Inhibition of Neurotoxic Secretory Phospholipases A(2) Enzymatic, Edematogenic, and Myotoxic Activities by Harpalycin 2, an Isoflavone Isolated from Harpalyce brasiliana Benth

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Harpalycin 2 inhibits the enzymatic and platelet aggregation activities of PrTX-III, a D49 phospholipase A(2) from Bothrops pirajai venom

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Extração, caracterização e atividades biológicas de proteínas da espécie cnidoscolus urens (L.) Arthur

The extraction, chemical and structural characterization of a wide variety of compounds derived from plants has been a major source of bioactive molecules. Several proteases have been isolated in the plant kingdom, with numerous pharmacological and biotechnological applications. Among the proteases isolated from plants, are the fibrinogenolytic, with relevant application in the treatment of disorders in the coagulation cascade, in addition to potential use as a tool in clinical laboratories. In this study, in addition to evaluating the effects of the protein extract of Cnidoscolus urens (L.) Arthur (Euphorbiaceae) in the coagulation cascade also investigates the presence of antimicrobial activity and characterizes the proteolytic activity detected in this extract, aiming to determine their potential pharmacological and biotechnological application. In this way, crude protein extracts obtained from the leaves of C. urens in Tris-HCl 0.05M, NaCl 0.15M, pH 7.5, were precipitated in different concentrations of acetone, and assessed for the presence of proteolytic activity in azocaseína and fibrinogen. The most active fraction (F1.0) in these tests was chosen for assessment of biological activity and biochemical characterization. The Aα chain and Bβ of fibrinogen were completely cleaved at a concentration of 0.18 μg/μL of protein fraction in 4 minutes. Fibrinogenolytic activity presented total inhibition in the presence of E-64 and partial in the presence of EDTA. The fraction demonstrated coagulant activity in plasm and reduced the APTT, demonstrating acting on the factors coagulation of the intrinsic pathway and common, not exerting effects on the PT. Fibrinolytic activity on plasma clot was detected only in SDS-PAGE in high concentrations of fraction, and there were no defibrinating. Although several proteases isolated from plants and venomous animals are classically toxic, the fraction F1.0 of C. urens not expressed hemorrhagic nor hemolytic activities. Fraction F1.0 also showed no antimicrobial activity. In proteolytic activity on the azocasein, the optimal pH was 5.0 and optimum temperature of 60ºC. The enzyme activity has been shown to be sensitive to the presence of salts tested, with inhibition for all compounds. The surfactant triton did not influence the enzyme activity, but the tween-20 and SDS inhibited the activity. In the presence of reducing agents increase in enzyme activity occurred, a typical feature of enzymes belonging to the class of cysteine proteases. Several bands with proteolytic activity were detected in zymogram, in the region of high-molecular-weight, which were inhibited by E-64. In this study, we found that C. urens presents in its constitution cysteine proteases with fibrinogenolytic and procoagulant activity, which may be isolated, with potential application in treatment of bleeding disorders, thrombolytic and clinical laboratory

Digestive and functional properties of a partially hydrolyzed cassava solid waste with high insoluble fiber concentration

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Arraste via fecal de nutrientes da ingestão produzido por bagaço de mandioca hidrolisado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sewage sludge as nitrogen and phosphorus source for cane-plant and first ratoon crops

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

The effects of ovalbumin as a protein source during the in vitro production of bovine embryos

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

Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

Growth and antimicrobial activity of lactic acid bacteria from rumen fluid according to energy or nitrogen source

Objetivou-se avaliar os efeitos da suplementação com diferentes fontes de energia e de compostos nitrogenados sobre o crescimento e produção de bacteriocinas de bactérias ácido-láticas in vitro. As incubações foram conduzidas utilizando-se fluido ruminal originado de um novilho Holandês-Zebu fistulado no rúmen. O animal foi mantido em pastagem de Brachiaria decumbens recebendo 200 g/dia de proteína bruta suplementar. Os substratos e o inóculo foram acondicionados em frascos de vidro considerando-se oito tratamentos: celulose, celulose e caseína, celulose e peptona de soja, celulose e ureia, amido, amido e caseína, amido e peptona de soja e amido e ureia. Incubações sucessivas foram conduzidas para seleção dos microrganismos em função das fontes energéticas e de compostos nitrogenados. O amido favoreceu o crescimento de bactérias ácido-láticas em comparação à celulose. A suplementação com proteína verdadeira (peptona de soja e caseína) estimulou o crescimento dessas bactérias em comparação ao controle (sem suplementação com compostos nitrogenados). A adição de ureia não estimulou o crescimento de bactérias ácido-láticas. Nenhuma atividade antimicrobiana foi detectada a partir das colônias de bactérias ácido-láticas isoladas. Fontes de proteína verdadeira incrementam a competição entre bactérias fermentadoras de carboidratos estruturais e não-estruturais.

Dried yeast (Saccharomyces cerevisae) as a protein source for horses

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Environmental adaptation of the source of the subbasin of Rico Stream, Monte Alto - SP, Brazil

Este trabalho teve como principal objetivo definir padrões fotográficos que representem o uso e a ocupação do solo da cabeceira de drenagem do Córrego Rico, localizada no Município de Monte Alto, Estado de São Paulo, para fins de adequação ambiental no que tange à legislação florestal brasileira. O mapeamento foi realizado utilizando técnicas de sensoriamento remoto e interpretação visual da imagem World View, seguida da digitalização da rede de drenagem e vegetações (naturais e agrícolas) no AutoCad, com auxílio de documentos e trabalho de campo. A área de estudo apresenta uma superfície de 2.141,53 ha, e os resultados permitiram constatar que a principal cultura é a cana-de-açúcar, com 546,34 ha, seguida de pastagens, com 251,22 ha, culturas perenes, com 191,71 ha, Eucalipto, com 57,31 ha, e a cebola, com 49,52 ha da área total, confirmando o avanço dos canaviais na região. A área possui 375,04 ha de áreas de preservação permanente (APPs), e desta superfície verificou-se que apenas 72,17 ha (19,24%) encontram-se compostos por vegetação arbórea ou mata, e destas 302,87 ha precisam ser enriquecidos e reflorestados com vegetação nativa da região, de acordo com legislação vigente. Os dados de área possibilitam futuras propostas de modelos de adequação ambiental para a microbacia, de acordo com a legislação ambiental vigente.

DNA bar-coding reveals source and patterns of Thaumastocoris peregrinus invasions in South Africa and South America

Thaumastocoris peregrinus is a recently introduced invertebrate pest of non-native Eucalyptus plantations in the Southern Hemisphere. It was first reported from South Africa in 2003 and in Argentina in 2005. Since then, populations have grown explosively and it has attained an almost ubiquitous distribution over several regions in South Africa on 26 Eucalyptus species. Here we address three key questions regarding this invasion, namely whether only one species has been introduced, whether there were single or multiple introductions into South Africa and South America and what the source of the introduction might have been. To answer these questions, bar-coding using mitochondrial DNA (COI) sequence diversity was used to characterise the populations of this insect from Australia, Argentina, Brazil, South Africa and Uruguay. Analyses revealed three cryptic species in Australia, of which only T. peregrinus is represented in South Africa and South America. Thaumastocoris peregrinus populations contained eight haplotypes, with a pairwise nucleotide distance of 0.2-0.9% from seventeen locations in Australia. Three of these haplotypes are shared with populations in South America and South Africa, but the latter regions do not share haplotypes. These data, together with the current distribution of the haplotypes and the known direction of original spread in these regions, suggest that at least three distinct introductions of the insect occurred in South Africa and South America before 2005. The two most common haplotypes in Sydney, one of which was also found in Brisbane, are shared with the non-native regions. Sydney populations of T. peregrinus, which have regularly reached outbreak levels in recent years, might thus have served as source of these three distinct introductions into other regions of the Southern Hemisphere.

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

Produção de bioetanol a partir de pedúnculo de caju (Anacardium occidentale L.) por fermentação submersa

Recently, global demand for ethanol fuel has expanded very rapidly, and this should further increase in the near future, almost all ethanol fuel is produced by fermentation of sucrose or glucose in Brazil and produced by corn in the USA, but these raw materials will not be enough to satisfy international demand. The aim of this work was studied the ethanol production from cashew apple juice. A commercial strain of Saccharomyces cerevisiae was used for the production of ethanol by fermentation of cashew apple juice. Growth kinetics and ethanol productivity were calculated for batch fermentation with different initial sugar (glucose + fructose) concentration (from 24.4 to 103.1 g.L-1). Maximal ethanol, cell and glycerol concentrations (44.4 g.L-1, 17.17 g.L-1, 6.4 g.L-1, respectively) were obtained when 103.1 g.L-1 of initial sugar concentration were used, respectively. Ethanol yield (YP/S) was calculated as 0.49 g (g glucose + fructose)-1. Pretreatment of cashew apple bagasse (CAB) with dilute sulfuric acid was investigated and evaluated some factors such as sulfuric acid concentration, solid concentration and time of pretreatment at 121°C. The maximum glucose yield (162.9 mg/gCAB) was obtained by the hydrolysis with H2SO4 0.6 mol.L-1 at 121°C for 15 min. Hydrolysate, containing 16 ± 2.0 g.L-1 of glucose, was used as fermentation medium for ethanol production by S. cerevisiae and obtained a ethanol concentration of 10.0 g.L-1 after 4 with a yield and productivity of 0.48 g (g glucose)-1 and 1.43 g.L-1.h-1, respectively. The enzymatic hydrolysis of cashew apple bagasse treated with diluted acid (CAB-H) and alkali (CAB-OH) was studied and to evaluate its fermentation to ethanol using S. cerevisiae. Glucose conversion of 82 ± 2 mg per g CAB-H and 730 ± 20 mg per g CAB-OH was obtained when was used 2% (w/v) of solid and loading enzymatic of 30 FPU/g bagasse at 45 °C. Ethanol concentration and productivity was achieved of 20.0 ± 0.2 g.L-1 and 3.33 g.L-1.h-1, respectively when using CAB-OH hydrolyzate (initial glucose concentration of 52.4 g.L-1). For CAB-H hydrolyzate (initial glucose concentration of 17.4 g.L-1), ethanol concentration and productivity was 8.2 ± 0.1 g.L-1 and 2.7 g.L-1.h-1, respectively. Hydrolyzates fermentation resulted in an ethanol yield of 0.38 g/g glucose and 0.47 g/g glucose, with pretreated CABOH and CAB-H, respectively. The potential of cashew apple bagasse as a source of sugars for ethanol production by Kluyveromyces marxianus CE025 was evaluated too in this work. First, the yeast CE025 was preliminary cultivated in a synthetic medium containing glucose and xylose. Results showed that it was able to produce ethanol and xylitol at pH 4.5. Next, cashew apple bagasse hydrolysate (CABH) was prepared by a diluted sulfuric acid pre-treatment. The fermentation of CABH was conducted at pH 4.5 in a batch-reactor, and only ethanol was produced by K. marxianus CE025. The influence of the temperature in the kinetic parameters was evaluated and best results of ethanol production (12.36 ± 0.06 g.L-1) was achieved at 30 ºC, which is also the optimum temperature for the formation of biomass and the ethanol with a volumetric production rate of 0.25 ± 0.01 g.L-1.h-1 and an ethanol yield of 0.42 ± 0.01 g/g glucose. The results of this study point out the potential of the cashew apple bagasse hydrolysate as a new source of sugars to produce ethanol by S. cerevisiae and K. marxianus CE025. With these results, conclude that the use of cashew apple juice and cashew apple bagasse as substrate for ethanol production will bring economic benefits to the process, because it is a low cost substrate and also solve a disposal problem, adding value to the chain and cashew nut production

Crystallization of SiO2-CaO-Na2O Glass Using Sugarcane Bagasse Ash as Silica Source

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