Cette thèse traite de la conception, la synthèse et la caractérisation de chromophores dipolaires D-π-A pour l’imagerie de fluorescence et l’imagerie photoacoustique.La première partie est consacrée à la synthèse et à l’étude de nouveaux fluorophores émettant à l’état solide dans le proche infrarouge, pour l’imagerie de fluorescence biphotonique. Afin d’améliorer les propriétés optiques à l’état solide, nous avons exploré différentes pistes en modifiant d’abord le groupement D donneur d’électron, puis le groupement A accepteur et enfin le pont π conjugué de nos structures dipolaires.La seconde partie concerne l’imagerie photoacoustique. Cette imagerie, basée sur une excitation optique et une détection acoustique, nécessite une forte absorption dans le proche infrarouge. Une des stratégies pour effectuer ce décalage bathochrome de l’absorption a été incorporation de groupements thiophènes à faible aromaticité, afin d’augmenter la délocalisation du système π. Parallèlement, nous avons aussi étudiés des hémicyanines, présentant de fortes absorptions au-delà de 650 nm.Enfin, les colorants ont été formulés en nanoparticules organiques afin d’acquérir une solubilité aqueuse pour leur application en imagerie. Deux types de nanoparticules ont été étudiés : la nanoprécipitation des colorants en présence d’un agent tensioactif, ainsi que l’encapsulation des colorants dans une matrice polymérique amphiphile stabilisée par une couche de silice. Les meilleurs chromophores ont permis l’acquisition d’images de cellules par microscopie biphotonique ainsi que d’images photoacoustiques de circuits microfluidiques et de la microvascularisation de souris in vivo. / This thesis is focused on the conception, synthesis and characterization of dipolar dyes D-π-A for fluorescence imaging and photoacoustic imaging.The first part aim to synthesize and to study new fluorophores emitting in the solid-state in the near-infrared for fluorescence imaging with two-photon excitation. In order to improve the optical properties in the solid-state, we first explored various electron donating groups D, then acceptors A and finally we modified the π bridge. In a second part, we focused on dyes for photoacoustic imaging. This technique is based on an optical excitation and an acoustic detection. It requires a strong absorption in the biological window. To red shift the absorption we incorporated a thiophene bridge with low aromaticity to increase the delocalization. We also studied several hemicyanines with strong absorption above 650 nm.Finally, dyes were formulated as water-soluble nanoparticles for their final application for imaging. Two types of nanoparticles were studied: nanoprecipitated dyes stabilized by a surfactant to increase the colloidal stability or encapsulated dyes in a amphiphilic polymer matrix stabilized with a silica shell. Those nanoparticles enabled cells imaging with biphotonic florescence imaging as well as photoacoustic imaging of a microfluidic chip and of the microvascularisation of mice ears in vivo.

Made available in DSpace on 2016-08-03T16:15:42Z (GMT). No. of bitstreams: 1 Leticia dos Santos Brito.pdf: 1000721 bytes, checksum: b2409a81caea0b5901db8e2d23ec5d47 (MD5) Previous issue date: 2013-04-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Most low income families assign to the school system the solutions to their social problems, seeing in the school apparatus the only chance for social climb. However, it is worth asking: what is the role of low-income families s children within the public shcool? After all, when they are inserted into the educational system, mostly they show great difficulty to move along, thus creating a major problem for the school, since children who can not suceed in the first years of elementary school end up dragging this failure throughout their school life. For these children, educational assistance programs were created. But in what extent do they help and where they end up stigmatizing students who participate in those initiatives? In attempting to answer this questions, this study make the analysis of some educational assistance programs who proposed changes in the educational reality, offering more opportunities of progress for low-income children. The research is willing to establish whether they actually complied with its purpose or ultimately stigmatize students who participate in these projects. / A maior parte das famílias de baixa renda atribui à escola a solução para os seus problemas sociais, vendo no aparelho escolar a única chance de ascensão social. Contudo, cabe aqui perguntar: Qual é o espaço das crianças pobres dentro da escola publica? Afinal, quando inseridas dentro do sistema educacional, em sua maioria demonstram grande dificuldade para avançar, criando assim um grande problema para instituição escolar, uma vez que crianças que não conseguem obter sucesso nos primeiros anos do ensino fundamental acabam arrastando esse fracasso ao longo de toda sua vida escolar. Para essas crianças, foram criados os programas de auxilio pedagógico. Mas até que ponto eles ajudam e onde acabam por estigmatizar os alunos que deles participam? Na tentativa de responder essa pergunta, este trabalho buscou fazer a análise de alguns projetos de auxilio pedagógico que propuseram a mudança da realidade educacional, dando mais oportunidades de avanços às crianças de baixa renda. A pesquisa procura saber se eles realmente cumpriram com a sua finalidade ou acabaram por estigmatizar os alunos que participaram destes projetos de auxilio pedagógico.

Oligonucleotides are synthetic strings of DNA or RNA used mostly for biochemical analysis and diagnostics. For them to be useful in these fields, a purity over 90% is most often required. However, when synthesizing these sequences, many “failures” (shorter sequences) are made in the step-wise process. The synthesized oligonucleotides need to therefore be purified. This is most often done with gel electrophoresis or liquid chromatography. These methods are, on the other hand, very time-consuming and laborious. Solid phase extraction (SPE) is a much faster purification method if optimized and it can be done with the standard cartridges as well as 96-well plates, that allow many samples to efficiently be run at the same time. With reversed phase (RP) SPE, the dimethoxytrityl (DMT) group, that is attached to the target at the final synthesis step, can be used for stronger retention to the bed sorbent and leaving only the target at the final eluting stage. The impurities without a DMT-on group, that do not adsorb to the sorbent, are washed away in earlier steps. The purpose of this study is to optimize an SPE method for purification of oligonucleotides. Two different cartridges, Clarity QSP (Phenomenex) and Glen-Pak (Glen Research) were used. The purity analysis and oligonucleotide identification were done using anion exchange - high performance liquid chromatography (AIE-HPLC) and time-of-flight mass spectrometry (TOF MS). To conclude, Clarity QSP achieved, at the most, a purity of 68.8% with the recommended SPE steps by Phenomenex. Alterations in the extraction procedure resulted in similar purity or lower. Glen-Pak reached a peak purity of 78.8% when doing a double salt wash of 5% ACN in 2 M sodium chloride and another double wash after detritylation with 1% acetonitrile. This method has to be further optimized in order to reach a purity of at least 90% to be useful in industrial settings.

Thesis--Würzburg. / Vita. Bibliographical references in "Einleitung": p. [7]-13.

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2011. / Submitted by Matheus Denezine (matheusdenezine@yahoo.com.br) on 2011-06-21T14:28:14Z No. of bitstreams: 1 2011_KarinaSilvaFiorillo.pdf: 753032 bytes, checksum: 3c44807f8daafe4bb38f2e86e87f16f2 (MD5) / Approved for entry into archive by Guilherme Lourenço Machado(gui.admin@gmail.com) on 2011-06-21T16:27:04Z (GMT) No. of bitstreams: 1 2011_KarinaSilvaFiorillo.pdf: 753032 bytes, checksum: 3c44807f8daafe4bb38f2e86e87f16f2 (MD5) / Made available in DSpace on 2011-06-21T16:27:04Z (GMT). No. of bitstreams: 1 2011_KarinaSilvaFiorillo.pdf: 753032 bytes, checksum: 3c44807f8daafe4bb38f2e86e87f16f2 (MD5) / A equideocultura está em constante expansão e a Anemia Infecciosa Equina (AIE) é um obstáculo ao seu desenvolvimento por ser uma doença transmissível e incurável. Com o objetivo de se conhecer a situação epidemiológica da AIE em haras no Estado de Minas Gerais foi estimada a prevalência em sete regiões do Estado e realizada a identificação de possíveis fatores de risco. Trata-se da segunda etapa de um estudo já iniciado com a caracterização epidemiológica da enfermidade em animais de serviço. Foram amostrados 7742 equídeos pertencentes a 717 haras, distribuídos em sete estratos regionais. O diagnóstico laboratorial foi feito em série, sendo realizado o teste ELISA como triagem e a Imunodifusão em Gel Ágar (IDGA) como teste confirmatório. A prevalência encontrada foi de 0,44% [intervalo de confiança (IC) 95% = 0,00 - 0,871] haras positivos e de 0,07% [IC= 0 - 0,251] animais positivos para a AIE. Os animais de haras apresentaram menor prevalência de AIE do que a que foi previamente estimada para animas de serviço, porque provavelmente o valor zootécnico e a necessidade de emissão de guia de trânsito para participação em eventos controlados levam à maior preocupação em promover o saneamento da propriedade com exames periódicos e sacrifício dos animais positivos. Os eqüídeos de haras também transitam mais e por isso estão sujeitos a maior controle dos órgãos oficiais de defesa sanitária. As maiores prevalências foram encontradas nos estratos 1 (messoregiões Norte/Nordeste de Minas), com 0,34% e 2 (Vale do Mucuri/Jequitinhonha) com 0,72%. O estrato 7 (Campo das Vertentes e Zona da Mata) apresentou prevalência de 0,08% e nas demais regiões não foi encontrado nenhum animal positivo. Este estudo demonstra que a prevalência da AIE em haras de Minas Gerais é muito baixa e que o risco de ocorrência da doença na região norte é mais alto que no centro e sul do estado. _________________________________________________________________________________ ABSTRACT / Horse breeding is expanding and, given the fact that it is a transmissible, incurable disease, Equine Infectious Anaemia (EIA), or swamp fever, hampers its development. With a view to determining the incidence of EIA, a survey was carried out at horse farms in the State of Minas Gerais, Brazil, in order to estimate prevalence and identifying potential risk factors. This was the second stage of an on-going study on the epidemiology of the disease, that started with horses used for animal traction. A sample of 7,742 horses was examined from 717 horse farms in seven regions within the state. Laboratory tests were carried out in series using ELISA tests for screening and agar gel immunodiffusion tests to confirm the diagnosis. The prevalence was estimated at 0.44% [confidence interval (CI) 95% = 0.00 – 0.871] for farms testing positive and 0.07% [CI = 0 – 0.251] for horses. Low incidence horse farms are likely to be explained by the higher zootechnical value of stable-bred animals, compliance with the slaughter of test-positive animals, in order to remain disease-free. Stable-bred horses move more frequently and are thus more subject to compulsory testing, compared to working animals, for which the prevalence was previously estimated at higher rates. Greater prevalence among stable-bred animals was recorded in regions 1 (North/Northeast of Minas Gerais) at 0.34% and 2 (Vale do Mucuri/Jequitinhonha) at 0.72%. Region 7 (Campo das Vertentes & Zona da Mata) showed a prevalence of 0.08% while no animals tested positive in the remaining regions. This study demonstrated that the prevalence of equine infectious anaemia in stable-horses is low and that the northern region has higher disease risk compared to the centre and south of Minas Gerais.

Avec le progrès scientifique et les besoins sociétaux, diverses méthodes de détection spécifiques et sensibles des métaux, des protéines et d’autres biomolécules sont largement utilisées dans la protection de l’environnement, la surveillance des maladies, la pharmacothérapie, la production agricole, l’industrie et d’autres domaines importants. Les sondes fluorescentes sont largement développées sur la base des phénomènes de transfert d’énergie (ICT, PET, FRET) et appliquées par exemple à la détection de contaminants ou à l'imagerie cellulaire. Mais l’effet d’agrégation (ACQ) atténue généralement l’intensité de la fluorescence et limite ainsi les applications de sondes organiques (souvent peu solubles dans l’eau) dans l’imagerie cellulaire et les systèmes vivants. Par conséquent, le concept d'« agregated induced emission » (AIE) représente une solution à ces problèmes d’agrégation et plusieurs glycoclusters, glyco-sondes et glyco-complexes fluorescents ont été conçus et reportés pour des applications en analyse biologique. Notre premier projet visait à concevoir et à synthétiser des glyco-polymères fluorescents pour le ciblage cellulaire et l’adressage de médicaments, tandis que la fluorescence permettrait la détection des cellules ciblées. Pour surmonter l'effet ACQ et les interférences provenant de la fluorescence biologique naturelle, nous avons conjugué le dicyanométhylène-4H-pyrane (DCM) et le tétraphényléthène (TPE) afin d'obtenir des sondes fluorescentes (AIE) émettant dans le proche infrarouge. Les glycosides ont fourni une bonne solubilité dans l'eau et l'auto-assemblage a conduit à des systèmes de détection et à une imagerie des cellules cancéreuses. Les glyco-polymères à base de TPE ont été synthétisés à partir de monomères de TPE incorporant deux monosaccharides par conjugaison azide-alcyne (CuAAC) et ces monomères ont été polymérisés par des réactions de CuAAC ou thiol-ène. Les glyco-polymères à base de TPE ne présentaient malheureusement pas une assez grande longueur de chaîne (généralement moins de 7 unités) et les propriétés fluorescentes attendues ne pouvaient donc pas être atteintes. Nous avons ensuite conçu et synthétisé des glyco-dots auto-assemblés par des sondes DCM et des glycoclusters à base de TPE. Les glyco-dots ont présenté une hydrosolubilité élevée et une réponse sélective au peroxynitrite (ONOO-) à la fois in vitro et dans des analyses cellulaires. Les glyco-dots pourraient détecter ONOO- endogène et exogène, mais sans reconnaissance cellulaire spécifique. Nous avons conçu et synthétisé des sondes fluorescentes AIE pouvant s'auto-assembler avec des glycoclusters à base de TPE. Les glyco-dots résultants étaient facilement solubles dans l'eau et présentaient une sensibilité et une sélectivité excellentes pour la détection du thiophénol in vitro et dans des échantillons d'eau environnementaux.Nous avons finalement combiné les deux fragments TPE et DCM pour synthétiser un nouveau fluorophore AIE (TPE-DCM) avec émission à longue longueur d'onde. Ensuite, la conjugaison avec des glycosides par CuAAC a conduit à des sondes AIE fluorescentes à émission de longue longueur d'onde, avec une excellente solubilité dans l'eau. Une application à la détection de glycosidases in vitro et dans des dosages cellulaires ou sur des modèles animaux a été possible avec ces sondes / With scientific and social progress, various methods for the specific and sensitive detection of metals, proteins and other biomolecules are widely utilized in environmental protection, disease surveillance, drug therapy, agricultural production, industry and other significant areas. Fluorescent probes are widely developed based on ICT, PET, FRET and other fluorescence mechanisms, and applied to the detection of contaminants or in cell imaging. But the ACQ effect usually quenched the fluorescence intensity and thus limited the applications of organic probes in cell imaging and living systems. Therefore, the concept of aggregated-induced emission (AIE) appears as a possible solution to these problems and several fluorescent glycoclusters, glyco-probes and glyco-complexes were designed and reported for biological analysis. Our first project aimed to design and synthesize fluorescent glyco-polymers with multiple glycosides for cell targeting and drug delivery while fluorescence will allow the detection of the targeted cells. To overcome the ACQ effect and interference from natural biological background fluorescence, we conjugated dicyanomethylene-4H-pyran (DCM) and tetraphenylethene (TPE) to obtain near-infrared AIE fluorescent probes. The glycosides provided good water solubility and self-assembly in water led to detection systems and imaging cancer cells. TPE-based glycopolymers were synthesized from TPE monomers incorporating two monosaccharides by CuAAC conjugation and these monomers were polymerized by either CuAAC or thiol-ene “click” reactions. The TPE-based glycopolymers did not display a large chain length (typically less than 7 units) and the expected fluorescent properties could not be reached. We then designed and synthesized glyco-dots self-assembled by DCM probes and TPE-based glycoclusters. The glyco-dots displayed high water-solubility and selective response to peroxynitrite (ONOO-) both in vitro and in cell assays. The glyco-dots could detect endogenous and exogenous ONOO- but no specific cell recognition. We designed and synthesized AIE fluorescent probes which could self-assemble with TPE-based glycoclusters. The resulting glyco-dots were readily water soluble and displayed excellent sensitivity and selectivity for thiophenol detection in vitro and in environmental water samples. We finally combined both TPE and DCM moieties to synthesize a novel AIE fluorophore (TPE-DCM) with long-wavelength emission. Then conjugation with glycosides through CuAAC led to AIE fluorescent probes with long-wavelength emission, excellent water-solubility. Application to the detection of glycosidases in vitro and in cell assays or animal models was possible with these probes

Nous proposons de développer une méthodologie théorique pour étudier l’influence de stimuli externes sur les cristaux organiques. On s’intéresse en particulier au mécanochromisme, aggregation-induced emission, et le transfet des protons. Ces phénomènes, récemment observés expérimentalement, sont encore mal compris et il n’existe pas à l’heure actuelle de méthodologie bien définie pour les comprendre. Cette thèse développera les outils de modélisation des couplages des propriétés optiques aux phénomènes d’aggregation, réactivité et de déformation á l'etat solide. / To develop a theoretical methodology to study the influence of external stimuli on fluorescent crystals. We are particularly interested in excited state reactivity, aggregation-enhanced fluorescence and responses to mechanical stimuli. Such phenomena are often not yet understood at a molecular level, a problem exacerbated by the absence of adequate and cost-efficient computational models. This thesis counters this problem by developing a method capable of describing the solid state environment in the context of excited-state calculations at the quantum level.

Disertacijoje nagrinėjama naftos ir jos produktų garavimo terminaluose problema. Pagrindinis disertacijos tikslas – įvertinti atmosferos taršą lakiaisiais organiniais junginiais (LOJ) naftos terminaluose, nustatyti meteorologinių elementų ir krovos indikatoriaus įtaką LOJ koncentracijos kitimui pažemės atmosferos sluoksnyje. Darbe sprendžiami keli pagrindiniai uždaviniai: ištirti LOJ pasiskirstymą pažemės atmosferos sluoksnyje už naftos terminalų teritorijos skirtingais metų sezonais atsižvelgiant į meteorologinius elementus ir krovos indikatorių; sudaryti skaitinį modelį, leidžiantį prognozuoti LOJ, i��siskyrimus iš esamų, ir rekonstruojamų terminalų, sklaidą uosto zonoje; remiantis tyrimų rezultatais ir jų analize, pasiūlyti technologinius sprendimus sumažinti ar išvengti LOJ emisijos į atmosferą naftos terminaluose. / This day environment pollution research and modern environment protection technology installation became a prior aspect not only in our country, but also in the whole world. The intensification and development of the industrial processes has a negative impact on human’s health and environment. As a result, it increases waste products accumulation. It also has a disbalance of natural processes and reckless waste of natural resources. All of the above can cause greenhouse effect formation. Because of intensive expansion of energy, in industrial and transportation sectors, there is significant increase in atmosphere pollution in last decade. The West part of Lithuania is one most polluted area by volatile organic compounds (VOCs), because there is a location of oil production and refinery industry, as well as intensive development of oil terminals. The problem is becoming more serious and relevant when these objects are located near urban areas.

This day environment pollution research and modern environment protection technology installation became a prior aspect not only in our country, but also in the whole world. The intensification and development of the industrial processes has a negative impact on human’s health and environment. As a result, it increases waste products accumulation. It also has a disbalance of natural processes and reckless waste of natural resources. All of the above can cause greenhouse effect formation. Because of intensive expansion of energy, in industrial and transportation sectors, there is significant increase in atmosphere pollution in last decade. The West part of Lithuania is one most polluted area by volatile organic compounds (VOCs), because there is a location of oil production and refinery industry, as well as intensive development of oil terminals. The problem is becoming more serious and relevant when these objects are located near urban areas. / Disertacijoje nagrinėjama naftos ir jos produktų garavimo terminaluose problema. Pagrindinis disertacijos tikslas – įvertinti atmosferos taršą lakiaisiais organiniais junginiais (LOJ) naftos terminaluose, nustatyti meteorologinių elementų ir krovos indikatoriaus įtaką LOJ koncentracijos kitimui pažemės atmosferos sluoksnyje. Darbe sprendžiami keli pagrindiniai uždaviniai: ištirti LOJ pasiskirstymą pažemės atmosferos sluoksnyje už naftos terminalų teritorijos skirtingais metų sezonais atsižvelgiant į meteorologinius elementus ir krovos indikatorių; sudaryti skaitinį modelį, leidžiantį prognozuoti LOJ, išsiskyrimus iš esamų, ir rekonstruojamų terminalų, sklaidą uosto zonoje; remiantis tyrimų rezultatais ir jų analize, pasiūlyti technologinius sprendimus sumažinti ar išvengti LOJ emisijos į atmosferą naftos terminaluose.

Le travail de cette thèse intitulée "Nanomatériaux luminescents pour des applications en diagnostic" consiste en la synthèse, la caractérisation et les applications en bioimagerie des nanomatériaux. Des nanoparticules de silicium ont été synthétisées et modifiées avec des groupements fonctionnels (amino, carboxylate, sucre et complexes de platine(II)) sur la surface et appliquées dans l'imagerie cellulaire avec des cellules HeLa. En outre, l'ensemble des nanoparticules de silicium modifiées avec des complexes de platine(II) présentent un effet d’émission d’agrégation induite (AIE) intéressant. Par ailleurs, une série de complexes d'iridium(III) ont été synthétisés permettant d’obtenir une émission allant du jaune au rouge. Ces complexes d'iridium(III) ont en outre été utilisés pour une application en électrochimiluminescence (ECL). L’ECL est un outil d'imagerie qui génère des émissions sans excitation lumineuse et qui attire de plus en plus l’attention dans les analyses biologiques. Des nanosytèmes de « carbon dots » et des nanoparticules de silicium modifiés avec des complexes métalliques ont également été étudiés pour des propriétés en ECL. / The work of this thesis titled “Luminescent Nanomaterials for diagnostic applications” is synthesis, characterization and bioimaging applications of nanomaterials. Silicon nanoparticles were synthesized and modified with different functional groups such as amino, carboxylate, sugar and platinum(II) complex on the surface, and applies for cellular imaging at HeLa. Moreover, the assembly platinum(II) complexes modified silicon nanoparticles exhibit an interesting aggregation induced emission (AIE) effect. In addition, a series of iridium(III) complexes were synthesized with tunable emission color from yellow to red. Those iridium(III) complexes were further used for electrochemiluminescence (ECL) application. ECL is an imaging tool that generate emission without light excitation and has gained more attention in many bioassays. Besides, nanosystem of metal complex modified carbon dots and silicon nanoparticles were also investigated the ECL properties.