O Brasil possui condições climáticas adequadas para o desenvolvimento de um grande número de frutas nativas e essa biodiversidade tem se tornado um caminho promissor para a descoberta de novos compostos bioativos capazes de ser utilizados na formulação de alimentos funcionais e medicamentos. Os compostos fenólicos apresentam ações específicas, podendo atuar como antioxidantes e anti-inflamatórios, assim prevenindo doenças crônicas. Diante do exposto, o objetivo deste trabalho foi avaliar o potencial antioxidante, antiinflamatório e a composição fenólica de dez frutas nativas brasileiras ainda pouco conhecidas: araçá-boi (Eugenia stipitata), cambuití-cipó (Sagerectia elegans), murici vermelho (Bysonima arthropoda), murici guassú (Byrsonima lancifolia), morango silvestre (Rubus rosaefolius), cambuci (Campomanesia phaea), jaracatiá-mamão (Jacaratia spinosa), juquirioba (Solanum alterno-pinatum), fruta-do-sabiá (Acnistus arborescens) e cajá (Spondias mombin L.). Os extratos etanólicos (80% v/v) das polpas foram analisados inicialmente quanto à capacidade de sequestro dos radicais ABTS&#8729+ e ROO&#8729. A atividade anti-inflamatória foi avaliada in vivo por meio do modelo de migração de neutrófilos induzida por carragenina, enquanto que a composição fenólica foi realizada por técnicas cromatográficas (CLAE-DAD e CG-EM). As 5 frutas com as maiores atividades biológicas foram ainda analisadas quanto à capacidade de sequestro de O2 &#8729-, HOCl e NO&#8729, atividade anti-inflamatória por meio do ensaio de ativação do fator nuclear-&kappa;B (NF-&kappa;B) e composição fenólica por espectrometria de massas de alta resolução (LC-ESI-QTOF-MS). Em relação ao sequestro do radical ABTS&#8729+ o cambuití-cipó apresentou a maior atividade (749,88 &mu;mol TE.g-1) e para o ROO&#8729 o murici vermelho apresentou a maior atividade antioxidante (559,09 &mu;mol TE.g-1). Os animais tratados com araçá-boi, cambuití-cipó, murici vermelho, cajá e morango silvestre apresentaram reduções no influxo de neutrófilos comparados ao grupo carragenina (p < 0,05). Por meio das técnicas de CLAE-DAD e CG-EM foi possível identificar compostos fenólicos pertencentes a classe dos flavonoides (catequina, epicatequina, rutina, quercetina glicosilada, kaempeferol glicosilado, quercetina, procianidina B1 e procianidina B2), sub-classe do ácido hidroxibenzóico (ácido gálico) e sub-classe dos ácidos hidrocinâmicos (ácido cumárico, ácido ferúlico e caféico). O araçá-boi, cambuití-cipó, murici vermelho, morango silvestre e cajá foram as cinco frutas que apresentaram as maiores atividades antioxidantes e/ou anti-inflamatórias, cujo perfil fenólico por LC-ESI-QTOF-MS indicou a presença de 18 compostos no araçá-boi, 32 no cambuitícipó, 26 no murici vermelho e 20 e 11 compostos no morango silvestre e cajá, respectivamente. Vários dos compostos fenólicos identificados foram encontrados pela primeira vez nessas espécies. O cambuiti-cipó e murici vermelho se destacaram em relação ao sequestro de HOCl (EC50 4,99 e 4,41 &mu;g mL-1, respectivamente) e o cambuití-cipó foi o mais ativo para desativar o radical O2 &#8729- (EC50 68,33 &mu;g mL-1) e NO&#8729 (EC50 0,78 &mu;g mL-1). Já os extratos de murici-vermelho, cambuití-cipó e morango silvestre inibiram significativamente a ativação do NF-&kappa;B. Portanto, as frutas nativas brasileiras são fontes de substâncias antioxidantes e anti-inflamatórias, bem como de uma grande diversidade de compostos fenólicos, os quais podem propiciar importantes benefícios para a saúde humana. / Brazil has favorable climatic conditions for the development of a large number of native fruits and this biodiversity has become a promising path towards the discovery of new bioactive compounds capable of being used in the formulation of functional foods and medicines. Phenolic compounds show specific action mechanisms, being able to act as antioxidants and anti-inflammatories, thus preventing chronic diseases. However, few Brazilian native fruits are well known and consumed by the population, undermining the investigation of chemical composition as well as the identification/quantification of bioactive compounds. In light of this, the objective of this work was to evaluate the antioxidant and anti-inflammatory potentials as well as the phenolic composition of ten underexploited Brazilian native fruits, namely: araçá-boi (Eugenia stipitata), cambuití-cipó (Sagerectia elegans), murici vermelho (Bysonima arthropoda), murici guassu (Byrsonima lancifolia), morango silvestre (Rubus rosaefolius), cambuci (Campomanesia phaea), jaracatiá-mamão (Jacaratia spinosa), juquirioba (Solanum alterno-pinatum), fruto-do-sabiá (Acnistus arborescens) and cajá (Spondias mombin L.). Pulps ethanolic extracts (80%, v/v) were initially analyzed regarding scavenging capacity of the ABTS&#8729+ and ROO&#8729 radicals. Antiinflammatory activity was evaluated in vivo using the carrageenan-induced neutrophil migration model, while phenolic composition was determined by chromatographic techniques (HPLC-PAD and GC-MS). The five fruits with the highest biological activities were analyzed for O2&#8729-, HOCl and NO&#8729 radicals scavenging capacities, for in vitro anti-inflammatory activity by nuclear factor-&kappa;B (NF-&kappa;B) activation, and for phenolic composition by High Resolution Mass Spectrometry (LC-ESI-QTOF-MS). In relation to ABTS&#8729+ radical scavenging cambuiticipó showed the highest activity (749.88 &mu;mol TE.g-1), while for ROO&#8729 scavenging, murici vermelho had the highest antioxidant activity (559.09 &mu;mol TE.g-1). The animals treated with araçá-boi, cambuití-cipó, murici vermelho, cajá and morango silvestre reported decreases in neutrophils influx compared to carrageenan group (p <0.05). It was possible to identify by HPLC-DAD and GC-MS techniques phenolic compounds belonging to the class of flavonoids (catechin, epicatechin, rutin, glycosylated quercetin, glycosylated kaempeferol, quercetin, procyanidin B1 and procyanidin B2), subclass of hydroxybenzoic acid (gallic acid) and subclass of hydrocinnamic acids (coumaric, ferulic and caffeic acids). Araçá-boi, cambuitícipó, murici vermelho, morango silvestre and cajá were the five fruits with the highest antioxidant and / or anti-inflammatory activities. The phenolic profile analysis by LC-ESIQTOF- MS pointed the presence of 18 compounds in araçá-boi, 32 in cambuití-cipó, 26 in murici vermelho, 20 in morango silvestre and 11 in cajá. Several of the identified phenolic compounds were found for the first time in these fruit species. Cambuiti-cipó and murici vermelho stood out in relation to HOCl scavenging (EC50 4.99 and 4.41 &mu;g.mL-1, respectively) and cambuití-cipó was the most active to deactivate both O2 &#8729- radical (EC50 68.33 &mu;g.mL-1) as NO&#8729 (EC 500.78 &mu;g.mL-1). Murici vermelho, cambuití-cipó and morango silvestre extracts significantly inhibited the activation of NF- &kappa;B.Therefore, Brazilian native fruits are sources of antioxidant and anti-inflammatory substances, as well as a great diversity of phenolic compounds, which can provide important benefits for human health.

Conteúdo: v. 1 Texto - v. 2 Mapas / Resumo: Este trabalho apresenta Mapas de Vulnerabilidade Ambiental ao Óleo simulando um vazamento de óleo no fundeadouro do Porto de Santos, próximo ao litoral de Praia Grande. Foram também elaboradas Cartas de Sensibilidade Ambiental ao Óleo (SAO) para o trecho do litoral paulista entre os municípios de Praia Grande e Cananéia. A metodologia utilizada para elaboração das Cartas SAO é baseada nas normas do Ministério do Meio Ambiente e para a elaboração dos Mapas de Vulnerabilidade foi utilizado o modelo SIMAP, desenvolvido pela empresa ASA, capaz de representar a mancha de óleo na superfície do mar, na linha de costa e na coluna d'água. A área apresenta trechos com intenso uso turístico, principalmente nos meses de verão e áreas com ambientes naturais preservados, com destaque à presença do sistema estuarino-lagunar de Cananéia-Iguape e Unidades de Conservação, que conferem à área grande importância ecológica. As Carta SAO foram geradas nas escalas tática (1:150.000) e operacional (1:50.000 e 1:25.000) e os Mapas de Vulnerabilidade foram gerados em escalas entre 1:100.000 e 1:750.000, de forma a abranger toda a área da mancha de óleo. Os resultados confirmam a necessidade de avançar na abordagem da sensibilidade da costa ao óleo, incorporando às cartas SAO informações sobre a probabilidade das áreas serem atingidas, estimativas do tempo de contato na costa e informações sobre a área da mancha que tem potencial tóxico à biota. / Abstract: This paper shows the Environmental Vulnerability Maps a simulated oil spill in the anchorage of the Santos port, close to Praia Grande shore. ESI Maps for the São Paulo shore between the municipalities of Praia Grande and Cananéia were also built. The methodology used to build ESI Maps is based on the Ministry of the Environment rules and for the construction of the Vulnerability charts used the SIMAP model, developed by the ASA. This is capable of modeling the oil spill on the surface of the sea, in the coast line and in the water column. The whole area shows intense tourist use, especially during the summer and areas with preserved natural environments include the presence of the estuarine-lagoon system of Cananéia-Iguape and Conservation Units that give the area great ecological importance. ESI Maps were generated in the tatic range (1:150.000) and operational (1:50.000 and 1:25.000) and the vulnerability maps were generated using 1:100.000 and 1:750.000 scales, to cover the entire area of oil stain. The results confirm the sensitivity of the coast to oil, use of the information above has been incorporated in the ESI Maps to estimate shore contact lead time and of the oil spill and this toxicity potential on the environment. / Orientador: Paulina Setti Riedel / Coorientador: João Carlos Carvalho Milanelli / Banca: Eduardo Ayres Yassuda / Banca: Gabriel Henrique da Silva / Banca: Gilberto José Garcia / Banca: Iris Regina Fernandes Poffo / Doutor

Conteúdo: v. 1. Texto - v. 2 Atlas / Resumo: Derrames de óleo causam recorrentes e graves prejuízos aos ecossistemas costeiros em todo o mundo. O mapeamento de sensibilidade ambiental é componente essencial e fonte de informação primária para o planejamento de contingência, que estabelece diretrizes de ações de resposta aos derrames. O objetivo deste trabalho foi definir áreas prioritárias de proteção e zonas de sacrifício por meio do mapeamento, em escala operacional, da sensibilidade a derrames de óleo dos ambientes costeiros dos municípios de São Vicente, Santos e Guarujá (SP), baseando-se na proposta elaborada pelo Ministério do Meio Ambiente, com utilização de um Sistema de Informação Geográfica. A região apresenta elevada complexidade e relevância econômica com alto risco de ocorrência de derrames pela presença do Porto de Santos. Possui grande variedade de ecossistemas costeiros e estuarinos sensíveis que abrigam comunidades de elevada diversidade biológica. Como resultados são apresentados 3 Cartas Táticas, 21 Cartas Operacionais e Quadros Descritivos com informações de cada segmento mapeado; além do mapa e das indicações das Áreas Prioritárias e Zonas de Sacrifício, sugeridas de acordo com critérios relevantes, baseados nas características de cada segmento, e considerados a partir do conhecimento detalhado adquirido no desenvolvimento deste trabalho, que constitui importante contribuição à gestão ambiental da zona costeira estudada / Abstract: Oil spills have caused recurrent and severe damages in coastal ecosystems all around the world. The environmental sensitivity mapping is an essential component and prime information source for the contingency planning, which establishes emergency response actions to oil spills. This study aimed to define high priority areas for protection and sacrifice zones by mapping, in detailed scales, the environmental sensitivity to oil spills of coastal habitats in São Vicente, Santos and Guarujá municipalities (São Paulo State). This mapping was done using a Geographic Information System and was based on the methodology and standards suggested by the Brazilian Environmental Ministry. The study area has high economic importance and complexity, with many risks of oil spills because of the presence of the "Porto de Santos". The region shows a wide variety of sensitive estuarine and coastal ecosystems that shelter high biodiversity. As result are presented 3 ESI maps in tactical scales; 21 ESI maps in detailed scales; Descriptive Tables with information from each coastal segment mapped; and the map with indicated areas for high priority protection and sacrifice zones. These areas were suggested according to relevant criteria, based on characteristics of each segment, and considered from the detailed knowledge acquired in the development of this work that represents an important contribution to the coastal environmental management in the study area / Orientador: Paulina Setti Riedel / Coorientador: João Carlos Carvalho Milanelli / Banca: Iris Regina Fernandes Poffo / Banca: Gilberto José Garcia / Acompanha 1 CD-ROM com o volume 2 (atlas) / Mestre

Mass spectrometry is a versatile, sensitive and fast technique with which to probe biophysical properties in biological systems and one of the most important analytical tools in the multidisciplinary field of proteomics. The study of nativestate proteins and their complexes in the gas-phase is well established and direct infusion electrospray ionisation mass spectrometry (DI-ESI-MS) techniques are becoming increasingly popular as a tool for screening and determining quantitative information on protein-protein and protein-ligand interactions. However, complexes retained by ESI-MS are not always representative of those in solution and care must be taken in interpreting purely gas-phase results. This thesis details modification and advancement of solution phase techniques devised by Gross et al. utilising ESI-MS and Fitzgerald et al. applying matrix assisted laser desorption ionisation (MALDI)-MS termed PLIMSTEX (protein-ligand interactions by mass spectrometry, titration and hydrogen-deuterium-exchange)[1] and SUPREX (Stability of unpurified proteins from rates of H/D exchange)[2] to quantify these interactions with regards to high throughput analysis. The first part of this thesis describes the different developmental stages of the devised HPLC-front ends and their optimisation with myoglobin and insulin. The successfully developed HPLC-front end in conjunction with PLIMSTEX and SUPREX and ESI-MS then gets tested with self expressed and purified cyclophilin A(CypA)- cyclosporin A (CsA) system, followed by a test screen with potential CypA binding ligands. Dissociation constants (Kd’s) within one order of magnitude to reported values are determined. In the third part of this thesis the application of the devised ESI-SUPREX methodology has been applied to anterior gradient 2 (AGr2) and the factor H complement control proteins module 19-20 (fH19-20) exhibiting binding potential to a taggedhexapeptide and a synthetic pentasaccharide, respectively, resulting in thermodynamical data for these protein-ligand interactions. For the AGr2 system another dimension of investigation has been added by temperature controlling the devised ESI-SUPREX approach, revealing a phase transition in the protein at higher temperatures. The final part of this thesis describes the application of the ESI-SUPREX methodology to probe folding properties of CypA in the presence of the self expressed and purified E. coli chaperonin groEL. Thereby the denaturing properties of groEL have been emphasised along with the stabilisation of a denatured CypA species.

No description available.

Cette thèse a permis de comparer les caractéristiques spectrales des supernovæ de type Ia en fonction du<br />décalage vers le rouge ("évolution"). Dans le cadre des collaborations Supernova Cosmology Project (SCP) et SuperNova Legacy Survey (SNLS), dont l'objectif scientique commun est l'étude de l'énergie noire à l'aide de supernovæ de type Ia à grand décalage vers le rouge, une part importante du travail de thèse est consacrée à la réduction des données spectrales,<br />étape nécessaire pour obtenir le spectre physiquement exploitable à partir de données observées. La réduction de l'ensemble des spectres SCP issus du spectrographe à échellettes Keck-ESI a permis d'obtenir des supernovæ de type Ia parmi les plus lointaines jamais observées. Dans l'expérience SNLS, l'identication spectroscopique est essentiellement réalisée avec le spectrographe longue fente FORS1 monté au foyer du VLT UT1. Pour le SNLS, il s'agit de réduire et d'identier une dizaine de spectres par lunaison pendant 5 ans. Dans le cadre de cette thèse, un logiciel d'identication en temps réel de SNIa a été developpé, il permet d'établir le type, le décalage vers le rouge et l'âge du candidat quasi automatiquement. Il évalue aussi la contamination<br />de la galaxie hôte (dont on peut aussi estimer la morphologie) dans le spectre. Le logiciel a été testé sur un échantillon de spectres analysés en détail.<br />Par ailleurs, pour certains d'entre eux, on a mesuré la vitesse du CaH&K (3945.12Å) dans la photosphère puis<br />on a comparé les résultats avec les mêmes mesures réalisées sur un lot de spectres proches. Ce résultat a permis de conrmer l'hypothèse de standardité des SNIa à grand décalage vers le rouge. C'est une hypothèse fondamentale pour mesurer les paramètres cosmologiques avec les supernovæ de type Ia.

<p>This thesis describes the current status and novel aspects of the analysis of the molecules of life, i.e. peptides and proteins, using capillary electrophoresis (CE) coupled to mass spectrometry (MS) via (sheathless) electrospray ionization (ESI). Early reports of sheathless CE-ESI-MS were plagued by limited lifetimes of the electrospray emitter. In this thesis, two new approaches, the Black Dust and the Black Jack methods, utilizing polymer-embedded graphite instead of noble metals are presented. These emitters have shown improved long-term stability and proven excellent for sheathless electrospray operation. Failure of an emitter is often caused by electrochemical reactions occurring at the emitter-liquid interface. The electrochemical properties of the graphite coated emitters were therefore evaluated by classical electrochemical methods, such as cyclic voltammetry and chronoamperometry. The graphite coated emitters showed excellent electrochemical stability and properties compared to noble metal and polymer configurations.</p><p>Analyte-wall interactions have long been known to cause problems in the CE analysis of biomolecules. This can be circumvented by internal modification of the capillary walls. Additionally, it is of outermost importance to have a stable and sufficiently high electroosmotic flow (EOF) to sustain the electrospray, when using a sheathless approach. New monomer and polymer coatings are presented for rapid and high-efficient CE-ESI-MS separations of peptides and proteins.</p><p>Furthermore, the use of CE-ESI coupled to Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) shows great potential for rapid proteomic probing of human cerebrospinal fluid. The results are comparable with more established techniques, such as liquid chromatography and two-dimensional gel electrophoresis coupled to MS. However, the CE-ESI-FTICRMS analysis has significantly lower sample consumption and faster analysis time compared to the other techniques. The applications and use of CE-ESI-MS is expected to have a bright future with continued growth as current trends of multidimensional hyphenation and microfabricated devices are further developed and explored.</p>

<p>Transition metals have played an important role in synthetic organic chemistry for more than a century, and offer catalytic transformations that would have been impossible with classical chemistry. One of the most useful and versatile of the transition metals is palladium, which over the years has catalyzed many important carbon-carbon forming reactions. Popular cross-coupling reactions such as the Suzuki, Stille and the Heck reaction are all catalyzed by palladium, or more correctly, by palladium in its ground state, Pd(0). </p><p>Recently, interest in palladium(II)-catalyzed transformations has started to grow, partly due to the development of the vinylic substitution reaction, commonly called the oxidative Heck reaction, presented in this thesis. This Pd(II)-catalyzed, ligand-modulated reaction occurs under air at room temperature, and for the first time a general protocol employing a wide range of olefins and arylboronic acids was obtained. Ligand screening showed that the bidentate nitrogen ligand, 2,9-dimethyl-1,10-phenanthroline (dmphen), was the most suitable ligand. Dmphen is believed to facilitate regeneration of active Pd(II), increase catalytic stability and improve the regioselectivity in the reaction. A mechanistic investigation was conducted using electrospray ionization mass spectrometry (ESI-MS), making it possible to observe cationic intermediates in a productive oxidative Heck arylation. The results obtained are in agreement with the previously proposed catalytic cycle.</p><p>The emerging discipline of high-speed synthesis is making contributions to society’s growing demand for new chemical entities. This inspired the development of two ultrafast, microwave-accelerated carbonylation reactions with dicobalt octacarbonyl acting both as an in situ carbon monoxide supplier and reaction mediator. A wide range of symmetrical benzophenones was produced in only 6 to 10 s, using aryl iodides as the substrate. The second carbonylation reaction provided symmetrical and unsymmetrical ureas in process times ranging from 10 s to 40 minutes using primary and secondary amines.</p>

La protéase du VIH-1 est une cible de choix dans le traitement du SIDA, car l’inhibition de son activité protéolytique contrecarre la réplication virale. Dans ce contexte, une approche de conception de novo d’inhibiteurs non peptidiques de cet enzyme, réalisée au sein de notre laboratoire, avait permis d’épingler une famille originale de complexes métallo-organiques au cuivre (II). Au cours de cette thèse, nous avons entrepris la caractérisation physico-chimique et structurale de ces composés afin d’élucider leur mode d’interaction avec la protéase, en nous appuyant sur diverses techniques expérimentales (DRX, RPE, ESI-MS) et théoriques(mécanique moléculaire, SIBFA). En particulier, nous avons montré que la stabilité thermodynamique de ces complexes en solution était indispensable à toute activité biologique. L’adaptation de la méthode SIBFA aux complexes au cuivre (II) nous a permis d’étudier les phénomènes énergétiques intervenant dans leur stabilité. Enfin, nous avons réuni les critères structuraux responsables de l’activité anti-protéolytique de ces complexes au sein d’un modèle pharmacophorique optimalisé./HIV-1 protease is a main target for the AIDS treatment, because its inhibition blocks the viral replication. De novo drug design, previously conducted in our laboratory, had pointed out several copper (II) chelates as a new family of non peptidic protease inhibitors. In order to provide a better understanding of their structure-activity relationships, we performed the physico-chemical characterization of these compounds using experimental (XRD, EPR, ESIMS)and theoretical (molecular mechanics, SIBFA method) techniques. We demonstrated that the thermodynamic stability of the complexes is an essential property to provide inhibitory activity. The SIBFA procedure adapted to copper (II) complexes helped us to study the energetics involved in the stability process. From our results, we derived a pharmacophore model describing the structural properties needed to achieve a good inhibition of the enzyme.

This thesis describes the current status and novel aspects of the analysis of the molecules of life, i.e. peptides and proteins, using capillary electrophoresis (CE) coupled to mass spectrometry (MS) via (sheathless) electrospray ionization (ESI). Early reports of sheathless CE-ESI-MS were plagued by limited lifetimes of the electrospray emitter. In this thesis, two new approaches, the Black Dust and the Black Jack methods, utilizing polymer-embedded graphite instead of noble metals are presented. These emitters have shown improved long-term stability and proven excellent for sheathless electrospray operation. Failure of an emitter is often caused by electrochemical reactions occurring at the emitter-liquid interface. The electrochemical properties of the graphite coated emitters were therefore evaluated by classical electrochemical methods, such as cyclic voltammetry and chronoamperometry. The graphite coated emitters showed excellent electrochemical stability and properties compared to noble metal and polymer configurations. Analyte-wall interactions have long been known to cause problems in the CE analysis of biomolecules. This can be circumvented by internal modification of the capillary walls. Additionally, it is of outermost importance to have a stable and sufficiently high electroosmotic flow (EOF) to sustain the electrospray, when using a sheathless approach. New monomer and polymer coatings are presented for rapid and high-efficient CE-ESI-MS separations of peptides and proteins. Furthermore, the use of CE-ESI coupled to Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) shows great potential for rapid proteomic probing of human cerebrospinal fluid. The results are comparable with more established techniques, such as liquid chromatography and two-dimensional gel electrophoresis coupled to MS. However, the CE-ESI-FTICRMS analysis has significantly lower sample consumption and faster analysis time compared to the other techniques. The applications and use of CE-ESI-MS is expected to have a bright future with continued growth as current trends of multidimensional hyphenation and microfabricated devices are further developed and explored.