Construção de modelos de interação in silico e in vitro do inibidor do tipo Kunitz de Adenanthera pavonina L. para as enzimas cisteínicas e serínicas

Serines proteinases inhibitors (PIs) are widely distributed in nature and are able to inhibit both in vitro and in vivo enzymatic activites. Seed PIs in than leguminous are classified in seven families, Bowman-Birk and Kunitz type families that most studied representing an important role in the first line of defense toward insects pests. Some Kunitz type inhibitors possess activities serine and cysteine for proteinases named bifunctional inhibitor, as ApTKI the inhibitor isolate from seed of Adenanthera pavonina. The A. pavonina inhibitor presenting the uncommon property and was used for interaction studies between proteinases serine (trypsin) and cysteine (papain). In order to determinate the in vitro interaction of ApTKI against enzymes inhibitor purification was carried cut by using chromatographic techniques and inhibition assays. The 3D model of the bifunctional inhibitor ApTKI was constructed SWISS-MODEL program by homology modeling using soybean trypsin inhibitor (STI, pdb:1ba7), as template which presented 40% of identity to A. pavonina inhibitor. Model quality was evaluated by PROCHECK program. Moreover in silico analyzes of formed complex between the enzymes and ApTKI was evaluated by HEX 4.5 program. In vitro results confirmed the inhibitory assays, where the inhibitor presented the ability to simultaneously inhibit trypsin and papain. The residues encountered in the inhibitor model of folder structural three-dimensional that make contact to enzymes target coud explain the specificity pattern against serine and cysteine proteinases

Determinação de motivos de ligação à quitina em vicilinas de Canavalia ensiformis e Vigna unguiculata através de métodos in silico e relação com suas toxicidades para o bruquídeo Callosobruchus maculatus (Coleoptera:Bruchidae)

Chitin is an important structural component of the cellular wall of fungi and exoskeleton of many invertebrate plagues, such as insects and nematodes. In digestory systems of insects it forms a named matrix of peritrophic membrane. One of the most studied interaction models protein-carbohydrate is the model that involves chitin-binding proteins. Among the involved characterized domains already in this interaction if they detach the hevein domain (HD), from of Hevea brasiliensis (Rubber tree), the R&R consensus domain (R&R), found in cuticular proteins of insects, and the motif called in this study as conglicinin motif (CD), found in the cristallography structure of the β-conglicinin bounded with GlcNac. These three chitin-binding domains had been used to determine which of them could be involved in silico in the interaction of Canavalia ensiformis and Vigna unguiculata vicilins with chitin, as well as associate these results with the WD50 of these vicilins for Callosobruchus maculatus larvae. The technique of comparative modeling was used for construction of the model 3D of the vicilin of V. unguiculata, that was not found in the data bases. Using the ClustalW program it was gotten localization of these domains in the vicilins primary structure. The domains R&R and CD had been found with bigger homology in the vicilins primary sequences and had been target of interaction studies. Through program GRAMM models of interaction ( dockings ) of the vicilins with GlcNac had been gotten. The results had shown that, through analysis in silico, HD is not part of the vicilins structures, proving the result gotten with the alignment of the primary sequences; the R&R domain, although not to have structural similarity in the vicilins, probably it has a participation in the activity of interaction of these with GlcNac; whereas the CD domain participates directly in the interaction of the vicilins with GlcNac. These results in silico show that the amino acid number, the types and the amount of binding made for the CD motif with GlcNac seem to be directly associates to the deleterious power that these vicilins show for C. maculatus larvae. This can give an initial step in the briefing of as the vicilins interact with alive chitin in and exert its toxic power for insects that possess peritrophic membrane

Biodegradação de hidrocarbonetos aromáticos policíclicos: prospecção metagenômica e modelagem computacional 3-D de proteínas

Knowledge of the native prokaryotes in hazardous locations favors the application of biotechnology for bioremediation. Independent strategies for cultivation and metagenomics contribute to further microbiological knowledge, enabling studies with non-cultivable about the "native microbiological status and its potential role in bioremediation, for example, of polycyclic aromatic hydrocarbons (HPA's). Considering the biome mangrove interface fragile and critical bordering the ocean, this study characterizes the native microbiota mangrove potential biodegradability of HPA's using a biomarker for molecular detection and assessment of bacterial diversity by PCR in areas under the influence of oil companies in the Basin Petroleum Geology Potiguar (BPP). We chose PcaF, a metabolic enzyme, to be the molecular biomarker in a PCR-DGGE detection of prokaryotes that degrade HPA s. The PCR-DGGE fingerprints obtained from Paracuru-CE, Fortim-CE and Areia Branca-RN samples revealed the occurrence of fluctuations of microbial communities according to the sampling periods and in response to the impact of oil. In the analysis of microbial communities interference of the oil industry, in Areia Branca-RN and Paracuru-CE was observed that oil is a determinant of microbial diversity. Fortim-CE probably has no direct influence with the oil activity. In order to obtain data for better understanding the transport and biodegradation of HPA's, there were conducted in silico studies with modeling and simulation from obtaining 3-D models of proteins involved in the degradation of phenanthrene in the transport of HPA's and also getting the 3-D model of the enzyme PcaF used as molecular marker in this study. Were realized docking studies with substrates and products to a better understanding about the transport mechanism and catalysis of HPA s

Ergonomia, sustentabilidade sócio-ambiental e atividade de pesca artesanal com jangadas: estudo de caso na Praia de Ponta Negra, Natal-RN

The fishing is made possible by the handiwork of a fisherman, not using appliances that achieve the same levels of industrial fishing productivity and not causing environmental impacts on fish stocks with the same intensity. The adoption of sustainability practices in this type of fishing has been requested for the achievement of environmental and socioeconomic improvements, since its production has been decreasing over the years and overfishing has led to reduced stocks also stand out environmental aggressions such as pollution. Thus, this research aims to analyze the interference of environmental aspects/impacts that permeate the activity performed by fisherman on the beach of Ponta Negra, Natal-RN, in order to propose recommendations for environmental sustainability. We used a methodology based on the ergonomic work analysis (WISNER, 1987; GUERIN, 2001; VIDAL, 2008) using techniques such as observational and interactional observation protocols, photographs, videos, conversational action, and listening to verbalizations. To understand the vulnerability of the environment in which the activity is carried fisherman and the interferences that it suffers, we used the study of environmental aspects/impacts (SÁNCHEZ 2008). As a result, it appears that from the observations located and reports raftsmen, changes in the sea, the search for new fishing grounds farther from the coast, reducing fishery production, waste generation and pollution of the workplace, and the difficulty in docking rafts, factors arising from the pressures both for society as Seen by fisherman, denoting the necessity of drawing up recommendations and actions proposed by management activity by promoting discussion among raftsmen and accountable institutions, and training through workshops on environmental education, aiming to positive changes in the activity fisherman from Ponta Negra and the environment in which raftsmen are inserted in order to seek a sustainable activity, preserving its characteristics

Estudo térmico e de materiais na construção de casas populares com blocos confeccionados a partir de um composto a base de cimento, gesso, eps e raspa de pneu

We built an experimental house on an UFRN´s land using blocks made by a composite consisting of cement, plaster, EPS, crushed rubber and sand. Several blocks were made from various compositions and we made preliminary tests of mechanical and thermal resistance, choosing the most appropriate proportion. PET bottles were used inside the block to provide thermal resistance. In this work, a second function was given to the bottles: to serve as a docking between the blocks, because the ends of the cylinders came out of each block on top as well as at the bottom, with the bottom cut, allowing to fit of the extremities of the upper cylinder of a block in the lower holes of the other one, which were formed by the cutting already mentioned. Minimum compression tests were performed according to ABNT standards for walls closing blocks (fence). With that house built, we did studies of thermal performance in order to ascertain conditions of comfort, checking external and internal temperatures in the walls and in the ambient, among other variables, such as wind speed and relative humidity. The resulting blocks provided adequate thermal insulation to the environment, where the walls presented differences up to 11.7 ºC between the outer and inner faces, getting the maximum temperature inside the house around 31 °C, within the so-called thermal comfort zone for warm climates. At the end of the experiments it was evident the effectiveness of that construction in order to provide thermal comfort in the internal environment of the house, as well as we could confirm the viability of building houses from recyclable materials, reducing the constructive costs, becoming a suitable alternative for low- incoming families. Moreover, besides the low cost, the proposal represents an alternative use of various recyclable materials, therefore considered an ecological solution

Limites e evolução geodinâmica do Sistema Jaguaribeano, Província Borborema, Nordeste do Brasil

The studied area is geologically located in the Northern Domain of the Borborema Province (Northeast Brazil), limited to the south by the Patos shear zone. Terranes of the Jaguaribeano system are dominant, flanked by the Piranhas (E and S sides) and Central Ceará (NE side) terranes. Its basement comprises gneiss -migmatite terrains of Paleoproterozoic to Archean age (2.6 to 1.9 Ga old), overprinted by neoproterozoic to cambrian tectonotherma l events. Narrow supracrustal belts ( schist belts) display a 1.6 to 1.8 Ga age, as shown by whole - rock Rb-Sr and zircon U-Pb and Pb/Pb dates in acid metavolcanics which dominate in the lower section of these sequences, and in coeval metaplutonics (granitic augen gneisses). From the stratigraphic point of view, three Staterian belts are recognized: 1. Orós Belt - made up by the Orós Group, subdivided in the Santarém (predominantly pure to impure quartzites, micaschists and metacarbonates) and Campo Alegre (metandesites, metabasalts, metarhyolites and metarhyodacites, interlayered with metatuffs and metasediments) formations, and by the Serra do Deserto Magmatic Suite (granitic augen gneisses). 2. Jaguaribe Belt - its lithostratigrahic-lithodemic framework is similar to the one of the Orós Belt, however with a greater expression of the volcano -plutonic components (Campo Alegre Formation and Serra do Deserto Magmatic Suite). The Peixe Gordo Sequence, separately described, is also related to this belt and contain s metasedimentary, metavolcanic (with subordinated volcanoclastics) and metaplutonic units. The first one correlated to the Orós Group and the latter the Serra do Deserto Magmatic Suite. 3. Western Potiguar Belt - represented by the Serra de São José Gro up, subdivided in the Catolezinho (biotite -amphibole gneisses with intercalations of metacarbonates, calcsilicate rocks, amphibolites and quartzite beds to the top) and Minhuins (quartzites, micaschists, metaconglomerates, calcsilicate rocks, acid to the b asic metavolcanics and metatuffs) formations. Its late Paleoproterozoic (Staterian) age was established by a Pb/Pb date on zircons from a granitic orthogneiss of the Catolezinho Formation. The petrographic characteristics and sedimentary structures of the Santarém Formation of the Orós Group point to deltaic to shallow marine depositional systems, overlain by deep water deposits (turbidites). The geodynamic setting of this region encompassed a large depositional basin, probably extending to the east of the Portalegre shear zone and west of the Senador Pompeu shear zone, with possible equivalents in the Jucurutu Formation of the Seridó Belt and in the Ceará Group of central Ceará. The Arneiróz Belt, west Ceará, displays some stratigraphic features and granito ids geochemically akin to the ones of the Orós Belt. The evolutionary setting started with an extensional phase which was more active in the eastern part of this domain (Western Potiguar and part of the Jaguaribe belts), where the rudite and psamite sedime ntation relates to a fluviatile rift environment which evolved to a prograding deltaic system to the west (Orós Group). The basaltic andesitic and rhyolitic volcanics were associated to this extensional phase. During this magmatic event, acid magmas also crystallized at plutonic depths. The Orós Group illustrates the environmental conditions in the western part of this domain. Later on, after a large time gap (1.6 to 1.1 Ga), the region was subjected to an extensional deformational episode marked by 900 Ma old (Sm-Nd data) basic rocks, possibly in connection with the deposition of the Cachoeirinha Group south of the Patos shear zone. In the 800 to 500 Ma age interval, the region was affected by important deformational and metamorphic events coupled with in trusion of granitic rocks of variable size (dykes to batholiths), related to the Brasiliano/Pan -African geotectonic cycle. These events produced structural blocks which differentiate, one from the other, according to the importance of anatectic mobilizatio n, proportion of high-grade supracrustals and the amount of neoproterozoic -cambrian granitoid intrusions. On this basis, a large portion of the Jaguaretama Block/Terrane is relatively well preserved from this late overprint. The border belts of the Jagua retama Block (Western Potiguar and Arneiroz) display kyanite-bearing (medium pressure) mineral associations, while in the inner part of the block there is a north-south metamorphic zoning marked by staurolite or sillimanite peak metamorphic conditions. Regarding the deformations of the Staterian supracrustal rocks, second and third phases were the most important, diagnosed as having developed in a progressive tectonic process. In the general, more vigorous conditions of PT are related to the interval tardi - phase 2 early-phase 3, whose radiometric ages and regional structuring indicators places it in the Brasiliano/Pan-African Cycle. In the Staterian geodynamic setting of Brazilian Platform , these sequences are correlated to the lower Espinhaço Supergroup (p.ex., Rio dos Remédios and Paraguaçu groups, a paleproterozoic rift system in the São Francisco Craton), the Araí and Serra da Mesa groups (north of Goiás, in the so -called Goiás Central Massif), and the Uatumã Group (in the Amazonian Craton). Granitic ( augen gneisses) plutonics are also known from these areas, as for example the A-type granites intrusive in the Araí and Serra da Mesa groups, dated at 1.77 Ga. Gravimetric and geological data place the limits of the Jaguaribeano System (terranes) along the Senador Pompeu Shear Zone (western border) and the Portalegre- Farias Brito shear zone (eastern and southern). However, the same data area not conclusive as regards the interpretation of those structures as suture of the terrane docking process. The main features of those shear zones and of involved lothological associations, appear to favour an intracontinental transpressional -transcurrent regime, during Neoproterozoic-Cambrian times, marking discontinuities along which different crustal blocks were laterally dispersed. Inside of this orogenic system and according to the magnetic data (total field map), the most important terrane boundary appears to be the Jaguaribe shear zone. The geochronological data, on some tectonostratigraphic associations (partly represented by the Ceará and Jucurutu groups), still at a preliminary level, besides the lack of granitic zonation and other petrotectonic criteria, do not allow to propose tectonic terrane assembly diagrams for the studied area

Análise de terrenos na porção setentrional da Província Borborema, NE do Brasil: integração de dados geológicos e gravimétricos

It is presently assumed that the Borborema Province resulted from a complex collisional process associated with the convergent movement of plates, possibly involving amalgamation and accretion of microplates. This process was consolidated at the end of the Brasiliano event. It is investigated the possible limits for the tectonostratigraphic terranes in the northern portion of the province based on an integrated study of geological and gravity data. The study area comprises the portion of the Borborema Province located north of the Patos Lineament, limited by longitudes 33º00 W and 43º29 44"W and latitudes 1º36 S and 8º00 S. A revision of the regional geology allowed to identify areas presenting contrasting geological attributes, possibly representing different terranes whose limits are always shear zones of Brasiliano-age. The Sobral-Pedro II shear zone is the only one undoubtedly presenting geological attributes of sutures zones. The other shear zones are very likely associated with a geodinymic context of accretion, involving oblique collisions (docking), transcurrent and/or transforming sutures, and deep intracrustal shear zones. The gravity data contributed as a tool to identify strong lateral contrasts of density inside the upper crust possibly associated with crustal blocks tectonically juxtaposed. The dominant long wavelength anomaly in the Bouguer anomaly map is an expressive gradient, grossly parallel to the continental margin, caused by density variation across the crust-mantle interface in the transition from the continental crust to the oceanic crust originated by the separation between South America and Africa. Medium to small wavelength anomalies are due to intracrustal heterogeneities such as different Precambrian crustal blocks, Brasiliano-age granites and Mesozoic sedimentary basins. A regional-residual separation of the Bouguer anomaly map was performed in order to enhance in the residual map the effect due to intracrustal heterogeneities. The methodology used for this separation was a robust polinomial fitting. The inversion of residual gravity field resulted in a density contrast map (Δρ), in an equivalent layer that provided more accurated anomalies contours and consolidated the model which the sources of residual anomalies are located in the upper part of the present crust. Based on the coincidence of gravity lineaments in the residual map and Brasiliano shear zones, and using additional geological information, the following shear zones are proposed as limits between terranes: Patos shear zone, Sobral-Pedro II shear zone, Picuí-João Câmara shear zone, Remígio-Pocinhos shear zone, Senador Pompeu shear zone, Tauá shear zone, and Portalegre shear zone. Based on the geological/geophysical information it is attributed a higher level of confidence to the first three proposed limits(Patos, Sobral Pedro II, and Picuí-João Câmara shear zones). From west to east, these shear zones individualize the following terranes: Northwest of Ceará terrane, Central Ceará terrane, Tauá terrane, Orós-Jaguaribe terrane, Seridó terrane, and São José de Campestre terrane. In our study, the Rio Piranhas and Patos terranes are questioned because their previously proposed limits do not present good geological and gravimetric evidences. On the other hand, the previously proposed Cearense terrane is now subdivided into Central Ceará and Tauá terranes. Two residual gravity profiles located in the Seridó belt were interpreted using 2 ½ D direct gravity modeling. The main result of the modeling process is that all anomalies, with the exception of one, can be explained by outcroppring bodies, therefore restricted to the upper part of the present crust

Novos protocolos teóricos utilizados nos estudos das ligações hidrogênio-hidrogênio, na estabilidade do tetraedrano, seus derivados e das reações de diels-alder e na quantificação da afinidade de fármacos

This thesis was performed in four chapters, at the theoretical level, focused mainly on electronic density. In the first chapter, we have applied an undergraduate minicourse of Diels-Alder reaction in Federal University of Rio Grande do Norte. By using computational chemistry tools students could build the knowledge by themselves and they could associate important aspects of physical-chemistry with Organic Chemistry. In the second chapter, we studied a new type of chemical bond between a pair of identical or similar hydrogen atoms that are close to electrical neutrality, known as hydrogen-hydrogen (H-H) bond. In this study performed with complexed alkanes, provides new and important information about their stability involving this type of interaction. We show that the H-H bond playing a secondary role in the stability of branched alkanes in comparison with linear or less branched isomers. In the third chapter, we study the electronic structure and the stability of tetrahedrane, substituted tetrahedranes and silicon and germanium parents, it was evaluated the substituent effect on the carbon cage in the tetrahedrane derivatives and the results indicate that stronger electron withdrawing groups (EWG) makes the tetrahedrane cage slightly unstable while slight EWG causes a greater instability in the tetrahedrane cage. We showed that the sigma aromaticity EWG and electron donating groups (EDG) results in decrease and increase, respectively, of NICS and D3BIA aromaticity indices. In addition, another factor can be utilized to explain the stability of tetra-tert-butyltetrahedrane as well as HH bond. GVB and ADMP were also used to explain the stability effect of the substituents bonded to the carbon of the tetrahedrane cage. In the fourth chapter, we performed a theoretical investigation of the inhibitory effect of the drug abiraterone (ABE), used in the prostate cancer treatment as CYP17 inhibitor, comparing the interaction energies and electron density of the ABE with the natural substrate, pregnenolone (PREG). Molecular dynamics and docking were used to obtain the CYP1ABE and CYP17-PREG complexes. From molecular dynamics was obtained that the ABE has higher diffusion trend water CYP17 binding site compared to the PREG. With the ONIOM (B3LYP:AMBER) method, we find that the interaction electronic energy of ABE is 21.38 kcal mol-1 more stable than PREG. The results obtained by QTAIM indicate that such stability is due a higher electronic density of interactions between ABE and CYP17

Construção de modelos de interação in silico e in vitro do inibidor do tipo Kunitz de Adenanthera pavonina L. para as enzimas cisteínicas e serínicas

Serines proteinases inhibitors (PIs) are widely distributed in nature and are able to inhibit both in vitro and in vivo enzymatic activites. Seed PIs in than leguminous are classified in seven families, Bowman-Birk and Kunitz type families that most studied representing an important role in the first line of defense toward insects pests. Some Kunitz type inhibitors possess activities serine and cysteine for proteinases named bifunctional inhibitor, as ApTKI the inhibitor isolate from seed of Adenanthera pavonina. The A. pavonina inhibitor presenting the uncommon property and was used for interaction studies between proteinases serine (trypsin) and cysteine (papain). In order to determinate the in vitro interaction of ApTKI against enzymes inhibitor purification was carried cut by using chromatographic techniques and inhibition assays. The 3D model of the bifunctional inhibitor ApTKI was constructed SWISS-MODEL program by homology modeling using soybean trypsin inhibitor (STI, pdb:1ba7), as template which presented 40% of identity to A. pavonina inhibitor. Model quality was evaluated by PROCHECK program. Moreover in silico analyzes of formed complex between the enzymes and ApTKI was evaluated by HEX 4.5 program. In vitro results confirmed the inhibitory assays, where the inhibitor presented the ability to simultaneously inhibit trypsin and papain. The residues encountered in the inhibitor model of folder structural three-dimensional that make contact to enzymes target coud explain the specificity pattern against serine and cysteine proteinases

Determinação de motivos de ligação à quitina em vicilinas de Canavalia ensiformis e Vigna unguiculata através de métodos in silico e relação com suas toxicidades para o bruquídeo Callosobruchus maculatus (Coleoptera:Bruchidae)

Chitin is an important structural component of the cellular wall of fungi and exoskeleton of many invertebrate plagues, such as insects and nematodes. In digestory systems of insects it forms a named matrix of peritrophic membrane. One of the most studied interaction models protein-carbohydrate is the model that involves chitin-binding proteins. Among the involved characterized domains already in this interaction if they detach the hevein domain (HD), from of Hevea brasiliensis (Rubber tree), the R&R consensus domain (R&R), found in cuticular proteins of insects, and the motif called in this study as conglicinin motif (CD), found in the cristallography structure of the β-conglicinin bounded with GlcNac. These three chitin-binding domains had been used to determine which of them could be involved in silico in the interaction of Canavalia ensiformis and Vigna unguiculata vicilins with chitin, as well as associate these results with the WD50 of these vicilins for Callosobruchus maculatus larvae. The technique of comparative modeling was used for construction of the model 3D of the vicilin of V. unguiculata, that was not found in the data bases. Using the ClustalW program it was gotten localization of these domains in the vicilins primary structure. The domains R&R and CD had been found with bigger homology in the vicilins primary sequences and had been target of interaction studies. Through program GRAMM models of interaction ( dockings ) of the vicilins with GlcNac had been gotten. The results had shown that, through analysis in silico, HD is not part of the vicilins structures, proving the result gotten with the alignment of the primary sequences; the R&R domain, although not to have structural similarity in the vicilins, probably it has a participation in the activity of interaction of these with GlcNac; whereas the CD domain participates directly in the interaction of the vicilins with GlcNac. These results in silico show that the amino acid number, the types and the amount of binding made for the CD motif with GlcNac seem to be directly associates to the deleterious power that these vicilins show for C. maculatus larvae. This can give an initial step in the briefing of as the vicilins interact with alive chitin in and exert its toxic power for insects that possess peritrophic membrane