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

Dinâmica de reprodução e comportamento reprodutivo de branchoneta Dendrocephalus brasiliensis Pesata, 1921 como incremento na produção de alimento vivo para peixes ornamentais

Reproductive behavior of Dendrocephalus brasiliensis Pesta, 1921(Crustacean: Anostracan). The reproductive behavior of fresh water Anostracan has been poorly investigated in carcinology specialized literature, specifically in relation to Dendrocephlaus brasiliensis Pesta, 1921, with abundant data just about the geographical distribution of that Anostracan. The objective of this work was to know the reproductive behavior of this Anostracan, in different seasons (dry and raine). For this, ripe individuals of both sexes were collected in four pond of the of Fish farming Station of Paulo Afonso (EPPA), through monthly captures in each ponds, from December 2004 to November 2005. The type of reproduction was observed after the placing in aquariums (a) of then with males and (b) other on individually, at the nauplii phase, where they stayed for 15 days until the reproductive age. The sexual proportion was calculated through the relative frequencies of males and females, every month, for the whole collection period during 10 days. the production of cysts was related to the size of the female. The male: female ratio in the study period was 1 male:1,07 female. The proportion male:female was from 48,25% : 51,75% along the year. Concerning the type of reproduction, it was observed that this specie is characteerized by sexual reproduction