La transplantation pulmonaire est une alternative thérapeutique ultime dans de nombreuses maladies pulmonaires sévères, en particulier chez des patients atteints de mucoviscidose (Cystic Fibrosis) , de fibrose pulmonaire idiopathique (IPF idiopathie pulmonary fibrosis), delymphangioléiomyomatose (LAM) ou d'hypertension pulmonaire. Cependant, les besoins en greffe dépassent largement le nombre de transplantations pratiquées en France. Les causes de ce déséquilibre incluent un trop faible nombre de donneurs potentiels en raison de critères actuels pour l'acceptation du greffon qui sont restreints car les biomarqueurs biologiques de qualité et de viabilité du greffon n'ont pas été décrits à ce jour. L'une des possibilités d'augmenter le nombre de donneurs, est de mettre en évidence des biomarqueurs de la qualité du greffon et d'étendre les critères d'acceptabilité du greffon en permettant les prélèvements à partir de donneurs à coeur arrêté est une possibilité. Les prélèvements pulmonaires après arrêt cardiaque sont effectués, en clinique expérimentale chez l'homme, en Espagne depuis trois ans. Il existe dans ce pays une législation très favorable et surtout une conscience collective qui rend le don d'organes extrêmement facilité. Il est donc essentiel d'optimiser l' utilisation de cette ressource. Pour cela, la mise en place de critères de validation de la qualité du greffon est une donnée clé pour palier à ce manque de transplantation pulmonaire. Les critères d'acceptabilité d'un greffon pulmonaires sont basés sur des données cliniques n'existant pas de biomarqueurs biologiques de qualité et de viabilité du greffon. Nous proposons ici l'utilisation de la métabolomique par spectroscopie en Résonance Magnétique Nucléaire à haute résolution par rotation à angle magique (RMN HRMAS) pour mettre en évidence des biomarqueurs de la qualité du greffon. La métabolomique par la spectroscopie RMN HRMAS, est une technique d'analyse rapide (20 minutes) et originale, caractérisée par l'analyse directe d'un tissu biologique intact sans nécessité d'extraction. Cette technique étudie les profils métaboliques dans le but de mettre en évidence des biomarqueurs métaboliques. La métabolomique a été largement utilisée dans des études en cancérologie pour la détermination de la malignité d'un tissu (Bertini 1. et coll. , Canser Res. 2012/ Griffin JL et coll ., Nat Rev Cancer, 2004/ Li M. et coll. , PLoSOne, 2011/ O'connell TM. Bioanalysis. 2012/ Ma Y. et coll. Mol Biol Rep. 2012). Cependant, très peu de publications dans la littérature s'intéressent au domaine de la transplantation et à la qualité du greffon (Rocha C. et coll. , Journal of Proteome Research, 2011/ RobertR. et coll. J Critical Ca re 2010/ Stenlund H. et coll., Chemometrics and Intelligent Laboratory Systems, 2009/ Du a rte F.L. et coll. , Anal.Chem.2005). Ce projet de thèse avait pour objectifs de :1. Etudier la faisabilité d'utiliser la métabolomique par la spectroscopie RMN HRMAS pour évaluer la qualité du greffon pulmonaire.2. Mettre en évidence de potentiels biomarqueurs de la qualité du greffon pulmonaire.3. Evaluer une éventuelle introduction de cette technique en pratique courante dans un environnement hospitalier. Pour répondre à ces objectifs, il a été entrepris :• Etudier les métabolomes pulmonaires de différentes espèces animales, puis de les comparer au métabolome du poumonhumain afin d'identifier le modèle expérimental le plus adapté à la transplantation pulmonaire.• Evaluer la qualité du greffon chez un modèle animal (porc Large White) pour la transplantation pulmonaire (modèle expérimental de préservation pulmonaire en in situ chez le donneur à coeur arrêté, modèle de perfusion pulmonaire en ex vivo sur machine ocs™).• Evaluer l'effet de la perfusion de deux solutions de conservation sur la qualité du greffon pulmonaire chez le porc. / Lung transplantation is a therapeutic alternative in many severe pulmonary diseases, especially in patients suffering from CF (CysticFibrosis), idiopathie pulmonary fibrosis (IPF), lymphangioleiomyomatosis (LAM) or pulmonary hypertension . However, the need fortransplantation far outweighs the number of transplants performed in France. The causes of this imbalance include an insufficient number of potential donors because of the current criteria for the acceptance of the graft that are restricted as biomarkers of quality and viability of the graft have not been described so far.One of the possibilities to increase the donor pool is to identify biomarkers of the quality of the graft and expand the criteria foracceptability of the graft allowing withdrawals from non-heart-beating donors. The lung taking were performed after cardiac arrest inclinical trials carried out on humans for three years in Spain.lt is therefore essential to optimize the use of this resource. For this, the establishment of criteria for validating the quality of the graft isgiven a key to solve this problem of lung transplantation. The criteria for acceptability of a lung transplant are based on clinical data inabsence of biomarkers of quality and viability for the lung graft.We propose the use of the metabolomics by high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMASNMR) to highlight potential biomarkers for the quality of the graft.The Metabolomics by NMR HRMAS spectroscopy is an original analytical technique characterized by a rapid analysis (20 minutes)performed on intact biopsy samples without extraction prior to analysis. This technique studies the metabolic profiles in arder to identifymetabolic biomarkers. Metabolomics has been widely used in studies in oncology for the determination of malignancy of a tissue (Bertini et al. , Canser Res. 2012/ Griffin JL et al., Nat Rev Cancer, 2004/ Li M. et al. , PLoS One, 2011/ O'connel! TM. Bioanalysis. 2012/ Ma Y. et al. Mol Biol Rep. 2012). However, very few papers in the literature combine the use of the metabolomics NMR HRMAS and the assessment of the quality of the graft (Rocha C. et al. , Journal of Proteome Research, 2011/ Robert R. et al. J Critical Care 2010/ Stenlund H. et al, Chemometrics and Intelligent Laboratory Systems, 2009/ Duarte F.L. et al. , Anal. Chem. 2005).The purposes of this research work were:1. Studying the feasibility of using the metabolomics by NMR HRMAS for the assessment of the quality of the lung graft2. Assess the metabolome of the lung in degradation conditions and highlight potential biomarkers of the quality of the graft3. Assess the possible use of the metabolomics by NMR HRMAS as a tool in clinical practice within a hospital environment.To answer to these purposes we made experimental experiences as follows:- Studying the lung metabolome of various animal species, and compare them to the human metabolome to identify the most suitableexperimental model for lung transplantation.- Assessing the quality of the graft in an animal model (pig Large White) for lung transplantation (experimental model of lung preservation in situ in the case of non-heart-beating donor, lung model for an ex vivo perfusion using OCS ™ Machine)- Evaluating the effect of perfusion with two preservation solutions on the quality of lung graft in pig model.

Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-06-23T19:03:26Z No. of bitstreams: 1 texto completo.pdf: 790178 bytes, checksum: f15ebd918e983ee92d3e70cf19f11647 (MD5) / Made available in DSpace on 2017-06-23T19:03:26Z (GMT). No. of bitstreams: 1 texto completo.pdf: 790178 bytes, checksum: f15ebd918e983ee92d3e70cf19f11647 (MD5) Previous issue date: 2004-11-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Com a elevação dos custos da adubação mineral, resíduos orgânicos produzidos pelo meio rural passaram a ser mais usados para melhorar as condições químicas e físicas dos solos. Porém, não se sabe ao certo quais as doses mais adequadas de cada tipo de resíduo orgânico para diferentes culturas nos diferentes ambientes edáficos e também pouco se sabe dos possíveis riscos ambientais decorrentes de seu uso. Desta maneira, visou-se avaliar o potencial fertilizante de camas de aviário quanto a P, dado ser este o nutriente mais limitante nos solos brasileiros. Assim, foram objetivos deste trabalho: obter uma estimativa das doses adequadas de camas de aviário para maximizar a produção de matéria seca de plantas de milho e avaliar a participação de formas orgânicas e inorgânicas de P destas camas de aviário e sua dinâmica de transformação, na ausência e na presença de solo. No experimento 1, foram avaliados cinco tipos de camas de aviário em doses correspondentes a 0, 20, 40, 80 e 160 t ha -1 , que foram aplicadas em dois Latossolos (um muito argiloso e o outro de textura média) em três repetições, incubados por 15 dias e realizados três cultivos sucessivos de milho, determinando-se a matéria seca produzida e o P acumulado na planta. No experimento xiv2, determinaram-se os teores de formas orgânicas de P (Po) e inorgânicas (Pi) de cinco camas de aviário em três repetições e as transformações que ocorreram nessas formas após incubação na ausência ou presença de solo, utilizando-se fracionamento químico e ressonância magnética nuclear de 31 P ( 31 P-RMN). O experimento 3 foi conduzido semelhantemente ao 1, porém com dose única de cama de maravalha de 80 t ha -1 , além de um tratamento-testemunha (ausência de aplicação de cama) visando avaliar a dinâmica das transformações das formas de P no solo com o tempo, por meio de fracionamento químico. Os resultados indicaram que para se obter a produção máxima acumulada em três cultivos, sem que ocorressem efeitos tóxicos às plantas, seria adequada a aplicação de uma dose de cama de aviário equivalente a 70 t ha -1 para o solo muito argiloso e em torno de 60 t ha -1 para o solo textura média. Verificou-se que os ortofosfatos de monoésteres foram as formas de Po predominantes nas camas de aviário (15,1 % do P extraído por água), porém, os ortofosfatos de diésteres estávam presentes em quantidades apreciáveis (4,7 % do P extraído por água) e disponibilizaram Pi mais rapidamente. A mineralização do Po foi relativamente rápida nos primeiros 15 dias, mas existiu diferença significativa nas taxas de transformação de Po em Pi entre tipos de camas. No solo, a cama de casca de café teve maior taxa de mineralização de Po (44,7 %), enquanto a menor taxa foi observada para a cama de maravalha (4,9 %). As maiores transformações do P nas diversas frações ocorreram durante o período de incubação (0 a 15 dias), sendo que no solo argiloso as frações de P de baixa disponibilidade (extraídas por hidróxido e residual) predominaram, mas no solo de textura média as formas mais lábeis de P (por resina e por bicarbonato) e o P extraído por água apresentaram maiores teores. O solo de textura média, comparativamente ao muito argiloso, poderia estar mais sujeito à perda de P solúvel em água por lixiviação quando fertilizado com camas de aviário. / Due to the rising costs of chemical fertilization, organic compounds produced in rural areas are being more frequently used to improve chemical and physical soil conditions. However, it is not well known the adequate doses of different types of organic residues for distinct crops and soils. Furthermore, little is known on the possible risks that the use of these organic residues may pose to the environment. This study sought to evaluate the potential of different poultry litters to supply phosphorus (P), the nutrient most limiting to crop growth in Brazilian soils. Thus, the objectives of this research were: to obtain an estimate of adequate doses of poultry litter to maximize growth of corn plants and evaluate the organic and inorganic P forms in the poultry litter and the dynamics of their transformation, in the absence and presence of soil. In experiment 1 it was evaluated five types of poultry litter in doses equivalent to 0, 20, 40, 80 and 160 t ha -1 , applied to two Oxisols (a clayey and a sandy-loam), with three replicates. The soils plus poultry litters mixtures were incubated for 15 days under greenhouse conditions and then corn was cultivated for three successive times. When plants were 25-day old they were harvested and the dry matter production and P uptake were determined. In experiment 2, it was determined the contents of organic (Po) and xviinorganic (Pi) phoshorus forms of the poultry litters and the transformations of these P forms after their incubation in the absence and presence of soil, using wet chemical fractionation and 31 P – Nuclear Magnetic Ressonance Spectroscopy ( 31 P-NMR). Experiment 3 was carried out similarly to experiment 1, except that only wood chip litter was used (0 and 80 t ha -1 ) with the objective of evaluate the transformations of P forms in the soil along the time of incubation using wet chemical fractionation. The results indicate that in order to obtain the maximum estimated growth (accumulated in the three cultivations) and no toxicity effect occurred to plants, a dose of poultry litter equivalent to 70 t ha -1 in the clayey soil and 60 t ha -1 in the sandy-loam soil would be adequate. It was found that monoester orthophosphates were the dominant Po forms in the poultry litter (15.1% of P extracted with water) but, the diester orthophosphate were present in substantial amount (4.7 % of P extracted with water) and mineralized more rapidly. The mineralization of Po was relatively rapid in the first 15 days, but there were significant differences in the Po and Pi transformation rates among poultry litter types. In the soil, the coffee cherry skin litter showed the greatest mineralization rate (44.7%), whereas the lowest rate was observed for the wood chip litter (4.9%). The majority of transformations of P from several forms occurred during the incubation period (0 to 15 days). In the clayey soil the low availability P fractions (residual and NaOH-extracted) were dominant, but in the sandy-loam soil more labile P fractions (resin and bicarbonate-extracted) and water-extractable P presented higher values. Compared to the clayey soil, the sandy-loam would probably be more prone to soluble P losses in the percolation water when fertilized with poultry litter.

NASCIMENTO, Raimundo Regivaldo Gomes do. Estudo Químico e Biológico de Margaritopsis carrascoana Wright (Rubiaceae). 2014. 275 f. Tese (Doutorado em química)- Universidade Federal do Ceará, Fortaleza-CE, 2014. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-06-02T20:19:14Z No. of bitstreams: 1 2014_tese_rrgnascimento.pdf: 13947616 bytes, checksum: 924a0eb7fa05854c8e13053f4965ef41 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-07-20T20:09:26Z (GMT) No. of bitstreams: 1 2014_tese_rrgnascimento.pdf: 13947616 bytes, checksum: 924a0eb7fa05854c8e13053f4965ef41 (MD5) / Made available in DSpace on 2016-07-20T20:09:26Z (GMT). No. of bitstreams: 1 2014_tese_rrgnascimento.pdf: 13947616 bytes, checksum: 924a0eb7fa05854c8e13053f4965ef41 (MD5) Previous issue date: 2014 / Margaritopsis carrascoana is a small shrub belonging to the Rubiaceae family and endemic from northeastern of Brazil flora growing in the sandy soils of the region of Ibiapaba and Araripe plateaus – Ceará state. The absence of reports of phytochemical studies related to this species, combined with occurrence of bioactive alkaloids in the genus, motivated us to perform chemical study. The plant specimen was collected in Araripe plateu, in Moreilândia-PE county. The phytochemical investigation of the ethanolic extract from the stems yielded the alkaloids calycosidine, hodgkinsine, N-8”-formyl-calycosidine and N-8”-methyl-N-1’-desmethylisocalycosidine, besides neolignan dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside, the flavonol luteolin 7-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl, the triterpenes lupeol and ursolic acid, and the mixture of β-sitosterol and stigmasterol steroids, as aglycones and glycosylated. From the ethanolic extract of the leaves were isolated the flavonoid luteolin 7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside, chrysoeriol 7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside, luteolin 7-O-{β-D-apiofuranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranosyl} and luteolin 7-O-{α-L-rhamnopyranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranosyl}. The alkaloids N-8"-formyl-calycosidine, N-8”-methyl-N-1’-desmethylisocalycosidine, luteolin 7-O-{β-D-apiofuranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranosyl} and luteolin 7-O-{α-L-rhamnopyranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyra-nosyl}, are being reported for the first time in the literature, and the other secondary metabolites are unprecedented in the genus Margaritopsis. The secondary metabolites were isolated using classical chromatography techniques; including adsorption chromatography on silica gel, exclusion chromatography on Sephadex LH-20, reverse phase chromatography (C-18), and high performance liquid chromatography (HPLC). For structural characterization were used infrared spectroscopy, mass spectrometry and nuclear magnetic resonance techniques including uni (1H NMR and 13C NMR and DEPT 135) and two-dimensional experiments (HMBC, HSQC, COSY and NOESY), and comparison with the literature data. In addition, the flavonoids flavonol luteolin 7-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl, luteolin 7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside, luteolin 7-O-{β-D-apiofuranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranosyl} and luteolin 7-O-{α-L-rhamnopyranosyl-(1→6)-[β-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranosyl, sho-wed antioxidant activity greater than the BHT and quercetin standards, while ethanol extracts of stems and leaves showed inhibitory activity on the acetylcholinesterase enzyme. On the other hand, hodgkinsine showed potent cytotoxic activity against ovary, glioblastoma and colon cancer cells lines. The ethanol extract of the leaves and its alkaloidal fraction were submitted to nociception test and yielded good results. The ethanolic extract of the leaves was subjected to gastric antiulcer activity test, leading to a significant reduction in gastric lesions induced by ethanol in mice. / Margaritopsis carrascoana é um pequeno arbusto pertencente à família Rubiaceae e endêmico da flora do Nordeste brasileiro, que cresce em solos arenosos do planalto da Ibiapaba e serra do Araripe - Ceará. A ausência de relatos acerca de estudos fitoquímicos relacionados à espécie, aliada a ocorrência de alcalóides bioativos no gênero, nos motivou ao seu estudo químico. Desta forma, o espécimen vegetal foi coletado na chapada do Araripe, município de Moreilândia-PE. A investigação fitoquímica do extrato etanólico dos talos resultou no isolamento dos alcalóides calicosidina, hodgkinsina, N-8”-formilcalicosidina e N-8”-metil-N-1’-desmetilisocalicosidina, da neolignana álcool 4-O-β-D-glicopiranosil-di-hidro-desidrodiconiferílico, do flavonóide 7-O-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil luteolina, dos triterpenos lupeol e o àcido ursólico, e da mistura de esteróides β-sitosterol e estigmasterol, como agliconas e nas formas glicosiladas. A partir do estudo do extrato etanólico das folhas foram isolados os flavonóides 7-O-[β-D-glicopiranosil-(1→6)-β-D-apiofuranosil] luteolina, 7-O-[β-D-glicopiranosil-(1→6)-β-D-apiofuranosil] crisoeriol, 7-O-{β-D-apiofuranosil-(1→6)-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina e 7-O-{α-L-ramnopiranosil - (1→6) - [α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina. Os alcalóides N-8”-formilcalicosidina e N-8”-metil-N-1’-desmetilisocalicosidina, e os flavonóides 7-O-{β-D-apiofuranosil-(1→6)-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina e 7-O-{α-L-ramnopiranosil-(1→6)-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina, estão sendo relatados pela primeira vez na literatura, enquanto todas as demais substâncias possuem caráter inédito no gênero Margaritopsis. O isolamento dos metabólitos secundários foi conduzido através de técnicas cromatográficas clássicas, incluindo cromatografia de adsorção em gel de sílica, cromatografia por exclusão molecular em Sephadex LH-20, cromatografia de fase reversa (C-18) e cromatografia líquida de alta eficiência (CLAE). Para a caracterização estrutural foram utilizadas técnicas espectroscópicas utilizando infravermelho, espectrometria de massas e ressonância magnética nuclear, incluindo técnicas uni (RMN 1H e RMN 13C e DEPT 135) e bidimensionais (HMBC, HSQC, COSY e NOESY), além de comparação com dados descritos na literatura. Em adição, os flavonóides 7-O-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil luteolina, 7-O-[β-D-glicopiranosil-(1→6)-β-D-apiofuranosil] luteolina, 7-O-{β-D-apiofuranosil-(1→6)-[α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina e 7-O-{α-L-ramnopiranosil-(1→6) - [α-L-ramnopiranosil-(1→2)-β-D-glicopiranosil} luteolina apresentaram atividade antioxidante maior que os padrões BHT e quercetina, enquanto os extratos etanólicos dos talos e folhas apresentaram atividade inibidora da enzima acetilcolinesterase. Por outro lado, o alcaloide hodgkinsina apresentou potencial citotóxico frente às células de ovário, glioblastoma e colon. No teste de nocicepção, realizado com o extrato etanólico das folhas e a fração alcalóidica, foram observados resultados positivos para ambas as frações. O extrato etanólico das folhas foi submetido a teste de atividade antiúlcera gástrica, levando a uma redução significativa da área de lesão gástrica induzida pelo etanol em camundongos.

Ce travail de thèse porte sur la synthèse de nanocristaux de phosphure d'indium (InP) et en particulier sur l'utilisation de précurseurs phosphorés tels que le phosphore blanc et les aminophosphines. Les nanocristaux d'InP sont des matériaux semi-conducteurs prometteurs dans le cadre d'applications biologiques et optoélectroniques grâce à leur faible toxicité et à leurs spectres d'absorption et de fluorescence dans le visible. En outre, le phosphore blanc, allotrope le plus réactif du phosphore, est un produit industriel fabriqué à très grande échelle. Il est en effet à l'origine de tous les produits phosphorés à bas degrés d'oxydation tels que les phosphines. Sa fonctionnalisation directe visant à former des espèces chimiques nouvelles ou valorisables est un domaine de recherche actif. Alors que les nanocristaux d'InP sont traditionnellement synthétisés en utilisant une silylphosphine comme précurseur phosphoré, une nouvelle méthodologie reposant sur l'utilisation d'aminophosphines a émergé. Les aminophosphines étant plus facilement accessibles et manipulables que les silylphosphines, il s'agit d'une avancée importante pour le développement des nanocristaux d'InP. C'est pourquoi nous avons étudié précisément le mécanisme de formation de ces nanocristaux, en nous appuyant notamment sur la RMN, la spectrométrie de masse et les calculs DFT. Cette compréhension fine du mécanisme a permis l'optimisation de la synthèse des nanocristaux d'InP.L'utilisation du phosphore blanc pour former des nanocristaux d'InP repose sur la formation de nanoparticules d'indium monodisperses puis sur l'incorporation du phosphore à l'intérieur des nanoparticules. En partant d'une méthodologie de synthèse de nanoparticules d'indium de la littérature, nous avons découvert le paramètre central de la synthèse : la présence d'une quantité bien précise d'eau dans le solvant. Les nanoparticules d'indium ainsi formées sont oxydées en surface et doivent être activées afin de réagir avec le phosphore blanc.Enfin, la fonctionnalisation moléculaire du phosphore blanc avec des borohydrures pour former des liaisons P-H et par voie radicalaire pour former des silylphosphines est présentée. / Ce travail de thèse porte sur la synthèse de nanocristaux de phosphure d'indium (InP) et en particulier sur l'utilisation de précurseurs phosphorés tels que le phosphore blanc et les aminophosphines. Les nanocristaux d'InP sont des matériaux semi-conducteurs prometteurs dans le cadre d'applications biologiques et optoélectroniques grâce à leur faible toxicité et à leurs spectres d'absorption et de fluorescence dans le visible. En outre, le phosphore blanc, allotrope le plus réactif du phosphore, est un produit industriel fabriqué à très grande échelle. Il est en effet à l'origine de tous les produits phosphorés à bas degrés d'oxydation tels que les phosphines. Sa fonctionnalisation directe visant à former des espèces chimiques nouvelles ou valorisables est un domaine de recherche actif. Alors que les nanocristaux d'InP sont traditionnellement synthétisés en utilisant une silylphosphine comme précurseur phosphoré, une nouvelle méthodologie reposant sur l'utilisation d'aminophosphines a émergé. Les aminophosphines étant plus facilement accessibles et manipulables que les silylphosphines, il s'agit d'une avancée importante pour le développement des nanocristaux d'InP. C'est pourquoi nous avons étudié précisément le mécanisme de formation de ces nanocristaux, en nous appuyant notamment sur la RMN, la spectrométrie de masse et les calculs DFT. Cette compréhension fine du mécanisme a permis l'optimisation de la synthèse des nanocristaux d'InP.L'utilisation du phosphore blanc pour former des nanocristaux d'InP repose sur la formation de nanoparticules d'indium monodisperses puis sur l'incorporation du phosphore à l'intérieur des nanoparticules. En partant d'une méthodologie de synthèse de nanoparticules d'indium de la littérature, nous avons découvert le paramètre central de la synthèse : la présence d'une quantité bien précise d'eau dans le solvant. Les nanoparticules d'indium ainsi formées sont oxydées en surface et doivent être activées afin de réagir avec le phosphore blanc.Enfin, la fonctionnalisation moléculaire du phosphore blanc avec des borohydrures pour former des liaisons P-H et par voie radicalaire pour former des silylphosphines est présentée.

Made available in DSpace on 2016-06-02T20:34:04Z (GMT). No. of bitstreams: 1 TeseGBA.pdf: 7751002 bytes, checksum: 6ec58a5eabb6147b50170358eceefa09 (MD5) Previous issue date: 2007-02-02 / This present work describes the chemotaxonomic analysis of lichens through the utilization of NMR spectroscopic techniques, HR-MAS and NMR in solution, and IR allied with chemometrics methods. This analysis were carried out with eleven species set of lichens divided in two families and six genera. The HR-MAS NMR spectra together the chemometric, obtained from intact vegetal material, present the discrimination between the families, genera and species and the classification percentage was much satisfactory (70,8%). The analyzed substances include the carbohydrates class of low molecular-weight, the polyols. The NMR in solution analysis, corresponding to crude extracts, present the discrimination between genera and species and the classification was excellent (88,9%). In this analysis, where the secondary metabolites of lichens are evaluated, was realized the identification of some substances in mixture. The analysis by infrared spectroscopy (IR), also obtained from intact material, was detached by the excellent power of samples classification (100% of correct prediction), with discrimination between genera and species. The comparative application of this techniques confirm the results obtained separately, and point for a great potentiality of IR in chemotaxonomic analysis of lichens, due its quickness and low price, however the NMR techniques are more attractive when desire the circumstantial analysis of spectral profile. / O presente trabalho descreve a análise quimiotaxonômica de liquens, a partir da utilização das técnicas espectroscópicas de RMN, HR-MAS e em solução, e IV aliadas aos métodos quimiométricos. As análises foram efetuadas empregando-se um conjunto de onze espécies de liquens, distribuídas em duas famílias e seis gêneros. Os espectros de RMN HR-MAS, obtidos a partir do material vegetal intacto, apresentou, juntamente com a quimiometria, a discriminação entre as famílias, gêneros e espécies e ainda um percentual de classificação muito satisfatório (70,8%). As substâncias analisadas compreendem a classe dos carboidratos de baixa massa molecular, os polióis. As análises de RMN em solução, correspondentes aos extratos brutos, apresentaram a discriminação de gêneros e espécies e a classificação foi excelente (88,9%). Nessa análise, a qual os metabólitos secundários de liquens são avaliados, foi realizada a identificação em mistura de algumas substâncias. A análise por espectroscopia na região do infravermelho (IV), obtidas também a partir do material intacto foi destacada pelo seu excelente poder de classificação de amostras (100% de predições corretas), com discriminação entre os gêneros e espécies. A aplicação comparativa dessas técnicas espectroscópicas corrobora os resultados obtidos separadamente, e aponta para uma grande potencialidade do IV em análises quimiotaxonômicas de liquens, dado a sua rapidez e baixo custo, embora as técnicas de RMN sejam muito atrativas quando se deseja uma análise minuciosa do perfil espectral.

Made available in DSpace on 2016-06-02T20:34:16Z (GMT). No. of bitstreams: 1 2111.pdf: 4098526 bytes, checksum: 934a5ed1df0d898498c78f0ae9e109af (MD5) Previous issue date: 2005-06-29 / Financiadora de Estudos e Projetos / The Biotechnology has made possible the development of genetically modified (GM) organisms, including soybean, main Brazilian agricultural product. These GM specimens has been produced by changing some original characteristics of plant in order to increase productivity and to reduce production costs. However, these novel organisms should be evaluated, in order to verify its safety for environment and human and animal consume. In this context, it is necessary to develop new analytical methodologies for substantial equivalence determination of these novel plants. NMR is an important analytical tool allowing acquisition of a great number of chemical information in a single experiment. Within NMR techniques, HR-MAS makes possible direct acquisition on intact samples (In situ), without any sample treatment. In addition, chemometric analysis is a powerful tool, which allows interpretation of high complex data, such as NMR spectra. In this work, HR-MAS NMR has been employed for acquisition of NMR spectra directly of soybeans and leaves. Application of chemometric analysis on NMR data, has made possible to discriminate samples according to cultivar, geographic origin and mainly genetic origin, as well as to predict the identity of unknown samples by means of classification models. For this, all methodology, including HR-MAS NMR spectra measurements and chemometric analysis, was developed and optimized. Chemometric analysis performed on HR-MAS NMR spectra has also allowed to determine differences on chemical composition among samples, mainly those between traditional and GM specimens, giving important information about substantial equivalence of GM soybean crops. In conclusion, it has been demonstrated that HR-MAS NMR technique together with chemometric analysis is an important tool to be used in metabolomic studies and in quality control of agricultural products. / A biotecnologia tem possibilitado o desenvolvimento de organismos geneticamente modificadas (OGMs), entre eles a soja, principal produto agrícola brasileiro. Permitindo assim, alterar as características da planta, visando aumentar a produtividade e reduzir custos. No entanto, é necessário avaliar os OGMs quanto a segurança alimentar e ambiental. Desta forma, são necessárias metodologias analíticas capazes de atuar na determinação da equivalência substancial destes novos organismos. Dentre as técnicas, a RMN possui um importante atrativo devido ao fato de fornecer um grande número de informações químicas sobre o sistema em estudo, em um único experimento. Além disso, a técnica de RMN HR-MAS possibilita a aquisição de medidas diretamente de materiais intactos (In situ), sem a necessidade de quaisquer etapas de preparo de amostra. Aliada a esta, a quimiometria é uma poderosa ferramenta que permite analisar e interpretar dados de alta complexidade, tais como, espectros de RMN. Neste trabalho, a RMN HR-MAS tem sido empregada para a aquisição de experimentos diretamente de sementes de soja e de folhas da planta. A aplicação de análises quimiométricas sobre os espectros de RMN HR-MAS possibilitou discriminar as amostras de acordo com a cultivar, origem geográfica e, principalmente, origem genética, bem como predizer a origem de amostras desconhecidas através da construção de modelos de classificação. Para isso, toda a metodologia desde a aquisição dos espectros de RMN HR-MAS às análises quimiométricas foi desenvolvida e otimizada. A análise quimiométrica dos espectros de RMN HR-MAS tem permitido também determinar as diferenças na composição química entre as amostras, principalmente aquelas entre espécimens GM e convencionais, fornecendo importantes informações sobre a equivalência substancial da soja GM. Desta forma, a RMN HR-MAS aliada a quimiometria tem se mostrado uma importante ferramenta para atuar em estudos metabolômicos e no controle de qualidade de produtos agrícolas.

Made available in DSpace on 2016-06-02T20:34:29Z (GMT). No. of bitstreams: 1 3943.pdf: 3672997 bytes, checksum: 976906627d883988e6f1df73a31d7614 (MD5) Previous issue date: 2011-11-24 / Financiadora de Estudos e Projetos / This work highlights the implementation, development and application of the Ultrafast Nuclear Magnetic Resonance (UFNMR) technique, proposed by Frydman and co-workers. Through this technique, it is possible to perform multidimensional experiments in a single scan, thus reducing the experiment time drastically, which can be a fraction of second. This study shows a practical protocol, which was optimized for the implementation of the first ultrafast 2D NMR COSY and HSQC experiments performed in Brazil and, to our knowledge, in the southern hemisphere. This process involves the setting of several parameters from the initial calibration of encoding gradients and pulses, to the setting of a specific acquisition scheme required by the technique. Furthermore, some important operational aspects are discussed in order to contribute, even more, to future implementations to be carried out safely and efficiently. Also, as part of this study, a software in C language was developed to obtain the real-time processing (1 second) of the UF-NMR data, and this was incorporated in the Bruker software for the acquisition and processing of NMR data - TopSpin® version 3.0. Finally, two studies were also performed using the UF-NMR experiments. The first one was the real-time identification of a mixture of flavonoids (epicatechin, naringenin and naringin), which underwent on-flow chromatographic separation, in a commercial HPLC-NMR system. UF-COSY experiments were performed during the chromatographic run, so that it was possible to obtain the 2D NMR spectra related to each flavonoid and to successfully follow their separation over the course of time. The second application was the monitoring of the hydrolysis reaction of the 2-(4-nitrophenyl)-1,3-dioxolane acetal. The hemiacetal intermediate of this reaction has a short lifetime, being difficult to identify by conventional NMR. Therefore, HSQC-UR experiments were performed during the reaction development, thus, it was possible to characterize the presence of the hemiacetal intermediate. This assignment was confirmed by quantum calculation of 1H and 13C NMR chemical shifts and of the NBOs (Natural Bonding Orbital). / O trabalho em questão destaca a implementação, desenvolvimento e aplicação da técnica de Ressonância Magnética Nuclear Ultrarrápida (RMN-UR), proposta por Frydman e colaboradores. Por meio desta técnica, é possível realizar experimentos multidimensionais numa única varredura, reduzindo dessa forma, drasticamente, o tempo de aquisição, que pode durar uma fração de segundo. É apresentado um protocolo prático, que foi otimizado para implementação dos primeiros experimentos de RMN 2D COSY e HSQC ultrarrápidos realizados no Brasil, e ao que se sabe no hemisfério Sul. Este processo envolve desde a calibração de pulsos e gradientes, até a configuração do esquema específico de aquisição relativo a esta técnica. Além disso, alguns aspectos operacionais importantes são discutidos visando contribuir ainda mais para que futuras implementações sejam realizadas de maneira segura e eficiente. Também como parte deste estudo foi desenvolvido um programa em linguagem de programação C, para o processamento em tempo real (1 segundo) dos dados de RMN-UR, e este foi incorporado ao programa computacional de aquisição e processamento dos dados de RMN da empresa Bruker - TopSpin® versão 3.0. Foram realizados ainda dois estudos aplicando os experimentos de RMN-UR. O primeiro trata-se da identificação em tempo real de uma mistura de 3 flavonoides (epicatequina, naringenina e naringina) submetida à separação cromatográfica, num sistema CLAE-RMN no modo fluxo contínuo. Experimentos de COSY-UR foram realizados ao longo de toda corrida cromatográfica, sendo possível obter os espectros 2D correspondentes de cada flavonoide e acompanhar com sucesso a separação destes no decorrer do tempo. A segunda aplicação foi o monitoramento da reação de hidrólise do acetal 2-(4-nitrofenil)-1,3-dioxolano. O intermediário hemiacetal desta reação possui curta duração, sendo portanto de difícil identificação via RMN tradicional. Por isso, foram realizados experimentos de HSQC-UR durante todo o desenvolvimento da reação e, por meio destes, foi possível caracterizar a presença do intermediário hemiacetal. A correlação atribuída a este foi confirmada através de cálculos quânticos de deslocamento químico de RMN de 1H e 13C e dos NBOs (Natural Bonding Orbital).

Les microARNs sont une classe de petits ARNs non codants qui régulent l'expression des gènes via un mecanisme d'interference par ARN. Les microARNs humains sont produits par une série de réactions enzymatiques. En particulier, dans le cytoplasme le precurseur de miRNA (pre-miRNA) est reconnu et clivé par un complexe contenant l'enzyme RNAse III Dicer et plusieurs cofacteurs protéiques. La proteine TRBP (HIV TAR RNA binding protein) est l'un de ces cofacteurs et augmente la stabilité du complexe, influe sur la cinétique, la position du clivage et a role potentiel dans la reconnaissance du substrat et dans le transfet du produit vers le complexe RISC (RNA-induced silencing complex) effecteur de l'interference par ARN. TRBP est composé de 3 domaines de liaison aux ARN doubles brin (dsRBDs). La région d'interaction de TRBP avec les ARNs est composé des deux premiers dsRBDs liés par une région interdomaine non charactérizée. La présente étude rapporte la caractérisation biophysique in vitro de la région d'interaction avec les ARNs de TRBP dans l'état apo de TRBP ou dans l'état lié avec les deux precurseurs cytoplasmique successifs du microARN oncogène miR-155 comprenant la tige boucle pre-miR-155 et le duplex miR-155/miR-155* résultat du clivage de pre-miR-155 par Dicer. L'étude montre que la région d'intéraction de TRBP avec les ARNs est monomerique, est composée de deux dsRBDs independants en solution et que la région interdomaine de 60 résidus est flexible. Le premier dsRBD, non caractérisé précédement en solution est le siège d'un equilibre plié/déplié integral dans une grande gamme de conditions physico-chimiques. Les deux premiers dsRBDs de TRBP peuvent interagir avec un même precurseur de microARN et deux régions d'interaction de TRBP avec les ARNs peuvent interagir avec un même precuseur. La région d'interaction de TRBP avec les ARNs interagit avec pre-miR-155 et le duplex miR-155/miR-155* avec des affinités très similaires. Dans le complexe avec une région d'interaction de TRBP avec les ARN liée à pre-miR-155 ou au duplexe miR-155/miR-155*, aucune indice de contact entre les deux dsRBDs n'a été detecté et la protéine interagit avec les deux precurseurs par la même surface d'interaction. Les informations récoltées suggèrent que TRBP peut jouer un rôle avant et après le clivage des pre-miARN par Dicer, notamment dans le complexe de chargement de RISC. / MicroRNAs (miRNA) are a class of small non-coding RNAs that regulate gene expression through RNA interference (RNAi). Human miRNAs are generated via a series of enzymatic processing steps. In particular, in the cytoplasm, the precursor miRNA (pre-miRNA) is recognized and cleaved by a complex containing the RNase III enzyme Dicer and several non-catalytic accessory proteins. HIV TAR element-binding protein (TRBP) is a constituent of the Dicer complex, which augments complex stability, has effect on the cleavage kinetics and on the cleavage site and potentially functions in substrate recognition and product transfer to the RNA-induced silencing complex (RISC). TRBP is composed of three double stranded RNA binding domains (dsRBDs). The RNA binding region of TRBP is composed of the first two dsRBDs and an uncharacterized interdomain region. The present study reports the in vitro biophysical characterization of the RNA binding region of TRBP in the apo state and in the RNA bound state with the two successive cytoplasmic precursors of the oncogenic human microRNA miR-155, the hairpin pre-miR-155 and the related Dicer product miR-155/miR-155* duplex. The study shows that the RNA binding region of TRBP is monomeric and comprises two independent double-stranded RNA-binding domains connected by a 60 residues flexible linker. The first dsRBD, uncharacterized previously in solution, undergoes a full folding/unfolding equilibrium in a wide range of physico-chemical conditions. The two first dsRBDs of TRBP can interact with one microRNA precursor and two RNA binding regions can interact with one precursor molecule. The RNA-binding region of TRBP interacts with both pre-miR-155 and miR-155/miR-155* duplex with similar affinities. In the complex with one RNA binding region of TRBP bound to either pre-miR-155 or miR-155/miR-155* duplex, no evidence of contact between the two dsRBDs were observed and the protein interacts with both precursors via the same protein binding surface. The data presented here suggest that the RNA binding region of TRBP can play a role before and after processing of pre-miRNAs by Dicer, including in the RISC loading complex.

Submitted by VIVIANE MAGRINI null (viviane_magrini@yahoo.com.br) on 2016-03-01T20:13:04Z No. of bitstreams: 1 Viviane Magrini.pdf: 5645437 bytes, checksum: 25fafce9b5405010cb429b17c172011c (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-03-03T19:58:52Z (GMT) No. of bitstreams: 1 magrini_v_me_araiq_par.pdf: 1328742 bytes, checksum: 4a2b2b27d644fb4d71ce706407afdc38 (MD5) / Made available in DSpace on 2016-03-03T19:58:52Z (GMT). No. of bitstreams: 1 magrini_v_me_araiq_par.pdf: 1328742 bytes, checksum: 4a2b2b27d644fb4d71ce706407afdc38 (MD5) Previous issue date: 2016-02-18 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O gênero Calycophyllum pertence à família Rubiaceae, conhecida pela diversidade de metabólitos secundários, e pela variedade de atividades biológicas: antifúngica, bactericida, antiviral e inseticida. A espécie vegetal Calycophyllum spruceanum Benth (Rubiaceae) é muito utilizada pelas populações da América do Sul no combate a doenças dermatológicas, estomacais, diabetes, parasitoses, câncer, entre outras, despertando o interesse pela investigação química deste táxon. A presente pesquisa objetivou o estudo bio-guiado dos extratos obtidos das folhas, galhos e de cascas do caule de C. spruceanum visando à identificação das substâncias bioativas isoladas, incluindo as inativas. A meta é identificar nas substâncias isoladas aquelas com propriedades cosméticas associadas ao uso popular desta espécie, conhecida na região Amazônica por esta propriedade. Diferentes extratos foram preparados e analisados por Cromatografia Líquida de Alta Eficiência (CLAE) e Espectroscopia de Ressonância Magnética Nuclear (RMN 1H), acompanhados por ensaios biológicos (ensaio de capacidade redutora dos radicais DPPH˙ e ABTS˙+) para determinação de atividade antioxidante. O direcionamento do estudo foi feito em função dos ensaios antioxidantes aplicados aos extratos, conduzindo o estudo experimental para o extrato hidroetanólico de folhas ao foco do estudo, por ter apresentado a maior atividade antioxidante. Por meio de métodos espectroscópicos e espectrométricos (CLAE-DAD, CLAE-EM, CLAE-EM/EM; RMN de 1H e 13C; TOCSY 1D; HMBC; EM-IES) deste extrato e respectivas frações, pode-se identificar algumas substâncias antioxidantes potencialmente ativas, sendo estas: ácido 3,5-di-O-E-cafeoilquínico; duas séries homólogas de proantocianidinas (tipo-B) com grau de polimerização de 1 a 4, e proantocianidinas monoglicosiladas (tipo-B O-glicosilada) com grau de polimerização de 1 a 3; ácido 1,4-di-O-E-cafeoilquínico, quercetina 3-O-α-arabinopiranosil (16) β-glucopiranosídeo, ácido cafeoilquínico e canferol 3-O-β-D-glicopiranosil (12) β-D-xilopiranosídeo. Espera-se que ao final deste estudo, as informações forneçam subsídios para uma possível aplicação dos resultados científicos sobre a espécie vegetal, visando um produto para a linha de cosmético funcional. / The Calycophyllum genus belongs to the Rubiaceae’s family, known for its high diversity of secondary metabolites, which have a wide range of biological activities (antifungal, antibacterial, antiviral and pesticide). It is widely used by the people of South America against skin diseases, stomach, diabetes, parasites, cancer, among others. It is therefore essential to study the chemical composition of these species used in folk medicine, aiming to validate the traditional use, with scientific data and thus contributing to the search for new bioproducts. This research aimed the bio-guided fractionation of the extracts from the leaves, branches and stem bark extracts of Calycophyllum spruceanum Benth (Rubiaceae), to identify the secondary metabolites in the extracts and fractions, including bioactive ones, for further evaluation of the cosmetic potential of this species, commonly known in the Amazon region. Different extracts were prepared and analyzed by High Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance spectroscopy (1H NMR), and screened by biological assays (assay of reducing capability of ABTS˙+ and DPPH˙radicals) to determine the antioxidant activity. The chemical investigation was carried out based on the antioxidant activity of the extracts. The experimental study focused on the hydroethanolic extract of leaves, that presented the higher antioxidant activity, which would support the traditional uses of this specie as cosmetic. By means spectroscopic and spectrometric analysis (HPLC-DAD, HPLC-MS, HPLC-MS/MS; 1H NMR and 13C; TOCSY 1D; HMBC, ESI-MS) of this extract and its fractions, some compounds were identified: the 3,5-di-O-E-caffeoylquinic acid; two homologous series of proanthocyanidins (B-type) with a degree of polymerization from 1 to 4 and monoglicosyl proanthocyanidins (B-type O-glycosylated) with degree of polymerization from 1 to 3; 1,4-di-O-E-caffeoylquinic acid, quercetin-3-O-α-arabinopyranosyl (16) β-glucopyranoside, caffeoylquinic acid and kaempferol 3-O-β-D-glucopyranosyl (12) β-D-xilopyranosil, which could be responsible for the activity. It is expected that the the complete study of this species provide chemical information, which will validate this plant specie as a potential source of antioxidant natural products that could be material for functional cosmetics. / CNPq: 166069/2014-0

Neste trabalho foram realizadas medidas dos tempos de relaxação nuclear do 19F, em três amostras ternárias, não estequiométricas e que apresentam estrutura fluorita. Na amostra Na0.4Y0.6F2.2 foram medidos o tempo de relaxação spin-spin (T2) em função da temperatura nas fases onde sua estrutura não é fluorita (600K à 900K), e o tempo de relaxação spin-rede (T1), no mesmo intervalo de temperaturas, nas freqüências de Larmor: 20.42 MHz e 34.24 MHz. Obtivemos para as medidas de T1, um comportamento similar ao observado em sistemas com dois sítios inequivalentes. Na amostra Pb0.84Bi0.16F2.16 foram feitas medidas de T2 no intervalo de temperaturas 300K à 830K, dentro de um ciclo térmico de aquecimento e resfriamento, afim de se obter a energia de formação de defeitos. Porém a diferença de energia obtida, de 0.08 eV entre a energia obtida durante o aquecimento e o resfriamento, parece estar associada a mudanças estruturais nos clusters. Na amostra K0.04Bi0.06 F2.2: 2% PbF2 foram realizadas medidas de T2 em um intervalo de temperatura de 300K a 800K, dentro de um ciclo térmico. Não se observou mudança na energia de ativação durante o ciclo. Mediu-se também, no mesmo intervalo de temperaturas, T1 nas freqüências de Larmor: 11.71 MHz, 20.42 MHz e 34.24 MHz. A análise das curvas de T1 parece indicar a existência de dois mecanismos de saltos no material. / The 19F NMR relaxation times T1 and T2 were measured is ternary and nonstoichiometric compounds with the fluorite-type structure. We have studied the Na0.4Y0.6F2.2 crystal in the temperature range 600K to 900K, where the crystal hás not the fluorite structure. The T1 values were measured in two Larmor frequencies: 20.42 MHz and 34.24 MHz. The results for T1 were seem to be qualitatively similar to those measured in the system with two inequivalent sublattices. The T2 measurement, in the Pb0.84Bi0.16F2.16 crystal, were made during temperature cycles in the range of 300K to 830K. The difference in activation energy between cooling and heating half cycles, found to be approximatly 0.08 eV, appear to be associated with the change in the clusters structure and not to the energy of defect formation. Finally, similar T2 measurements during the temperature cycling was made in K0.04Bi0.06 F2.2: 2% PbF2 crystal in the temperature range 300K to 800K, but in this case no difference in the cooling and heating results was observed. We also measured, in the same temperature range, the T1 relaxation time in 3 Larmor frequencies: 11.71 MHz, 20.42 MHz and 34.24 MHz. This results appear to indicate the existence of two hopping mechanism.