Les nanoparticules polymériques fluorescentes apparaissent comme des outils importants pour l'imagerie en temps réel des processus biologiques au niveau moléculaire et cellulaire. L’objectif de mon projet de doctorat a été d’optimiser les nanoparticules polymériques fluorescentes pour l’imagerie biologique. Premièrement, nous avons pu, en faisant varier la chimie des polymères, obtenir un très bon contrôle de leur taille. Ceci a permis de mettre en évidence l’importance de la taille des NPs pour des applications intracellulaires avec une taille maximale de 23 nm pour une distribution dans tout le cytosol. Deuxièmement, nous avons pu montrer que la simple adsorption d’un amphiphile PEGylé de type Pluronic permet la stabilisation des nanoparticules dans des milieux biologiques. Le nombre de molécules incorporées et leur stabilité ont été étudiés en combinant des techniques de FRET et de FCS. Les meilleures formulations résultent en une stabilité des nanoparticules in vivo, ce qui a permis leur imagerie en tant que particules individuelles dans les vaisseaux sanguins du cerveau de souris. Troisièmement, le transfert d’énergie entre différents fluorophores encapsulés dans les NPs a été étudié et optimisé. / Fluorescent polymeric nanoparticles appear as important tools for real-time imaging of biological processes at the molecular and cellular level. The objective of my PhD project was to optimize fluorescent polymeric nanoparticles for biological imaging. First, by varying the chemistry of the polymers, we have been able to obtain a very good control of their size. This made it possible to highlight the importance of NPs size for intracellular applications with a maximum size of 23 nm for optimal distribution throughout the cytosol. Secondly, we have shown that simple adsorption of a PEGylated amphiphiles pluronic family allows the stabilization of nanoparticles in biological media. The number of incorporated molecules and their stability has been studied by combining FRET and FCS techniques. The best formulations result in nanoparticle stability in vivo, which allowed their imaging as individual particles in the blood vessels of the mouse brain. Third, energy transfer among different fluorophores encapsulated in NPs has been studied and optimized.

Nos travaux ont porté sur le développement d'une méthode contrôlée de complexation de polyélectrolytes. La complexation est un processus spontané, sous contrôle cinétique et irréversible dans le cas de polysaccharides tels que le chitosane et les polysulfates, essentiellement le sulfate de dextrane ou l'héparine. Une conséquence de ce contrôle cinétique est que l'obtention d'objets de taille colloïdale requiert de travailler à fortes dilutions. De plus, les nanovecteurs obtenus ne sont pas toujours compatibles avec des conditions d'utilisation dans des milieux physiologiques. Le contrôle de l'association de polysaccharides se fait par écrantage des interactions électrostatiques attractives en présence de chlorure de sodium à la concentration au moins égale à 2 mol.L-1. L'élimination du sel par dialyse induit la formation d'hydrogels dont les caractéristiques et les propriétés dépendent principalement du rapport de charges n+/n- et de la cinétique d'élimination du sel. Ainsi, l'on peut former des hydrogels massifs ou des systèmes dispersés à des concentrations en polymères jusqu'à 30 fois plus élevées que par les méthodes sous contrôle cinétique. De plus, cette technologie permet l'encapsulation des principes actifs dans les particules qui peuvent aussi être fonctionnalisées par des biomolécules d'adressage. Le résultat majeur de ce travail réside en la maîtrise des associations entre polysaccharides de charges opposées, permettant d'obtenir des systèmes colloïdaux et massifs à fort potentiels d'applications biomédicales / Our work dealt with the development of a controlled method of polyelectrolyte complexation. The complexation is a spontaneous process, under kinetic control and irreversible in the case of polysaccharides such as the chitosan and polysulfates, essentially dextran sulfate or heparin. A consequence of this kinetic control is the requirement to work at high dilution to obtain objects of colloidal size. Moreover, the obtained nanovectors were not always adapted for use in physiological media. The control of the association of polysaccharides was achieved by screening the attractive electrostatic interactions in the presence of sodium chloride at concentration at least equal to 2 mol. L-1. Removal of salt by dialysis resulted in the formation of hydrogels, whose characteristics and properties depended mainly on the charge ratio n +/ n- and the kinetics of the salt elimination. Thus, massive or dispersed hydrogels were formed at polymer concentrations up to 30 times higher than by the methods under kinetic control. Furthermore, this technology allowed the encapsulation of active ingredients in the particles that could also be functionalized with biomolecules for targeting. The major result of this work was the control over the associations between oppositely charged polysaccharides which provided colloidal and massive systems of high potentialities in biomedical applications

International Telemetering Conference Proceedings / October 18-21, 2004 / Town & Country Resort, San Diego, California / In the past few years, an evolution has been occurring in test range network topologies. With the proliferation of IP-based networks at the desktop, range officers are seeking ways to extend IP-based networks to the test range, to derive the cost and operational benefits offered with IP technology. This transition is not without its own set of problems. The operational transition from the traditional, ATM-based ranges to IP-based ranges must be addressed. In many cases, it is desired to maintain the ATM range, and add IP capabilities as time and budget permits. The net result is that frequently a mixed protocol network emerges. Terawave Communications has been developing telemetry transport solutions for both ATM and IP-based networks, along with technology to enable convergence of additional services such as video, voice, and data across test ranges. Terawave has developed a solution for various network topologies from ATM-only and IP-only to mixed protocol implementations, which supports end-to-end interworking of telemetry, video, and additional services over mixed protocol networks. In this paper, Terawave will detail the implementation decisions made in the course of application development, and share a framework for enabling seamless intra- and inter- range communication of telemetry and mixed services.

Cancer metastasis directly accounts for an estimated 90% of all cancer related deaths and is correlated with the presence of malignant cells in systemic circulation. This observed relationship has prompted efforts to develop a fluid biopsy, with the goal of detecting these rare cells in patient peripheral blood as surrogate markers for metastatic disease as a partial replacement or supplement to tissue biopsies. Numerous platforms have been designed, yet these have generally failed to support a reliable fluid biopsy due to poor performance parameters such as low throughput, low purity of enriched antigen positive cells, and insufficiently low detection thresholds to detect poor expressed surface markers of target cell populations. This work describes the development of a rapid cell sorting technology called Antigen Specific Lysis (ASL) based on photo-crosslinked polymer encapsulation to isolate tumor cells in suspension. In the first study, we characterize the chemical and structural properties of the surface-initiated polymer films formed directly on mammalian cell surfaces. Coated populations are shown to remain highly viable after coating formation. Biomolecular transport is examined though film coatings on cellular substrates using fluorescent, time-resolved confocal microscopy and diffusivity estimates are generated for these materials. In the next study, a lysis-based cell isolation platform is described in which marker positive cells can be specifically coated in a heterogeneous cell suspension. Anionic surfactants lyse virtually 100% of uncoated cells while fully encapsulated cells remain protected, and are then easily collected by centrifugation. We report that purified cells are released from polymeric coatings to yield viable and functional populations. We monitor cell response throughout the isolation process by multiple techniques, and report viability >80% after the sorting process. Lastly, we examine the response of process yield on the level of photoinitiator loading on target populations. Streptavidin-fluorochrome loading was quantitatively assessed on a panel of markers, both epithelial and mesenchymal, on representative model breast and lung cancer cells. We report that ASL is fundamentally capable of achieving 50-60% yield which is promising for fluid biopsy applications. Finally, both EpCAM and metastatic targeting strategies are then compared to covalently biotinylated samples to inform future robust targeting strategies.

ITC/USA 2015 Conference Proceedings / The Fifty-First Annual International Telemetering Conference and Technical Exhibition / October 26-29, 2015 / Bally's Hotel & Convention Center, Las Vegas, NV / There is great potential in using Best Source Selectors (BSS) to improve link availability in aeronautical telemetry applications. While the general notion that diverse data sources can be used to construct a consolidated stream of "better" data is well founded, there is no standardized means of determining the quality of the data streams being merged together. Absent this uniform quality data, the BSS has no analytically sound way of knowing which streams are better, or best. This problem is further exacerbated when one imagines that multiple vendors are developing data quality estimation schemes, with no standard definition of how to measure data quality. In this paper, we present measured performance for a specific Data Quality Metric (DQM) implementation, demonstrating that the signals present in the demodulator can be used to quickly and accurately measure the data quality, and we propose test methods for calibrating DQM over a wide variety of channel impairments. We also propose an efficient means of encapsulating this DQM information with the data, to simplify processing by the BSS. This work leads toward a potential standardization that would allow data quality estimators and best source selectors from multiple vendors to interoperate.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: A new class of nanocolorants is described for the use as a colorant in hot-melt ink application for ink-jet printing technology. An inverse miniemulsion polymerization process was utilized successfully as a one-step encapsulation process to encapsulate the highly hydrophilic water-soluble fluorescent Rhodamine B dye (RhB) by the hydrophilic water-soluble poly(acrylamide) (PAAm). Three types of Rhodamine B-based nanocolorants, PAAm/RhB, crosslinked-PAAm/RhB and poly(AAm-co-Sty)/RhB, were synthesized using inverse miniemulsion polymerization. The PAAm/RhB nanocolorants exhibited solid dark nanoparticles morphology, while crosslinked-PAAm/RhB and poly(AAm-co-Sty)/RhB showed a core-shell type of morphology. The nanocolorants showed improved light and dye migration fastness as well as high thermal stability, especially, nanocolorants with core-shell morphology. The synthesis of polymerizable RhB-based nanocolorants is described. Poly(AAm-co-RhB) nanocolorants were successfully synthesized for the first time via inverse miniemulsion polymerization. RhB dye was first functionalized by esterification reaction to introduce an acrylate polymerizable group. The RhB-acrylate dye was copolymerized with AAm monomer in an inverse miniemulsion polymerization to produce nanocolorants with superior light and migration fastness. Crosslinked-poly(AAm-co-RhB) nanocolorants could be obtained based on the incorporation of a crosslinking agent. Poly(AAm-co-RhB) and crosslinked-poly(AAm-co-RhB) nanocolorants exhibited a morphology of dark solid and core-shell particles, respectively. In both nanocolorants, the RhB-acrylate dye was completely integrated by copolymerization into the polymer matrix, and by that, the dye migration was completely supressed. Both poly(AAm-co-RhB) and crosslinked-poly(AAm-co-RhB) nanocolorants showed high thermal stability as well as high Tg values. The syntheses of PAAm/RhB nanocolorants-based solid inks were carried out successfully via inverse miniemulsion polymerization. An in situ inverse miniemulsion polymerization, with the paraffin wax as the organic phase, was utilized in making a crosslinked-PAAm/RhB nanocolorants-based solid ink. A crosslinked-poly(AAm-co-RhB) nanocolorants-based solid ink was prepared by the direct mixing of the readymade crosslinked-PAAm/RhB nanocolorants (suspended in cyclohexane) with paraffin wax at temperature above the melting temperature of the wax until all the cyclohexane evaporated. The obtained solid inks appeared as a solid homogenous waxy material with a deep bright colour reflecting that the nanocolorants were well dispersed in the wax. DSC thermograms showed that the solid inks have one sharp melting transition indicating the applicability of our nanocolorants for hot-melt ink applications. / AFRIKAANSE OPSOMMING: ‘n Nuwe reeks nonokleursels word beskryf vir die gebruik in ink-smelt drukker tegnologie. Inverse minie-emulsie polymerisasie was suksesvol gebruik om die kleurstof Rhodamine B (RhB) in die water oplosbare poly(akrielamied) (PAAm) te enkapsuleer. Die roete is gebruik om drie tipes kleurstof te produseer. Elk van die kleurstowwe was gebaseer op Rhodamine B en ‘n PAAm, naamlik PAAm/RhB, kruisgebonde PAAm/RhB en poli(akrielamied-ko-stireen)/RhB. PAAm/RhB nanokleursel was in die vorm van soliede donker nanopartikels. Die kruisgebonde PAAm/RhB en poli(akrielamied-ko-stireen) het bestaan uit nanoparikels met ‘n kern en skil morfologie. Die nanokleursels het ‘n verbetering in terme van lig en hitte stabiliteit getoon. Die migrasie van kleursel uit die nanopartikels, veral die met kern en skil morfologie, was baie minder. Die sintese van ‘n polimeeriseerbare nanokleursel gebaseer op RhB word beskryf. Poly(AAm-ko-RhB) nanokleursels was vir die eerste keer suksesvol gesintetiseer met behulp van ‘n inverse minie-emulsie polimerisasie. RhB kleursel was eers gefunksionaliseer deur middel van ‘n esterifikasie reaksie om ‘n polimeeriseerbare akrilaat groep te verkry. Die RhB-akrilaat kleursel was gekopolimeeriseer met AAm monomeer in ‘n inverse minie-emulsie polimerisasie om nanokleursels met verbeterde lig en migrasie stabiliteit te verkry. Kruisgebonde poli(AAm-ko-RhB) nanokleursels was verkry deur ‘n geskikte verbinding in die reaksie mengsel by te voeg. Beide poli(AAm-ko-RhB) and kruisgebonde poly(AAm-ko-RhB) nanokleursels was verkry as donker partikels met ‘n kern en skil morfologie. In beide gevalle was die RhB-akrilaat kleursel deeglik geintegreer in die matriks en sodoende was die migrasie van die kleursel heeltemal onderdruk. In albei gevalle het poli(AAm-ko-RhB) en kruisgebonde poli(AAm-ko-RhB) nanokleursels hoë hitte stabiliteit en hoë Tg waardes getoon. Die sintese van nanokleursels gebaseer op PAAm/RhB was sukselvol uit gevoer via inverse minie-emulsie polimerisasie. ‘n In situ inverse minie-emulsie polimerisasie met paraffin waks as die organiese fase was gebruik om soliede ink te produseer wat opgemaak is uit kruisgebonde PAAm/RhB nanokleursel. Die kruisgebonde poli(AAm-ko-RhB) soliede ink was voorberei deur die kruisgebonde PAAm/RhB nanokleursels (in suspensie met sikloheksaan) direk met die paraffin waks te meng by ‘n temperatuur hoër as die smeltpunt van die waks todat al die sikloheksaan verdamp het. Die soliede ink was verkry as ‘n homogene waksagtige materiaal met ‘n diep en helder kleur wat ‘n aanduiding was dat die nanokleursels goed versprei was in die waks. DSC termogramme het bewys dat die ink slegs een skerp smelt punt oorgang het wat beteken dat die materiaal geskik is om te gebruik in ink-smelt drukkers.

<p>Nowadays there are several contaminated buildings that receive a new field of application. These buildings used to have industrial activities which contaminated the buildings with both organic and inorganic pollutants that can cause serious health problems. Now these buildings get new functions like office work, school buildings or daycare centers. But before these buildings can be used as for instance schools some kind of remediation has to be carried out to remove the contamination. The aim with this report was to make an inventory of available methods for remediation of contaminated buildings and to evaluate the result of some performed remediations. The buildings that were investigated in this report were contaminated with organic pollutants. This report includes a summary of the legislation that concerns contaminated buildings, health problems that solvents and oils can cause and information of available methods for remediation of polluted buildings. A survey was made and property holders of remediated contaminated buildings answered it. According to the survey it emerged that air-regulated floors and new floor constructions were the most commonly used methods for remediation. The choice of method was often a balance between time and economy. After the remediation it’s important to do inspections to make sure that the remediation had removed the contamination. Inspection can be made by measuring the indoor air. Another way to inspect a remediation is to send out surveys to those people who stay in the building. The most important aspect during the remediation is to guarantee peoples health. Results from the survey showed that air-regulated floors are a good remediation method and often used with good results. To make the remediation work easier generic guideline values for different building materials should be developed to be used in the analysis of the degree of the remediation.</p>

Le cytosquelette des cellules est une structure composite et versatile qui leur confère des propriétés mécaniques extrêmement complexes. En particulier, le cortex d'actine qui s'assemble de manière dynamique sous la membrane cellulaire fournit la force nécessaire aux déformations et au mouvement de la cellule : la polymérisation de l'actine permet aux filaments en formation de pousser la membrane et les moteurs moléculaires génèrent des forces contractiles. L'utilisation de systèmes biomimétiques permet d'isoler des modules particuliers du cytosquelette pour les étudier indépendamment de façon simplifiée. Une expérience de reconstitution du cortex d'actine in vitro a été mise au point dans ce but. Les protéines et métabolites nécessaires pour la polymérisation de l'actine sont ainsi introduits dans un liposome et la réaction est localisée à la membrane, en y greffant l'activateur de la polymérisation de l'actine, sur le modèle du cortex cellulaire. Une fois la polymérisation déclenchée, nous sommes arrivés à reproduire un gel d'actine à la membrane, formant une coque. Les propriétés mécaniques de ce système simplifié sont alors étudiées par des expériences qui caractérisent leur dynamique d'étalement sur des surfaces. Les résultats sont comparés à ceux obtenus sur des cellules, et reproduisent une bonne partie des comportements cellulaires. On utilise également ces liposomes dans une situation physiologique: l'internalisation de la toxine de Shiga dans les cellules et nous montrons que la toxine est internalisée dans un système aussi épuré que des liposomes comportant un cortex reconstitué, prouvant le rôle important de l'actine dans ce processus.

Cell-based therapies involving adipose-derived stem cells (ASCs) have shown promise in stimulating cardiovascular regeneration, including in the treatment of myocardial infarction (MI) and ischemic heart disease. However, previous studies involving the delivery of ASCs following MI have indicated that therapeutic efficacy has been limited by low survival and/or poor retention of the transplanted cells at the site of injury. To address these limitations, the goal of this thesis was to develop a more effective delivery strategy incorporating an injectable biomaterial combined with chemotactic growth factor delivery to enhance ASC retention within the gel. Working towards future in vivo analysis in a rat model, multilineage characterization studies confirmed that ASCs isolated from the epididymal fat pad of male Wistar rats could differentiate in vitro along the adipogenic, osteogenic, and chondrogenic lineages. Subsequently, the chemotactic response of the rat ASCs (rASCs) to varying concentrations of stromal derived factor-1 α (SDF-1α) and hepatocyte growth factor (HGF) was analyzed using a modified Boyden chamber assay. The results demonstrated that SDF-1α and HGF, at 20, 50, and 100 ng/mL elicited significant migratory responses under normoxic (21%) and hypoxic (5%) culture conditions. RT-PCR analysis was conducted to assess the expression of the two chemotactic growth factors and their associated receptors in the rASCs, and secreted SDF-1α protein expression was quantified by ELISA. Moving towards the development of the biomaterials-based delivery approach, the viability of rASCs encapsulated by photopolymerization in methacrylated glycol chitosan (MGC) hydrogels modified with various degrees of arginine-glycine-aspartic acid (RGD)-peptide modification was examined. More specifically, rASCs were encapsulated in MGC hydrogels with 0%, 4%, and 7% RGD modification and cultured for up to 14 days. Viability staining results indicated that rASC viability was enhanced in the 4% and 7% RGD-modified MGC hydrogels in comparison to the MGC hydrogels with no peptide modification. Pre-loading the gels with 50 ng/mL of SDF-1α had no significant effects on cell viability over 14 days. Overall, the results demonstrate that peptide modification to promote cell adhesion within the MGC hydrogels is key to improving cell viability and thereby improving the therapeutic potential of ASCs. / Thesis (Master, Chemical Engineering) -- Queen's University, 2012-10-24 23:54:37.126

In this paper a novel stimuli responsive hydrogel material, methacrylated sodium alginate beta-cyclodextrin (Alg-MA-β-CD), was used in combination with a microfluidic device to create microspheres. Currently there is no reliable method for fabricating homogeneous stimuli-responsive microspheres, in-house microfluidic devices are not reliable in manufacture quality or long-term use. Alginate hydrogels have many attractive characteristics for bioengineering applications and are commonly used to mimic the features and properties of the extracellular matrix (ECM). Human mesenchymal stem cells (hMSCs) are of top interest to tissue engineers. hMSCs are widely available and can be harvested and cultured directly out of human bone marrow. hMSCs have the ability to differentiate into osteoblasts, adipocytes, chondrocytes, muscle cells, and stromal fibroblasts depending on mechanical signals transmitted through surrounding ECM. The biomechanical properties of alginate based stimuli-responsive hydrogels can be tuned to match those of different types of tissues. When trying to transport and control the differentiation of hMSCs into generating new tissues or regenerating damaged tissues, it is highly beneficial to encapsulate the cells inside a microsphere made from these hydrogels. The proposed research objectives are: 1) To optimize fabrication techniques and create functional microfluidic devices; 2) Analyze the effects of flow parameters on microsphere production; and 3) Encapsulate viable hMSCs inside multi-stimuli responsive alginate microspheres using the fabricated microfluidic devices (MFDs). In this study, photolithography microfabrication methods were used to create flow-focusing style MFDs. The hydrogel materials were characterized via rheological methods. Syringe pumps controlled flow rates of fluids through the devices. Active droplets formation was monitored through a camera attached to an inverted microscope, where images were analyzed. Microsphere production was analyzed optically and characterized. Alg-MA-β-CD polymer solutions containing hMSCs were encapsulated, and a live/dead florescence assay was preformed to verify cell viability. Using a modified fabrication process it was possible to manufacture Alg-MA-β-CD microspheres and encapsulate and maintain viable hMSCs inside.