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

Distribuição espacial, formação de grupos, uso do hábitat e ecologia comportamental de jovens de tubarão limão, negaprion brevirostris (poey 1868), no arquipélago de fernando de noronha

Lemon sharks, Negaprion brevirostris, are common in the Fernando de Noronha Archipelago, but detailed information about the species in this site is lacking. The aim of this study was to describe the spatial distribution, grouping behavior, habitat use and behavioral ecology of juvenile lemon sharks in the archipelago, and their interaction with some environmental and ecological factors. During 2006 and 2007, the presence and spatial distribution of juvenile sharks were quantified through scuba diving and snorkeling at several sites of the archipelago. In 2008 the habitat use of juvenile sharks was quantified through visual census while snorkeling along 300 x 8 m strip transects. During these transects the grouping behavior of lemon sharks was quantified by ad libitum. Results indicate that Fernando de Noronha Archipelago is used as a nursery area for lemon sharks, and the parturition occurs from November to April. Juveniles preferred using shallower areas available by the tide variation and formed groups only in the presence of adult conspecifics. This preference for shallower habitats and the group behavior probably are anti-predatory tactics used by juvenile lemon sharks, in response to the low availability of shelter and high predation risk of the studied areas. Quantifications of prey availability and predation risk of juveniles showed that, in general, lemon sharks are trading-off food by security and investing in sites with higher possibility of energetic return. Behavioral observations enabled to record juvenile carangid fishes following juvenile lemon sharks, remora host-parasite and juvenile sharks foraging on schools of herrings and octopuses. We also recorded the behavior of juvenile sharks following conspecifics of similar size, circling with two or three individuals and smaller individuals giving way to larger juveniles. When adults are present, juvenile lemon sharks are more social than solitary, indicating that predation is one of the factors that contribute to social behaviors of the species. Results also suggest that when grouped the juveniles have a hierarchical organization according to body size. Furthermore, observation of large adult females with several fresh mating bites and scars in the same habitats used by juvenile lemon sharks, indicates that Fernando de Noronha Archipelago is used as nursery and mating grounds by this species