Comparative structural studies on Lys49-phospholipases A(2) from Bothrops genus reveal their myotoxic site

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mode of Peroxisome Proliferator-Activated Receptor gamma Activation by Luteolin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mecanismos celulares e moleculares que controlam o desenvolvimento e o crescimento muscular

O músculo estriado esquelético é formado pela associação de fibras musculares com a matriz extracelular. Esse tecido possui alta plasticidade e o conhecimento das características morfológicas, da miogênese, e da dinâmica do crescimento é importante para o entendimento da morfofisiologia bem como para a seleção de animais visando a melhoria na produção de carne. A maioria dos músculos estriados originam-se de células precursoras do mesoderma a partir dos somitos do embrião e o controle da diferenciação ocorre pela ação de fatores indutores ou inibidores. Um grupo de fatores transcricionais, pertencentes à família MyoD tem um papel central na diferenciação muscular. Coletivamente chamados de Fatores de Regulação Miogênica (MRFs), são conhecidos quatro tipos: MyoD, myf-5, miogenina e MRF4. Esses fatores ligam-se à seqüências de DNA conhecidas como Ebox (CANNTG) na região promotora de vários genes músculo-específicos, levando à expressão dos mesmos. As células embrionárias com potencial para diferenciação em células musculares (células precursoras miogênicas) expressam MyoD e Myf-5 e são denominadas de mioblastos. Essas células proliferam, saem do ciclo celular, expressam miogenina e MRF4, que regulam a fusão e a diferenciação da fibra muscular. Uma população de mioblastos que se diferencia mais tardiamente, as células miossatélites, são responsáveis pelo crescimento muscular no período pós natal, que pode ocorrer por hiperplasia e hipertrofia das fibras. As células satélites quiescentes não expressam os MRFs, porém, sob a ação de estímulos como fatores de crescimento ou citocinas, ocorre a ativação desse tipo celular que prolifera e expressa os MRFs de maneira similar ao que ocorre com as células precursoras miogênicas durante a miogênese. Os mecanismos de crescimento muscular são regulados pela expressão temporal dos (MRFs), que controlam a expressão dos genes relacionados com o crescimento muscular.

Video target tracking by using competitive neural networks

A target tracking algorithm able to identify the position and to pursuit moving targets in video digital sequences is proposed in this paper. The proposed approach aims to track moving targets inside the vision field of a digital camera. The position and trajectory of the target are identified by using a neural network presenting competitive learning technique. The winning neuron is trained to approximate to the target and, then, pursuit it. A digital camera provides a sequence of images and the algorithm process those frames in real time tracking the moving target. The algorithm is performed both with black and white and multi-colored images to simulate real world situations. Results show the effectiveness of the proposed algorithm, since the neurons tracked the moving targets even if there is no pre-processing image analysis. Single and multiple moving targets are followed in real time.

O projeto para o pavilhão brasileiro na expo' 92 em sevilha e a chamada arquitetura paulista

This article discusses the controversy surrounding the winning project of the competition organized by the Brazilian Ministry of Foreign Affairs for the Brazilian pavilion at the Expo'92 in Seville, Spain; the views of architecture critics made at the time of the results, and, their implications for Brazilian architecture. At the beginning of the 90s, this competition was a kind of confrontation of ideas, between the architects who were in favour of a renewal of architecture and those who defended the resumption of national architectural traditions, buildings with large spans, constructed in reinforced concrete. These architects were the heirs of the so called Paulista architecture, which was characterised by the work undertaken from the 60's by important architects such as Vilanova Artigas and Paulo Mendes da Rocha. The modern references adopted by the winning project, from the architects Angelo Bucci, Alvaro Puntoni and José Oswaldo Vilela, sparked controversies because of the difficulty of resuming the teachings of the old modern masters when faced with new times. These controversies were related to the end of the military dictatorship in Brazil and the process of opening markets, and by the relevance of a re-evaluation of the so-called Paulista architecture.

Estruturação do Eumenine Mastroparano por dinâmica molecular em misturas de TFE-água

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Uso de uma rede unidimensional harmônica com o potencial de Rosen-Morse on site para modelar o DNA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modelagem molecular da interação entre a proteína de fusão do vírus sincicial respiratório humano e inibidores da ação viral. -

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Características reológicas do exopolissacarídeo botiosferana produzido pelo Botryosphaeria rhodina MAMB-05 em três fontes de carbono: glucose, frutose e sacarose

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Liberdade sem sustos, nem inquietações: significados e sentidos do fundo de emancipação no Grão-Pará (1871-1888)

A segunda metade do século XIX no Brasil foi marcada por reflexões e debates sobre o processo de encaminhamento da emancipação que, para a sociedade como um todo, transformou-se em um verdadeiro dilema a ser resolvido: o problema do elemento servil. Esses debates eram sustentados pelos diferentes setores sociais que encaminhariam um processo de libertação de forma controlada e dirigida por meio do controle do Estado e sobre determinados escravos. O projeto vencedor desse debate foi a Lei do Ventre Livre de 1871 que permitiu uma emancipação indenizatória e de controle sobre a população de libertos por meio do Fundo de Emancipação. Nesse sentido, o projeto expressava a necessidade de se manter as relações sociais da escravidão, cujo principal tema em jogo era a perda do controle sobre a “propriedade escrava”. Esse controle sobre a propriedade, no entanto, não significou que os sujeitos sociais diretamente atingidos pela política de emancipação do Estado não construíssem suas respostas para enfrentar os desafios na busca de suas liberdades. As ações diante à justiça, aos relacionamentos cotidianos costurados, às ações junto à produção das matrículas ou das listas de classificação de escravos que seriam libertos pelo Fundo de Emancipação se constituíram, entre outras formas de intervenção escrava pautadas na própria legislação emancipacionista que garantiria ao escravo o caminho da liberdade.

Estudos da lesão ao DNA por corantes têxteis e da capacidade protetora de flavonóides empregando biossensor eletroquímico: Carolina Venturini Uliana. -

Pós-graduação em Química - IQ

Mode of Peroxisome Proliferator-Activated Receptor gamma Activation by Luteolin

The peroxisome proliferator-activated receptor gamma (PPAR gamma) is a target for treatment of type II diabetes and other conditions. PPAR gamma full agonists, such as thiazolidinediones (TZDs), are effective insulin sensitizers and anti-inflammatory agents, but their use is limited by adverse side effects. Luteolin is a flavonoid with anti-inflammatory actions that binds PPAR gamma but, unlike TZDs, does not promote adipocyte differentiation. However, previous reports suggested variously that luteolin is a PPAR gamma agonist or an antagonist. We show that luteolin exhibits weak partial agonist/antagonist activity in transfections, inhibits several PPAR gamma target genes in 3T3-L1 cells (LPL, ORL1, and CEBP alpha) and PPAR gamma-dependent adipogenesis, but activates GLUT4 to a similar degree as rosiglitazone, implying gene-specific partial agonism. The crystal structure of the PPAR gamma ligand-binding domain (LBD) reveals that luteolin occupies a buried ligand-binding pocket (LBP) but binds an inactive PPAR gamma LBD conformer and occupies a space near the beta-sheet region far from the activation helix (H12), consistent with partial agonist/antagonist actions. A single myristic acid molecule simultaneously binds the LBP, suggesting that luteolin may cooperate with other ligands to bind PPAR gamma, and molecular dynamics simulations show that luteolin and myristic acid cooperate to stabilize the Omega-loop among H2', H3, and the beta-sheet region. It is noteworthy that luteolin strongly suppresses hypertonicity-induced release of the pro-inflammatory interleukin-8 from human corneal epithelial cells and reverses reductions in transepithelial electrical resistance. This effect is PPAR gamma-dependent. We propose that activities of luteolin are related to its singular binding mode, that anti-inflammatory activity does not require H12 stabilization, and that our structure can be useful in developing safe selective PPAR gamma modulators.

Molecular architecture of fur binding to iron-induced and - repressed genes in Helicobacter pylori

The ferric uptake regulator protein Fur regulates iron-dependent gene expression in bacteria. In the human pathogen Helicobacter pylori, Fur has been shown to regulate iron-induced and iron-repressed genes. Herein we investigate the molecular mechanisms that control this differential iron-responsive Fur regulation. Hydroxyl radical footprinting showed that Fur has different binding architectures, which characterize distinct operator typologies. On operators recognized with higher affinity by holo-Fur, the protein binds to a continuous AT-rich stretch of about 20 bp, displaying an extended protection pattern. This is indicative of protein wrapping around the DNA helix. DNA binding interference assays with the minor groove binding drug distamycin A, point out that the recognition of the holo-operators occurs through the minor groove of the DNA. By contrast, on the apo-operators, Fur binds primarily to thymine dimers within a newly identified TCATTn10TT consensus element, indicative of Fur binding to one side of the DNA, in the major groove of the double helix. Reconstitution of the TCATTn10TT motif within a holo-operator results in a feature binding swap from an holo-Fur- to an apo-Fur-recognized operator, affecting both affinity and binding architecture of Fur, and conferring apo-Fur repression features in vivo. Size exclusion chromatography indicated that Fur is a dimer in solution. However, in the presence of divalent metal ions the protein is able to multimerize. Accordingly, apo-Fur binds DNA as a dimer in gel shift assays, while in presence of iron, higher order complexes are formed. Stoichiometric Ferguson analysis indicates that these complexes correspond to one or two Fur tetramers, each bound to an operator element. Together these data suggest that the apo- and holo-Fur repression mechanisms apparently rely on two distinctive modes of operator-recognition, involving respectively the readout of a specific nucleotide consensus motif in the major groove for apo-operators, and the recognition of AT-rich stretches in the minor groove for holo-operators, whereas the iron-responsive binding affinity is controlled through metal-dependent shaping of the protein structure in order to match preferentially the major or the minor groove.

Da alleati vittoriosi ad acerrimi rivali: i processi di ri-allineamento dopo i conflitti egemonici. Teorie a confronto e studio di casi

Why some powers manage to coordinate their security efforts while others confront each other as rivals is still one of the most relevant and debated questions in the field of IR theory. The dissertation wants to give a contribution to this important debate. In particular, the main goal of the research is to analyse the dynamics of great power interactions after the end of hegemonic conflicts, that is to understand why, following the defeat of the common enemies, some of the winning allies continue to cooperate, while others begin to engage in political and military competition. In order to understand this difference, the study compares the explanatory value of two rival theoretical perspectives: neorealism, in its main version of the balance of power framework, and a liberal approach focused on domestic politics. The thesis is divided in two sections. In the first, I do summarize the main assumptions and predictions of the theories, from which I derive two different sets of hypotheses on the evolution of post-war great power relations. In the second part, I test the hypotheses by focusing on two cases of post-war alignment dynamics: 1) the relations among Austria, Prussia, Russia, Great Britain and France after the Napoleonic wars; 2) the relations among the US, the UK, France and Italy after the end of WWI. The historical cases disconfirm the logic of the balance of power and confirm the liberal hypotheses, seeing that the results of the analysis show changes in the domestic structures of the great powers had a much larger impact on the emergence of new alliances and rivalries than did the international distribution of power. In the conclusion of the dissertation, I provide the reader with a discussion of the main theoretical implications of the empirical findings.

Tetracycline-inducible RNAi Knockdown of SCL (Stem Cell Leukaemia) in Mice

RNAi (RNA interference) is a powerful technology for sequence-specific targeting of mRNAs. This thesis was aimed at establishing conditions for conditional RNAi-mediated silencing first in vitro and subsequently also in transgenic mice. As a target the basic helix-loop-helix transcription factor encoding gene SCL (stem cell leukaemia also known as Tal-1 or TCL5) was used. SCL is a key regulator for haematopoietic development and ectopic expression of SCL is correlated with acute T-lymphoblastic leukaemias. Loss of SCL function studies demonstrated that ab initio deletion of SCL resulted in embryonic lethality around day E9 in gestation. To be able to conditionally inactivate SCL, RNAi technology was combined with the tetracycline-dependent regulatory system. This strategy allowed to exogenously control the induction of RNAi in a reversible fashion and consequently the generation of a completely switchable RNAi knockdown. First a suitable vector allowing for co-expression of tetracycline-controlled shRNAs (small hairpin RNAs) and constitutively active EGFP (enhanced green fluorescent protein) was generated. This novel vector, pRNAi-EGFP, was then evaluated for EGFP expression and tetracycline-mediated expression of shRNAs. Four sequences targeting different regions within the SCL mRNA were tested for their efficiency to specifically knockdown SCL. These experiments were performed in M1 murine leukaemia cells and subsequently in the HEK 293 cell line, expressing an engineered HA-tagged SCL protein. The second assay provided a solid experimental method for determining the efficiency of different SCL-siRNA knockdown constructs in tissue culture. Western blotting analyses revealed a down regulation of SCL protein for all four tested SCL-specific target sequences albeit with different knockdown efficiencies (between 25% and 100%). Furthermore, stringent tetracycline-dependent switchability of shRNA expression was confirmed by co-transfecting the SCL-specific pRNAi-EGFP vector (SCL-siRNA) together with the HA-tagged SCL expression plasmid into the HEK 293TR /T-REx cell line constitutively expressing the tetracycline repressor (TetR). These series of experiments demonstrated tight regulation of siRNA expression without background activity. To be able to control the SCL knockdown in vivo and especially to circumvent any possible embryonic lethality a transgenic mouse line with general expression of a tetracycline repressor was needed. Two alternative methods were used to generate TetR mice. The first approach was to co-inject the tetracycline-regulated RNAi vector together with a commercially available and here specifically modified T-REx expression vector (SCL-siRNA T-REx FRT LoxP mouse line). The second method involved the generation of a TetR expressor mouse line, which was then used for donating TetR-positive oocytes for pronuclear injection of the RNAi vector (SCL-siRNA T-REx mouse line). As expected, and in agreement with data from conditional Cre-controlled adult SCL knockout mice, post-transcriptional silencing of SCL by RNAi caused a shift in the maturation of red blood cell populations. This was shown in the bone marrow and peripheral blood by FACS analysis with the red blood cell-specific TER119 and CD71 markers which can be used to define erythrocyte differentiation (Lodish plot technique). In conclusion this study established conditions for effective SCL RNAi-mediated silencing in vitro and in vivo providing an important tool for further investigations into the role of SCL and, more generally, of its in vivo function in haematopoiesis and leukaemia. Most importantly, the here acquired knowledge will now allow the establishment of other completely conditional and reversible knockdown phenotypes in mice.