Bicamadas de lípides sintéticos como o brometo de dioctadecildimetilamônio (DODAB) ou dihexadecilfosfato de sódio (DHP) foram avaliadas como carreadores em potencial de miconazol (MCZ), uma droga altamente hidrofóbica. A solubilização e/ou estabilização do MCZ foi avaliada através de métodos de espectrofotometria no UV-visível, determinações de tamanho por espalhamento de luz dinâmico, determinações de potencial zeta e cinéticas de agregação da droga por medidas turbidez em presença ou ausência das dispersões dos anfifílicos. Os resultados mostram que há uma notável capacidade de solubilização de MCZ em fragmentos de membrana (BF) de dispersões de DODAB, o que não ocorre para vesículas grandes (LV) dos mesmos anfifilicos. Ainda, demonstrou-se uma evidente estabilização coloidal do particulado de droga em dispersão aquosa por deposição de bicamadas aniônicas de DHP sobre os grânulos catiônicos do miconazol. Em elevadas proporções molares drogai lípide sintético observou-se um aumento na estabilidade coloidal do particulado de droga que foi atribuído ao recobrimento do particulado catiônico de droga com bicamadas aniônicas. Posteriormente, foram avaliadas as possíveis interações existentes entre células de Candida albicans e fragmentos e/ou vesículas de DODAB. Isotermas de adsorção foram obtidas em função do método de dispersão, concentração do anfifílico e densidade numérica de células. Para os fragmentos de bicamada, as isotermas mostraram-se de alta afinidade (adsorção não-reversivel) não havendo desorção, ao contrário do resultado obtido para as vesículas grandes, onde foi observada desorção a partir da superficie celular. A desorção é favorecida pelo aumento da quantidade de vesículas livres no sobrenadante, sugerindo assim que existe uma significante interação intervesicular de natureza hidrofóbica entre DODAB adsorvido e as vesículas livres. Experimentos de mobilidade eletroforética (ME) mostram a mesma tendência. Dois tipos de interações puderam ser notados. As interações de natureza eletrostática estão presentes em ambas condições (interação das células com fragmento e/ou vesícula). A interação ocorre entre a cabeça carregada do anfifilico e a membrana do fungo. Interações hidrofóbicas possivelmente ocorrem somente em fragmentos de bicamada devido a interações extra-atrativas entre regiões hidrofóbicas da parede celular e as bordas hidrofóbicas dos fragmentos. / Bilayer of synthetic lipids such as sodium dioctadecyldimethylarnrnoniurn bromide (DODAB) or dihexadecylphosphate (DHP) were assessed as potential carriers for miconazol (MCZ), a very hydrophobic drug. The solubilization anel/or stabilization of the drug were tested by UV-visible spectrophotometry methods, size deterrninations by dynamic light scattering, zeta-potential detenninations and drug aggregations kinetics by turbidity changes in the presence or absence of amphiphiles dispersions. Our results show an outstanding capacity solubilization of MCZ in bilayer fragrnents (BF) of DODAB dispersions, what doesn\'t occur with large vesicles (LV) from the same amphiphiles. Still, we have demonstrated a colloidal stabilization of drug particles in water dispersion by deposition of anionic bilayers of DHP on the cationic MCZ granules. In high molar ratio drug/synthetic lipid we observed an increasing colloidal stabilization of drug particles that was attributed to the covering of the drug cationic particles with anionic bilayers. Subsequently, were evaluated the possible interactions that existed between Candida Albicans cells and DODAB fragrnents and/or vesicles. Adsorption isotherrns were obtained as a function of the method of dispersion, concentration of the amphiphiles and cells number density. For the bilayer fragrnents, the isotherrns showed high affinity (irreversible adsorption) without desorption, in contrast with the isotherrns obtained for large vesicles, where we observed desorption of these vesicles over the cellular surface. The desorption was favored by the increase of free vesicles in the supematant, thus suggesting that there is a significant inter-vesicle interaction ofhydrophobic nature between adsorbed DODAB and the free vesicles. Electrophoretic mobility (ME) experiments showed the same tendency. Two types of interactions could be observed. The electrostatic nature interactions were present in both conditions (cellular interaction with fragrnent and/or vesicle). The interaction occurs between the charged head of the amphiphilic and the fungus membrane. Hydrophobic interactions possibly only occur with bilayer fragrnents due the extraattractive interaction existing between hydrophobic regions of the cell wall and the hydrophobic borders of the fragrnents.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A avaliação da qualidade dos solos e a redução da Mata Atlântica tem sido alvo de diversas pesquisas no Brasil e no mundo, principalmente quando estes estão atrelados ao recorte espacial de Unidades de Conservação. No entanto, tem sido difícil para os gestores dessas unidades a avaliação dos impactos ambientais gerados ao longo dos anos nas bacias hidrográficas, principalmente pela falta de investimentos. Esta dissertação teve por objetivo geral avaliar o atual estágio de degradação da bacia hidrográfica do Córrego da Caçada pertencente à Área de Proteção Ambiental Federal de Cairuçu, no município de Paraty RJ, analisando quantitativamente e qualitativamente a redução dos fragmentos de Mata Atlântica e estabelecendo relações com a degradação física e química dos solos dentro e fora dos fragmentos florestais. A metodologia utilizada para a redução ou avanço dos fragmentos de Mata Atlântica baseou-se no uso de fotografias aéreas do ano de 1956 e imagens de satélite de 2012, onde possibilitou a avaliação espaço-temporal do uso e cobertura das terras, através da produção de um mapa temático final. Além disso, foram elaborados mapas temáticos de reconhecimento da área de estudo, como o de hipsometria, de declividade, de orientação e forma das encostas, além da geração de perfis topográficos. Para a avaliação da qualidade física, química e biológica dos solos foram determinadas as curvas de distribuição granulométrica, a densidade relativa dos grãos sólidos e a densidade aparente, porosidade total, os limites de liquidez e plasticidade, a estabilidade dos agregados em água, análises morfológicas, a saturação de bases, a capacidade de troca catiônica (CTC), a saturação por alumínio, fósforo, pH e o carbono orgânico. Para tal, foi realizada a abertura de três perfis, sendo um em área de fragmento florestal e dois em áreas de pasto. O resultado das análises permitiu, segundo a Sociedade Brasileira de Ciência do solo, a classificação de dois tipos de solos na bacia, sendo: Cambissolo Háplico Tb Distrófico Típico em área fragmento florestal e em área de pasto, e um Latossolo Amarelo Tb Distrófico Típico em área de pasto. Os resultados de laboratório mostraram que os solos avaliados têm baixa fertilidade e valores variados nos resultados de física do solo. No entanto, além do histórico de uso do solo caracterizado pelas práticas rudimentares do manejo empregado pelos Caiçaras, o clima predominante na região possibilita um regime pluviométrico anual que passa dos 2.000mm de chuva/ano, caracterizando solos muito lixiviados e pobres quimicamente. Portanto, conclui-se que a relação das propriedades físicas e químicas avaliadas junto ao manejo inadequado ao longo dos anos tem apresentado um cenário de grandes dificuldades para a recuperação florestal na bacia hidrográfica do Córrego da Caçada, o que mostra a importância da avaliação dos impactos ambientais não só pelo recorte de bacias hidrográficas, como contextualizar seu posicionamento dentro de Unidades de Conservação, com legislações e objetivos específicos. / Evaluation of soil quality and the reduction of the Atlantic Forest has been the subject of several studies in Brazil and abroad, especially when these are linked to the spatial area of protected areas. However, it has been difficult for managers of these units the assessment of environmental impacts over the years in river basins, mainly by lack of investment. This work had as main objective to assess the current watershed degradation stage Hunt Stream belonging to the Federal Environmental Protection Area Cairuçu in the municipality of Paraty - RJ, analyzing qualitatively and quantitatively the reduction of Atlantic Forest fragments and establishing relationships with physical and chemical degradation of soil in and out of the forest fragments. The methodology used for the reduction or advancement of Atlantic Forest fragments was based on air use of photographs of the 1956 and 2012 satellite images, which enabled the evaluation space-use and coverage of land, by producing a final thematic map. In addition, thematic maps were prepared to recognize the study area, such as hypsometry, slope, orientation and shape of the slopes, as well as generating topographic profiles. For assessing the physical, chemical and biological soil were determined particle size distribution curves, the relative density of the solid grains and bulk density, total porosity, the limits of liquidity and plasticity, the stability of water in aggregate, morphological analysis , base saturation, the cation exchange capacity (CEC), saturation of aluminum, phosphorus, pH and organic carbon. To do this, the opening of three profiles was conducted, one in forest fragment area and two in pasture areas. The results of the analysis allowed, according to the Brazilian Society of Soil Science, the classification of two types of soils in the basin, as follows: Cambisol Tb dystrophic Typical in forest fragment area and pasture area, and an Oxisoil Tb dystrophic Typical in pasture area. The laboratory results showed that reviews have low soil fertility and various amounts of soil in physical results. However, besides the history of land use characterized by rudimentary management practices employed by Caiçaras, the prevailing mood in the region provides an annual rainfall of rain passing 2.000mm / year, featuring very leached soils and poor chemically. Therefore, it is concluded that the relationship of the physical and chemical properties evaluated by the inadequate management over the years has presented a scene of great difficulty for forest restoration in the basin of Stream of Hunt, which shows the importance of impact assessment environment not only for crop watersheds, as contextualize its positioning within protected areas, with legislation and specific objectives.

La méthode de conception de médicaments à partir de molécules « fragments » (connue sous le nom de « Fragment-Based Drug Design ») a été proposée au milieu des années 90, et a depuis été reconnue comme une alternative tangible aux techniques plus classiques de recherche de médicaments telles que le criblage à haut débit par exemple. La méthode des fragments consiste à cribler un petit nombre (< 10000) de composés organiques de faible poids moléculaire (< 300 Da) afin de détecter ceux qui se lient à la cible (protéine ou acides nucléiques). Du fait de leur faible complexité, les fragments présentent une affinité faible pour la cible, et la détection s'effectue généralement grâce à une technique biophysique (en particulier, résonance magnétique nucléaire (RMN), cristallographie aux rayons X, résonance plasmonique de surface). Les fragments « hits » sont ensuite modifiés par addition de nouvelles fonctions chimiques, ou par liaison de deux fragments, afin d'élaborer, étape par étape, une molécule capable d'établir des interactions plus nombreuses avec la cible, et d'améliorer ainsi l'affinité. Comparée aux méthodes classiques de criblage haut débit, la méthode des fragments offre divers avantages, notamment une meilleure exploration de l'espace chimique, une meilleure efficacité de liaison des molécules « hits », et une plus grande facilité d'optimisation des hits en molécules plus affines. Dans le cadre de ce projet de thèse, plusieurs aspects de la méthode des fragments ont été abordés : dans une première partie, nous étudions un cas concret d'application de la méthode des fragments à la recherche d'un inhibiteur de la peroxiredoxine 5 humaine, en utilisant la RMN comme outil de criblage des fragments ainsi que comme outil d'étude des interactions protéine-fragment. La découverte d'un inhibiteur de cette enzyme représente une avancée importante, qui devrait permettre de mieux comprendre son fonctionnement. Les autres parties de ce projet de thèse abordent des aspects plus méthodologiques de la méthode des fragments : les fragments conservent-ils leur site de liaison, leur efficacité de liaison et leur mode d'interaction au cours de leur élaboration en inhibiteur ? Les fragments peuvent-ils être spécifiques d'une protéine ? D'un site de liaison particulier ? Ces questions, rarement traitées, sont pourtant essentielles à la compréhension du comportement des molécules fragments, et sont abordées d'une part en défragmentant plusieurs inhibiteurs de la protéine Bcl-xL et en étudiant par RMN le comportement de ces fragments vis-à-vis de la protéine en termes d'affinité et de site de liaison, d'autre part en réalisant le criblage par RMN d'une série de fragments sur cinq protéines différentes (peroxiredoxine 5 humaine, sérum albumine humaine et trois protéines homologues de la famille Bcl-2). De manière générale, ce projet de thèse vise à étudier des aspects peu abordés de la méthode des fragments et à proposer des pistes permettant de mieux comprendre le comportement des fragments vis-à-vis de leur cible, au cours du criblage initial comme lors de leur optimisation / Fragment-Based Drug Design (FBDD) has been proposed in 1996 and has since been recognized as a tangible alternative to the more classical drug discovery methods such as High-Throuput Screening. FBDD consists of screening a small number (< 10 000) of low-molecular weight (< 300 Da) compounds and detect those that bind to the target (protein or nucleic acids). Because of their low complexity, fragment molecules usually display low affinities for their target, hence detecting fragment-protein interactions is mostly achieved using a biophysical technique (mostly Nuclear Magnetic Resonance (NMR), X-ray crystallography or Surface Plasmon Resonance). “Hit” fragments are then modified by addition of chemical substituents, or linked together, so as to elaborate a more complex molecule, forming more interactions with the target and hence displaying an improved affinity. As compared to the more classical High Throughput Screening method, fragment screening provides several advantages, including a better exploration of chemical space, highly ligand-efficient hits and easier optimization of the hits into more affine molecules. In this PhD project, several aspects of FDBB have been addressed : first, FBDD approaches were applied to the research of an inhibitor of the human peroxiredoxin 5 protein, using NMR not only as a screening method but also for the characterization of the protein-fragment interactions. The discovery of an inhibitor against this enzyme would allow to better understand its function. Next, methodological aspects of the FBDD method were addressed : Do fragments conserve their binding site, binding efficiency and mode of interaction upon optimization? Can the fragments display specificity towards a given target? Towards a given binding site? These issues, rarely studied, are yet essential to the understanding of the behavior of fragment molecules, and will be addressed firstly by defragmentating several Bcl-xL inhibitors into fragments and studying their behavior towards the protein in terms of a_nity and binding mode, secondly by screening a set of fragments against five different proteins (human peroxiredoxin 5, human serum albumin and three homologous proteins of the Bcl-2 family of proteins). More generally, this PhD project aims at studying less characterized aspects of the fragment methodology and proposing answers to better understand the behavior of fragment molecules towards their targets, both in the initial screening step and then during their optimization

As bactérias fototróficas freqüentemente apresentam florescimentos em lagoas de estabilização utilizadas no tratamento de esgoto sanitário, formando uma camada de cor púrpura na sua superfície. Portanto, o estudo das condições que propiciam tais florescimentos, a diversidade microbiana, o potencial de remoção da matéria orgânica e o estabelecimento das relações entre tais conhecimentos, permitem compreender o metabolismo do sistema. Nesse sentido, o objetivo deste trabalho foi avaliar a diversidade de bactérias (domínio Bacteria), bactérias fototróficas púrpuras e bactérias redutoras de sulfato (BRS) em lagoas de estabilização do Vale do Ribeira (Cajati, SP). Para tal, foram realizadas coletas sazonais (primavera, verão, outono e inverno) na sub-superfície, camada intermediária e interface água-sedimento, em dois horários (14:00 h e 02:00 h), nas lagoas anaeróbia e facultativa. Para analisar os diferentes grupos de microrganismos, utilizou-se a técnica de PCR/DGGE, com primers específicos. Nas análises de filogenia realizou-se o seqüenciamento parcial do gene RNAr 16S e da subunidade M do centro de reação fotossintético das bactérias fototróficas púrpuras. Análises físico-químicas, tais como sulfato, DQO, sólidos, nitrogênio e fósforo foram realizadas, além da determinação da concentração de oxigênio dissolvido, pH, temperatura e radiação solar fotossinteticamente ativa incidente. No outono observou-se maior diversidade de microrganismos do domínio Bacteria, bactérias fototróficas púrpuras e BRS, enquanto na primavera foi verificada a menor diversidade desses microrganismos para as duas lagoas. Na lagoa facultativa foi observada maior diversidade do domínio Bacteria e das BRS em relação à lagoa anaeróbia. Verificou-se maior diversidade de bactérias fototróficas púrpuras na lagoa anaeróbia, caracterizada por duas populações predominantes nas quatro estações e nas diferentes profundidades. A concentração de matéria orgânica (DQO) variou de 60,3 mg/L (inverno) a 298,0 mg/L (primavera) e a maior concentração de sulfato observada foi de 51,0 mg/L (inverno). Bacilo curvo Gram negativo, semelhante à bactéria fototrófica púrpura não sulfurosa, presente em amostra proveniente da sub-superfície da lagoa anaeróbia foi purificado e apresentou 92% de similaridade com Rhodopseudomonas palustris. Em ambas as lagoas foram identificadas bactérias semelhantes a Chromobacterium suttsuga (95%), Clostridium sp. (99%), Rhodobacter sphaeroides (99%), Rhodopseudomonas palustris (99%), Lampropedia hyalina (97%), Campylobacter fetus (99%), Desulfovibrio vulgaris (95%), Rhodospirillum rubrum (95%) e diferentes bactérias não cultivadas. / The phototrophic bacteria frequently blossom in the stabilization lagoons that are used in sanitary sewer treatment, forming a purple layer on its surface. Therefore, the study of the conditions that propitiate such blooms, the microbial diversity, the removal of the organic matter and the establishment of the relations between them permit to understand the metabolism of the system. The objective of this work was to evaluate the diversity of the bacteria (Bacteria domain), purple phototrophic bacteria and sulfate reducing bacteria (SRB) in stabilization lagoons of Vale do Ribeira (Cajati - SP). For this, it was made seasonal collects (spring, summer, autumn and winter) from the sub-surface, intermediate layer and interface water-sediment, at two times (14:00 h and 02:00 h) of the anaerobic and facultative lagoons. To analyze the different groups of microorganisms it was used the PCR/DGGE technique, with specific primers; for the phylogenic analysis it was realized the DNA partial sequencing of the 16S RNAr gene and of the subunit M of the photosynthetic center of reaction of the purple photosynthetic bacteria. It was determined: the concentration of dissolved oxygen, pH, temperature and photosynthetically active incident solar radiation, and the physical-chemistry analysis as: COD, solids, nitrogen and phosphorus. In the autumn it was observed greater diversity of microorganisms of the Bacteria domain, the group of the purples phototrophic bacteria and SRB, while in the spring it was verified minor diversity of these microorganisms in the two lagoons studied. In the facultative lagoon it was observed greater diversity of the Bacteria domain and of the SRB with respect to the anaerobic lagoon. It was verified greater diversity of the purple phototrophic bacteria in the anaerobic lagoon, of what in the facultative lagoon, which was characterized by the two predominant populations in the four seasons and in the different points of collect. The concentration of the organic matter (COD) varied from 60,3 mg/L (winter) to 298,0 mg/L (spring) and the greater concentration of sulfate observed was of 51,0 mg/L (winter). Arched bacillus Gram-negative similar to purple not sulfurous bacteria, from a sample of the sub-surface of the anaerobic lagoon was purified and presented 92% of similarity with Rhodopseudomonas palustris. In both lagoons it was identified bacteria similar to Chromobacterium suttsuga (95%), Clostridium sp. (99%), Rhodobacter sphaeroides (99%), Rhodopseudomonas palustris (99%), Lampropedia hyalina (97%), Campylobacter fetus (99%), Desulfovibrio vulgaris (95%), Rhodospirillum rubrum (95%).

This study investigated the utility to vertebrates of upland linear riparian rainforest fragments on the Atherton and Evelyn Tablelands in the Australian Wet Tropics region, north Queensland. Similar linear fragments were selected, that varied in forest age and their connectivity to large areas of continuous forest:- (connected primary (N=6), isolated primary (N=5), connected secondary (N=6) and isolated secondary (N=7)). Primary sites had either never been cleared or only subject to selective logging, while secondary forest had been completely cleared and allowed to regenerate for at least 30 years. These linear fragments were contrasted with riparian sites within continuous forest sites (N=6 to 7), which were situated in State Forest or National Parks, and sites within the cleared matrix (pasture, N=6). Vertebrates surveyed were birds, ground-dwelling mammals and reptiles, particularly leaf-litter skinks. All surveys were conducted between September and December in 2001 and/or 2000. Chapter 2 investigates the effects of forest age, isolation and structural vegetation features on bird assemblages within linear riparian fragments of rainforest. Bird surveys and structural vegetation assessments were conducted within connected and isolated primary and secondary linear fragments, and compared with those of continuous forest habitat (N=6) and pasture. There were strong effects of forest age; all three types of primary rainforest had higher values than secondary rainforest for most measured attributes of vegetation structure (including canopy height and cover; and frequency of large-diameter trees, lianes, epiphytes, strangler figs; and woody debris), but lower frequencies of tree ferns and thorny scramblers. Sites within primary rainforest also had a greater frequency of many bird species across different guilds of habitat, feeding and movement. Assemblages of rainforest-dependent birds showed an effect of isolation, although its strength was less than that of forest age. Isolated fragments of primary rainforest differed significantly from continuous primary rainforest in their rainforest-dependent bird species assemblages (and had lower species richness), and isolated fragments of secondary rainforest differed from those that were connected. There was a significant association between the species composition of rainforest birds and some measured vegetation parameters across all sites, but not within primary or secondary sites. Vegetation differences did not explain the lowered frequency of several species in isolated fragments. Limited dispersal seems unlikely to be a main cause, and causal processes probably vary among species. Specialist rainforest species endemic to the Wet Tropics region showed stronger responses to present-day rainforest age and fragmentation than those not endemic. Variation in nest depredation levels associated with rainforest fragmentation (edge effects) is examined in Chapter 3. Artificial nests were placed in the forest understorey at seven edge sites where continuous forest adjoined pasture, seven interiors (about one kilometre from the edge), and six primary linear riparian forest remnants (50-100 m wide) that were connected to continuous forest. Four nest types were compared, representing different combinations of two factors; height (ground, shrub) and shape (open, domed). At each site, four nests of each type, containing one quail egg and two model plasticine eggs, were interspersed about 15 m apart within a 160 m transect. Predators were identified from marks on the plasticine eggs. The overall depredation rate was 66.5% of 320 nests' contents damaged over a three-day period. Large rodents, especially the rat Uromys caudimaculatus, and birds, especially the spotted catbird Ailuroedus melanotis, were the main predators. Mammals comprised 56.5% and birds 31.0% of identified predators, with 12.5% of unknown identity. The depredation rate did not vary among site-types, or between open and domed nests, and there were no statistically significant interactions. Nest height strongly affected depredation rates by particular types of predator; depredation rates by mammals were highest at ground nests, whereas attacks by birds were most frequent at shrub nests. These effects counterbalanced so that overall there was little net effect of nest height. Mammals accounted for 78.4% of depredated ground nests and birds for at least 47.4% of shrub nests (and possibly up to 70.1%). The main predators were species characteristic of rainforest, rather than habitat generalists, open-country or edge specialists. For birds that nest in the tropical rainforest understorey of the study region, it is unlikely that edges and linear remnants presently function as ecological population sinks due to mortality associated with increased nest depredation. The use of linear riparian remnants by small ground-dwelling mammals and reptiles (mainly leaf litter skinks), is reported in Chapter 4. Site types were continuous rainforest, connected and isolated linear fragments of both uncleared primary rainforest and secondary regrowth rainforest. Mammals were also surveyed in pasture sites. Neither reptile species richness nor abundance varied significantly among site types. Although mammal species richness varied significantly between site types, with isolated primary sites containing highest species richness, overall mammal abundance did not differ significantly among site types. Pasture sites differed significantly from all rainforest sites in their mammal species composition, and were dominated by the introduced house mouse (Mus musculus). This species was absent from all rainforest sites, which were characterised by moderate abundances of bush rat/Cape York rat Rattus fuscipes/leucopus, fawn-footed melomys Melomys cervinipes and giant white-tailed rat Uromys caudimaculatus. None of these species varied significantly in abundance among site types, although the giant white-tailed rat showed a trend (P=0.09) for reduced abundance in isolated secondary sites. A single reptile species, the prickly forest skink Gnypetoscincus queenslandiae, occurred in sufficient numbers for individual analysis, and its abundance varied significantly among the forested site types, being less abundant in all linear fragments than in continuous forest sites. The utility of linear riparian rainforest for vertebrates appears to be species-specific and involves many factors. However, overall, species endemic to the Wet Tropics (which are hence of the highest conservation significance) appear to be the most sensitive to fragmentation. These species were most likely to show altered abundances or frequencies of occurrence due to isolation, forest age, and habitat linearity. The ecology of species within this group warrants further investigation within fragmented and non-fragmented regions of the Tablelands. For many other vertebrates examined in this study, there appears to be sufficient functional connectedness between remnants on the Tablelands to minimise the effects of fragmentation. Nevertheless, the lower density of many of these species in pasture may indicate that their long-term persistence within the fragmented rainforest areas could benefit from the maintenance or establishment of habitat linkages. Certainly, if the current rainforest vegetation cover were further reduced, or if the land use in the matrix became more intensive, the establishment of specific habitat linkages could become more important as existing dispersal routes could be lost. It also appears that nest depredation levels are unlikely to limit the value of linear rainforest remnants and other small rainforest remnants as breeding habitat for birds (at least for understorey-nesting species), relative to more intact rainforest, in the study region.

La géologie de l'Equateur se distingue de celle du reste des Cordillères andines par la présence de terrains océaniques qui s'accrètent successivement à la marge depuis la fin du Crétacé jusqu'au Paléocène. Ces terrains exotiques sont constitués de fragments de deux plateaux océaniques du Crétacé inférieur (123 Ma) et supérieur (90 Ma) et de plusieurs arcs insulaires (100 à 40 Ma). Dans la Cordillère occidentale, le plateau crétacé inférieur et les terrains océaniques du crétacé supérieur (plateau et arcs insulaires) s'accrètent respectivement à 85-80 Ma et 70-65 Ma. Le plateau océanique daté à 123 Ma est constitué: (i) de cumulats basiques et ultrabasiques (série de San Juan) et (ii) de basaltes en coussins et de dolérites massives recoupés par des petites intrusions de gabbro sub-doléritique. Les cumulats forment une série continue depuis des dunites et wehrlites jusqu'à des gabbros à clinopyroxène. Basaltes, dolérites et gabbros présentent tous les caractères de basalte de plateau océanique (OPB; MgO = 7%; spectres de terres rares plats, pas d'anomalie négative en Nb et Ta). Les compositions isotopiques (Nd et Pb) des cumulats et de l'ensemble basalte-dolérite sont identiques et démontrent que toutes ces roches dérivent d' une source mantellique enrichie. Les picrites et les basaltes enrichis ou non en MgO, associés à des dolérites et des gabbros, représentent des témoins accrétés du plateau océanique crétacé supérieur caraïbe. Les picrites (MgO -20 %) sont appauvries en terres rares légères, peu radiogéniques en Pb et caractérisées par des eNd élevés (+8 à +10). Les basaltes riches en MgO (9-10%) sont enrichis en terres rares légères. La géochimie en éléments traces des basaltes pauvres en MgO est identique à celle d'OPB. Par contre, les rapports isotopiques du Pb des basaltes, riches en MgO ou non, sont très élevés indiquant que toutes ces laves dérivent d'une source enrichie de type HIMU, caractère commun à tous les basaltes du plateau caraïbe .

Das Lektin aus <i>Pisum sativum</i>, der Gartenerbse, ist Teil der Familie der Leguminosenlektine. Diese Proteine haben untereinander eine hohe Sequenzhomologie, und die Struktur ihrer Monomere, ein all-ß-Motiv, ist hoch konserviert. Dagegen gibt es innerhalb der Familie eine große Vielfalt an unterschiedlichen Quartärstrukturen, die Gegenstand kristallographischer und theoretischer Arbeiten waren. Das Erbsenlektin ist ein dimeres Leguminosenlektin mit einer Besonderheit in seiner Struktur: Nach der Faltung in der Zelle wird aus einem Loop eine kurze Aminosäuresequenz herausgeschnitten, so dass sich in jeder Untereinheit zwei unabhängige Polypeptidketten befinden. Beide Ketten sind aber stark miteinander verschränkt und bilden eine gemeinsame strukturelle Domäne. Wie alle Lektine bindet Erbsenlektin komplexe Oligosaccharide, doch sind seine physiologische Rolle und der natürliche Ligand unbekannt. In dieser Arbeit wurden Versuche zur Entwicklung eines Funktionstests für Erbsenlektin durchgeführt und seine Faltung, Stabilität und Monomer-Dimer-Gleichgewicht charakterisiert. Um die spezifische Rolle der Prozessierung für Stabilität und Faltung zu untersuchen, wurde ein unprozessiertes Konstrukt in <i>E. coli</i> exprimiert und mit der prozessierten Form verglichen. <br /> <br /> Beide Proteine zeigen die gleiche kinetische Stabilität gegenüber chemischer Denaturierung. Sie denaturieren extrem langsam, weil nur die isolierten Untereinheiten entfalten können und das Monomer-Dimer-Gleichgewicht bei mittleren Konzentrationen an Denaturierungsmittel auf der Seite der Dimere liegt. Durch die extrem langsame Entfaltung zeigen beide Proteine eine apparente Hysterese im Gleichgewichtsübergang, und es ist nicht möglich, die thermodynamische Stabilität zu bestimmen. Die Stabilität und die Geschwindigkeit der Assoziation und Dissoziation in die prozessierten bzw. nichtprozessierten Untereinheiten sind für beide Proteine gleich. Darüber hinaus konnte gezeigt werden, dass auch unter nicht-denaturierenden Bedingungen die Untereinheiten zwischen den Dimeren ausgetauscht werden.<br /> <br /> Die Renaturierung der unprozessierten Variante ist unter stark nativen Bedingungen zu 100 % möglich. Das prozessierte Protein dagegen renaturiert nur zu etwa 50 %, und durch die Prozessierung ist die Faltung stark verlangsamt, der Faltungsprozess ist erst nach mehreren Tagen abgeschlossen. Im Laufe der Renaturierung wird ein Intermediat populiert, in dem die längere der beiden Polypeptidketten ein Homodimer mit nativähnlicher Untereinheitenkontaktfläche bildet. Der geschwindigkeitsbestimmende Schritt der Renaturierung ist die Assoziation der entfalteten kürzeren Kette mit diesem Dimer. / The lectin from <i>Pisum sativum</i> (garden pea) is a member of the family of legume lectins. These proteins share a high sequence homology, and the structure of their monomers, an all-ß-motif, is highly conserved. Their quaternary structures, however, show a great diversity which has been subject to cristallographic and theoretical studies. Pea lectin is a dimeric legume lectin with a special structural feature: After folding is completed in the cell, a short amino acid sequence is cut out of a loop, resulting in two independent polypeptide chains in each subunit. Both chains are closely intertwined and form one contiguous structural domain. Like all lectins, pea lectin binds to complex oligosaccharides, but its physiological role and its natural ligand are unknown. In this study, experiments to establish a functional assay for pea lectin have been conducted, and its folding, stability and monomer-dimer-equilibrium have been characterized. To investigate the specific role of the processing for stability and folding, an unprocessed construct was expressed in <i>E. coli</i> and compared to the processed form.<br /> <br /> Both proteins have the same kinetic stability against chemical denaturant. They denature extremely slowly, because only the isolated subunits can unfold, and the monomer-dimer-equilibrium favors the dimer at moderate concentrations of denaturant. Due to the slow unfolding, both proteins exhibit an apparent hysteresis in the denaturation transition. Therefore it has not been possible to determine their thermodynamic stability. For both proteins, the stability and the rates of association and dissociation into processed or unprocessed subunits, respectively, are equal. Furthermore it could be shown that even under non-denaturing conditions the subunits are exchanged between dimers.<br /> <br /> Renaturation of the unprocessed variants is possible under strongly native conditions with 100 % yield. The processed protein, however, can be renatured with yields of about 50 %, and its refolding is strongly decelerated. The folding process is finished only after several days. During renaturation, an intermediate is populated, in which the longer of the two polypeptide chains forms a homodimer with a native-like subunit interface. The rate limiting step of renaturation is the association of the unfolded short chain with this dimer.

The heaviest fragments, recoils, have been studied in proton and deuteron induced 28Si reactions and proton-20Ne reactions at 100-300 MeV per nucleon. Inclusive charge and angular distributions and coincidences between He nuclei and recoils have been compared to two theoretical models, Dubna Cascade Model and JAERI Quantum Molecular Dynamics. The overall agreement was good for the reactions with 28Si, however the angular distributions of He fragments could not be reproduced. For the 20Ne reactions the recoil angular distributions were only reproduced for large angles. There was a significant underestimation at small angles and low recoil charge. α-clustering in the bombarding nucleus is a possible explanation for the deviations. In the 100 MeV per nucleon reactions all assumptions of the models may not be valid and the agreement was worst for these reactions. In proton-natXe reactions intermediate mass fragments have been studied from 200 to 1400 MeV. Slow ramping was used to scan the energy. Charge distributions and a caloric curve have been compared to Cascade Fragmentation Evaporation Model. Charge distributions showed good agreement for fragments with Z&lt;8 but the heavier fragments were underestimated.

The Equation of State (EoS) of asymmetric nuclear matter has been explored through the study of mid-rapidity fragment dynamics from the 35 MeV/u $^{70}$Zn $^{70}$Zn, $^{64}$Zn $^{64}$Zn, and $^{64}$Ni $^{64}$Ni systems. The experimental data was collected at the Texas A and M Cyclotron Institute using the 4 NIMROD-ISiS array, which provided both event characterization and excellent isotopic resolution of charged particles. The transverse collective flow was extracted for proton, deuteron, triton, 3He, alpha, and 6He particles. Isotopic and isobaric effects were observed in the transverse flow of the fragments. In both cases, the transverse flow was shown to decrease with an increasing neutron content in the fragments. The (N/Z)sys dependence of the transverse flow and the difference betwen the triton and 3He flow were shown to be sensitive to the density dependence of the symmetry energy using the stochastic mean-field model. A stiff parameterization of Esym(p) was found to provide better agreement with the experimental data. The transverse flow for intermediate mass fragments (IMFs) was investigated, providing a new probe to study the nuclear EoS. A transition from the IMF flow strongly depending on the mass of the system, in the most violent collisions, to a dependence on the charge of the system, for the peripheral reactions, was observed. Theoretical simulations were used to show that the relative differences in the IMF flow are sensitive to the density dependence of the symmetry energy. The best agreement between the experiment and theory was achieved with a stiff Esym(p). A new method was developed in which correlations between the projectile-like and mid-rapidity fragments were examined using a scaled flow. Theoretical simulations were used to show that the scaled flow of the particles was connected to their average order of emission. The experimental results suggest that the mid-rapidity region is preferentially populated with neutron-rich light charged particles and the Z=3-4 IMFs at a relatively early stage in the collision. This work presents additional constraints on the nuclear EoS and insight into the mid-rapidity dynamics observed in Fermi energy heavy-ion collisions.

Prostate cancer remains one of the most commonly diagnosed cancers in men and is a leading cause of cancer death. While great success has been achieved at curing early stage prostate cancer, limited success has been obtained when treating late-stage hormone independent prostate cancer. This is due to the increased propensity for skeletal and non-skeletal metastases. Thus development of novel effective therapeutic modalities against late stage prostate cancer is of critical importance. / Towards these objectives, I have focused my attention on the role of prostate secretory protein (PSP-94) which is expressed in normal individuals and in patients with early stage prostate cancer. Using our well established in vivo models of prostate cancer, I have evaluated the ability of PSP-94 and its amino acids 31-45 required (PCK3145) to decrease tumor growth and skeletal metastases in vivo and evaluated the potential mechanism(s) associated with PCK3145 anti-cancer actions. / Prostatic cancer can also develop as a result of epigenetic activation of tumor promoting genes. To evaluate the role of methylation in prostate cancer, late stage prostate cancer cells were treated with the universal methylating agent S-adenosylmethionine (SAM) and an anti-sense oligonucleotide directed against MBD2 (AS). Scrambled oligonucleotide was included as a control (S). Both SAM and MBD2-AS resulted in inhibition in uPA, MMP-2 and VEGF production leading to decreased tumor cell invasive capacity. However, SAM and MBD2-AS were not able to either further repress partially methylated genes (GSTP1) or reactivate already methylated genes (AR). Furthermore, SAM and MBD2-AS treatment resulted in significant reduction in tumor growth in vivo . Immunohistochemical and RT-PCR analyses carried out on SAM and MBD2-AS tumors revealed decreased protein and mRNA expression of uPA and MMP-2 which was partially due to increased methylation of the respective promoters even after 10 weeks post in vitro treatment as analyzed by bisulfate sequencing. In addition decreased levels of angiogenesis and tumor survival markers were observed. / Collectively, these studies are aimed at the development of novel reliable approached to diagnose and treat advanced, hormone refractory prostate cancer to reduce tumor associated morbidity and mortality.