Análise computacional dos determinantes de atividades meio- e neurotóxicas de fosfolipases A2 do veneno de serpentes

A superfamília das fosfolipases A2 (FLA2) é composta por proteínas dotadas de propriedades diferenciadas que as tornam capazes de apresentar distintas atividades biológicas, além de sua atividade catalítica. Esta diversidade funcional é intrigante devido à alta similaridade de seqüência primária e de estrutura entre essas proteínas. O principal objetivo deste trabalho é o desenvolvimento de uma metodologia para a predição de atividades biológicas específicas em FLA2 de peçonha de serpentes a partir da análise de seqüências primárias. A metodologia desenvolvida compreende: a) seleção de seqüências anotadas quanto à função ou atividade biológica que desempenham; b) detecção e validação estatística de motivos de seqüência relacionados à atividade estudada; e c) construção de Modelos Ocultos de Markov (MOMs) representando cada motivo. MOM consiste em uma modelagem estatística que tem sido aplicada com sucesso em diversas áreas onde se faz necessária a detecção e representação de padrões de informação; por sua base matemática robusta e formal, pode ser utilizada na automação deste tipo de processo. A metodologia foi testada para duas atividades de FLA2 de peçonha de serpente: neurotoxicidade e miotoxicidade. Para as FLA2 neurotóxicas, foram detectados seis motivos conservados, dos quais três foram validados estatisticamente como sendo adequados na discriminação de seqüências neurotóxicas de não neurotóxicas. Para as FLA2 miotóxicas, foram detectados seis motivos conservados, dos quais quatro foram validados. Os MOMs dos motivos validados podem ser usados na predição de atividade neurotóxica e miotóxica. As relações entre seqüência, estrutura, função e evolução das FLA2s são discutidas. Os dados obtidos foram coerentes com a hipótese apresentada por Kini (2003), da existência de diferentes sítios farmacológicos na superfície das FLA2, interagindo independente ou cooperativamente com diferentes receptores, para gerar as diversas funções biológicas observadas. Por não haver, até o momento, qualquer ferramenta automatizada para a predição de função biológica de FLA2, os resultados deste trabalho foram a base para a construção de uma ferramenta (disponível em www.cbiot.ufrgs.br/bioinfo/phospholipase) para a identificação de miotoxicidade e neurotoxicidade em FLA2.

The determinantss of foreign direct investment distribution among the Brazilian states

O Investimento Estrangeiro Direto (IED) tem desempenhado um papel importante no esforço do Brasil para tornar-se uma economia orientada para o mercado. De 1995 a 2012 o Brasil recebeu $ 511.5 bilhões de dólares em IED. Em 2012, o Brasil foi o segundo país em desenvolvimento que mais recebeu IED e o quarto no mundo (UNCTAD).Devido à concentração geográfica, os estados brasileiros que são consideravelmente menos desenvolvidos e mais pobres, são aqueles que mais precisam de investimentos e que no entanto, não têm sido receptores relevantes de IED. Em 2010, os estados com os maiores estoques de IED foram São Paulo, com 42,3 por cento do total ($ 99,9 bilhões de dólares), Rio de Janeiro com 13,3 por cento ($ 31,4 bilhões de dólares) e Minas Gerais com 10,6 por cento do total ($ 25,1 bilhões de dólares). Como pode ser observado, apenas três dos vinte e sete estados brasileiros receberam cerca de 66 por cento do total de IED destinado ao Brasil.Dada tal diferenciação na distribuição de IED entre os estados brasileiros, o presente estudo busca explicar se o benefício tributário também é determinante para o fluxo de IED, além das demais variáveis já consideradas como determinantes em outros estudos. Dada a limitação de dados, realizamos duas análises econométricas com dados em painel: 1. Usando seis variáveis chaves: tamanho do mercado consumidor, a qualidade da mão de obra, infraestrutura, custo da mão de obra, carga tributária e benefício tributário (por macro regiões), nos anos de 1995, 2000, 2005 and 2010; 2. Usando cinco variáveis: as mesmas do primeiro modelo, excluindo o custo da mão de obra (por falta de dados) e utilizando os dados de benefício tributário por estado, nos anos de 2010, 2011 e 2012.

Início da função gonadal no sagui comum (Callithrix jacchus): diferenças entre gêneros

Os primatas não humanos, como Callithrix jacchus vêm sendo utilizados com maior frequência em pesquisa biomédica e, recentemente, nosso laboratório propôs uma nova classificação para monitorar com mais detalhes as modificações comportamentais e hormonais relacionadas ao desenvolvimento. O presente trabalho monitorou os níveis de hormônios gonadais de 4 machos e 6 fêmeas de C. jacchus durante as fases juvenil I e II e subadulta (dos 5 aos 15 meses). Os resultados apontaram que os níveis de progesterona nas fêmeas e andrógenas nos machos foram estatisticamente mais elevados nos estágios juvenil II e subadulto, respectivamente, em relação ao estágio juvenil I. Estes achados indicam que, de acordo com a nova classificação, foi possível observar que o funcionamento gonadal se instala mais cedo nas fêmeas

Modulação pela progesterona da sensibilidade dolorosa a estímulos mecânicos e isquêmicos em mulheres saudáveis e jovens

BRUNO, S. S. ; SOUSA, M. B. C. . Modulação pela progesterona da sensibilidade dolorosa a estímulos mecânicos e isquêmicos em mulheres saudáveis e jovens. RBGO. Revista Brasileira de Ginecologia e Obstetrícia , v. 30, p. 306-311, 2008

Dramaturgia do aqui e agora: presença, recepção e sentido no corpo do ator

The actor is an actor during all the phases of creation and development of his character. (S) He goes beyond the moment of interpreting. In this sense, in her (his) daily life, the actor may use her (his) potential to extract from reality the necessary elements for the work of creation. This is a theoretical research which focus on the concept of presence in the actor s work, stretching the concept beyond the scene, encompassing different components such as the body, the word, the silence, the technique and the acting as the actor s stance in regards to his own reality. The objective of such stance is to integrate him (her) self in the environment, balancing his (hers) inner life with the outside life flow. To reach this objective, the research drew theoretical resources from the concept of presence in the actor s work according to BROOK, BARBA, GROTOWSKI and MNOUCHKINE and the studies on reception theory in ISER, 1996; GUMBRECHT, 2010. Thus, the dialogue between reader and actor high lights both as receivers in this work. A practical description of a street theater Kamchàtka show, featured by the Kamchàtka Company is used as an example of the research on the presence. The elements localized in this show are: play, listening, word, silence, meaning, relationship with here and now and the effects of the said presence

Avaliação do efeito citotóxico da lectina da esponja marinha Cliona varians contra células de leucemia mielóide crônica

In this study, a BCR-ABL expressing human chronic myelogenous leukaemia cell line (K562) was used to investigate the antitumoral potential of a novel lectin (CvL) purified from the marine sponge Cliona varians. CvL inhibited the growth of K562 cells with an IC50 value of 70 g/ml, but was ineffective to normal human peripheral blood lymphocytes in the same range of concentrations tested (180 g/ml). Cell death occurred after 72 h of exposure to the lectin and with sign of apoptosis as analysed by DAPI staining. Investigation of the possible effectors of this process showed that cell death occurred in the presence of Bcl-2 and Bax expression, and involved a caspase-independent pathway. Confocal fluorescence microscopy indicated a major role for the lysosomal protease cathepsin B in mediating cell death. Accordingly, pre-incubation of K562 cells with the cathepsin inhibitor L-trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64) abolished the cytotoxic effect of CvL. Furthermore, we found upregulation of tumor necrosis factor receptor 1 (TNFR1) and down-modulation of p65 subunit of nuclear factor kappa B (NFB) expression in CvL-treated cells. These effects were accompanied by increased levels of p21 and downmodulation of pRb, suggesting that CvL is capable of cell cycle arrest. Collectively, these findings suggest that cathepsin B acts as death mediator in CvL-induced cytotoxicity possibly in a still uncharacterized connection with the membrane death receptor pathway

Avaliação do extrato do fungo caripia montagnei e de agonistas de PPAR - α no processo inflamatório

The mushrooms have been object of intense research in view of its potential raising of application in different sectors of the pharmacology and alimentary industry. Among diverse bioactive composites of polyssacharides nature that exist in the fungus the glucans are much searched. These are polymers of glucose and classified as the type of glicosidic linking [α, β]. Peroxisome proliferator-activated receptors (PPARs), ranscription factors belonging to the family of nuclear receptors that bind themselves o specific agonists, have shown their importance in controlling the inflammatory process. The aim of this study was to perform a chemical characterization of extract rom the mushroom Caripia montagnei, assess its antiinflammatory and antibacterial effect and determine if this effect occurs via PPAR. This mushroom is composed of carbohydrates (63.3±4.1%), lipids (21.4l±0.9%) and proteins (2.2± 0.3%). The aqueous solution resulting from the fractionation contained carbohydrates (98.7±3.3%) and protein (1.3±0.25%). Analyses of infrared spectrophotometry and of nuclear magnetic esonance demonstrated that the extract of mushroom C. montagnei is rich in β-glucans. In hioglycolate-induced peritonitis, the C. montagnei glucans (50 mg/kg) educed the inflammatory process in 65.5±5.2% and agonists, pharmacological igands, for PPAR: Wy-14643 (49.3±6.1%), PFOA (48.9±3.8%) and clofibrate in 45.2±3.2%. Sodium diclofenac showed a reduction of 81.65±0.6%. In the plantar edema, the glucans from C. montagnei (50 mg/kg) and L-NAME reduced the edema to a similar degree 91.4±0.3% and 92.8±0,5 %, respectively. In all the groups tested, nitric oxide (NO), an inflammation mediator, showed a significant reduction in the nitrate/nitrite levels when compared to the positive control (P<0.001). The C. montagnei glucans did not show cytotoxicity in the concentrations tested (2.5, 5.0, 10.0, 20.0 and 40.0 µg/100 µL). Antibacterial activity demonstrated that, unlike total extract, there was no inhibition of bacterial growth. The C. montagnei glucans show great potential for antiinflammatory applications. This effect suggests that it is mediated by PPAR activation and by COX and iNOS inhibition

Uma Lectina D-lactose específica da esponja marinha Cinachyrella apion: purificação, caracterização e interação com Leishmania chagasi

Four different sponge species were screened using Ouchterlony agarose gel and immunodiffusion tests to identify cross-reactivity with the polyclonal antibody IgG anti-deglicosilated CvL, a lectin from Cliona varians. Crude extract from the sponge Cinachyrella apion showed cross-reactivity and also a strong haemmaglutinating activity towards human erythrocytes of all ABO groups. Thus, it was submitted to acetone fractionation, IgG anti-deglicosilated CvL Sepharose affinity chromatography, and Fast Protein Liquid Chromatography (FPLC-AKTA) gel filtration on a Superose 6 10 300 column to purify a novel lectin. C. apion lectin (CaL) agglutinated all types of human erythrocytes with preference for papainized type A and O erythrocytes. The haemagglutinating activity is independent of Ca2+, Mg2+ and Mn2+ ions, and it was strongly inhibited by the disaccharide D-lactose, up to a minimum concentration of 6.25 mM. CaL molecular mass determined by FPLC-AKTA gel filtration on a Superose 12 10 300 column and SDS gel electrophoresis was approximately 124 kDa, consisting of eight subunits of 15.5 kDa, assembled by hydrophobic interactions. The lectin was relatively heat- and pH-stable. Leishmania chagasi romastigotes were agglutinated by CaL, indicating that lactose receptors could be presented in this parasite stage. These findings are indicative of the physiological defense roles of CaL and its possible use in the antibiosis of pathogenic protozoa

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Caracterização funcional de dois cDNAs homólogos à ciclofilina e à proteína reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para floração em tomateiro

Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

Atividades antinociceptiva e anti-inflamatória de uma fração rica em heterogalactana sulfatada extraída da alga codium isthmocladum (vickers 1905)

Sulfated polysaccharides comprise a complex group of macromolecules with a range of several biological activities, including antiviral activity, anticoagulant, antiproliferative, antiherpética, antitumor, anti-inflammatory and antioxidant. These anionic polymers are widely distributed in tissues of vertebrates, invertebrates and algae. Seaweeds are the most abundant sources of sulfated polysaccharides in nature. The green algal sulfated polysaccharides are homo or heteropolysaccharides comprised of galactose, glucose, arabinose and/or glucuronic acid. They are described as anticoagulant, anti-inflammatory, antiviral, anti-angiogenic, antitumor compounds. However, there are few studies about elucidation and evaluation of biological/pharmacological effects of sulfated polysaccharides obtained from green algae, for example, there is only one paper reporting the antinociceptive activity of sulfated polysaccharides of these algae. Therefore this study aimed to obtain sulfated polysaccharides of green seaweed Codium isthmocladum and evaluates them as potential antinociceptive agents. Thus, in this study, the total extract of polysaccharides of green alga C. isthmocladum was obtained by proteolytic digestion, followed by fractionation resulting in five fractions (F0.3, F0.5, F0.7, F0.9 and F1.2) by sequential precipitation with acetone. Using the test of abdominal contractions we observed that the fraction F0.9 was the most potent antinociceptive aompound. F0.9 consists mainly of a sulfated heterogalactana. More specific tests showed that Fo.9 effect is dose and time dependent, reaching a maximum at 90 after administration (10 mg / kg of animal). F0.9 is associated with TRPV1 and TRPA1 receptors and inhibits painful sensation in animals. Furthermore, F0.9 inhibits the migration of lymphocytes induced peritonitis test. On the other hand, stimulates the release of NO and TNF-α. These results suggest that F0.9 has the potential to be used as a source of sulfated galactan antinociceptive and anti-inflammatory

Avaliação demográfica, clinico-laboratorial e genética de indivíduos com lupus eritematoso sistêmico e artrite reumatóide residentes em região tropical

The aetiology of autoimmunes disease is multifactorial and involves interactions among environmental, hormonal and genetic factors. Many different genes may contribute to autoimmunes disease susceptibility. The major histocompatibility complex (MHC) genes have been extensively studied, however many non-polymorphic MHC genes have also been reported to contribute to autoimmune diseases susceptibility. The aim of the present study was to evaluate the influence of SLC11A1 gene in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Ninety-six patients with SLE, 37 with RA and 202 controls enrolled in this case-control study, were evaluated with regard to demographic, genetic, laboratorial and clinical data. SLE mainly affects females in the ratio of 18 women for each man, 88,3% of the patients aged from 15 to 45 years old and it occurs with similar frequency in whites and mulattos. The rate of RA between women and men was 11:1, with 77,1% of the cases occurring from 31 to 60 years. The genetic analysis of the point mutation -236 of the SLC11A1 gene by SSCP did not show significant differences between alleles/genotypes in patients with SLE or RA when compared to controls. The most frequent clinical manifestations in patients with SLE were cutaneous (87%) and joint (84.9%). In patients with RA, the most frequent out-joint clinical manifestation were rheumatoid nodules (13,5%). Antinuclear antibodies were present in 100% of the patients with SLE. There was no significant relation between activity of disease and presence of rheumatoid factor in patients with RA, however 55,6% of patients with active disease presented positive rheumatoid factor. Significant association between alleles/genotypes of point mutation -236 and clinical manifestations was not found

Filtro kl/dvs para atenuação do ruído "ground roll"

In geophysics there are several steps in the study of the Earth, one of them is the processing of seismic records. These records are obtained through observations made on the earth surface and are useful for information about the structure and composition of the inaccessible parts in great depths. Most of the tools and techniques developed for such studies has been applied in academic projects. The big problem is that the seismic processing power unwanted, recorded by receivers that do not bring any kind of information related to the reflectors can mask the information and/or generate erroneous information from the subsurface. This energy is known as unwanted seismic noise. To reduce the noise and improve a signal indicating a reflection, without losing desirable signals is sometimes a problem of difficult solution. The project aims to get rid of the ground roll noise, which shows a pattern characterized by low frequency, low rate of decay, low velocity and high amplituds. The Karhunen-Loève Transform is a great tool for identification of patterns based on the eigenvalues and eigenvectors. Together with the Karhunen-Loève Transform we will be using the Singular Value Decomposition, since it is a great mathematical technique for manipulating data