Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Fucana ativa via das Map quinases e inibe a proliferação de células de ovário de hamster chinês (CHO)

Fucan is a term used to denominate L-fucose rich sulfated polysaccharides. The fucans have been studied due their pharmacological activities like antithrombotic, antiproliferative and antioxidant. We have extracted three fucan fractions from the brown seaweed Spatoglossum schröederi. These fucans were denominated Fuc B 1, Fuc B 1.5 and Fuc B 2. The chemical analyzes show that the fucans have very similar composition as demonstrated by agarose electrophoresis gel, sugar and sulfate content. The antiproliferative effect was determined by MTT and BrdU methodologies in CHO cells. The inhibition of proliferation effect of the three fractions was about 40%. Therefore this we proceed just with the Fuc B 2 due the higher yield. There is no apoptosis indication using the anexin V/propidium iodide test. We found a cell cycle phase G1 arrest. The western blotting show that the PKC; pFAK; pERK 1/2 are activated when the cells were treated with fucans. The treatement with inhibitor of MAPK PD98059 extinguished the fucan effect. These results indicates that fucan act by the ERK pathway inducing the cell death.

Indução de morte celular em células de adenocarcinoma cervical humano (HeLa) pela lectina da esponja Cinachyrella apion (CaL)

Cancer is a term used to represent a set of more than 100 diseases, including malignant tumors from different locations. The malignancies are the second leading cause of death in the population, representing approximately 17% of deaths of known cause. Strategies that induce differentiation have had limited success in the treatment of established cancers. In this work, a lectin purified from the marine sponge Cinachyrella apion (CaL) was evaluated due to its hemolytic, cytotoxic and antiproliferative properties, besides the ability to induce cell death via apoptosis in tumor cells. The antiproliferative activity of CaL was tested against cell lines, with the highest inhibition of tumor growth for HeLa, reducing cell growth at a dose dependent manner, with a concentration of 10 μg/mL. The hemolytic activity and toxicity against peripheral blood cells were tested using the concentration of IC50 for both trials and twice the IC50 for analysis in flow cytometry, indicating that CaL is not toxic to these cells. To assess the mechanism of cell death caused by CaL in HeLa cells, we performed flow cytometry and western blotting. The results showed the lectin probably induces cell death by apoptosis activation by pro-apoptotic protein Bax, promoting mitochondrial membrane permeabilization, cell cycle arrest in S phase, with accumulation of cells of approximately 57% in this phase, and acting as both dependent and/or independent of caspases pathway. These results suggest that CaL has the potential to be used as drug treatment against cancer.

Análise de falhas em martelos oscilantes dos desfibradores da indústria sucroalcooleira da Região Norte-Nordeste

The failure of materials is always an unwelcome event for several reasons: human lives are put in danger, economic losses, and interference in the availability of products and services. Although the causes of failures and behaviour of materials can be known, the prevention of such a condition is difficult to be guaranteed. Among the failures, wear abrasion by the low voltage is the kind of failure that occurs in more equipment and parts industry. The Plants Sucroalcooleiras suffer significant losses because of such attrition, this fact that motivated their choice for the development of this work. For both, were considered failures in the swing hammers desfibradores stopped soon after the exchange provided in accordance with tonnage of cane processed, then were analyzed by the level of wear testing of rubber wheel defined by the standard ASTM G65-91.The failures were classified as to the origin of the cause and mechanism, moreover, were prepared with samples of welding procedures according to ASME code, sec. IX as well, using the technique of thermal spraying to analyze the performance of these materials produced in laboratories, and compares them with the solder used in the plant. It was observed that the bodies-of-proof prepared by the procedure described as welding, and the thermal spraying the results of losing weight have been minimized significantly compared to the preparations in the plant. This is because the use of techniques more appropriate and more controlled conditions of the parameters of welding. As for the thermal spraying, this technique has presented a satisfactory result, but requires the use of these coatings in the best condition for real affirmation of the results

Adição de vermiculita expandida mícron em pastas de baixa densidade para cimentação de poços de petróleo

The northeastern region of Brazil has a large number of wells producing oil using a method of secondary recovery steam injection, since the oil produced in this region is essentially viscous. This recovery method puts the cement / coating on thermal cycling, due to the difference in coefficient of thermal expansion between cement and metal coating causes the appearance of cracks at this interface, allowing the passage of the annular fluid, which is associated with serious risk socioeconomic and environmental. In view of these cracks, a correction operation is required, resulting in more costs and temporary halt of production of the well. Alternatively, the oil industry has developed technology for adding new materials in cement pastes, oil well, providing high ductility and low density in order to withstand the thermo-mechanical loads generated by the injection of water vapor. In this context, vermiculite, a clay mineral found in abundance in Brazil has been applied in its expanded form in the construction industry for the manufacture of lightweight concrete with excellent insulation and noise due to its high melting point and the presence of air in their layers lamellar. Therefore, the vermiculite is used for the purpose of providing low-density cement paste and withstand high temperatures caused by steam injection. Thus, the present study compared the default folder containing cement and water with the folders with 6%, 8% and 10% vermiculite micron conducting tests of free water, rheology and compressive strength where it obtained the concentration of 8 % with the best results. Subsequently, the selected concentration, was compared with the results recommended by the API standard tests of filtered and stability. And finally, analyzed the results from tests of specific gravity and time of thickening. Before the study we were able to make a folder with a low density that can be used in cementing oil well in order to withstand the thermo-mechanical loads generated by steam injection

Ambiente para avaliação de controladores fuzzy aplicados ao método de elevação artificial por bombeio centrífugo submerso

From their early days, Electrical Submergible Pumping (ESP) units have excelled in lifting much greater liquid rates than most of the other types of artificial lift and developed by good performance in wells with high BSW, in onshore and offshore environments. For all artificial lift system, the lifetime and frequency of interventions are of paramount importance, given the high costs of rigs and equipment, plus the losses coming from a halt in production. In search of a better life of the system comes the need to work with the same efficiency and security within the limits of their equipment, this implies the need for periodic adjustments, monitoring and control. How is increasing the prospect of minimizing direct human actions, these adjustments should be made increasingly via automation. The automated system not only provides a longer life, but also greater control over the production of the well. The controller is the brain of most automation systems, it is inserted the logic and strategies in the work process in order to get you to work efficiently. So great is the importance of controlling for any automation system is expected that, with better understanding of ESP system and the development of research, many controllers will be proposed for this method of artificial lift. Once a controller is proposed, it must be tested and validated before they take it as efficient and functional. The use of a producing well or a test well could favor the completion of testing, but with the serious risk that flaws in the design of the controller were to cause damage to oil well equipment, many of them expensive. Given this reality, the main objective of the present work is to present an environment for evaluation of fuzzy controllers for wells equipped with ESP system, using a computer simulator representing a virtual oil well, a software design fuzzy controllers and a PLC. The use of the proposed environment will enable a reduction in time required for testing and adjustments to the controller and evaluated a rapid diagnosis of their efficiency and effectiveness. The control algorithms are implemented in both high-level language, through the controller design software, such as specific language for programming PLCs, Ladder Diagram language.

Bioprospecção de extratos metanólicos de macroalgas marinhas do litoral do Rio Grande do Norte: avaliação das atividades anticoagulante, antioxidante e antiproliferativa

Seaweeds are organisms known to exhibit a variety of biomolecules with pharmacological properties. The coast of Rio Grande do Norte has over 100 species of seaweeds, most of them not yet explored for their pharmacological potential. Sugars and phenolic compounds are the most studied of these being assigned a range of biological properties, such as anticoagulant , antiinflammatory, antitumor and antioxidant activities. In this work, we obtained methanolic extracts from thirteen seaweeds of the coast of Rio Grande do Norte (Dictyota cervicornis; Dictiopterys delicatula; Dictyota menstruallis; D. mertensis; Sargassum filipendula; Spatoglossum schröederi; Acanthophora specifera; Botryocladia occidentalis; Caulerpa cupresoides; C. racemosa; C. prolifera; C. sertularioides e Codium isthmocladum). They were evaluated as anticoagulant and antioxidant drugs, as well as antiproliferative drugs against the tumor cell line HeLa. None of the methanolic extracts showed anticoagulant activity, but when they were evaluated as antioxidant drugs all of extracts showed antioxidant activity in all tests performed (total antioxidant capacity, sequestration of superoxide and hydroxyl radicals, ferric chelation and reductase activity), especially the algae D. mentrualis, D. cilliolata and C. prolifera, who had the greatest potential to donate electrons.In addition, the ability of iron ions chelation appears as the main antioxidant mechanism of the methanolic extracts of these seaweeds mainly for the extract of the C. racemosa seaweed, which reached almost 100% activity. In the MTT assay, all extracts showed inhibitory activity at different levels againts HeLa cells. Moreover, D. cilliolata (MEDC) and D. menstrualis (MEDM) extracts showed specific activity to this cell line, not inhibiting the viability of 3T3 normal cell line, so they were chosen for detailing the antiproliferative mechanism of action. Using flow cytometry, fluorescence microscopy and in vitro assays we demonstrated that MEDC and MEDM induced apoptosis in HeLa cells by activation of caspases 3 and 9 and yet, MEDC induces cell cycle arrest in S phase. Together, these results showed that the methanolic extracts of brown seaweed D. menstrualis and D. cilliolata may contain agents with potential use in combatting cells from human uterine adenocarcinoma. This study also points to the need for more in-depth research on phytochemical and biological context to enable the purification of biologically active products of these extracts

Síntese e avaliação da atividade antitumoral de nanogéis de fucana A da alga marrom Spatoglossum schöederi (C. Agardh) Kützing

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Modelagem de dados de falhas de equipamentos de sub-superfície em poços de petróleo da Bacia Potiguar

This master´s thesis presents a reliability study conducted among onshore oil fields in the Potiguar Basin (RN/CE) of Petrobras company, Brazil. The main study objective was to build a regression model to predict the risk of failures that impede production wells to function properly using the information of explanatory variables related to wells such as the elevation method, the amount of water produced in the well (BSW), the ratio gas-oil (RGO), the depth of the production bomb, the operational unit of the oil field, among others. The study was based on a retrospective sample of 603 oil columns from all that were functioning between 2000 and 2006. Statistical hypothesis tests under a Weibull regression model fitted to the failure data allowed the selection of some significant predictors in the set considered to explain the first failure time in the wells

Modelo de gerenciamento de estoques de peças de reposição para empresas de transporte urbano de passageiros

This work searches to offer a model to improve spare parts stock management for companies of urban passenger transport by bus, with the consequent progress in their maintenance management. Also known as MRO items (Maintenance, Repair and Operations), these spare parts, according their consumption and demand features, cost, criticity to operation, lead-time, quantity of suppliers, among other parameters, shouldn´t have managed their inventory like normal production items (work in process e final products), that because their features, are managed by more predictable models based, for example, in economic order quantity. In the case specifically of companies of urban passenger transport by bus, items MRO have significant importance in their assets and a bad management of these inventories can cause serious losses to company, leading it even bankrupticy business, in more severe situations which missing spare part provokes vehicles shutdown indefinitely. Given slight attention to the issue, which translates in little literature available about it when compared to that literature about normal items stocks, and due the fact that MRO items be critical to bus urban transport of passengers companies´, it is necessary, so, deepen in this theme searching to give technical and scientific subsidies to companies that work, in many times, empirically, with these so decisive inputs to their business. As a typical portfolio problem, in which there are n items, separated into critical and noncritical, while competing for the same resource, it was developed a new algorithm to aid in a better inventory management of spare parts used only in corrective maintenance (whose failures are unpredictable and random), by analyzing the cost-benefit ratio, which compares the level of service versus cost of each item. The model was tested in a company of urban passenger transport by bus from the city of Natal, who anonymously provided their real data to application in this work

Controle on-line do número de não-conformidades por itens em uma produção finita

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Proposta de algoritmo para detecção de vazamentos em oleodutos utilizando análise freqüencial de sinais de pressão

Pipeline leak detection is a matter of great interest for companies who transport petroleum and its derivatives, in face of rising exigencies of environmental policies in industrialized and industrializing countries. However, existing technologies are not yet fully consolidated and many studies have been accomplished in order to achieve better levels of sensitivity and reliability for pipeline leak detection in a wide range of flowing conditions. In this sense, this study presents the results obtained from frequency spectrum analysis of pressure signals from pipelines in several flowing conditions like normal flowing, leakages, pump switching, etc. The results show that is possible to distinguish between the frequency spectra of those different flowing conditions, allowing recognition and announce of liquid pipeline leakages from pressure monitoring. Based upon these results, a pipeline leak detection algorithm employing frequency analysis of pressure signals is proposed, along with a methodology for its tuning and calibration. The proposed algorithm and its tuning methodology are evaluated with data obtained from real leakages accomplished in pipelines transferring crude oil and water, in order to evaluate its sensitivity, reliability and applicability to different flowing conditions

Sistema para detecção e diagnóstico de falhas em motores elétricos de indução utilizando lógica fuzzy

Induction motors are one of the most important equipment of modern industry. However, in many situations, are subject to inadequate conditions as high temperatures and pressures, load variations and constant vibrations, for example. Such conditions, leaving them more susceptible to failures, either external or internal in nature, unwanted in the industrial process. In this context, predictive maintenance plays an important role, where the detection and diagnosis of faults in a timely manner enables the increase of time of the engine and the possibiity of reducing costs, caused mainly by stopping the production and corrective maintenance the motor itself. In this juncture, this work proposes the design of a system that is able to detect and diagnose faults in induction motors, from the collection of electrical line voltage and current, and also the measurement of engine speed. This information will use as input to a fuzzy inference system based on rules that find and classify a failure from the variation of thess quantities

Controlador preditivo não linear aplicado ao controle de golfadas em processos de produção de petróleo

Slugging is a well-known slugging phenomenon in multiphase flow, which may cause problems such as vibration in pipeline and high liquid level in the separator. It can be classified according to the place of its occurrence. The most severe, known as slugging in the riser, occurs in the vertical pipe which feeds the platform. Also known as severe slugging, it is capable of causing severe pressure fluctuations in the flow of the process, excessive vibration, flooding in separator tanks, limited production, nonscheduled stop of production, among other negative aspects that motivated the production of this work . A feasible solution to deal with this problem would be to design an effective method for the removal or reduction of the system, a controller. According to the literature, a conventional PID controller did not produce good results due to the high degree of nonlinearity of the process, fueling the development of advanced control techniques. Among these, the model predictive controller (MPC), where the control action results from the solution of an optimization problem, it is robust, can incorporate physical and /or security constraints. The objective of this work is to apply a non-conventional non-linear model predictive control technique to severe slugging, where the amount of liquid mass in the riser is controlled by the production valve and, indirectly, the oscillation of flow and pressure is suppressed, while looking for environmental and economic benefits. The proposed strategy is based on the use of the model linear approximations and repeatedly solving of a quadratic optimization problem, providing solutions that improve at each iteration. In the event where the convergence of this algorithm is satisfied, the predicted values of the process variables are the same as to those obtained by the original nonlinear model, ensuring that the constraints are satisfied for them along the prediction horizon. A mathematical model recently published in the literature, capable of representing characteristics of severe slugging in a real oil well, is used both for simulation and for the project of the proposed controller, whose performance is compared to a linear MPC

Validação de uma metodologia de ensaio de resistência ao cisalhamento para avaliação da aderência de interfaces revestimento metálico-bainha de cimento aplicada a poços de petróleo

Improving the adherence between oilwell metallic casing and cement sheath potentially decrease the number of corrective actions present/y necessary for Northeastern wells submitted to steam injection. In addition to the direct costs involved in the corrective operations, the economic impact of the failure of the primary cementing aIso includes the loss in the production of the well. The adherence between casing and cement is current/y evaluated by a simple shear tests non standardized by the American Petroleum Institute (API). Therefore, the objective of the present is to propose and evaluate a standardized method to assess the adherence of oilwell metallic casing to cement sheath. To that end, a section of a cemented oilwell was simulated and used to test the effect of different parameters on the shear stress of the system. Surface roughness and different cement compositions submitted or not to thermal cycling were evaluated. The results revealed that the test geometry and parameters proposed yielded different values for the shear stress of the system, corresponding to different adherent conditions between metallic casing and cement sheath