ABSTRACT
It has been reported that the immune response due to alpha-Gal epitopes is an important factor in tissue valve failure. The elimination of the interaction between the natural anti-Gal antibodies and alpha-gal epitopes on the xenografts is a prerequisite to the success of xenografts in humans. Previously, we reported that the green coffee bean alpha-galactosidase could remove all alpha-Gal epitopes from cell surface of porcine aortic valve and pericardial tissue, but it has limitations on cost effectiveness. In this study we wanted to know whether the recently produced recombinant human alpha-galactosidase A has the same effective enzymatic activity as green coffee bean alpha-galactosidase in removing alpha-Gal epitopes from the same tissues. After treating fresh porcine aortic valve and pericardial tissue with recombinant alpha-galactosidase A, each sample was stained with Griffonia simplicifolia type I isolectin B4 indirect immunoperoxidase avidin-biotin technique. We then examined whether the alpha-Gal epitopes were reduced or abolished in each consecutive concentration of recombinant alpha-galactosidase A by comparing the degree of the Griffonia simplicifolia isolectin B4 staining. As a result, the recombinant alpha-galactosidase A could remove cell surface alpha-Gals on porcine aortic valve and pericardial tissue as effectively as green coffee bean alpha-galactosidase.
Subject(s)
Adolescent , Animals , Child , Humans , Aortic Valve/chemistry , Coffea/enzymology , Epitopes/immunology , Heart Valve Prosthesis Implantation , Pericardium/chemistry , Recombinant Proteins/genetics , Swine , Transplantation, Heterologous/immunology , alpha-Galactosidase/geneticsABSTRACT
Generalmente, las plantas responden de manera diferenciada a las técnicas de cultivo in vitro, dependiendode la respuesta inicial del cultivo y su capacidad embriogénica del genotipo, el estado fisiológico de la planta yel explante, la edad de la planta donante, los factores físicos, los reguladores del crecimiento y las condiciones nutricionales, así como el cambio estacional. De aquí que el presente estudio se realizó con el objetivo de evaluar la influencia de la época del año en que fue tomada la fuente de los explantes en la respuesta in vitro de genotipos de Robusta. Se realizaron muestreos durante todos los meses del año, los explantes foliares fueron cultivados en un medio contentivo de las sales minerales de Murashige y Skoog (1962) y 0,5 y 2,0 mg.L-1 de 2,4-D y kinetina, respectivamente. Se observó que la época del año en que se tomó la muestra ejerció un marcado efecto en la respuesta de los explantes de los genotipos evaluados. Para las condiciones estudiadas resultó favorable la toma de las muestras foliares en los períodos mayo-junio, enero-febrero y noviembre-diciembre, dados los bajos índices de actividad enzimática peroxidasa, oxidación fenólica y contaminación fúngica, así como un elevado porcentaje de formación de callos y mayor contenido de proteínas totales presentes en los mismos. Se determinó que la actividad enzimática peroxidasa pudiera constituir un importante marcador en la etapa inicial de selección de las muestras.
Plants generally respond in different ways to in vitro cultivation techniques, depending on the crops initial response and the embryogenic capacity of its genotype, the physiological state of the plant and the explant, the age of the donor plant, physical factors, growth regulators, nutritional conditions and seasonal change. The present study was thus aimed at evaluating the influence of the time of year during which the source of explants was taken on the in vitro response of Robusta genotypes. Samples were taken during each month ofthe year, foliar explants were cultured in a medium containing Murashige and Skoog mineral salts (1962) and 0,5 and 2,0 mg.L-1 2,4-D and kinetine, respectively. It was observed that the time of the year when a sample was taken exercised as marked effect on the response of the explants from the genotypes evaluated. Taking foliar samples during May-June, January-February and November-December proved favourable in the conditions studied here given the low indices of peroxidase enzymatic activity, phenol oxidation and fungal contamination, as well as the high percentage of callus formation and their greater total protein content. It was determined that peroxidase enzymatic activity could constitute an important marker during the initial stage of selecting samples.