The thrombin-like enzyme from Bothrops barnetti named barnettobin was purified. We report some biochemical features of barnettobin including the complete amino acid sequence that was deduced from the cDNA. Snake venom serine proteases affect several steps of human hemostasis ranging from the blood coagulation cascade to platelet function. Barnettobin is a monomeric glycoprotein of 52 kDa as shown by reducing SDS-PAGE, and contains approx. 52% carbohydrate by mass which could be removed by N-glycosidase. The complete amino acid sequence was deduced from the cDNA sequence. Its sequence contains a single chain of 233 amino acid including three N-glycosylation sites. The sequence exhibits significant homology with those of mammalian serine proteases e.g. thrombin and with homologous TLEs. Its specific coagulant activity was 251.7 NIH thrombin units/mg, releasing fibrinopeptide A from human fibrinogen and showed defibrinogenating effect in mouse. Both coagulant and amidolytic activities were inhibited by PMSF. N-deglycosylation impaired its temperature and pH stability. Its cDNA sequence with 750 bp encodes a protein of 233 residues. Indications that carbohydrate moieties may play a role in the interaction with substrates are presented. Barnettobin is a new defibrinogenating agent which may provide an opportunity for the development of new types of anti-thrombotic drugs.
En este artículo se presentan los resultados de una Revisión Sistemática de la Literatura realizada para identificar y caracterizar estudios primarios que incorporan elementos de gamificación, tanto para los procesos vinculados con el ámbito de la Educación en Ingeniería de Software, como con los relacionados con la industria del software. Los resultados obtenidos muestran que existen diversos trabajos en los que los principios de la gamificación han sido utilizados como estrategia para promover mejoras; no obstante, la mayoría se encuentran en el ámbito de la educación. En el caso de los elementos de gamificación utilizados con mayor frecuencia en los estudios analizados, los tres más citados son: puntos, niveles, e insignias. Por otro lado, pudimos también observar que muchas de las iniciativas reportadas, incorporan el uso de una aplicación software para apoyar la dinámica instruccional.
The immunogenicity of Bothrops atrox, “jergón”, venom was studied using ELISA and Western Blot methods, as well as cross-reactivity patterns against venoms of Bothrops brazili, Lachesis muta and Crotalus durissus. For this purpose, New Zealand white rabbits (2 kg aprox) were immunized with four 500 μg doses of B. atrox venom in a period of 90 days. Antibody production was followed using ELISA technique, and title of hiper-immune serum was determined at the end of immunization protocol. Additionally, electrophoretic patterns of venoms were analyzed by SDS-PAGE and venom reactivity against obtained serum by ELISA and Western Blot. Immunization schedule allowed a pronounced antibody production since day 20 of protocol. At the end of process, serum title was 256000, which demonstrated both efficacy and usefulness of the developed procedure. On the other hand, studied venoms showed a heterogenic protein composition according to their electrophoretic patterns, whereas cross-reactivity values of 23,7%, 4,0% and 1,8% were obtained between B. atrox venom and B. brazili, L. muta and C. durissus venoms, respectively, using immunoenzymatic methods. According to our results, this procedure constitutes an initial step for further assays directed to optimization in immunoserum production for envenoming treatment and development of kits for diagnosis and species identification of snakes.
Un grupo de 90 mujeres con antecedente de 2 o más abortos espontáneos de causa desconocida, sin historia de partos anteriores, con estudio inmunológico que determinó ausencia de respuesta aloinmune y un grupo de control de 31 mujeres con antecedentede fertilidad comprobada, se estudiaron para anticuerpos a antígenos nucleares, anticoagulante lúpico y fosfolípidos. Demostramos serológicamente un incremento en la respuesta autoinmune a partir del segundo aborto. Los anticuerpos antifosfolípidos y los anticuerpos antinucleares aparecen presentes ya como epifenómenos ya como factores etiológicos en pacientes con pérdida recurrente del embarazo PRE.La importancia de los hallazgos mencionados y la novedad del presente traba,jo consiste en la identificación del incremento en la respuesta autoinmune a partir del segundo aborto.
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