Verificação formal automatizada para sistemas de raciocínio procedural (PRS) utilizando redes de petri coloridas (RPC)

Este trabalho apresenta uma técnica de verificação formal de Sistemas de Raciocínio Procedural, PRS (Procedural Reasoning System), uma linguagem de programação que utiliza a abordagem do raciocínio procedural. Esta técnica baseia-se na utilização de regras de conversão entre programas PRS e Redes de Petri Coloridas (RPC). Para isso, são apresentadas regras de conversão de um sub-conjunto bem expressivo da maioria da sintaxe utilizada na linguagem PRS para RPC. A fim de proceder fia verificação formal do programa PRS especificado, uma vez que se disponha da rede de Petri equivalente ao programa PRS, utilizamos o formalismo das RPCs (verificação das propriedades estruturais e comportamentais) para analisarmos formalmente o programa PRS equivalente. Utilizamos uma ferramenta computacional disponível para desenhar, simular e analisar as redes de Petri coloridas geradas. Uma vez que disponhamos das regras de conversão PRS-RPC, podemos ser levados a querer fazer esta conversão de maneira estritamente manual. No entanto, a probabilidade de introdução de erros na conversão é grande, fazendo com que o esforço necessário para garantirmos a corretude da conversão manual seja da mesma ordem de grandeza que a eliminação de eventuais erros diretamente no programa PRS original. Assim, a conversão automatizada é de suma importância para evitar que a conversão manual nos leve a erros indesejáveis, podendo invalidar todo o processo de conversão. A principal contribuição deste trabalho de pesquisa diz respeito ao desenvolvimento de uma técnica de verificação formal automatizada que consiste basicamente em duas etapas distintas, embora inter-relacionadas. A primeira fase diz respeito fias regras de conversão de PRS para RPC. A segunda fase é concernente ao desenvolvimento de um conversor para fazer a transformação de maneira automatizada dos programas PRS para as RPCs. A conversão automática é possível, porque todas as regras de conversão apresentadas seguem leis de formação genéricas, passíveis de serem incluídas em algoritmos

Desenvolvimento de nanosistemas farmacêuticos para terapia gênica

Gene therapy is one of the major challenges of the post-genomic research and it is based on the transfer of genetic material into a cell, tissue or organ in order to cure or improve the patient s clinical status. In general, gene therapy consists in the insertion of functional genes aiming substitute, complement or inhibit defective genes. The achievement of a foreigner DNA expression into a population of cells requires its transfer to the target. Therefore, a key issue is to create systems, vectors, able to transfer and protect the DNA until it reaches the target. The disadvantages related to the use of viral vectors have encouraged efforts to develop emulsions as non-viral vectors. In fact, they are easy to produce, present suitable stability and enable transfection. The aim of this work was to evaluate two different non-viral vectors, cationic liposomes and nanoemulsions, and the possibility of their use in gene therapy. For the two systems, cationic lipids and helper lipids were used. Nanoemulsions were prepared using sonication method and were composed of Captex® 355; Tween® 80; Spam® 80; cationic lipid, Stearylamine (SA) or 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and water (Milli-Q®). These systems were characterized by average droplet size, Polidispersion Index (PI) and Zeta Potential. The stability of the systems; as well as the DNA compaction capacity; their cytotoxicity and the cytotoxicity of the isolated components; and their transfection capacity; were also evaluated. Liposomes were made by hydration film method and were composed of DOTAP; 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), containing or not Rhodaminephosphatidylethanolamine (PE- Rhodamine) and the conjugate Hyaluronic Acid DOPE (HA-DOPE). These systems were also characterized as nanoemulsions. Stability of the systems and the influence of time, size of plasmid and presence or absence of endotoxin in the formation of lipoplexes were also analyzed. Besides, the ophthalmic biodistribution of PE-Rhodamine containing liposomes was studied after intravitreal injection. The obtained results show that these systems are promising non-viral vector for further utilization in gene therapy and that this field seems to be very important in the clinical practice in this century. However, from the possibility to the practice, there is still a long way

Modelagem molecular na caracterização eletrônica de oligopeptídeos e na descrição quântica da interação fármaco-receptor

In this dissertation, the theoretical principles governing the molecular modeling were applied for electronic characterization of oligopeptide α3 and its variants (5Q, 7Q)-α3, as well as in the quantum description of the interaction of the aminoglycoside hygromycin B and the 30S subunit of bacterial ribosome. In the first study, the linear and neutral dipeptides which make up the mentioned oligopeptides were modeled and then optimized for a structure of lower potential energy and appropriate dihedral angles. In this case, three subsequent geometric optimization processes, based on classical Newtonian theory, the semi-empirical and density functional theory (DFT), explore the energy landscape of each dipeptide during the search of ideal minimum energy structures. Finally, great conformers were described about its electrostatic potential, ionization energy (amino acids), and frontier molecular orbitals and hopping term. From the hopping terms described in this study, it was possible in subsequent studies to characterize the charge transport propertie of these peptides models. It envisioned a new biosensor technology capable of diagnosing amyloid diseases, related to an accumulation of misshapen proteins, based on the conductivity displayed by proteins of the patient. In a second step of this dissertation, a study carried out by quantum molecular modeling of the interaction energy of an antibiotic ribosomal aminoglicosídico on your receiver. It is known that the hygromycin B (hygB) is an aminoglycoside antibiotic that affects ribosomal translocation by direct interaction with the small subunit of the bacterial ribosome (30S), specifically with nucleotides in helix 44 of the 16S ribosomal RNA (16S rRNA). Due to strong electrostatic character of this connection, it was proposed an energetic investigation of the binding mechanism of this complex using different values of dielectric constants (ε = 0, 4, 10, 20 and 40), which have been widely used to study the electrostatic properties of biomolecules. For this, increasing radii centered on the hygB centroid were measured from the 30S-hygB crystal structure (1HNZ.pdb), and only the individual interaction energy of each enclosed nucleotide was determined for quantum calculations using molecular fractionation with conjugate caps (MFCC) strategy. It was noticed that the dielectric constants underestimated the energies of individual interactions, allowing the convergence state is achieved quickly. But only for ε = 40, the total binding energy of drug-receptor interaction is stabilized at r = 18A, which provided an appropriate binding pocket because it encompassed the main residues that interact more strongly with the hygB - C1403, C1404, G1405, A1493, G1494, U1495, U1498 and C1496. Thus, the dielectric constant ≈ 40 is ideal for the treatment of systems with many electrical charges. By comparing the individual binding energies of 16S rRNA nucleotides with the experimental tests that determine the minimum inhibitory concentration (MIC) of hygB, it is believed that those residues with high binding values generated bacterial resistance to the drug when mutated. With the same reasoning, since those with low interaction energy do not influence effectively the affinity of the hygB in its binding site, there is no loss of effectiveness if they were replaced.

Condicionamento estrutural das zonasa de cisalhamento da região de Forquilha, domínio Ceará Central: uma abordagem integrada de sensoriamento remoto e geologia estrutural

The studied region, named Forquilha and localized in northwestern Central Ceará domain (northern portion of Borborema Province), presents a lithostratigraphic framework constituted by paleoproterozoic metaplutonics, metasedimentary sequences and neoproterozoic granitoids. The metasedimentary rocks of Ceará group occupy most part of the area. This group is subdivided in two distinct units: Canindé and Independência. Canindé unit is represented basically by biotite paragneisses and muscovite paragneisses, with minor metabasic rocks (amphibolite lens). Independência sequence is composed by garnetiferous paragneisses, sillimanite-garnet-quartz-muscovite schists and quartz-muscovite schists, pure or muscovite quartzites and rare marbles. At least three ductile deformation events were recognized in both units of Ceará group, named D1, D2 and D3. The former one is interpreted as related to a low angle tangential tectonics which mass transport is southward. D2 event is marked by the development of close/isoclinal folds with a N-S oriented axis. Refolding patterns generated by F1 and F2 superposition are found in several places. The latest event (D3) corresponds to a transcurrent tectonics, which led to development of mega-folds and several shear zones, under a transpressional regime. The mapped shear zones are Humberto Monte (ZCHM), Poço Cercado (ZCPC) and Forquilha (ZCF). Digital image processing of enhanced Landsat 7-ETM+ satellite images, combined with field data, demonstrate that these penetrative structures are associated with positive and negative geomorphologic patterns, distributed in linear and curvilinear arrangements with tonal banding, corresponding to the ductile fabric and to crests. Diverse color composites were tested and RGB-531 and RGB-752 provided the best results for lineament analysis of the most prominent shear zones. Spatial filtering techniques (3x3 and 5x5 filters) were also used and the application of Prewitt filters generated the best products. The integrated analysis of morphological and textural aspects from filtered images, variation of tonalities related to the distribution of geologic units in color composites and the superposition over a digital elevation model, contributed to a characterization of the structural framework of the study area. Kinematic compatibility of ZCHM, ZCPC, ZCF shear zones, as well as Sobral-Pedro II (ZCSPII) shear zone, situated to the west of the study area, was one of the goal of this work. Two of these shear zones (ZCHM, ZCPC) display sinistral movements, while the others (ZCSPII, ZCF) exhibit dextral kinematics. 40Ar/39Ar ages obtained in this thesis for ZCSPII and ZCPC, associated with other 40Ar/39Ar data of adjacent areas, indicate that all these shear zones are related to Brasiliano orogeny. The trend of the structures, the opposite shear senses and the similar metamorphic conditions are fitted in a model based on the development of conjugate shear zones in an unconfined transpression area. A WNW-ESE bulk shortening direction is infered. The geometry and kinematic of the studied structures suggest that shortening was largely accommodated by lateral extrusion, with only minor amounts of vertical stretch

Estudo geológico e geofísico da falha de carnaubais, Bacia Potiguar-RN, e implicações neotectônicas

The segment of Carnaubais Fault located in the southeasthern portion of Guamaré Graben (Potiguar Basin) was studied. Several structures were detected and some of them strongly suggest that the last movements in Carnaubais Fault are of Neotectonic age. The study comprises an integrated interpretation of geologic, geomorphologic and geophysical data (gravimetry, magnetometry, resistivity, and self potential methods). According to the size of the studied areas, two approaches were used in this research. The first approach is of a regional nature and was conducted in an area, hereafter named Regional Area, having approximately 6,000 km2 and localized in the northern portion of Rio Grande do Norte state, around Macau city. The second approach comprises detailled studies of two small areas inside the Regional Area: the Camurupim and São Bento areas. Gravimetric and topographic data were used in the Regional Area. A separation into regional and residual components were conducted both on gravimetric and topographic data. The interpretation of the residual component of the gravimetric data allows a precise mapping of the borders of the Guamaré Graben. The regional component features of the topographic data are controlled by the pair of conjugate faults composed by the Carnaubais Fault (NE direction) and the Afonso Bezerra Fault (NW direction). On the other hand, the residual component of the topographic data shows that river valleis of NW direction are sharply interrupted where they intersect Carnaubais Fault. This fact is interpreted as an evidency that the last significant moviments occured in the Carnaubais Fault. Geologic, geomorphologic and geophysical data (magnetometry, resistivity, and self potential methods) were used in the Camurupim Area. The geologic mapping allows to identify five lithophacies unities. The first two unities (from base to top) were interpreted as composing a marine (or transitional) depositional sequency while the other were interpreted is composing a continental depositional sequence. The two sequences are clearly separated of an erosional discordance. The unities grouped in the marine sequence are composed by calcarenites (Unity A) and mudstones (Unity B). Unity A was deposited in a shalow plataform while Unity B, in a tidal flat. The unities grouped in the continental sequence are composed of conglomerate (Unity C) and sandstones (Unities D and E). Unities C and D are fluvial deposits while unity E is an eolian deposit. Unities A and B can be stratigraphycally correlated with Guamaré Formation. Unities C and D present three possible correlations. They may be correlated with Tibau Formation; or with Barreiras Formation; or with a clastic sediment deposit, commonly found in some rivers of Rio Grande do Norte state, and statigraphycally positioned above Barreiras Formation. Based on the decrease of the grain sizes from base to top both on unities C and D, it is proposed that these unities are correlated with the clastic sediment above mentioned. In this case, these unities would have, at least, Pleistocenic age. Finally, it is proposed that Unity E represent an eolian deposit that sufferred recent changes (at least in the Quaternary). The integrated interpretation of hydrographic, morphologic and geophysical data from Camurupim Area shows that Carnaubais Fault is locally composed by a system of several paralel subvertical faults. The fault presenting the larger vertical slip controls the valley of Camurupim river and separates the area in two blocks; in the nothern block the top of the Jandaira limestone is deeper than in the southern block. In addition, at least one of the faults in the northern block is cutting the whole sedimentary section. Because unities C , D, and/or E may be of Quaternary age, tectonic moviments possibly occured in Carnaubais Fault during this period. Detailled geologic mapping were conducted in beachrocks found in São Bento Area. This area is located at the intersection of the coast line with the Carnaubais Fault. The detected structures in the beachrocks are very similar to those caused by fragile deformations. The structures mapped in the beachrocks are consistent with a stress field with maximun compressional stress in E-W direction and extensional stress in the N-S direction. Since the Carnaubais Fault has a NE direction, it is optimally positioned to suffer tectonic movements under the action of such stress field. In addition, the shape of the coastal line appear to be controlled by the Carnaubais Fault. Furthemore, the observed structures in Camurupim Área are consistent with this stress field. These facts are interpreted as evidences that Carnaubais Fault and beachrocks suffered coupled tectonic movements. These moviments are of Neotectonic age because the beachrocks present ages less than 16,000 years

Estudo teórico de intermediários tetraédricos acidez / basicidade e estereosseletividade enzimáticos

The present work aimed first, the theoretical study of tetrahedral intermediate stability formed from carbonyl addition reactions using the second (MP2) and third (MP3) order Møller–Plesset perturbation theory. Linear correlations between electronic energy difference of reactions with Wiberg Indexes and C-O bond lengths were obtained, and was observed that the stability of adducts formed depends directly of electronic density involved between these atoms. The knowing of electronic parameters of these structures has an important hole due to the large use on reactions that in his course forms this tetrahedral intermediate. Employing the ONIOM (B3LYP:AMBER) methodology, was evaluated the stereoselectivity of a enzymatic reaction between CAL B enzyme and a long chain ester. In this study, were obtained the electronic energies of ground state and intermediate state of transesterification rate-determing step from two possible proquirals faces Re and Si. The objective was study the enantioselectivity of CAL B and rationalizes it using quantum theory of atoms in molecules (QTAIM). A theoretical study employing inorganic compounds was performed using ab initio CBS-QB3 method aiming to find a link between thermodynamic and equilibrium involving acids and bases. The results observed showed an excellent relationship between difference in Gibbs free energy, ΔG of acid dissociation reaction and ΔG of hydrolysis reaction of the corresponding conjugate base. It was also observed, a relationship between ΔG of hydrolysis reaction of conjugate acids and their corresponding atomic radius showing that stability plays an important role in hydrolysis reactions. The importance of solvation in acid/base behavior when compared to theoretical and experimental ΔG´s also was evaluated.

Bioquímica quântica na diferenciação dos níveis de ativação de receptores AMPA por agonistas parciais Wilardina

In the central nervous system (CNS) of mammalian, fast synaptic transmission between nerve cells is performed primarily by α-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid (AMPA) receptors, an ionotropic glutamate receptor that is related with learning, memory and homeostasis of the nervous system. Impairments in their functions are correlated with development of many brain desorders, such as epilepsy, schizophrenia, autism, Parkinson and Alzheimer. The use of willardiine analogs has been shown a powerful tool to understanding of activation and desensitization mechanisms of this receptors, because the modification of a single ligand atom allows the observation of varying levels of efficacy. In this work, taking advantage of Fluorine Willardiine (1.35Å), Hydrogen Willardiine (1.65Å), Bromine Willardiine (1.8Å) and Iodine Willardiine (2.15Å) structures co-crystalized with GluA2 with codes 1MQI, 1MQJ, 1MQH and 1MQG, we attempted to energetically differentiate the four ligands efficacy. The complexes were submitted to energetic calculations based on density functional theory (DFT), under the optics of molecular fractionation with conjugate caps (MFCC) method. Obtained results show a relationship between the energetic values and willardiines efficacy order (FW> HW > BrW > IW), also show the importance of E705, R485, Y450, S654, T655, T480 e P478 as the amino acids that contribute most strongly with the interaction of four partial agonists. Furthermore, we outlined the M708 behaviour, attracted by FW and HW ligands, and repels by BrW and IW. With the datas reported on this work, it is possible for a better understanding of the AMPA receptor, which can serve as an aid in the development of new drugs for this system.

Metodologia númerico-experimental para avaliação da espessura de filmes de nitreto de titânio obtidos por processamento a plasma

In recent decades, changes in the surface properties of materials have been used to improve their tribological characteristics. However, this improvement depends on the process, treatment time and, primarily, the thickness of this surface film layer. Physical vapor deposition (PVD) of titanium nitrate (TiN) has been used to increase the surface hardness of metallic materials. Thus, the aim of the present study was to propose a numerical-experimental method to assess the film thickness (l) of TiN deposited by PVD. To reach this objective, experimental results of hardness (H) assays were combined with a numerical simulation to study the behavior of this property as a function of maximum penetration depth of the indenter (hmax) into the film/substrate conjugate. Two methodologies were adopted to determine film thickness. The first consists of the numerical results of the H x hmax curve with the experimental curve obtained by the instrumental indentation test. This methodology was used successfully in a TiN-coated titanium (Ti) conjugate. A second strategy combined the numerical results of the Hv x hmax curve with Vickers experimental hardness data (Hv). This methodology was applied to a TiN-coated M2 tool steel conjugate. The mechanical properties of the materials studied were also determined in the present study. The thicknesses results obtained for the two conjugates were compatible with their experimental data.

Estudo do balanço de umidade por meio de modelos regionais para o clima do passado e do futuro sobre a América do Sul

In the context of climate change over South America (SA) has been observed that the combination of high temperatures and rain more temperatures less rainfall, cause different impacts such as extreme precipitation events, favorable conditions for fires and droughts. As a result, these regions face growing threat of water shortage, local or generalized. Thus, the water availability in Brazil depends largely on the weather and its variations in different time scales. In this sense, the main objective of this research is to study the moisture budget through regional climate models (RCM) from Project Regional Climate Change Assessments for La Plata Basin (CLARIS-LPB) and combine these RCM through two statistical techniques in an attempt to improve prediction on three areas of AS: Amazon (AMZ), Northeast Brazil (NEB) and the Plata Basin (LPB) in past climates (1961-1990) and future (2071-2100). The moisture transport on AS was investigated through the moisture fluxes vertically integrated. The main results showed that the average fluxes of water vapor in the tropics (AMZ and NEB) are higher across the eastern and northern edges, thus indicating that the contributions of the trade winds of the North Atlantic and South are equally important for the entry moisture during the months of JJA and DJF. This configuration was observed in all the models and climates. In comparison climates, it was found that the convergence of the flow of moisture in the past weather was smaller in the future in various regions and seasons. Similarly, the majority of the SPC simulates the future climate, reduced precipitation in tropical regions (AMZ and NEB), and an increase in the LPB region. The second phase of this research was to carry out combination of RCM in more accurately predict precipitation, through the multiple regression techniques for components Main (C.RPC) and convex combination (C.EQM), and then analyze and compare combinations of RCM (ensemble). The results indicated that the combination was better in RPC represent precipitation observed in both climates. Since, in addition to showing values be close to those observed, the technique obtained coefficient of correlation of moderate to strong magnitude in almost every month in different climates and regions, also lower dispersion of data (RMSE). A significant advantage of the combination of methods was the ability to capture extreme events (outliers) for the study regions. In general, it was observed that the wet C.EQM captures more extreme, while C.RPC can capture more extreme dry climates and in the three regions studied.

Desenvolvimento de nanosistemas farmacêuticos para terapia gênica

Gene therapy is one of the major challenges of the post-genomic research and it is based on the transfer of genetic material into a cell, tissue or organ in order to cure or improve the patient s clinical status. In general, gene therapy consists in the insertion of functional genes aiming substitute, complement or inhibit defective genes. The achievement of a foreigner DNA expression into a population of cells requires its transfer to the target. Therefore, a key issue is to create systems, vectors, able to transfer and protect the DNA until it reaches the target. The disadvantages related to the use of viral vectors have encouraged efforts to develop emulsions as non-viral vectors. In fact, they are easy to produce, present suitable stability and enable transfection. The aim of this work was to evaluate two different non-viral vectors, cationic liposomes and nanoemulsions, and the possibility of their use in gene therapy. For the two systems, cationic lipids and helper lipids were used. Nanoemulsions were prepared using sonication method and were composed of Captex® 355; Tween® 80; Spam® 80; cationic lipid, Stearylamine (SA) or 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and water (Milli-Q®). These systems were characterized by average droplet size, Polidispersion Index (PI) and Zeta Potential. The stability of the systems; as well as the DNA compaction capacity; their cytotoxicity and the cytotoxicity of the isolated components; and their transfection capacity; were also evaluated. Liposomes were made by hydration film method and were composed of DOTAP; 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), containing or not Rhodaminephosphatidylethanolamine (PE- Rhodamine) and the conjugate Hyaluronic Acid DOPE (HA-DOPE). These systems were also characterized as nanoemulsions. Stability of the systems and the influence of time, size of plasmid and presence or absence of endotoxin in the formation of lipoplexes were also analyzed. Besides, the ophthalmic biodistribution of PE-Rhodamine containing liposomes was studied after intravitreal injection. The obtained results show that these systems are promising non-viral vector for further utilization in gene therapy and that this field seems to be very important in the clinical practice in this century. However, from the possibility to the practice, there is still a long way