O interesse pela área de análise e caracterização de materiais biomédicos cresce, devido a necessidade de selecionar de forma adequada, o material a ser utilizado. Dependendo das condições em que o material será submetido, a caracterização poderá abranger a avaliação de propriedades mecânicas, elétricas, bioatividade, imunogenicidade, eletrônicas, magnéticas, ópticas, químicas e térmicas. A literatura relata o emprego da técnica de árvores de decisão, utilizando os algoritmos SimpleCart(CART) e J48, para classificação de base de dados (dataset), gerada a partir de resultados de artigos científicos. Esse estudo foi realizado afim de identificar características superficiais que otimizassem a atividade celular. Para isso, avaliou-se, a partir de artigos publicados, o efeito de tratamento de superfície do titânio na atividade celular in vitro (células MC3TE-E1). Ficou constatado que, o emprego do algoritmo SimpleCart proporcionou uma melhor resposta em relação ao algoritmo J48. Nesse contexto, o presente trabalho tem como objetivo aplicar, para esse mesmo estudo, os algoritmos CHAID (Chi-square iteration automatic detection) e CHAID Exaustivo, comparando com os resultados obtidos com o emprego do algoritmo SimpleCart. A validação dos resultados, mostraram que o algoritmo CHAID Exaustivo obteve o melhor resultado em comparação ao algoritmo CHAID, obtendo uma estimativa de acerto de 75,9% contra 58,6% respectivamente, e um erro padrão de 7,9% contra 9,1% respectivamente, enquanto que, o algoritmo já testado na literatura SimpleCart(CART) teve como resultado 34,5% de estimativa de acerto com um erro padrão de 8,8%. Com relação aos tempos de execução apurados sobre 22 mil registros, evidenciaram que o algoritmo CHAID Exaustivo apresentou os melhores tempos, com ganho de 0,02 segundos sobre o algoritmo CHAID e 14,45 segundos sobre o algoritmo SimpleCart(CART). / The interest for the area of analysis and characterization of biomedical materials as the need for selecting the adequate material to be used increases. However, depending on the conditions to which materials are submitted, characterization may involve the evaluation of mechanical, electrical, optical, chemical and thermal properties besides bioactivity and immunogenicity. Literature review shows the application decision trees, using SimpleCart(CART) and J48 algorithms, to classify the dataset, which is generated from the results of scientific articles. Therefore the objective of this study was to identify surface characteristics that optimizes the cellular activity. Based on published articles, the effect of the surface treatment of titanium on the in vitro cells (MC3TE-E1 cells) was evaluated. It was found that applying SimpleCart algorithm gives better results than the J48. In this sense, the present study has the objective to apply the CHAID (Chi-square iteration automatic detection) algorithm and Exhaustive CHAID to the surveyed data, and compare the results obtained with the application of SimpleCart algorithm. The validation of the results showed that the Exhaustive CHAID obtained better results comparing to CHAID algorithm, obtaining 75.9 % of accurate estimation against 58.5%, respectively, while the standard error was 7.9% against 9.1%, respectively. Comparing the obtained results with SimpleCart(CART) results which had already been tested and presented in the literature, the results for accurate estimation was 34.5% and the standard error 8.8%. In relation to execution time found through the 22.000 registers, it showed that the algorithm Exhaustive CHAID presented the best times, with a gain of 0.02 seconds over the CHAID algorithm and 14.45 seconds over the SimpleCart(CART) algorithm.

O interesse pela área de análise e caracterização de materiais biomédicos cresce, devido a necessidade de selecionar de forma adequada, o material a ser utilizado. Dependendo das condições em que o material será submetido, a caracterização poderá abranger a avaliação de propriedades mecânicas, elétricas, bioatividade, imunogenicidade, eletrônicas, magnéticas, ópticas, químicas e térmicas. A literatura relata o emprego da técnica de árvores de decisão, utilizando os algoritmos SimpleCart(CART) e J48, para classificação de base de dados (dataset), gerada a partir de resultados de artigos científicos. Esse estudo foi realizado afim de identificar características superficiais que otimizassem a atividade celular. Para isso, avaliou-se, a partir de artigos publicados, o efeito de tratamento de superfície do titânio na atividade celular in vitro (células MC3TE-E1). Ficou constatado que, o emprego do algoritmo SimpleCart proporcionou uma melhor resposta em relação ao algoritmo J48. Nesse contexto, o presente trabalho tem como objetivo aplicar, para esse mesmo estudo, os algoritmos CHAID (Chi-square iteration automatic detection) e CHAID Exaustivo, comparando com os resultados obtidos com o emprego do algoritmo SimpleCart. A validação dos resultados, mostraram que o algoritmo CHAID Exaustivo obteve o melhor resultado em comparação ao algoritmo CHAID, obtendo uma estimativa de acerto de 75,9% contra 58,6% respectivamente, e um erro padrão de 7,9% contra 9,1% respectivamente, enquanto que, o algoritmo já testado na literatura SimpleCart(CART) teve como resultado 34,5% de estimativa de acerto com um erro padrão de 8,8%. Com relação aos tempos de execução apurados sobre 22 mil registros, evidenciaram que o algoritmo CHAID Exaustivo apresentou os melhores tempos, com ganho de 0,02 segundos sobre o algoritmo CHAID e 14,45 segundos sobre o algoritmo SimpleCart(CART). / The interest for the area of analysis and characterization of biomedical materials as the need for selecting the adequate material to be used increases. However, depending on the conditions to which materials are submitted, characterization may involve the evaluation of mechanical, electrical, optical, chemical and thermal properties besides bioactivity and immunogenicity. Literature review shows the application decision trees, using SimpleCart(CART) and J48 algorithms, to classify the dataset, which is generated from the results of scientific articles. Therefore the objective of this study was to identify surface characteristics that optimizes the cellular activity. Based on published articles, the effect of the surface treatment of titanium on the in vitro cells (MC3TE-E1 cells) was evaluated. It was found that applying SimpleCart algorithm gives better results than the J48. In this sense, the present study has the objective to apply the CHAID (Chi-square iteration automatic detection) algorithm and Exhaustive CHAID to the surveyed data, and compare the results obtained with the application of SimpleCart algorithm. The validation of the results showed that the Exhaustive CHAID obtained better results comparing to CHAID algorithm, obtaining 75.9 % of accurate estimation against 58.5%, respectively, while the standard error was 7.9% against 9.1%, respectively. Comparing the obtained results with SimpleCart(CART) results which had already been tested and presented in the literature, the results for accurate estimation was 34.5% and the standard error 8.8%. In relation to execution time found through the 22.000 registers, it showed that the algorithm Exhaustive CHAID presented the best times, with a gain of 0.02 seconds over the CHAID algorithm and 14.45 seconds over the SimpleCart(CART) algorithm.

This master’s thesis dissertate the possibilities of the linkage between Asterisk softswitch and the 4th generation private branch exchange. This should create a new generation’s network, so-called NGN, by the convergence of existing telecommunication networks with an IP computer network. This master’s thesis is divided into several chapters. In introduction is described the evolution of the private branch exchanges as well as the principles of the voice digitizing, codecs and signaling commonly used in both TDM and VoIP networks. The main aim of this project is the configuration of Asterisk software exchange for connection with PBX Alcatel 4400 as well as public phone network PSTN. Another goal of this master’s thesis was the configuration of Alcatel PBX and diagnostics of CCS and CAS signaling on E1 interface. In conclusion there are summarized advantages of NGN networks and their utilization in the future.

Lors de l'infection par le virus herpès simplex de type 1 (VHS-1), quatre types de capsides nucléaires sont créés : les procapsides et les capsides A, B, et C. Sur les quatre capsides, seules les capsides C contiennent de l'ADN viral et deviendront des particules infectieuses. Un niveau de régulation se produit lors de la sortie du noyau qui favorise la sortie d’es capsides C du noyau. Le mécanisme qui sous-tend ce phénomène est actuellement inconnu. Les recherches actuelles suggèrent que l'interaction entre la protéine virale pUL25 modifie la conformation de la couche hexamérique plane du complexe de sortie nucléaire (NEC) pour y introduire des pentamères et donc causer un arrondissement de la membrane et le bourgeonnement des capsides. Cependant, des questions subsistent quant à la manière dont les capsides A, B et C sont différenciées au sein du noyau pour assurer une sortie spécifique de la capside C puisque pUL25 se retrouve dans tous les types de capsides. Nous étudions ici comment les protéines de l'hôte peuvent agir dans la sortie nucléaire des capsides C. En se basant sur une étude précédente du laboratoire où la protéine hôte poly(C)-binding protein 1 (PCBP1) a été trouvée spécifiquement sur les capsides C par spectrométrie de masse, nous explorons le rôle de la PCBP1 dans l'infection par le VHS-1. À l'aide d’essaies de plaques, nous montrons que la PCBP1 est importante pour l'infection virale, car en son absence, les titres diminuent et lorsque la PCBP1 est sur-exprimée, les titres augmentent. Ce résultat ne semble pas être dû au fait que les PCBP1 affectent l'expression génique de sous-ensembles de gènes viraux immédiats précoces, précoces ou tardifs, ni qu'ils affectent la réplication du génome ou son encapsidation. La réduction des PCBP1 ne provoque pas d'accumulation de capsides ou de particules matures tel qu’évalué par la microscopie électronique, mais elle augmente le nombre de capsides B enveloppées dans l'espace périnucléaire (PNS). L'inhibition de PCBP1 diminue également le niveau de protéine pUL24, une protéine virale importante pour la sortie du virus du noyau. Nos résultats démontrent que la PCBP1 pourrait réguler l’activité de pUL24, de sorte que lorsque la PCBP1 est épuisée, pUL24 permet à plus de capsides B de se rendre dans l'espace périnucléaire. Cette recherche constitue un point de départ pour une analyse plus approfondie du mécanisme exact des PCBP1 dans les infections à HSV-1. En outre, elle pourrait fournir des indices importants pour élucider comment le pUL24 favorise la sortie du nucléaire. / During herpes simplex virus type 1 (HSV-1) infection, four types of nuclear capsids are made: procapsids and A-, B- and C-capsids. Of the four capsids, only C-capsids contain the viral DNA and will become infectious progeny. A level of regulation occurs during nuclear egress that ensures only C-capsids exit the nucleus. The mechanism that underlies this phenomenon is presently unknown. Current research suggests the viral protein pUL25 alters the conformation of the viral nuclear egress complex (NEC) that forms a flat hexameric coat on nuclear membranes by the introduction of pentamers and therefore the induction of membrane rounding and viral budding. However, questions remain for how A-, B-, and C-capsids are differentiated within the nucleus to ensure C-capsid specific egress since pUL25 is found on all capsid types. Here we investigate how host proteins may play a role in nuclear egress of C-capsids. Based on the lab’s previous study where host protein poly(C)-binding protein 1 (PCBP1) was found specifically on C-capsids via mass spectrometry, we explore the role of PCBP1 in HSV-1 infection. Using plaque assays we show that PCBP-1 is important for viral infection, as in its absence titers decrease and when PCBP1 is over expressed titers increase. This result does not seem to be due to PCBP1 affecting gene expression of immediate early, early, or late viral gene subsets, nor does it seem to affect genome replication or encapsidation. PCBP1 knockdown does not cause an accumulation of capsids or mature particles as assessed by electron microscopy, but it does increase the number of enveloped B-capsids observed in the perinuclear space (PNS). Depletion of PCBP1 also decreases the level of pUL24, a viral protein implicated in viral nuclear egress. Our results suggest that PCBP1 could be regulating pUL24 for proper activity in nuclear egress, such that when PCBP1 is depleted, more B-capsids are able to bud through the PNS. This research constitutes a starting point for further analysis into the exact mechanism of PCBP1 in HSV-1 infections. In addition, it may provide important clues to elucidate how pUL24 supports nuclear egress.

Venezuelan equine encephalitis virus (VEEV) is an alphavirus that causes disease in humans and equines eliciting both an agricultural and public health threat. In humans, the disease typically presents as a febrile illness with common signs of fever and malaise. Four to fourteen percent of Venezuelan equine encephalitis (VEE) cases are associated with severe neurological complications due to encephalitis caused by VEEV's propensity to infect the brain. Public health concerns are exacerbated by VEEV's aerosolization capabilities, low infectious dose and affordability to mass produce. These qualities drove interest in the pathogen as a bioweapon by the US and the former Soviet Union during the cold war. As a precautionary response to VEEV's notoriety as a biothreat, the National Institute of Allergies and Infectious Diseases has classified VEEV as a category B priority pathogen, and the Human Health Services and United States Department of Agriculture list live virulent strains of VEEV as a select agent and require the pathogen to be manipulated in highly regulated biosafety level 3 (BSL3) facilities. There are currently no FDA approved vaccines or antivirals to target VEEV or other closely related alphaviruses associated with clinical disease in humans. The research performed in this dissertation aimed to elucidate new antiviral targets and treatments to help bridge gaps in current understanding of alphaviruses. The current market lacks available antibodies for E1 specific isolation. In response, a recombinant VEEV TC-83 was produced with a V5 tag at the C-terminal of the E1 sequence to enable VEEV E1 detection. Sequencing was used to verify V5 insertion in the plasmid and immunoprecipitation was used to verify V5 insertion within the E1 glycoprotein. Replication kinetics experiments verified the virus replicated similarly to the parental VEEV TC-83 strain, while passaging experiments verified the tag was highly stable for up to 10 passages. This research produced a cost-effective and highly efficient means to probe and isolate the E1 glycoprotein without modifying the viability of the virus. Knowledge of host protein interactions with VEEV E1 glycoprotein has been limited, with most E1 research focusing on its fusion capabilities. Utilizing 293-T cells infected with E1-V5 TC-83, co-immunoprecipitation was performed to isolate E1 and associated interactors. A total of 486 host and 5 viral protein interactors of E1 were discovered after normalization to the negative control. The top peptide spectrum matches (PSMs) revealed a number of chaperone proteins and ubiquitin proteins as top interactors of VEEV E1. These results effectively revealed a number of previously unknown alphavirus interactions that can be targeted by antivirals and explored further for implications in viral replication. LC-MS/MS results showed that protein disulfide isomerase family A member 6 (PDIA6) interacted with E1. High PSMs, presence in all 3 replicates, similar cellular localization to E1 and known associations between other viruses and protein disulfide isomerase (PDI) family members made this protein an optimum target for further analysis. Co-immunoprecipitation and co-localization experiments were used to validate the LC-MS/MS results. Involvement of PDIs in VEEV replication were explored utilizing two known PDI inhibitors, LOC14 and Nitazoxanide. LOC14, a non-FDA approved broad-spectrum PDI inhibitor, showed broad-spectrum alphavirus antiviral potential, decreasing titers of VEEV TC-83, VEEV Trinidad Donkey strain, eastern equine encephalitis virus (EEEV), chikungunya virus (CHIKV) and Sindbis (SINV) virus in a dose dependent manner. Nitazoxanide, an FDA approved drug known to inhibit PDIA3, was shown to have minimal toxicity and effectively reduced VEEV TC-83 and EEEV titers at concentrations with 100% cell viability. Time of addition assays, E1 expression time course studies, and early event assays showed PDI inhibition with these drugs effects early viral production events. RNA quantification, confocal microscopy and biotin switch assay experiments show that the drugs also prevented proper folding of the E1 glycoprotein and decreased expression of E1 on the peripheral membrane. With no current treatments for alphaviruses, these data provide an effective broad-spectrum target that affects viral replication at multiple stages in-vitro. Nitazoxanide also presents as a promising, non-toxic drug that could be repurposed to combat a number of clinically relevant alphaviruses. Valosin containing protein (VCP) was also shown to interact with the E1 glycoprotein. Exploration of VCP's interaction with alphavirus E1 has never been explored, yet it was previously shown to be involved in alphavirus replication. Co-localization and co-immunoprecipitation experiments were performed validating the interaction between VCP and E1. siRNA knockdown of VCP in 293-T cells and U87-MG cells showed a significant reduction in VEEV TC-83 titers. The allosteric VCP inhibitor, NMS-873, also reduced VEEV TC-83 titers, but was shown to be less effective against CHIKV, SINV and EEEV, suggesting the NMS-873 mechanism is more selective for VEEV. Mechanism experiments showed that reduction of VCP with NMS-873 inhibits early events of VEEV replication. These results elucidate VCP's association with E1 and show that VCP can be targeted to decrease VEEV viral replication. / Doctor of Philosophy / Venezuelan equine encephalitis virus (VEEV) causes disease in humans, as well as horses, donkeys and other closely related animals. In humans, the virus causes a flu-like disease and sometimes swelling of the brain. This can be associated with symptoms such as light sensitivity, confusion and sometimes coma. Prior to the Cold War, VEEV was researched by the US and previous Soviet Union's militaries in hopes to deploy the virus as a bioweapon. Current treaties prevent active production of such weapons, yet allows for defensive research to continue in preparation for a worst-case scenario. Currently no FDA approved medications or vaccines exist to combat the virus further exacerbating concerns. In order to protect laboratorians and prevent unintentional or intentional introduction of the virus into the community, the virus is only manipulated in highly secure facilities with barriers that separate the virus from personnel and the outside environment. A component of the virus called E1, allows for the virus to be released from a structure, called an endosome, that transports the virus into the cell. Currently, E1 is mostly known for this function, yet our research found that E1 interacts with 486 protein components of the host cell, suggesting a more elaborate role of E1 than previously understood. This list of interactors provides numerous new targets for potential medications to combat VEEV and other closely related viruses. Discovered E1 interactors, protein disulfide isomerase family A member 6 (PDIA6) and valosin containing protein (VCP), were validated through extensive experimentation and their function in viral replication was further explored. Protein disulfide isomerases (PDI), such as PDIA6, play an important role in folding proteins, which are cellular components made of organic building blocks called amino acids. PDIs do so by creating organic pillars, called disulfide bonds, between two cysteine amino acid residues. These disulfide bonds contribute to the 3D shape of the proteins they fold which are essential for the protein's function. E1 of VEEV has a total of eight disulfide bonds within its structure, highlighting that disulfide bonds are likely essential for the protein's structure, and therefore, function. We verified that E1 could not properly fold without PDI function by using two compounds that prevented PDI from forming or breaking disulfide bonds, specifically LOC14 and FDA approved drug nitazoxanide. Cells treated with one of either compound before and after infection with VEEV, were found to produce E1 protein with significantly less disulfide bonds therefore producing less viable virus. Further experiments also showed that the compounds also affected early stages in the virus production cycle. These two mechanisms explain the significant reduction in production of VEEV and related viruses when PDI is inhibited. These results provide a new VEEV drug target, PDIs, as well as two compounds that can potentially be used to combat VEEV and other related viruses that have no current treatment options. Another host interactor, VCP, functions throughout the cell and is known for unfolding of numerous substrates, including proteins. It is involved in numerous cellular functions thus making this interactor a promising target for drug treatment. Cells with reduced VCP function were shown to produce less progeny VEEV. Cells treated with NMS-873, a compound that reduces VCP function was also shown to reduce VEEV production. NMS-863 inhibition of VCP was shown to effect early events in VEEV replication. These results further emphasize the E1 interactors discovered are invaluable novel targets for VEEV drug treatment.

Neste trabalho analisamos e propomos a utilização de TDM (Time Division Multiplexing) nativo canalizado/estruturado em redes PON (Passive Optical Network) com padrão GPON (Gigabit Passive Optical Network), com ênfase na estrutura de transmissão do legado das redes de telefonia. O objetivo principal é obter um aumento na eficiência de banda transmitida através da fragmentação de sinais E1 sem que seja necessário o uso de técnicas de emulação de circuito (que reduzem a eficiência de banda devido à adição de cabeçalhos). Inicialmente, é descrito o transporte TDM em redes GPON, como efetuado pelos equipamentos comerciais atuais através de duas técnicas: CES - Circuit Emulation Service e TDM nativo não estruturado. Em seguida, é introduzido o conceito de comutação digital visando sua aplicação no transporte TDM nativo estruturado em redes GPON. Nesta etapa, é proposta uma solução para este transporte, é descrito o protocolo utilizado bem como seu funcionamento. Por fim, como prova de conceito, é apresentada uma implementação em HDL (Hardware Description Language) para FPGA (Field Programmable Gate Array). / In this work we analyze and propose the use of native channeled /structured TDM (Time Division Multiplexing) in GPON (Gigabit Passive Optical Network), with emphasis on the structure for transmission of the telephone network legacy. The main target is to achieve an increase in transmitted bandwidth efficiency by fragmenting E1 signals, thus avoiding the use of circuit emulation techniques (which reduce the bandwidth efficiency due to overhead addition). Initially, it is described in TDM transport in GPON networks, as it is performed in present commercial equipment by two techniques: CES - Circuit Emulation Service and Native TDM - unstructured. Next, we introduce the concepts of digital switching aiming its application on the transport of native and structured TDM in GPON. At this stage, we propose a transport solution, describe its protocol and functionalities. Finally, for concept proof, we present an implementation in HDL (Hardware Description Language) meant to FPGA (Field Programmable Gate Array) application.

Neste trabalho analisamos e propomos a utilização de TDM (Time Division Multiplexing) nativo canalizado/estruturado em redes PON (Passive Optical Network) com padrão GPON (Gigabit Passive Optical Network), com ênfase na estrutura de transmissão do legado das redes de telefonia. O objetivo principal é obter um aumento na eficiência de banda transmitida através da fragmentação de sinais E1 sem que seja necessário o uso de técnicas de emulação de circuito (que reduzem a eficiência de banda devido à adição de cabeçalhos). Inicialmente, é descrito o transporte TDM em redes GPON, como efetuado pelos equipamentos comerciais atuais através de duas técnicas: CES - Circuit Emulation Service e TDM nativo não estruturado. Em seguida, é introduzido o conceito de comutação digital visando sua aplicação no transporte TDM nativo estruturado em redes GPON. Nesta etapa, é proposta uma solução para este transporte, é descrito o protocolo utilizado bem como seu funcionamento. Por fim, como prova de conceito, é apresentada uma implementação em HDL (Hardware Description Language) para FPGA (Field Programmable Gate Array). / In this work we analyze and propose the use of native channeled /structured TDM (Time Division Multiplexing) in GPON (Gigabit Passive Optical Network), with emphasis on the structure for transmission of the telephone network legacy. The main target is to achieve an increase in transmitted bandwidth efficiency by fragmenting E1 signals, thus avoiding the use of circuit emulation techniques (which reduce the bandwidth efficiency due to overhead addition). Initially, it is described in TDM transport in GPON networks, as it is performed in present commercial equipment by two techniques: CES - Circuit Emulation Service and Native TDM - unstructured. Next, we introduce the concepts of digital switching aiming its application on the transport of native and structured TDM in GPON. At this stage, we propose a transport solution, describe its protocol and functionalities. Finally, for concept proof, we present an implementation in HDL (Hardware Description Language) meant to FPGA (Field Programmable Gate Array) application.

Cette thèse transdisciplinaire réalisée en collaboration avec le laboratoire SPCTS (Sciences des Procédés Céramiques et Traitement de Surface) et l’EA 3842 (Homéostasie cellulaire et pathologies) de l’université de Limoges est un projet de recherche à l’interface entre la biologie et la chimie et a été consacrée à l’étude de l’influence des propriétés physico-chimiques de biocéramiques de phosphate de calcium sur leur comportement biologique in vitro.L’exploration des processus d’interaction entre matériaux et cellules reste une problématique scientifique de premier plan tant d’un point de vue fondamental qu’appliqué pour la mise au point de biomatériaux performants. L’objectif final est d’optimiser l’efficacité thérapeutique des céramiques phosphocalciques comme matériaux de substitution pour la régénération osseuse. La première partie de la thèse est une revue bibliographique générale présentant la problématique actuelle abordée en lien avec les besoins cliniques et les limitations des études actuelles. Les connaissances sur la biologie du tissu osseux sain ainsi que les aspects de régulation du processus de remodelage osseux ont également été abordés dans ce chapitre. Ce chapitre se termine par une synthèse bibliographique sur les biomatériaux et la régénération osseuse. Le chapitre 2 est relatif à la synthèse puis à la caractérisation physico-chimique des matériaux céramiques. Des céramiques de trois compositions chimiques : HA (hydroxyapatite : Ca10(PO4)6(OH)2 , SiHA (hydroxyapatite silicatée : Ca10(PO4)5,6(SiO4)0,42(OH)1,6 et CHA (hydroxyapatite carbonatée : Ca9,5(PO4)5,5(CO3)0,48(OH)1,08(CO3)0,23 , chacune avec deux microstructures différentes : dense ou poreuse, ont été élaborées et rigoureusement caractérisées (porosité, topographie de surface, mouillabilité, potentiel zêta, taille des grains, distribution et taille des pores, surface spécifique). Le chapitre 3 décrit l’approche expérimentale employée pour l’évaluation biologique des interactions matériaux/cellules explorées dans ce travail. Les analyses biologiques ont été réalisées avec deux lignées cellulaires différentes. La lignée cellulaire pré-ostéoblastique MC3T3-E1 et la lignée cellulaire de monocytes/macrophages, précurseurs des ostéoclastes RAW 264.7, (très importantes pour les aspects osseux, mais moins souvent explorées que les lignées ostéoblastiques dans la littérature). Enfin, le chapitre 4 reporte et commente les résultats biologiques obtenus dans ce travail. Tous les biomatériaux évalués dans cette étude sont biocompatibles, néanmoins, le biomatériau poreux CHA s’est avéré le plus prometteur des six variantes de biomatériaux testés. / This transdisciplinary thesis, carried out in collaboration with the SPCTS laboratory (sciences of ceramic processes and surface treatment) and EA 3842 (Cellular homoeostasis and pathologies) of the University of Limoges, is a research project at the interface between biology and chemistry and was devoted to the study of the influence of the physico-chemical properties of calcium phosphate bioceramics on their biological behavior in vitro.The exploration of the processes of interaction between materials and cells remains a major scientific issue, both from a fundamental and applied point of view for the development of highperformance biomaterials. The ultimate objective is to optimize the therapeutic efficiency of phosphocalcic ceramics as substitute materials for bone regeneration.The first part of the thesis is a general bibliographic review presenting the current issues tackled with the clinical needs and limitations of current studies. Knowledge of the biology of healthy bone tissue as well as the regulatory aspects of the bone remodeling process was also discussed in this chapter. It includes also a bibliographic overview of biomaterials and bone regeneration.Chapter 2 relates to the synthesis and the physico-chemical characterization of ceramic materials. HA (hydroxyapatite: Ca10 (PO4) 6 (OH) 2, SiHA (silicated hydroxyapatite: Ca10 (PO4) 5.6 (SiO4) 0.42 (OH) 1.6 and CHA (carbonated hydroxyapatite: Ca9.5 (PO4) 5.5 (CO3) 0.48 (OH) 1.08 (CO3) 0.23, ceramics each with two different microstructures : dense or porous, have been elaborated and thoroughly characterized (porosity, surface topography, wettability, zeta potential, grain size, pore size and distribution, specific surface area). Chapter 3 describes the experimental approach used for the biological evaluation of the interactions between materials and cells. Biological analyzes were performed with two different cell lines. The pre-osteoblastic MC3T3-E1 cell line and the RAW 264.7cell line of monocytes / macrophages, precursors of the steoclasts, (very important for the bone aspects, but less often explored than the osteoblastic lines in the literature). Finally, Chapter 4 reports and comments on the biological results obtained in this work. All biomaterials evaluated are biocompatible, nevertheless, the porous CHA biomaterial was the most promising of the six variants of biomaterials tested.

Scintillation of Global Positioning Systems (GPS) signals have been extensively studied at low and high latitude regions of the Earth. It has been shown in past studies that amplitude scintillation is severe at low latitudes and phase scintillation is severe at high latitudes. Unlike low and high latitude regions, mid-latitude scintillation has not been extensively studied. Further, it has been suggested that mid-latitude scintillation is negligible. The purpose of this research is to challenge this belief. A multi-constellation and multi-frequency receiver, that tracks American, Russian, and European satellites, was used to monitor scintillation activity at the Virginia Tech Space Center. Analysis was performed on collected data from various days and compared to past research done at high, mid, and low latitudes. The results are discussed in this thesis. / Master of Science

The disposal of waste from agricultural activities has been recognised as a source of environmental contamination by endocrine disrupting chemicals (EDCs). The New Zealand dairy industry produces a large volume of dairy farm effluent, which contains EDCs in the form of estrogens. Most of this dairy farm effluent is applied onto the land for disposal. Groundwater and soil contamination by estrogens following waste application on the land have been reported overseas, but our understanding of the processes and factors governing the fate of estrogens in the soil is poor. Therefore the main goal of the present study was to better understand the fate and transport of estrogens, in particular 17β-estradiol (E2) and estrone (E1) in soil. In order to quantify E1 and E2 in drainage water and soil samples, chemical analysis by gas-chromatography mass-spectrometry (GC-MS) was carried out. This included sample extraction, sample clean-up through silica gel and gel permeation chromatography, and sample extract derivatisation prior to analysis. In order to develop a reliable method to extract estrogens from soil, research was conducted to optimise E1 and E2 extraction conditions by adjusting the number of sonication and shaking events, as well as the volume and type of solvent. Among five solvents and solvent mixtures tested, the best recovery on spiked and aged soil was obtained using an isopropanol/water (1:1) mix. A microcosm experiment was carried out to determine the dissipation rates of E2 and E1, at 8°C and at field capacity, in the Templeton soil sampled at two different depths (5-10 cm and 30-35 cm). The dissipation rates decreased with time and half-life values of 0.6-0.8 d for E1 and 0.3-0.4 d for E2 were found for the two depths studied. A field transport experiment was also carried out in winter, over three months, by applying dairy farm effluent spiked with estrogens onto undisturbed Templeton soil lysimeters (50 cm in diameter and 70 cm deep). The hormones were applied in dairy farm effluent at 120 mg m⁻² for E2 and 137 mg m⁻² for E1. The results of the transport experiment showed that in the presence of preferential/macropore flow pathways 0.3-0.7% of E2 and 8-13% of E1 was recovered in the leachate at the bottom of the lysimeters after 3 months, and 1-7% of the recovered E2 and 3-54% of the recovered E1 was leached within 2 days of application. These results suggest that leaching of estrogens via preferential/macropore flow pathways is the greatest concern for groundwater contamination. In the absence of preferential/macropore flow pathways, a significant amount (> 99.94%) of both hormones dissipated in the top 70 cm of soil, due to sorption and rapid biodegradation. Surprisingly, in all cases, estrogen breakthrough occurred before that of an inert tracer (bromide). This could not be explained by the advection-dispersion transport of estrogens, nor by their presence as antecedent concentrations in the soil. It was therefore suggested that colloidal enhanced transport of estrogens was responsible for the earlier breakthrough of estrogens and caused the leaching of a fraction of the applied estrogens to a soil depth of 70 cm. A two-phase model, adapted from a state-space mixing cell model, was built to describe the observed estrogen transport processes under transient flow. The model takes into account 3 transport processes namely, advection-dispersion, preferential/macropore flow and colloidal enhanced transport. This model was able to successfully describe the estrogen transport observed from the lysimeters.