1000 resultados para João Manuel dos Santos Cunha
Resumo:
Proteinases are enzymes distributed widely founded in several organisms and perform many different functions, from maintaining homeostasis to the worsening of some diseases such as cancer, autoimmune diseases and infections. The proteins responsible of controlling the action of these enzymes are the inhibitors, that are classified based on their target proteases and are founded since simple organisms, such as bacteria, to higher organisms, such as larger plants and mammals. Plant proteinase inhibitors act by reducing or inactivating the activity of target proteases, thus, these proteins have been studied as potential tools in the treatment of diseases related to protease activities. In this context, an inhibitor of chymotrypsin from Erythrina velutina, called EvCI was previously purified and it was observed that this protein plays in vitro anticoagulant activity and anti-inflammatory activity in in vivo model. Aiming to reduce the environmental impact caused by the purification EvCI in high amounts and to facilitate the process of obtaining this protein, the recombinant chymotrypsin inhibitor from Eryhrina velutina was produced after cloning and expression in Escherichia coli. The bacteria were grown in LB medium and after induction of the expression this material was subjected to procedures for cell lysis and the product was applied on Nickel-affinity column. The proteins adsorbed were digested by thrombin and applied on Chymotrypsin-Sepharose affinity column, obtaining the purified inhibitor, named recEvCI. After electrophoresis, the recombinant inhibitor showed an approximately molecular mass of 17 kDa, and reduced the chymotrypsin and elastase activities in vitro. The recombinant inhibitor was sequenced and was found similar amino acids residues when compared to other inhibitors deposited in the database, with some modifications. recEvCI showed high stability under pH variations and reducing conditions, maintaining its activity around 80%. This protein increased the blood coagulation time in vitro by acting on the intrinsic pathway and did not show cytotoxicity against strains of mouse 3T3 fibroblasts and RAW 264.7 macrophages. recEvCI showed microbicide activity related to release of nitric oxide and consequently the activation of macrophages, futhermore having proinflammatory effects assessed by increased release of TNF-α. These results indicate that recEvCI can be biotechnologically used as a new tool in the control of coagulation-related diseases as well as can be an activating agent of the immune system in immunosuppressed individuals
Resumo:
Esta tese incide sobre o desenvolvimento de modelos computacionais e de aplicações para a gestão do lado da procura, no âmbito das redes elétricas inteligentes. É estudado o desempenho dos intervenientes da rede elétrica inteligente, sendo apresentado um modelo do produtor-consumidor doméstico. O problema de despacho económico considerando previsão de produção e consumo de energia obtidos a partir de redes neuronais artificiais é apresentado. São estudados os modelos existentes no âmbito dos programas de resposta à procura e é desenvolvida uma ferramenta computacional baseada no algoritmo de fuzzy-clustering subtrativo. São analisados perfis de consumo e modos de operação, incluindo uma breve análise da introdução do veículo elétrico e de contingências na rede de energia elétrica. São apresentadas aplicações para a gestão de energia dos consumidores no âmbito do projeto piloto InovGrid. São desenvolvidos sistemas de automação para, aquisição monitorização, controlo e supervisão do consumo a partir de dados fornecidos pelos contadores inteligente que permitem a incorporação das ações dos consumidores na gestão do consumo de energia elétrica; SMART GRIDS - COMPUTATIONAL MODELS DEVELOPMENT AND DEMAND SIDE MANAGMENT APPLICATIONS Abstract: This thesis focuses on the development of computational models and its applications on the demand side management within the smart grid scope. The performance of the electrical network players is studied and a domestic prosumer model is presented. The economic dispatch problem considering the production forecast and the energy consumption obtained from artificial neural networks is also presented. The existing demand response models are studied and a computational tool based on the fuzzy subtractive clustering algorithm is developed. Energy consumption profiles and operational modes are analyzed, including a brief analysis of the electrical vehicle and contingencies on the electrical network. Consumer energy management applications within the scope of InovGrid pilot project are presented. Computational systems are developed for the acquisition, monitoring, control and supervision of consumption data provided by smart meters allowing to incorporate consumer actions on their electrical energy management.
Resumo:
Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.
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In Brazil, accidents with scorpions are considered of medical importance, not only by the high incidence, but also for the potentiality of the venom from some species in determining severe clinical conditions. Tityus stigmurus is a widely distributed scorpion species in Northeastern Brazil and known to cause severe human envenomations, inducing pain, hyposthesia, edema, erythema, paresthesia, headaches and vomiting. The present study uses a transcriptomic approach to characterize the molecular repertoire from the non-stimulated venom gland of Tityus stigmurus scorpion. A cDNA library was constructed and 540 clones were sequenced and grouped into 37 clusters, with more than one EST (expressed sequence tag) and 116 singlets. Forty-one percent of ESTs belong to recognized toxin-coding sequences, with antimicrobial toxins (AMP-like) the most abundant transcripts, followed by alfa KTx- like, beta KTx-like, beta NaTx-like and alfa NaTx-like. Our analysis indicated that 34% include other possible venom molecules , whose transcripts correspond to anionic peptides, hypothetical secreted peptides, metalloproteinases, cystein-rich peptides and lectins. Fifteen percent of ESTs are similar to cellular transcripts. Sequences without good matches corresponded to 11%. This investigation provides the first global view of cDNAs from Tityus stigmurus. This approach enables characterization of a large number of venom gland component molecules, which belong either to known or atypical types of venom peptides and proteins from the Buthidae family
Resumo:
Sugarcane (Saccharum spp.) is a plant from Poaceae family that has an impressive ability to accumulate sucrose in the stalk, making it a significant component of the economy of many countries. About 100 countries produce sugarcane in an area of 22 million hectares worldwide. For this reason, many studies have been done using sugarcane as a plant model in order to improve production. A change in gravity may be one kind of abiotic stress, since it generates rapid responses after stimulation. In this work we decided to investigate the possible morphophysiological, biochemical and molecular changes resulting from microgravity. Here, we present the contributions of an experiment where sugarcane plants were submitted to microgravity flight using a vehicle VSB-30, a sounding rocket developed by Aeronautics and Space Institute teams, in cooperation with the German Space Agency. Sugarcane plants with 10 days older were submitted to a period of six minutes of microgravity using the VSB-30 rocket. The morphophysiological analyses of roots and leaves showed that plants submitted to the flight showed changes in the conduction tissues, irregular pattern of arrangement of vascular bundles and thickening of the cell walls, among other anatomical changes that indicate that the morphology of the plants was substantially influenced by gravitational stimulation, besides the accumulation of hydrogen peroxide, an important signaling molecule in stress conditions. We carried out RNA extraction and sequencing using Illumina platform. Plants subjected to microgravity also showed changes in enzyme activity. It was observed an increased in superoxide dismutase activity in leaves and a decreased in its activity in roots as well as for ascorbate peroxidase activity. Thus, it was concluded that the changes in gravity were perceived by plants, and that microgravity environment triggered changes associated with a reactive oxygen specie signaling process. This work has helped the understanding of how the gravity affects the structural organization of the plants, by comparing the anatomy of plants subjected to microgravity and plants grown in 1g gravity
Resumo:
The stereographic projection is a bijective smooth map which allows us to think the sphere as the extended complex plane. Among its properties it should be emphasized the remarkable property of being angle conformal that is, it is an angle measure preserving map. Unfortunately, this projection map does not preserve areas. Besides being conformal it has also the property of projecting spherical circles in either circles or straight lines in the plane This type of projection maps seems to have been known since ancient times by Hipparchus (150 BC), being Ptolemy (AD 140) who, in his work entitled "The Planisphaerium", provided a detailed description of such a map. Nonetheless, it is worthwhile to mention that the property of the invariance of angle measure has only been established much later, in the seventeenth century, by Thomas Harriot. In fact, it was exactly in that century that the Jesuit François d’Aguilon introduced the terminology "stereographic projection" for this type of maps, which remained up to our days. Here, we shall show how we create in GeoGebra, the PRiemannz tool and its potential concerning the visualization and analysis of the properties of the stereographic projection, in addition to the viewing of the amazing relations between Möbius Transformations and stereographic projections.
Resumo:
Complex functions, generally feature some interesting peculiarities, seen as extensions real functions, complementing the study of real analysis. However, the visualization of some complex functions properties requires the simultaneous visualization of two-dimensional spaces. The multiple Windows of GeoGebra, combined with its ability of algebraic computation with complex numbers, allow the study of the functions defined from ℂ to ℂ through traditional techniques and by the use of Domain Colouring. Here, we will show how we can use GeoGebra for the study of complex functions, using several representations and creating tools which complement the tools already provided by the software. Our proposals designed for students of the first year of engineering and science courses can and should be used as an educational tool in collaborative learning environments. The main advantage in its use in individual terms is the promotion of the deductive reasoning (conjecture / proof). In performed the literature review few references were found involving this educational topic and by the use of a single software.
Resumo:
The aim of this workshop to present some of the strategies studied to use GeoGebra in the analysis of complex functions. The proposed tasks focus on complex analysis topics target for students of the 1st year of higher education, which can be easily adapted to pre-university students. In the first part of this workshop we will illustrate how to use the two graphical windows of GeoGebra to represent complex functions of complex variable. The second part will present the use of the dynamic color Geogebra in order to obtain Coloring domains that correspond to the graphic representation of complex functions. Finally, we will use the threedimensional graphics window in GeoGebra to study the component functions of a complex function. During the workshop will be provided scripts orientation of the different tasks proposed to be held on computers with Geogebra version 5.0 or high.
Resumo:
Esta dissertação desenvolve uma plataforma de controlo interactiva para edifícios inteligentes através de um sistema SCADA (Supervisory Control And Data Acquisition). Este sistema SCADA integra diferentes tipos de informações provenientes das várias tecnologias presentes em edifícios modernos (controlo da ventilação, temperatura, iluminação, etc.). A estratégia de controlo desenvolvida implementa um controlador em cascada hierárquica onde os "loops" interiores são executados pelos PLC's locais (Programmable Logic Controller), e o "loop" exterior é gerido pelo sistema SCADA centralizado, que interage com a rede local de PLC's. Nesta dissertação é implementado um controlador preditivo na plataforma SCADA centralizada. São apresentados testes efectuados para o controlo da temperatura e luminosidade de salas com uma grande área. O controlador preditivo desenvolvido tenta optimizar a satisfação dos utilizadores, com base nas preferências introduzidas em várias interfaces distribuídas, sujeito às restrições de minimização do desperdício de energia. De forma a executar o controlador preditivo na plataforma SCADA foi desenvolvido um canal de comunicação para permitir a comunicação entre a aplicação SCADA e a aplicação MATLAB, onde o controlador preditivo é executado. ABSTRACT: This dissertation develops an operational control platform for intelligent buildings using a SCADA system (Supervisory Control And Data Acquisition). This SCADA system integrates different types of information coming from the several technologies present in modem buildings (control of ventilation, temperature, illumination, etc.). The developed control strategy implements a hierarchical cascade controller where inner loops are performed by local PLCs (Programmable Logic Controller), and the outer loop is managed by the centralized SCADA system, which interacts with the entire local PLC network. ln this dissertation a Predictive Controller is implemented at the centralized SCADA platform. Tests applied to the control of temperature and luminosity in hugearea rooms are presented. The developed Predictive Controller tries to optimize the satisfaction of user explicit preferences coming from several distributed user-interfaces, subjected to the constraints of energy waste minimization. ln order to run the Predictive Controller at the SCADA platform a communication channel was developed to allow communication between the SCADA application and the MATLAB application where the Predictive Controller runs.
Resumo:
The Vitamin E consists of eight chemically homologous forms, designated alpha, beta, gamma and delta tocopherols and tocotrienols. Biologically, the alpha-tocopherol (α-TOH) is the most important. Commercially, are found two types of α-TOH a natural (RRR-alpha-tocopherol) and another synthetic (all-rac-alpha-tocopherol). Both forms are absorbed in the intestine, the liver is a preference in favor of forms 2R, due to transfer protein α-TOH. It has higher affinity to these stereoisomers. Newborns are considered high risk for vitamin E deficiency, mainly premature, these have breast milk as a food source for maintenance of serum α-TOH. Clinical signs such as thrombocytosis, hemolytic anemia, retrolental fibroplasia, intraventricular hemorrhage, bronchopulmonary dysplasia and spinocerebellar degeneration can be found in case of a low intake of α-TOH. Thus, maternal supplementation on postpartum with α-TOH can be an efficient way to increase levels of vitamin E in breast milk and thus the consequently increase the supply of micronutrient for the newborn. However, most studies with vitamin E supplementation have been conducted in animals and little is known about the effect of maternal supplementation in humans, as well as on its efficiency to increase levels of α-TOH in human milk, depending on the shape natural or synthetic. The study included 109 women, divided into three groups: control without supplementation (GC) (n=36), supplemented with natural capsule (GNAT) (n=40) and the synthetic capsule (GSINT) (n=33). Blood samples were collected for determination of maternal nutritional status, and colostrums at initial contact and after 24 hours post-supplementation. Analyses were performed by High Performance Liquid Chromatography. Values of α-TOH in serum below 499.6mg/dL were considered deficient. We used the Kruskal-Wallis test and Tukey test to confirm the increase of alpha-tocopherol in milk and efficiency of administered capsules. Daily consumption of α-TOH was based on daily intake of 500 mL of colostrum by the newborn and compared with the nutritional requirement for children from 0 to 6 months of age, 4 mg / day. The mothers had mean concentration of serum α-TOH in 1016 ± 52, 1236 ± 51 and 1083 ± 61 mg / dL, in CG, GNAT and GSINT respectively. There were no women with deficiiency. The GC did not change the concentrations of α-TOH in colostrum. While women supplemented with natural and synthetic forms increased concentrations of α-TOH colostrum in 57.6% and 39%, respectively. By comparing supplemented groups, it was observed a significant difference (p=0.04), the natural capsule more efficient than the synthetic, approximately 49.6%. Individually, 21.1% of the women provided below 4mg/day of α-TOH, after supplementation for this index declined4.1%. Thus, maternal supplementation postpartum raised the levels of alpha-tocopherol in colostrum, and increased efficiency was observed with the natural form
Resumo:
Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites
Resumo:
Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism
Resumo:
The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response
Resumo:
A actual crise social que afecta o país atingiu as PME de uma forma global, provocando uma redução drástica do volume dos seus negócios, em particular as empresas ligadas à gestão de energia, que constituem um exemplo ilustrativo de uma crise mais geral, dado que dependem muito da realização de obras de grande vulto, nomeadamente em edifícios públicos, como escolas, hospitais, universidades, ou outros organismos públicos e privados, as quais estão, de momento, paralisadas. O trabalho a desenvolver centrar-se-á no estudo de um conjunto de empresas ligadas a este sub-sector (players), com análise das atuais dificuldades, e procura de informação relevante sobre a estratégia empresarial com vista à diversificação de produtos para a sua instalação em novos segmentos de mercado ou avaliação de uma estratégia de inovação ou de internacionalização e qual o mercado a desenvolver.. Através deste estudo pretende-se: • Identificar as actuais estratégias empresariais, sobretudo como irão atuar as empresas no mercado nos próximos anos; • Analisar processos que proporcionem a diversificação do produto final, com garantia de sustentabilidade económica do modelo e ampla aceitação das entidades públicas e privadas; • Conhecer as intenções de internacionalização, e as razões de preferência por determinados mercados em detrimento de outros.
Resumo:
Este estudo procurou, por um lado, caracterizar as bibliotecas das Universidades públicas portuguesas em termos de recolha de dados estatísticos, seu tratamento e posterior utilização na gestão e tomada de decisão nessas bibliotecas. Por outro lado, procurou auscultar a satisfação do staff e a satisfação dos utilizadores dessas mesmas bibliotecas. Além desta competente mais descritiva, este estudo incluiu também uma componente explicativa baseada num modelo teórico que sugere, primeiramente, a existência de uma relação entre a utilização de informação estatística (IE) na gestão e tomada de decisão e a satisfação do staff e a satisfação dos utilizadores. E, seguidamente, a presença de uma relação de interdependência entre o grau de satisfação do staff e o grau de satisfação dos utilizadores, tal como é sugerido pela teoria da cadeia de lucro na prestação de serviços (Heskett et al., 1994). Foi possível apurar a existência de uma cultura de recolha de dados estatísticos, tratamento e utilização dessa informação para a gestão e tomada de decisão entre as bibliotecas participantes. O estudo também revela que a recolha e utilização de IE tem um impacto positivo na satisfação dos utilizadores, mas afecta negativamente a satisfação do staff com o ambiente de trabalho. / ABSTRACT; This study characterizes the Portuguesa Higher Education Libraries with respect to the collection of data, its statistical analysis and subsequent use in the management and decision making of these Libraries. ln addition, the study also tried to obtain information on the level of satisfaction of staff and users in the same Libraries. Besides this descriptive component, this study includes an explicative component based on a theoretical model. This model first suggests the existence of a positive relationship between the use of statistical information in management and decision making in a Library and the level of satisfaction of users and staff. A second prediction of the model is the existence of an interdependence relationship between the level of satisfaction of the staff and the level of satisfaction of the users, as it is suggesting by the service-profit-chain theory (Heskett et al., 1994). The results reveal the existence of a culture of data collection, statistical analysis of the data and subsequent use in management and decision making among the participant libraries. The study also shows that the collection and use of statistical information has a positive impact on the level of satisfaction of the users but a negative impact on the level of satisfaction of staff regarding the job environment.