Purification, characterization, crystallization and theoretical molecular modeling of gyroxin fraction from Crotalus durissus terrificus venom (Laurenti, 1768)

The venom of Crotalus durissus terrificus snakes presents various substances, including a serine protease with thrombin-like activity, called gyroxin, that clots plasmatic fibrinogen and promote the fibrin formation. The aim of this study was to purify and structurally characterize the gyroxin enzyme from Crotalus durissus terrificus venom. For isolation and purification, the following methods were employed: gel filtration on Sephadex G75 column and affinity chromatography on benzamidine Sepharose 6B; 12% SDS-PAGE under reducing conditions; N-terminal sequence analysis; cDNA cloning and expression through RT-PCR and crystallization tests. Theoretical molecular modeling was performed using bioinformatics tools based on comparative analysis of other serine proteases deposited in the NCBI (National Center for Biotechnology Information) database. Protein N-terminal sequencing produced a single chain with a molecular mass of similar to 30 kDa while its full-length cDNA had 714 bp which encoded a mature protein containing 238 amino acids. Crystals were obtained from the solutions 2 and 5 of the Crystal Screen Kit (R), two and one respectively, that reveal the protein constitution of the sample. For multiple sequence alignments of gyroxin-like B2.1 with six other serine proteases obtained from snake venoms (SVSPs), the preservation of cysteine residues and their main structural elements (alpha-helices, beta-barrel and loops) was indicated. The localization of the catalytic triad in His57, Asp102 and Ser198 as well as S1 and S2 specific activity sites in Thr193 and Gli215 amino acids was pointed. The area of recognition and cleavage of fibrinogen in SVSPs for modeling gyroxin B2.1 sequence was located at Arg60, Arg72, Gln75, Arg81, Arg82, Lis85, Glu86 and Lis87 residues. Theoretical modeling of gyroxin fraction generated a classical structure consisting of two alpha-helices, two beta-barrel structures, five disulfide bridges and loops in positions 37, 60, 70, 99, 148, 174 and 218. These results provided information about the functional structure of gyroxin allowing its application in the design of new drugs.

Ovine leptospirosis in Brazil

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

Luz e sombra: uma interpretação de suas significações imaginárias nas imagens do cinema expressionista alemão e do cinema noir americano

The present investigation proposes an approach of light and shadow and their imaginary significations in the images of both German expressionist movie and the American noir movie, whose aesthetic experience is expressed through a contrasting bright/dark photography, loaded with symbolism. The interpretation of the imaginary significations of this imagistic material is based on Gilbert Durand´s imaginary anthropological theories that deal with a myriad of symbols gathered according to their semantic isomorphism and linked to more general structures named as Daily and nightly Image Regime . There come to gravitate, around such regimes, symbols attached to division and purification, ascensional and spectacular, with imaginary significations homological to Light and Good, and symbols associated to night, fall and animalism, isomorphic of Shadow and Evil

Eram realmente pitagórico(a)s os homens e mulheres catalogado(a)s por Jâmblico em sua obra Vida de Pitágoras?

Pythagoras was one of the most important pre-Socratic thinkers, and the movement he founded, Pythagoreanism, influenced a whole thought later in religion and science. Iamblichus, an important Neoplatonic and Neopythagorean philosopher of the third century AD, produced one of the most important biographies of Pythagoras in his work Life of Pythagoras. In it he portrays the life of Pythagoras and provides information on Pythagoreanism, such as the Pythagorean religious community which resembled the cult of mysteries; the Pythagorean involvement in political affairs and in the government in southern Italy, the use of music by the Pythagoreans (means of purification of healing, use of theoretical study), the Pythagorean ethic (Pythagorean friendship and loyalty, temperance, self-control, inner balance); justice; and the attack on the Pythagoreans. Also in this biography, Iamblichus, almost seven hundred years after the termination of the Pythagorean School, established a catalog list with the names of two hundred and eighteen men and sixteen women, supposedly Pythagoreans of different nationalities. Based on this biography, a question was raised: to what extent and in what ways, can the Pythagoreans quoted by Iamblichus really be classified as Pythagoreans? We will take as guiding elements to search for answers to our central problem the following general objectives: to identify, whenever possible, which of the men and women listed in the Iamblichus catalog may be deemed Pythagorean and specific; (a) to describe the mystery religions; (b) to reflect on the similarities between the cult of mysteries and the Pythagorean School; (c) to develop criteria to define what is being a Pythagorean; (d) to define a Pythagorean; (e) to identify, if possible, through names, places of birth, life, thoughts, work, lifestyle, generation, etc.., each of the men and women listed by Iamblichus; (f) to highlight who, in the catalog, could really be considered Pythagorean, or adjusting to one or more criteria established in c, or also to the provisions of item d. To realize these goals, we conducted a literature review based on ancient sources that discuss the Pythagoreanism, especially Iamblichus (1986), Plato (2000), Aristotle (2009), as well as modern scholars of the Pythagorean movement, Cameron (1938), Burnet (1955), Burkert (1972), Barnes (1997), Gorman (n.d.), Guthrie (1988), Khan (1999), Mattéi (2000), Kirk, Raven and Shofield (2005), Fossa and Gorman (n.d.) (2010). The results of our survey show that, despite little or no availability of information on the names of alleged Pythagoreans listed by Iamblichus, if we apply the criteria and the definition set by us of what comes to be a Pythagorean to some names for which we have evidence, it is possible to assume that Iamblichus produced a list which included some Pythagoreans

Use of repellents for honeybees (Apis mellifera L.) in vitro in the yellow passion-fruit (Passiflora edulis Deg) crop and in confined beef cattle feeders

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

Caracterização de um isolado de Bidens mosaic virus proveniente de alface

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

Concepção e estudo de um biofiltro para tratamento de compostos orgânicos voláteis COVs

In the last decade, biological purification of gaseous waste has become an important alternative to many conventional methods of exhaust air treatment. More recently, biofiltration has proved to be an effective and inexpensive method for the treatment of air contaminated with volatile organic compounds (VOCs). A biofilter consists in a reactor packed with a porous solid bed material, where the microorganisms are fixed. During the biofiltration process, polluted air is transported through the biofilter medium where the contaminant is degraded. Within the biofilm, the pollutants in the waste gases are energy and carbon sources for microbial metabolism and are transformed into CO2, water and biomass. The bed material should be characterized by satisfactory mechanical and physical properties as structure, void fraction, specific area and flow resistance. The aim of this research was the biofilter construction and study of the biological degradation of ethanol and toluene, as well as the modeling of the process. Luffa cylindrica is a brazilian fiber that was used as the filtering material of the present work. The parameters and conditions studied were: composition of nutrients solution; effect of microflorae strains, namely Pseudomanas putida and Rhodococcus rhodochrous; waste gas composition; air flow rate; and inlet load of VOCs. The biofilter operated in diffusion regime and the best results for remotion capacity were obtained when a microorganisms consortion of Pseudomanas putida and Rhodococcus rhodochrous,were used, with a gas flow rate of 1 m3.h-1 and molar ratio nitrogene/phosphore N/P=2 in the nutrients solution. The maximum remotion capacity for ethanol was around 90 g.m-3.h-1 and 50 g.m-3.h-1 to toluene. It was proved that toluene has inhibitory effect on the ethanol remotion When the two VOCs were present in the same waste gas, there was a decrease of 40% in ethanol remotion capacity. Luffa cylindrica does not present considerable pressure drop. Ottengraf and van Lith models were used to represent the results obtained for ethanol and toluene, respectively. The application of the transient model indicated a satisfactory approximation between the experimental results obtained for ethanol and toluene vapors biofiltration and the ones predicted it

Variação da qualidade da água de chuva no início da precipitação

One of the most evident and actual concern, not only in the scientific means, but also of the public knowledge in general, is the lack problem of the potable water, that come increasing more each time, motivated mainly for the po llution of the sources, the bad use of the water and the increase of the population. In such a way, the necessity of looking for new water sources and the development of techniques to use sources minus explored is becoming even more important and urgent. T he rainwater comes being used since a long time like a supplying source, but, due the few knowledge of its characteristics, the generated preconception and the discrimination around its consumption and to the bad use of the technique of collection and stor age, it comes being little used, wasting consequently a significant parcel of this source. Trying to develop the knowing of some characteristics of the rainwater, the present work looks for to define the curve of variation of the quality of the rainwater i n three points with distinct characteristics of the city of Natal -RN, in the course of the precipitation, in some situations of time and space. For describe the curve of variation of the water s quality, some variables must be analyzed, and to be identifie d when they modify themselves in the endurance of rain, showing in which moment the purification of the water is more or less quickly. The pH, the Turbidity and the Electric Conductivity can be related with a big part of the physicist -chemistries variables found in the water and, like its analyses don't spend any material, they have easy access and measurement. The present work analyzes the curves of decline of these three variables, in three points with distinct characteristics in the city of Natal -RN, being these points: one next to the sea, another one in region with great buildings concentration and the last point in a less polluted area. For the studied region, it was during the five first millimeters of rain that occurs the biggest reduction of the exi sting impurities in the atmosphere, mainly between the first and the second millimeter, and after the five first millimeters the values of the variables stabilize. With exception of the University Campus, where initially the rainwater already has very good quality, the values of Turbidity and Electric Conductivity suffer a brusque reduction after the first millimeter of rain

Métodos e estratégias em proteômica e suas aplicações na área vegetal

A implementação da espectrometria de massa (MS) para as análises de peptídeos (MS) e de aminoácidos (MS em tandem ou MS/MS) tornou possível a identificação de centenas de proteínas em experimentos únicos. Uma grande variedade de estratégias está disponível atualmente para o fracionamento e a purificação de amostras, a identificação de proteínas, a quantificação, a análise de modificações pós-traducionais (MPT's) e os estudos de interação. Dessa forma, a proteômica abre novas perspectivas na biologia de plantas com ênfase nos estudos de variabilidade genética, estresses fisiológicos e desenvolvimento de plantas.