Análisis electroforético de proteínas del veneno de Bothrops atrox “jergón” (Ofidia: Viperidae) de distintas zonas geográficas de la Amazonia Peruana

Análisis electroforético de proteínas del veneno de Bothrops atrox “jergón” (Ofidia: Viperidae) de distintas zonas geográficas de la Amazonia Peruana Electrophoretic analysis of proteins in the venom of Bothrops atrox “lancehead” (Ofidia: Viperidae) from different geographical zones to the Peruvian Amazon Rommel Rojas, Roberson Ramírez, Marianela Cobos y Juan Castro Centro de Investigaciones de Recursos Naturales-CIRNA Universidad Nacional de la Amazonia Peruana-UNAP. Apartado postal 496S Facultad de Ciencias Biológicas. DOI: https://doi.org/10.33017/RevECIPeru2011.0050/ RESUMEN Los venenos de serpientes son secreciones endógenas constituidas en su mayor parte por proteínas, que pueden causar diversos cuadros sintomatológicos, y en algunos casos la muerte. Las serpientes fueron recolectadas en el campo por muestreos herpetológicos utilizando la técnica del revelamiento por encuentros casuales [1] y el veneno recolectado por ordenamiento manual. Usando la técnica de electroforesis discontinua en gel de poliacrilamida [2] al 7.5% y 12% con dodecilsulfato de sodio (PAGE-SDS) se analizaron los perfiles electroforéticos de 5 especímenes de Bothrops atrox “jergón” provenientes de 5 zonas geográficas de la Amazonia Peruana, siendo evaluadas en condiciones reductoras utilizando veneno centrifugado, asimismo se realizó la cuantificación de las proteínas por el método espectrofotométrico utilizando el método de Biuret [3]. Los resultados muestran la presencia de proteínas en los rangos de 14 kD a 116 kD, obteniéndose variaciones en la cantidad de bandas proteínas, 9,11,8,7,8 para las poblaciones de Moena Caño, Isla Iquitos, Panguana, Genaro Herrera y Nina Rumi, mientras que la cuantificación muestra la existencia de una mayor cantidad de proteínas en el veneno de la serpiente proveniente de Isla Iquitos (67.41 mg/ml) y una menor cantidad en el espécimen de Genaro Herrera (32.13 mg/ml). Se concluye mencionando la existencia de variaciones en los perfiles electroforéticos y cantidades de las proteínas presentes en el veneno de Bothrops atrox de la Amazonia Peruana. Descriptores: venenos, proteínas, electroforesis, poliacrilamida, cuantificación. ABSTRACT The snake´s venoms are endogenous secretions constituted for proteins and can produce different symptomatology and in some cases the dead. Using the electrophoretic technique in polyacrylamide discontinuous gel of 7.5% and 12% whit sodium dodecylsulphate (PAGE-SDS) in reductions conditions were carried on a protein analysis of the venom in 5 Bothrops atrox snake´s from 5 different geographical zones to the Peruvian Amazon using centrifuge crude venom, additionally the proteins were quantification for using an spectrophotometer following the Biuret method. The results evidence the presents of different bands in the proteins, in ranges of 14 kD to 116 kD, finding 9, 11,8,7,8 bands of proteins for the populations of Moena Caño, Isla Iquitos, Panguana, Genaro Herrera and Nina Rumi, while the quantification show the presents to major quantity of protein in the snake from Isla Iquitos (67.41 mg/ml) and a less quantity in the specie from Genaro Herrera (32.13 mg/ml). We conclude mentioning the existence of variations in the electrophoretic profile and quantification to proteins from the venom of Bothrops atrox to the Peruvian Amazon. Keywords: venoms, proteins, electrophoretic, polyacrylamide, quantification.

POLYMER-ASSISTED DEPOSITION OF HYDROXYAPATITE COATINGS USING ELECTROPHORETIC TECHNIQUE

Hydroxyapatite (HA) finds use as powder, scaffold, paste, and coatings for orthopedic and dental applications. Plasma spraying is the most commonly used technique to coat HA on metallic implants. However, undesirable phase changes at high temperatures encourage to adopt ambient temperature deposition techniques such as dip coating, electrophoretic, and physical vapor deposition (PVD). Electrophoretic technique is being used extensively to deposit HA, however sintering is required after the deposition to enhance adhesion of coatings to the substrate. In the present work, polyethylene glycol (PEG) modified HA was deposited on 316L Stainless Steel plates using electrophoretic deposition (EPD), which improved the binding strength of the HA to the substrate with increased packing density of HA particles without the need of sintering. PEG is a biocompatible and soluble polymer that helps HA to bond well with the substrate and in addition, prevents the agglomeration and precipitation of HA. Phase identification and crystal structure of the coatings were determined using X-ray diffraction (XRD). The stability of the coatings was assessed by Fourier transform infrared spectroscopy (FTIR), whereas scanning electron microscopy (SEM) was utilized in order to investigate the morphological properties of the deposited coatings. The mechanical properties of the coatings were investigated using the indentation testing that depicted an enhanced level of adhesion of coatings to the substrate.