decepción

La interferometría con imágenes de radar de apertura sintética (SAR: Synthetic Aperture Radar) desde satélite es una técnica que permite obtener información altimétrica del relieve terrestre, siendo especialmente útil en áreas remotas. Este trabajo muestra una aplicación de esta técnica en las islas Shetland del Sur (Antártida). Para ello se han utilizado imágenes SAR obtenidas por los satélites ERS (European Remote Sensing) de la Agencia Espacial Europea (ESA: European Space Agency) y un proceso interferométrico desarrollado entre el Departament de Geodinàmica i Geofísica de la Universitat de Barcelona y el Institut Cartogràfic de Catalunya.

A digital holography technique for generating beams with arbitrary polarization and shape

A method for generating beams with arbitrary polarization and shape is proposed. Our design requires the use of a Mach-Zehnder set-up combined with translucent liquid crystal displays in each arm of the interferometer; in this way, independent manipulation of each transverse beam components is possible. The target of this communication is to develop a numerical procedure for calculating the holograms required for dynamically encode any amplitude value and polarization state in each point of the wavefront. Several examples demonstrating the capabilities of the method are provided.

One-sided jet at milliarcsecond scales in LS I +61˚ 303

We present Very Long Baseline Interferometry (VLBI) observations of the high mass X-ray binary LS I +61˚303, carried out with the European VLBI Network (EVN). Over the 11 hour observing run, performed ~10 days after a radio outburst, the radio source showed a constant flux density, which allowed sensitive imaging of the emission distribution. The structure in the map shows a clear extension to the southeast. Comparing our data with previous VLBI observations we interpret the extension as a collimated radio jet as found in several other X-ray binaries. Assuming that the structure is the result of an expansion that started at the onset of the outburst, we derive an apparent expansion velocity of 0:003 c, which, in the context of Doppler boosting, corresponds to an intrinsic velocity of at least 0:4 c for an ejection close to the line of sight. From the apparent velocity in all available epochs we are able to establish variations in the ejection angle which imply a precessing accretion disk. Finally we point out that LS I +61˚303, like SS 433 and Cygnus X-1, shows evidence for an emission region almostorthogonal to the relativistic jet

Multifrequency VLBI Observations of Selected Active Galactic Nuclei

In this dissertation, active galactic nuclei (AGN) are discussed, as they are seen with the high-resolution radio-astronomical technique called Very Long Baseline Interferometry (VLBI). This observational technique provides very high angular resolution (_ 10−300 = 1 milliarcsecond). VLBI observations, performed at different radio frequencies (multi-frequency VLBI), allow to penetrate deep into the core of an AGN to reveal an otherwise obscured inner part of the jet and the vicinity of the AGN’s central engine. Multi-frequency VLBI data are used to scrutinize the structure and evolution of the jet, as well as the distribution of the polarized emission. These data can help to derive the properties of the plasma and the magnetic field, and to provide constraints to the jet composition and the parameters of emission mechanisms. Also VLBI data can be used for testing the possible physical processes in the jet by comparing observational results with results of numerical simulations. The work presented in this thesis contributes to different aspects of AGN physics studies, as well as to the methodology of VLBI data reduction. In particular, Paper I reports evidence of optical and radio emission of AGN coming from the same region in the inner jet. This result was obtained via simultaneous observations of linear polarization in the optical and in radio using VLBI technique of a sample of AGN. Papers II and III describe, in detail, the jet kinematics of the blazar 0716+714, based on multi-frequency data, and reveal a peculiar kinematic pattern: plasma in the inner jet appears to move substantially faster that that in the large-scale jet. This peculiarity is explained by the jet bending, in Paper III. Also, Paper III presents a test of the new imaging technique for VLBI data, the Generalized Maximum Entropy Method (GMEM), with the observed (not simulated) data and compares its results with the conventional imaging. Papers IV and V report the results of observations of the circularly polarized (CP) emission in AGN at small spatial scales. In particular, Paper IV presents values of the core CP for 41 AGN at 15, 22 and 43 GHz, obtained with the help of the standard Gain transfer (GT) method, which was previously developed by D. Homan and J.Wardle for the calibration of multi-source VLBI observations. This method was developed for long multi-source observations, when many AGN are observed in a single VLBI run. In contrast, in Paper V, an attempt is made to apply the GT method to single-source VLBI observations. In such observations, the object list would include only a few sources: a target source and two or three calibrators, and it lasts much shorter than the multi-source experiment. For the CP calibration of a single-source observation, it is necessary to have a source with zero or known CP as one of the calibrators. If the archival observations included such a source to the list of calibrators, the GT could also be used for the archival data, increasing a list of known AGN with the CP at small spatial scale. Paper V contains also calculation of contributions of different sourced of errors to the uncertainty of the final result, and presents the first results for the blazar 0716+714.

Electrophysiological measurements of spectral sensitivities: a review

Spectral sensitivities of visual systems are specified as the reciprocals of the intensities of light (quantum fluxes) needed at each wavelength to elicit the same criterion amplitude of responses. This review primarily considers the methods that have been developed for electrophysiological determinations of criterion amplitudes of slow-wave responses from single retinal cells. Traditional flash methods can require tedious dark adaptations and may yield erroneous spectral sensitivity curves which are not seen in such modifications as ramp methods. Linear response methods involve interferometry, while constant response methods involve manual or automatic adjustments of continuous illumination to keep response amplitudes constant during spectral scans. In DC or AC computerized constant response methods, feedback to determine intensities at each wavelength is derived from the response amplitudes themselves. Although all but traditional flash methods have greater or lesser abilities to provide on-line determinations of spectral sensitivities, computerized constant response methods are the most satisfactory due to flexibility, speed and maintenance of a constant adaptation level

The effect of lysobisphosphatidic acid (LBPA) on α-tocopherol transfer protein (α-TTP) binding to lipid membranes

The a-tocopherol transfer protein (a-TTP) is responsible for the retention of the atocopherol form of vitamin E in living organisms. The detailed ligand transfer mechanism by a-TTP is still yet to be fully elucidated. To date, studies show that a-TTP transfers a-tocopherol from late endosomes in liver cells to the plasma membrane where it is repackaged into very low density lipoprotein (VLDL) and released into the circulation. Late endosomes have been shown to contain a lipid known as lysobisphosphatidic acid (LBP A) that is unique to this cellular compartment. LBPA plays a role in intracellular trafficking and controlling membrane curvature. Taking these observations into account plus the fact that certain proteins are recruited to membranes based on membrane curvature, the specific aim of this project was to examine the effect of LBP A on a-TTP binding to lipid membranes. To achieve this objective, dual polarization interferometry (DPI) and a vesicle binding assay were employed. Whilst DPI allows protein binding affinity to be measured on a flat lipid surface, the vesicle binding assay determines protein binding affinity to lipid vesicles mimicking curved membranes. DPI analysis revealed that the amount of a-TTP bound to lipid membranes is higher when LBPA is present. Using the vesicle binding assay, a similar result was seen where a greater amount of protein is bound to large unilamellar vesicles (LUV s) containing LBP A. However, the effect of LBP A was attenuated when small unilamellar vesicles (SUVs) were replaced with LUVs. The outcome of this project suggests that aTTP binding to membranes is influenced by membrane curvature, which in turn is induced by the presence of LBP A.

Mechanism of tocopherol transfer by human α-tocopherol transfer protein (α-hTTP)

Vitamin E is a well known fat soluble chain breaking antioxidant. It is a general tenn used to describe a family of eight stereoisomers of tocopherols. Selective retention of a-tocopherol in the human circulation system is regulated by the a -Tocopherol Transfer Protein (a-TIP). Using a fluorescently labelled a-tocopherol (NBD-a-Toc) synthesized in our laboratory, a fluorescence resonance energy transfer (FRET) assay was developed to monitor the kinetics of ligand transfer by a-hTTP in lipid vesicles. Preliminary results implied that NBD-a-Toe simply diffused from 6-His-a-hTTP to acceptor membranes since the kinetics of transfer were not responsive to a variety of conditions tested. After a series of trouble shooting experiments, we identified a minor contaminant, E coli. outer membrane porin F (OmpF) that co-purified with 6-His-a-hTTP from the metal affinity column as the source of the problem. In order to completely avoid OmpF contamination, a GST -a-hTTP fusion protein was purified from a glutathione agarose column followed by an on-column thrombin digestion to remove the GST tag. We then demonstrated that a-hTTP utilizes a collisional mechanism to deliver its ligand. Furthennore, a higher rate of a-tocopherol transfer to small unilamellar vesicles (SUV s) versus large unilamellar vesicles (LUV s) indicated that transfer is sensitive to membrane curvature. These findings suggest that ahTTP mediated a-Toc transfer is dominated by the hydrophobic nature of a-hTTP and the packing density of phospholipid head groups within acceptor membranes. Based on the calculated free energy change (dG) when a protein is transferred from water to the lipid bilayer, a model was generated to predict the orientation of a-hTTP when it interacts with lipid membranes. Guided by this model, several hydrophobic residues expected to penetrate deeply into the bilayer hydrophobic core, were mutated to either aspartate or alanine. Utilizing dual polarization interferometry and size exclusion vesicle binding assays, we identified the key residues for membrane binding to be F 165, F 169 and 1202. In addition, the rates of ligand transfer of the u-TTP mutants were directly correlated to their membrane binding capabilities, indicating that membrane binding was likely the rate limiting step in u-TTP mediated transfer of u-Toc. The propensity of u-TTP for highly curved membrane provides a connection to its colocalization with u-Toc in late endosomes.

The Ligand and Membrane-binding Behaviour of the Phosphatidylinositol Transfer Proteins (PITPa & PITPb)

Human Class I phosphatidylinositol transfer proteins (PITPs) exists in two forms: PITPα and PITPβ. PITPs are believed to be lipid transfer proteins based on their capacity to transfer either phosphatidylinositol (PI) or phosphatidylcholine (PC) between membrane compartments in vitro. In Drosophila, the PITP domain is found to be part of a multi-domain protein named retinal degeneration B (RdgBα). The PITP domain of RdgBα shares 40 % sequence identity with PITPα and has been shown to possess PI and PC binding and transfer activity. The detailed molecular mechanism of ligand transfer by the human PITPs and the Drosophila PITP domain remains to be fully established. Here, we investigated the membrane interactions of these proteins using dual polarization interferometry (DPI). DPI is a technique that measures protein binding affinity to a flat immobilized lipid bilayer. In addition, we also measured how quickly these proteins transfer their ligands to lipid vesicles using a fluorescence resonance energy transfer (FRET)-based assay. DPI investigations suggest that PITPβ had a two-fold higher affinity for membranes compared to PITPα. This was reflected by a four-fold faster ligand transfer rate for PITPβ in comparison to PITPα as determined by the FRET assay. Interestingly, DPI analysis also demonstrated that PI-bound human PITPs have lower membrane affinity compared to PC-bound PITPs. In addition, the FRET studies demonstrated the significance of membrane curvature in the ligand transfer rate of PITPs. The ligand transfer rate was higher when the accepting vesicles were highly curved. Furthermore, when the accepting vesicles contained phosphatidic acid (PA) which have smaller head groups, the transfer rate increased. In contrast, when the accepting vesicles contained phosphoinositides which have larger head groups, the transfer rate was diminished. However, PI, the favorite ligand of PITPs, or the presence of anionic lipids did not appear to influence the ligand transfer rate of PITPs. Both DPI and FRET examinations revealed that the PITP domain of RdgBα was able to bind to membranes. However, the RdgBα PITP domain appears to be a poor binder and transporter of PC.

Role of Phosphorylation of the Oxysterol Binding Protein (OSBP) in (PI(4)P) and Sterol-Containing Membrane Recognition and Binding

Studies have demonstrated that the oxysterol binding protein (OSBP) acts as a phosphatidylinositol phosphate (PIP)-sterol exchanger at membrane contact sites (MCS) of the endoplasmic reticulum (ER) and Golgi. OSBP is known to pick up phosphatidylinositol-4-phosphate (PI(4)P) from the ER, transfer it to the trans-Golgi in exchange for a cholesterol molecule that is then transferred from the trans-Golgi to the ER. Upon further examination of this pathway by Ridgway et al. (1), it appeared that phosphorylation of OSBP played a role in the localization of OSBP. The dephosphorylation state of OSBP was linked to Golgi localization and the depletion of cholesterol at the ER. To mimic the phosphorylated state of OSBP, the mutant OSBP-S5E was designed by Ridgway et al. (1). The lipid and sterol recognition by wt-OSBP and its phosphomimic mutant OSBP-S5E were investigated using immobilized lipid bilayers and dual polarization interferometry (DPI). DPI is a technique in which the protein binding affinity to immobilized lipid bilayers is measured and the binding behavior is examined through real time. Lipid bilayers containing 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and varying concentrations of PI(4)Ps or sterols (cholesterol or 25-hydroxycholesterol) were immobilized on a silicon nitride chip. It was determined that wt-OSBP binds differently to PI(4)P-containing bilayers compared to OSBP-S5E. The binding behavior suggested that wt-OSBP extracts PI(4)P and the change in the binding behavior, in the case of OSBP-S5E, suggested that the phosphorylation of OSBP may prevent the recognition and/or extraction of PI(4)P. In the presence of sterols, the overall binding behavior of OSBP, regardless of phosphorylation state, was fairly similar. The maximum specific bound mass of OSBP to sterols did not differ as the concentration of sterols increased. However, comparing the maximum specific bound mass of OSBP to cholesterol with oxysterol (25-hydroxycholesterol), OSBP displayed nearly a 2-fold increase in bound mass. With the absence of the wt-OSBP-PI(4)P binding behavior, it can be speculated that the sterols were not extracted. In addition, the binding behavior of OSBP was further tested using a fluorescence based binding assay. Using 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3β-ol (22-NBD cholesterol), wt-OSBP a one site binding dissociation constant Kd, of 15 ± 1.4 nM was determined. OSBP-S5E did not bind to 22-NBD cholesterol and Kd value was not obtained.

Spectro-imagerie optique à faible flux et comparaison de la cinématique Ha et HI d'un échantillon de galaxies proches

Un nouveau contrôleur de EMCCD (Electron multiplying Charge Coupled Device) est présenté. Il permet de diminuer significativement le bruit qui domine lorsque la puce EMCCD est utilisé pour du comptage de photons: le bruit d'injection de charge. À l'aide de ce contrôleur, une caméra EMCCD scientifique a été construite, caractérisée en laboratoire et testée à l'observatoire du mont Mégantic. Cette nouvelle caméra permet, entre autres, de réaliser des observations de la cinématique des galaxies par spectroscopie de champ intégral par interférométrie de Fabry-Perot en lumière Ha beaucoup plus rapidement, ou de galaxies de plus faible luminosité, que les caméras à comptage de photon basées sur des tubes amplificateurs. Le temps d'intégration nécessaire à l'obtention d'un rapport signal sur bruit donné est environ 4 fois moindre qu'avec les anciennes caméras. Les applications d'un tel appareil d'imagerie sont nombreuses: photométrie rapide et faible flux, spectroscopie à haute résolution spectrale et temporelle, imagerie limitée par la diffraction à partir de télescopes terrestres (lucky imaging), etc. D'un point de vue technique, la caméra est dominée par le bruit de Poisson pour les flux lumineux supérieurs à 0.002 photon/pixel/image. D'un autre côté, la raie d'hydrogène neutre (HI) à 21 cm a souvent été utilisée pour étudier la cinématique des galaxies. L'hydrogène neutre a l'avantage de se retrouver en quantité détectable au-delà du disque optique des galaxies. Cependant, la résolution spatiale de ces observations est moindre que leurs équivalents réalisés en lumière visible. Lors de la comparaison des données HI, avec des données à plus haute résolution, certaines différences étaient simplement attribuées à la faible résolution des observations HI. Le projet THINGS (The HI Nearby Galaxy Survey a observé plusieurs galaxies de l'échantillon SINGS (Spitzer Infrared Nearby Galaxies Survey). Les données cinématiques du projet THIGNS seront comparées aux données cinématiques obtenues en lumière Ha, afin de déterminer si la seule différence de résolution spatiale peut expliquer les différences observées. Les résultats montrent que des différences intrinsèques aux traceurs utilisées (hydrogène neutre ou ionisé), sont responsables de dissemblances importantes. La compréhension de ces particularités est importante: la distribution de la matière sombre, dérivée de la rotation des galaxies, est un test de certains modèles cosmologiques.

Développement et caractérisation de sources de neutres réactifs pour l’étude des interactions plasmas-surfaces

L’objectif de ce mémoire de maîtrise est de développer et de caractériser diverses sources de neutres réactifs destinées à des études fondamentales des interactions plasmas-surfaces. Ce projet s’inscrit dans le cadre d’une vaste étude de la physique des interactions plasmas-parois mises en jeu dans les procédés de gravure par plasma des matériaux de pointe. Une revue de la littérature scientifique sur les diverses méthodes permettant de générer des faisceaux de neutres réactifs nous a permis de sélectionner deux types de sources. La première, une source pyrolitique, a été caractérisée par spectrométrie de masse en utilisant le C2F6 comme molécule mère. Nous avons montré que le C2F6 était dissocié à plus de 90% à 1000ºC et qu’il formait du CF4, lui-même dissocié en CF2 vers 900ºC. Ces résultats ont été validés à l’aide d’un modèle basé sur des calculs d’équilibres chimiques, qui a aussi prédit la formation de F à 1500ºC. La seconde source, un plasma entretenu par une onde électromagnétique de surfaces, a été caractérisée par spectroscopie optique d’émission et par interférométrie haute fréquence. Dans le cas du plasma d’argon créé par un champ électromagnétique (>GHz), nos travaux ont révélé une distribution en énergie des électrons à trois températures avec Te-low>Te-high

Analyse cinématique de l'hydrogène ionisé et étude du gaz ionisé diffus de trois galaxies du Groupe Sculpteur : NGC253, NGC300 et NGC247

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

A study of the large-scale structure of the wind of WR 134

Diverses méthodes ont été utilisées pour étudier les étoiles Wolf-Rayet (WR) dans le but de comprendre les phénomènes physiques variés qui prennent place dans leur vent dense. Pour étudier la variabilité qui n'est pas strictement périodique et ayant des caractéristiques différentes d'une époque à l'autre, il faut observer pendant des périodes de temps suffisamment longues en adopter un échantillonnage temporel élevé pour être en mesure d'identifier les phénomènes physiques sous-jacents. À l'été 2013, des astronomes professionnels et amateurs du monde entier ont contribué à une campagne d'observation de 4 mois, principalement en spectroscopie, mais aussi en photométrie, polarimétrie et en interférométrie, pour observer les 3 premières étoiles Wolf-Rayet découvertes: WR 134 (WN6b), WR 135 (WC8) et WR 137 (WC7pd + O9). Chacune de ces étoiles est intéressante à sa manière, chacune présentant une variété différente de structures dans son vent. Les données spectroscopiques de cette campagne ont été réduites et analysées pour l'étoile présumée simple WR 134 pour mieux comprendre le comportement de sa variabilité périodique à long terme dans le cadre d'une étude des régions d'interactions en corotation (CIRs) qui se retrouvent dans son vent. Les résultats de cette étude sont présentés dans ce mémoire.

Investigations on nanosecond laser produced plasma in air from the multi-component material YBa2Cu3O7

The laser produced plasma from the multi-component target YBa2CU3O7 was analyzed using Michelson interferometry and time resolved emission spectroscopy. The interaction of 10 ns pulses of 1.06 mum radiation from a Q-switched Nd:YAG laser at laser power densities ranging from 0.55 GW cm-2 to 1.5 GW cm-2 has been studied. Time resolved spectral measurements of the plasma evolution show distinct features at different points in its temporal history. For a time duration of less than 55 ns after the laser pulse (for a typical laser power density of 0.8 GW cm-2, the emission spectrum is dominated by black-body radiation. During cooling after 55 ns the spectral emission consists mainly of neutral and ionic species. Line averaged electron densities were deduced from interferometric line intensity measurements at various laser power densities. Plasma electron densities are of the order of 1017 cm-3 and the plasma temperature at the core region is about 1 eV. The measurement of plasma emission line intensities of various ions inside the plasma gave evidence of multiphoton ionization of the elements constituting the target at low laser power densities. At higher laser power densities the ionization mechanism is collision dominated. For elements such as nitrogen present outside the target, ionization is due to collisions only.