Avaliação de dois clones de Escherichia coli recombinante quanto ao crescimento e expressão de antígenos de Leishmania chagasi (kmp11 e P36)

Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37°C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies

Morphomolecular characterization of Pleurotus ostreatus (Jacq. Fr.) kummer strains in relation to luminosity and temperature of frutification

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

Mineral supplementation and productivity of the Shiitake mushroom on eucalyptus logs

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

Estudo do mecanismo de produção de oligossacarídeos com atividades nutracêuticas a partir da quitosana por hidrólise enzimática com processo fermentativo simultâneo

The obtaining of the oligosaccharides from chitosanase, has showed interest of the pharmaceutical area in the last years due their countless functional properties. Although, the great challenge founded out is how to keep a constant and efficient production. The alternative proposed by this present work was to study the viability to develop an integrated technology, with reduced costs. The strategy used was the obtaining of the oligomers through enzymatic hydrolysis using chitosanolitic enzymes obtained straight from the fermented broth, eliminating this way the phases involved in the enzymes purification. The two chitosanases producing strains chosen for the work, Paenibacillus chitinolyticus and Paenibacillus ehimensis, were evaluated according to the behavior in the culture medium with simple sugar and in relation to the pH medium variations. The culture medium for the chitosanases induction and production was developed through addition of soluble chitosan as carbon source. The soluble chitosan was obtained using hydrochloric acid solution 0.1 M and afterwards neutralization with NaOH 10 M. The enzymatic complexes were obtained from induction process in culture medium with 0.2% of soluble chitosan. The enzymes production was verified soon after the consumption of the simple sugars by the microorganisms and the maximum chitosanolitic activity obtained in the fermented broth by Paenibacillus chitinolyticus was 249 U.L-1 and by Paenibacillus ehimensis was 495U.L-1. These two enzymatic complexes showed stability when stored at 20°C for about 91 days. The enzymes in the fermented broth by Paenibacillus chitinolyticus, when exposed at temperature of 55°C and pH 6.0, where the activity is maximum, showed 50% lost of activity after 3 hours Meanwhile, for the complex produced by Paenibacillus ehimensis, after 6 days of exposure, it was detected 100% of the activity. The chito-oligosaccharides obtained by the hydrolysis of a 1% chitosan solution, using the enzymatic complex produced by Paenibacillus chitinolyticus showed larger quantity after 9 hours hydrolysis and using the complex produced by Paenibacillus ehimensis after 20 minutes was observed the chito-ligosacharides with polymerization degree between 3 and 6 units. Evaluating these results, it was verified that the production of chitosan-oligosaccharides is possible, using a simultaneous process

Normas jurídicas para a restauração ecológica: uma barreira a mais a dificultar o êxito das iniciativas?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influência das condições de cultivo na produção de antígenos recombinantes de Leishmania i. chagasi utilizando Escherichia coli M15 cultivada em incubador rotativo e biorreator

Escherichia coli has been one of the most widely used hosts in recombinant protein production, in both laboratory and industrial scale since the advent of recombinant DNA technology. Despite the substantial progress of studies on the molecular biology and immunology of infections, there is currently no medication-based prophylaxis capable of preventing leishmaniasis. As such, there is a great need to identify specific antigens for the development of vaccines and diagnostic kits against visceral leishmaniasis. Thus, the primary goal of the present study is to assess the influence of cultivation conditions on the production of Leishmania chagasi antigens, carried out in a rotating incubator and bioreactor. To that end, several assays were conducted to evaluate the kinetic behavior of antigens (648, 503) of Leishmania. i. chagasi in two different compositions of media (2xTY, TB), with and without an inducer. In order to improve expression, assays were performed in a benchtop bioreactor using the best conditions obtained in a rotating incubator, in addition to assessing the influence of stirring speed. Results show that high complexity of the cultivation medium favored kinetic growth of clones (648, 503). However, in assays submitted to induction by IPTG, this elevated complexity did not promote the expression of recombinant proteins. Expression of antigens 648 and 503 exhibited behavior associated with growth and, in terms of location, proteins 648 and 503 are intracellularly stored. Lactose may be the most adequate inducer in protein expression, when considering factors, cost, toxicity and stability. Elevated stirring may increase cell growth in clone 53, although it may not result in high concentrations for the protein of interest. On the other hand, positive results were obtained for all recombinant clones (648, 503) tested, confirmed by the electrophoretic profile

Obtenção de novos derivados de ß-naftol e cardanol hidrogenado e avaliação dos seus efeitos antioxidativos em gasolina automotiva

The chemical nature of the hidrocarbons found in gasoline directly affects the formation of oxidation complexes known as gums. Such complexes are slightly soluble in gasoline and their formation is delayed with the use of inhibitors, known as antioxidants. In this study, a synthetic compound (βnaphtol) and a natural subproduct (hydrogenated cardanol, derived from cashewnut shell liquid CNSL) have been used in order to generate novel antioxidant substances. These compounds were submitted to chemical reactions including alkylation, nitration and reduction, with the purpose of forming the following derivatives: 6(Nethyl,Nethylamino)βnaphtol (AO1); 6(Nethyl,Ndiethylamino)βnaphtol (AO2); aminoβnaphtol (AO3); 2(Nethyl,Nethylamino)pentadecylphenol (AOC1), 2(Nethyl,Ndiethylamino)pentadecylphenol (AOC2) and aminopentadecylphenol (AOC3). The derivatives were subjected to accelerated oxidative stability assays (Potential Gum and Induction Period) and to storage assays (Washed Gum and ASTM Color) during six months, with naphtha provided by the petroleum refinery RPBC (Refinaria Presidente Bernardes de Cubatão, in Brazil). The results for the derivatives were compared to those for commercial additives [DBPC (2,6ditbutyl4methylpcresol) and PDA (N,N disecbutylpphenylenediamine)], which were also added to the naphtha produced at RPBC at the moment of sampling. From all tested antioxidants, the novel antioxidant AOC1 (derived from hydrogenated cardanol) yielded a better global performance. During the period of time in which the naphtha was stored, an examination of this material was carried out in parallel, using the mass spectrometry technique. This study allowed to monitor the formation of a triolefinic compound, as well as the observation of subsequent formation and rupture of the olefinic constituents. As an eventual result from these experimental investigations, a reaction route leading to gum formation has been suggested

Paradigmas, valores e educação

O texto levanta os perfis epistemológico e socianalítico da questão paradigmática. Mauss evidenciara o moule affectif das noções científicas de força e causa. Posteriormente Baudouin falaria na indução arquetípica das noções e a antropologia do imaginário de Durand concluiria pela indução arquetipal do conceito pela imagem. Chegava-se, assim, ao desvendamento do substrato inconsciente das ideações, de um substrato regido pela catexis vetorializada, traduzindo-se nos valores como cerne das ideações. É o famoso a priori emotivo. Portanto, no texto, questionam-se dois mitos, esteios da ciência clássica: o mito da objetividade científica e o da neutralidade axiológica. Destaca, assim, a falácia da existência de uma ruptura epistemológica entre ciência e ideologia. A partir daí, as ideações tornam-se ideologias, sobretudo nas ciências do homem e nas ciências da educação que, ademais, tornam-se suporte de uma disfarçada luta ideológica, na qual, num colonialismo cognitivo, as estratégias de conhecimento dissimulam as de preconceito. Entretanto, assumir a realidade desse suporte fantasmanalítico e ideológico propicia uma tarefa educativa salutar: os paradigmas tornam-se fantasias e, nessa relativização crítica, podem ser usados como um campo de objetos transicionais coletivos num ludismo cultural e educativo. No policulturalismo da sociedade contemporânea, o politeísmo de valores de Weber transforma-se num politeísmo epistemológico, regido pelo relativismo ontológico de Feyerabend e por uma ética do pragmatismo. Articulando cultura, organização e educação, a antropologia das organizações educativas e a culturanálise de grupos de Paula Carvalho traduzem as heurísticas dessa dialética transicional.

Economia verde: a reiteração de ideias à espera de ações

A Iniciativa Economia Verde, numa perspectiva de análise teórica, é uma reiteração de velhas ideias; não é propriamente um novo conceito, mas sim a proposta de um conjunto de instrumentos para o alcance do desenvolvimento sustentável. Uma novidade importante dessa inciativa é a defesa do ativismo de políticas de indução às mudanças tecnológicas ambientais, o que revela sua aproximação com a economia evolucionária. No plano da economia política internacional, o potencial de conflito Norte-Sul sobre a Iniciativa Economia Verde se vincula aos impasses registrados nas negociações sobre liberalização do comércio de bens e serviços ambientais na Rodada Doha da OMC.

Resistência e performance na arte de João Cota

Researching about the art of tell and the tales told by João Cota is somehow revisiting the oral tradition and social practice in the story-telling art. It takes into consideration the resistance this art still exerts, mainly by using the performance of oral transmission and receptiveness to tales. The study of this practice contributes to the vivacious and dynamcis permanence of this authentic and traditional storyteller of his repertory and of his form to tellin our culture. Story-telling is part of the humankind living heritage communicated by means of popular wisdom. Despite the risk of vanishing into thin air, along with their narrators, the tales still manage to resist the contemporary mass culture model. How long further will stories like the ones narrated by João Cota be able to resist to strong and stronger structures dictated by writing and other communication means? João Cota s practice in story-telling will be studied not only as a proposal to identify the presence of this practice and the oral cultural resistance but also, through the performance prospective, to identify the oral transmission and receptiveness to the tales that are part of this storyteller s repertoire. In other words: what he tells, how he tells it, and why he tells it. The advent of new technologies such as internet, through which people can easily communicate with others in different parts of the world, and the greater and greater expansion in the writing skill concept interfere the maintenance of the oral tradition elements present in João Cota s narratives and inserted in the Brazilian culture. This has become more visible in the latest decades although we still notice the living tradition and permanence of the story-telling practice in several parts of the country through their wise storytellers. Our research target will require - in each of its study stages reference to works by several theoreticians namely Paul Zumthor, Mikhail Bakhtin, Câmara Cascudo, theoreticians from the receptiveness aesthetic, from the written culture history, from oral cultures and reading practices, from tradition and the Brazilian Culture of oral story-telling. In order to get to know and draw a profile of this storyteller, we ve chosen to use the comprehensive interview method by French Sociologist Jean Claude Kaufmann. The originality in the method consists of the qualitative data put together in situ , concentrated on the storyteller s narratives/speeches recorded on tape, which will be the focal point of this study. Our analysis method is based on tireless sessions of listening to interviews out of which we gathered information related to the storyteller, his practice in telling the tales, and his repertoire

Ação do extrato metanólico e etanólico de Davilla elliptica St. Hill. (Malpighiaceae) na resposta imune

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

Os argumentos transcendentais: kant e o problema de hume

This work whose title is "The transcendental arguments: Kant Andy Hume's problem" has as its main objective to interpret Kant's answer to Hume's problem in the light of the conjunction of the causality and induction themes which is equivalent to skeptical- naturalist reading of the latter. In this sense, this initiative complements the previous treatment seen in our dissertation, where the same issue had been discussed from a merely skeptical reading that Kant got from Hume thought and was only examined causality. Among the specific objectives, we list the following: a) critical philosophy fulfills three basic functions, a founding, one negative and one would argue that the practical use of reason, here named as defensive b) the Kantian solution of Hume's problem in the first critisism would fulfill its founding and negative functions of critique of reason; c) the Kantian treatment of the theme of induction in other criticisms would will fulfill the defense function of critique of reason; d) that the evidence of Kant's answer to Hume's problem are more consistent when will be satisfied these three functions or moments of criticism. The basic structure of the work consists of three parts: the first the genesis of Hume's problem - our intention is to reconstruct Hume's problem, analyzing it from the perspective of two definitions of cause, where the dilution of the first definition in the second match the reduction of psychological knowledge to the probability of following the called naturalization of causal relations; whereas in the second - Legality and Causality - it is stated that when considering Hume in the skeptic-naturalist option, Kant is not entitled to respond by transcendental argument A􀁴B; A⊢B from the second Analogy, evidence that is rooted in the position of contemporary thinkers, such as Strawson and Allison; in third part - Purpose and Induction - admits that Kant responds to Hume on the level of regulative reason use, although the development of this test exceeds the limits of the founding function of criticism. And this is articulated in both the Introduction and Concluding Remarks by meeting the defensive [and negative] function of criticism. In this context, based on the use of so-called transcendental arguments that project throughout the critical trilogy, we provide solution to a recurring issue that recurs at several points in our submission and concerning to the "existence and / or the necessity of empirical causal laws. In this light, our thesis is that transcendental arguments are only an apodictic solution to the Hume s skeptical-naturalist problem when is at stake a practical project in which the interest of reason is ensured, as will, in short, proved in our final considerations

Analysis of the molecular association of rifampicin with hydroxypropyl-²-cyclodextrin

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

Propriedades magnéticas de nanopartículas de ferro em substratos antiferromagnéticos

The effect of finite size on the magnetic properties of ferromagnetic particles systems is a recurrent subject. One of the aspects wide investigated is the superparamagnetic limit where the temperature destroys the magnetic order of ferromagnetic small particles. Above the block temperature the thermal value of the magnetic moment of the particle vanishes, due to thermal fluctuations. The value of the blocking temperature diminishes when the size of the particle is reduced, reflecting the reduction of the anisotropy energy barrier between the uniform states along the uniaxial axis. The increasing demand for high density magnetic media has recently attracted great research interest in periodic arrangements of nanometric ferromagnetics particles, approach in the superparamagnetic limit. An interesting conjecture is the possibility of stabilization of the magnetic order of small ferromagnetic particles (F) by interface coupling with antiferromagnetic (AF) substrate. These F/AF systems may also help to elucidate some details of the effect of exchange bias, because the effect of interface roughness and the paper of domain walls, either in the substrate or the particle, are significantly reduced. We investigate the magnetic phases of small ferromagnetic particles on a antiferromagnetic substrate. We use a self-consistent local field method, incorporating the interface field and the dipole interaction between the spins of the ferromagnetic particle. Our results indicate that increasing the area of the interface favors the formation of the uniform state. Howere above a critical height value appears a state non-uniform is formed where the spins of in the particle s free surface are rotated with respect to the interface spins direction. We discuss the impact of the competition between the dipolar and interface field on the magnetic charge, that controls the field of flux leakage of the particle, and on the format of the hysteresis curves. Our results indicate that the liquid magnetic charge is not a monotonically increasing function of the height of the particle. The exchange bias may display anomalous features, induced for the dipolar field of the spins near the F/AF interface

Formação estelar desencadeada pela colisão entre nuvens moleculares magnetizadas

We use a finite diference eulerian numerical code, called ZEUS 3D, to do simulations involving the collision between two magnetized molecular clouds, aiming to evaluate the rate of star formation triggered by the collision and to analyse how that rate varies depending on the relative orientations between the cloud magnetic fields before the shock. The ZEUS 3D code is not an easy code to handle. We had to create two subroutines, one to study the cloud-cloud collision and the other for the data output. ZEUS is a modular code. Its hierarchical way of working is explained as well as the way our subroutines work. We adopt two sets of different initial values for density, temperature and magnetic field of the clouds and of the external medium in order to study the collision between two molecular clouds. For each set, we analyse in detail six cases with different directions and orientations of the cloud magnetic field relative to direction of motion of the clouds. The analysis of these twelve cases allowed us to conform analytical-theoretical proposals found in the literature, and to obtain several original results. Previous works indicate that, if the cloud magnetic fields before the collision are orthogonal to the direction of motion, then a strong inhibition of star formation will occur during a cloud-cloud shock, whereas if those magnetic fields are parallel to the direction of motion, star formation will be stimulated. Our treatment of the problem confirmed numerically those results, and further allowed us to quantify the relative star forming efficiencies in each case. Moreover, we propose and analyse an intermediate case where the field of one of the clouds is orthogonal to the motion and the field of the other one is parallel to the motion. We conclude that, in this case, the rate at which the star formation occurs has a value also intermediate between the two extreme cases we mentioned above. Besides that we study the case in which the fields are orthogonal to the direction of the motion but, instead of being parallel to each other, they are anti-parallel, and we obtained for this case the corresponding variation of the star formation rate due to this alteration of the field configuration. This last case has not been studied in the literature before. Our study allows us to obtain, from the simulations, the rate of star formation in each case, as well as the temporal dependence of that rate as each collision evolves, what we do in detail for one of the cases in particular. The values we obtain for the rate of star formation are in accordance with those expected from the presently existing observational data