Nanotechnological delivery systems for the oral administration of active molecules: Polymeric microparticles and microemulsions applied to anti-inflammatory and anti-infectious drugs

This thesis was devoted to the development of innovative oral delivery systems for two different molecules. In the first part, microparticles (MPs) based on xylan and Eudragit® S- 100 were produced and used to encapsulate 5-aminosalicylic acid for colon delivery. Xylan was extracted from corn cobs and characterized in terms of its physicochemical, rheological and toxicological properties. The polymeric MPs were prepared by interfacial cross-linking polymerization and spray-drying and characterized for their morphology, mean size and distribution, thermal stability, crystallinity, entrapment efficiency and in vitro drug release. MPs with suitable physical characteristics and satisfactory yields were prepared by both methods, although the spray-dried systems showed higher thermal stability. In general, spraydried MPs would be preferable systems due to their thermal stability and absence of toxic agents used in their preparation. However, drug loading and release need to be optimized. In the second part of this thesis, oil-in-water microemulsions (O/W MEs) based on mediumchain triglycerides were formulated as drug carriers and solubility enhancers for amphotericin B (AmB). Phase diagrams were constructed using surfactant blends with hydrophiliclipophilic balance values between 9.7 and 14.4. The drug-free and drug-loaded MEs presented spherical non-aggregated droplets around 80 and 120 nm, respectively, and a low polydispersity index. The incorporation of AmB was high and depended on the volume fraction of the disperse phase. These MEs did not reduce the viability of J774.A1 macrophage-like cells for concentrations up to 25 μg/mL of AmB. Therefore, O/W MEs based on propylene glycol esters of caprylic acid may be considered as suitable delivery systems for AmB

Desenvolvimento de sistemas micro e nanoestruturados de quitosana/MDI para aplicações cosméticas

Currently, studies in the area of polymeric microcapsules and nanocapsules and controlled release are considerably advanced. This work aims the study and development of microcapsules and nanocapsules from Chitosan/MDI, using a new technique of interfacial polycondensation combined to spontaneous emulsification, for encapsulation of BZ-3. It was firstly elaborated an experimental design of 23 of the particle in white without the presence of BZ-3 and Miglyol, where the variables were the concentrations of MDI, chitosan and solvent. Starting from the data supplied by the experimental design was chosen the experiment with smaller particle diameter and only added like this BZ-3 and Miglyol. The suspension containing concentrations of 6.25 mg/mL, 12.5 mg/mL, 18.75 mg/mL, 25 mg/mL of BZ-3 were prepared, nevertheless, during the storage time, these formulations presented drug precipitates in the suspensions of 18.75 mg/mL and 25 mg/mL of BZ-3. This apparition of precipitate was attributed to the diffusion of BZ-3 for the aqueous phase without any encapsulation, suggesting so the use of the smaller concentrations of the BZ-3. The suspension containing 6.25mg/mL of BZ3 presented average size of 1.47μm, zeta potential of 61 mV, pH 5.64 and this sample showed an amount of BZ-3 and drug entrapment of 100 %. The suspension containing 12.5mg/mL of BZ-3 presented average size of 1.76μm, zeta potential of 47.4 mV, pH 5.71 and this sample showed an amount of BZ-3 and drug entrapment of 100 %. Then, showing such important characteristics, these two formulations were chosen for futher continuity to the study. These formulations were also characterized by the morphology, FTIR, stability for Turbiscan, DSC and a study of controlled release of the BZ-3 was elaborated in different receiving means

Comportamento social de botos-cinza, sotalia guianensis, na praia de Pipa, RN, Brasil: dinâmica, sequência, sincronia e respostas ao turismo de observação

Social behavior of Guiana dolphins, Sotalia guianensis, at Pipa Beach, RN, Brazil: dynamics, sequence, breathing synchrony, and responses to dolphin watching. Social animals form groups that can range from temporary to permanent. Depending on the nature of the social relationships developed between individuals, groups present a particular social organization and the effect of these interactions shapes the activity patterns of these animals. This study investigates: (i) fission-fusion dynamics of Guiana dolphins, through the analysis of three dimensions of the social system (variation in spatial cohesion, variation in size and composition of groups), (ii) sequence, routine and behavioral stability, (iii) breathing intervals in synchronized groups and (iv) behavioral responses of the animals to dolphin watching. Systematic observations of Guiana dolphins were made from a platform located in cliffs about 25 m above sea level that surround Madeiro Bay, Pipa Beach. Sampling occurred from December 2007 to February 2009 between 0600 h and 1600 h, and the groups of Guiana dolphins were investigated according to their size (alone and group) and composition (adults, adults and juveniles, and adults and calves). According to the analysis of fission-fusion dynamics, Guiana dolphin groups frequently changed their composition, modifying their patterns of spatial grouping and cohesion every 20 minutes on average. More than 50% of the individuals maintained a distance of up to 2 m from other group members and new individuals were attracted to the group, especially during feeding, leaving it for foraging. Large groups were more unstable than small, while groups containing only adults were more stable than groups of adults and juveniles. According to the Z-score analysis to investigate the sequence and behavioral routine, lone individuals were more ! .7! ! involved in foraging and feeding, while resting was more common in groups. Foraging and feeding were more common in homogeneous groups (individuals of the same age class), while heterogeneous groups (different age classes) were often involved in socialization, displaying a broader behavioral repertoire. Foraging and resting behavior presented higher stability (continuous duration in minutes) than the other behaviors. The analysis of breathing intervals in synchronized groups showed significant differences depending on type of behavior, composition and area preference. During resting, breathing intervals were of longer duration, and groups with calves showed shorter breathing intervals than groups without calves. Lone individuals also preferred areas called corral , often used for the entrapment of fishes. The Markov chain analysis revealed behavioral changes in the presence of boats, according to the type of group composition. Groups composed of adults presented decreased resting and increased in traveling during the presence of boats. Groups of adults and juveniles showed a massive reduction of socialization, while the behavior transition probability traveling-traveling was higher in groups of adults and calves. In the presence of the boats, stability of resting was reduced by one third of its original duration and traveling more than doubled. The behavioral patterns analyzed are discussed in light of socio-ecological models concerning costs and benefits of proximity between individuals and behavioral optimization. Furthermore, significant changes in behavioral patterns indicate that Guiana dolphins, at Pipa Beach, have suffered the effects of tourism as a result of violation of rules of conduct established for the study area

Estudo da imobilização de proteases para a síntese de oligolisinas

Enzymatic synthesis of peptides using proteases has attracted a great deal of attention in recent years. One key challenge in peptide synthesis is to find supports for protease immobilization capable of working in aqueous medium at high performance, producing watersoluble oligopeptides. At present, few reports have been described using this strategy. Therefore, the aim of this thesis was to immobilize proteases applying different methods (Immobilization by covalent bound, entrapment onto polymeric gels of PVA and immobilization on glycidil metacrylate magnetic nanoparticles) in order to produce water-soluble oligopeptides derived from lysine. Three different proteases were used: trypsin, α-chymotrypsin and bromelain. According to immobilization strategies associated to the type of protease employed, trypsin-resin systems showed the best performance in terms of hydrolytic activity and oligopeptides synthesis. Hydrolytic activities of the free and immobilized enzymes were determined spectrophotometrically based on the absorbance change at 660 nm at 25 °C (Casein method). Calculations of oligolysine yield and average degree of polymerization (DPavg) were monitored by 1H-NMR analysis. Trypsin was covalently immobilized onto four different resins (Amberzyme, Eupergit C, Eupergit CM and Grace 192). Maximum yield of bound protein was 92 mg/g, 82 mg/g and 60 mg/g support for each resin respectively. The effectiveness of these systems (Trypsin-resins) was evaluated by hydrolysis of casein and synthesis of water-soluble oligolysine. Most systems were capable of catalyzing oligopeptide synthesis in aqueous medium, albeit at different efficiencies, namely: 40, 37 and 35% for Amberzyme, Eupergit C and Eupergit CM, respectively, in comparison with free enzyme. These systems produced oligomers in only 1 hour with DPavg higher than free enzyme. Among these systems, the Eupergit C-Trypsin system showed greater efficiency than others in terms of hydrolytic activity and thermal stability. However, this did not occur for oligolysine synthesis. Trypsin-Amberzyme proved to be more successful in oligopeptide synthesis, and exhibited excellent reusability, since it retained 90% of its initial hydrolytic and synthetic activity after 7 reuses. Trypsin hydrophobic interactions with Amberzyme support are responsible for protecting against strong enzyme conformational changes in the medium. In addition, the high concentration of oxirane groups on the surface promoted multi-covalent linking and, consequently, prevented the immobilized enzyme from leaching. The aforementioned results suggest that immobilized Trypsin on the supports evaluated can be efficiently used for oligopeptides synthesis in aqueous media

Evolução tectono-estrutural do Campo de Xaréu (Sub-bacia de Mundaú, Bacia do Ceará - NE do Brasil: abordagem multiescala e pluriferramental

The Xaréu Oil Field, located in the center-southern portion of the Mundaú Sub-Basin (eastern portion of the Ceará Basin), is characterized by a main Iramework of NW-trending and NE-dipping faults. The faults in the Xaréu Oil Field, among which the Xaréu Fautt stands out, are arranged according to an extensional-listriclan, rooted on a detachment surface corresponding to the Mundaú Fault, the border fautt of Mundaú Sub-Basin. During the tectonic-structural evolution of the Xaréu Oil Field and the Mundaú Sub-Basin, the Mundaú Fault played a crucial role on the control of the geometry of both compartments. The main carbonatic unit in the Xaréu Oil Field, named the Trairí Member(Paracuru Formation of Late Aptian to Early Albian age), contains the largest oil volume in the field, concentrated in structurally-controlled accumulations. The Trairí Member is composed by a variety of carbonatic rocks (massive, bedded or laminated calcilutites, ostracodites, calcarenites and carbonatic rudites, all of them presenting variable degrees of dolomitization). The carbonatic rocks are interbedded into thick packages of black shales and marls, besides local beds of siliciclastic conglomerates, sandstones, siltnes and argillites. From the spatial association and the genetic relationships between the carbonatic and siliciclastic units, it is possible to group them in three lithofacies associations (Marginal Plain, Ramp and Lacustrine Interior) that, together, were developed in a lacustrine system associated to a marginal sabkha. Structural studies based on drill coresthat sample the Trairí Member in the Xaréu Oil Field allowed to characterize two generations of meso- to microscale structures: the D1 group presents a typical hydroplastic character, being characterized by intra/interstratal to oblique-bedding shear zones. The hydroplastic character related to these structures allowed to infer their development at an early-lithilication stage of the Trairí Member, leading to infer an Early Cretaceous age to them. The second group of structures identified in the drill cores, nominated D2 and ascribed to a Neogene age, presents a strictly brttle character, being typilied by normal faults and slickenfibers of re-crystallized clayminerals, ali olthem displaying variable orientations. Although the present faults in the Xaréu Oil Field (and, consequently, in the Mundaú Sub-Basin) were classically relerred as struetures of essentially normal displacement, the kinematics analysis of the meso-to microscaie D1 struetures in the drill cores led to deline oblique displacements (normal with a clockwise strike-slip component) to these faults, indicating a main tectonic transport to ENE. These oblique movements would be responsible for the installation of a transtensive context in the Mundaú Sub-Basin, as part of the transcurrent to translormant opening of the Atlantic Equatorial Margin. The balancing of four struetural cross-sections ofthe Xaréu Oil Field indicates that the Mundaú Fault was responsible for more than 50% of the total stretching (ß factor) registered during the Early Aptian. At the initial stages of the "rifting", during Early Aptianuntil the Holocene, the Mundaú Sub-Basin (and consequently the Xaréu Oil Fleld) accumulated a total stretching between 1.21 and 1.23; in other words, the crust in this segment of the Atlantic Equatorial Margin was subjeeted to an elongation of about 20%. From estimates of oblique displacements related to the faults, it ws possible to construct diagrams that allow the determination of stretching factors related to these displacements. Using these diagrams and assuming the sense 01 dominant teetonictransport towards ENE, it was possible to calculate the real stretching lactors related to the oblique movement 0 of the faults in the Mundaú Sub-Basin. which reached actual values between 1.28 and 1.42. ln addnion to the tectonic-structural studies in the Xaréu Oil Field, the interpretation of remote sensing products, coupled wnh characterization of terrain analogues in seleeted areas along the northern Ceará State (continental margins of the Ceará and Potiguar basins), provided addnional data and constraints about the teetonic-structural evolution of the oil lield. The work at the analogue sites was particularly effective in the recognition and mapping, in semidetail scale, several generations of struetures originated under a brittle regime. Ali the obtained information (from the Xaréu Oil Field, the remote sensor data and the terrain analogues) were jointly interpreted, culminating with the proposnion of an evolutionary model lor this segment of the Atlantic Equatorial Margin; this model that can be applied to the whole Margin, as well. This segmentof the Atlantic Equatorial Margin was delormedin an early E-W (when considered lhe present-day position of the South American Plate) transcurrent to transform regime with dextral kinematics, started Irom, at least, the Early Aptian, which left its record in several outcrops along the continental margin of the Ceará State and specilically in the Xaréu off Field. The continuous operation of the regime, through the Albian and later periods, led to the definitive separation between the South American and African plates, with the formation of oceanic lithosphere between the two continental blocks, due to the emplacement off spreading centers. This process involved the subsequent transition of the transcurrent to a translorm dextral regime, creating lhe Equatorial Atlantic Oceano With the separation between the South American and African plates already completed and the increasing separation between lhe continental masses, other tecton ic mechanisms began to act during the Cenozoic (even though the Cretaceous tectonic regime lasted until the Neogene), like an E-W compressive stress líeld (related to the spreading olthe oceanic floor along lhe M id-Atlantic Ridge and to the compression of the Andean Chain) effective Irom the Late Cretaceous, and a state of general extension olthe horizontal surface (due to the thermal uplift ofthe central portion of Borborema Province), effective during the Neogene. The overlap of these mechanisms during the Cenozoic led to the imprint of a complex tectonic framework, which apparently influenced the migration and entrapment 01 hydrocarbon in the Ceará Basin

Caracterização macro, meso e microscópica das estruturas frágeis do corpo arenítico conglomerático da região de Santana do Acaraú (CE) e seu embasamento circundante

The structural framework of the sedimentary basins usually plays an important role in oil prospects and reservoirs. Geometry, interconectivity and density of the brittle features developed during basin evolution could change the permo-porous character of the rocks involved in generation, migration and entrapment of fluid flow. Once the structural characterization of the reservois using only sub-surface data is not an easy task, many studies are focused in analogous outcrops trying to understand the main processes by which brittle tectonic is archieved. In the Santana do Acaraú region (Ceará state, NE Brazil) a pack of conglomeratic sandstone (here named CAC) has its geometry controlled mainly by NE trending faults, interpreted as related to reactivation of a precambrian Sobral Pedro II Lineament (LSP-II). Geological mapping of the CAC showed a major NE-SW trending synform developed before its complete lithification during a dextral transpression. This region was then selected to be studied in details in order of constrain the cretaceous deformation and so help the understanding the deformation of the basins along the brazilian equatorial margin. In order to characterize the brittle deformation in different scales, I study some attributes of the fractures and faults such as orientation, density, kinematic, opening, etc., through scanlines in satellite images, outcrops and thin sections. The study of the satellite images showed three main directions of the macrostructures, N-S, NE-SW and E-W. Two of theses features (N-S and E-W) are in aggreement with previous geophysical data. A bimodal pattern of the lineaments in the CAC´s basement rocks has been evidenciated by the NE and NW sets of structures obtained in the meso and microscale data. Besides the main dextral transpression two others later events, developed when the sediments were complety lithified, were recognized in the area. The interplay among theses events is responsible for the compartimentation of the CAC in several blocks along within some structural elements display diferents orientations. Based on the variation in the S0 orientation, the CAC can be subdivided in several domains. Dispite of the variations in orientations of the fractures/faults in the diferents domains, theses features, in the meso and microscopic scale, are concentrated in two sets (based on their trend) in all domains which show similar orientation of the S0 surface. Thus the S0 orientation was used to group the domains in three major sets: i) The first one is that where S0 is E-W oriented: the fractures are oriented mainly NE with the development of a secondary NW trending; ii) S0 trending NE: the fractures are concentrated mainly along the trend NW with a secondary concentration along the NE trend; iii) The third set, where S0 is NS the main fractures are NE and the secondary concentration is NW. Another analized parameter was the fault/fracture length. This attribute was studied in diferent scales trying to detect the upscale relationship. A terrain digital model (TDM) was built with the brittlel elements supperposed. This model enhanced a 3D visualization of the area as well as the spatial distribution of the fault/fractures. Finally, I believe that a better undertanding of the brittle tectonic affecting both CAC and its nearby basement will help the future interpretations of the tectonic envolved in the development of the sedimentary basins of the brazilian equatorial margin and their oil reservoirs and prospects, as for instance the Xaréu field in the Ceará basin, which subsurface data could be correlated with the surface ones