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      El estadio amastigota precede la evolución del epimastigota durante la epimastigogénesis in vitro de Trypanosoma cruzi Translated title: The amastigote stadium precedes epimastigote evolution during in vitro Trypasnosoma cruzi epimastigogenesis

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          Abstract

          La transformación de tripomastigotas sanguíneos de Trypanosoma cruzi en epimastigotas, ocurre naturalmente en el intestino del insecto y se identifica como epimastigogénesis. Aquí reportamos que durante la epimastigogénesis in vitro, los tripomastigotas se transforman en formas redondeadas, biológica y antigénicamente equivalentes al estadio amastigota; con un tiempo de persistencia dependiente de la altura del medio de cultivo. Formas tripomastigotas procedentes de cultivo de tejidos infectados se incubaron en envases con diferentes alturas de medio LITB (3 y 83 mm), y se compararon las cinéticas de transformación hacia epimastigotas. Para la obtención de antígenos se colectaron masas de parásitos a diferentes tiempos de diferenciación. Los cambios morfológicos, incremento del inóculo y resistencia a complemento, se estudiaron por microscopía de contraste de fases y extendidos coloreados con Giemsa. Los cambios antigénicos se analizaron mediante Western Blot usando anticuerpo IgY anti-amastigotas. Los resultados mostraron que las formas redondeadas resisten la lisis por complemento, expresan antígenos amastigota-específicos y la velocidad de transformación hacia epimastigotas depende de la altura del medio sobrenadante en el cultivo. Las evidencias sugieren que además de caída de la temperatura, una baja tensión de oxígeno disuelto y una alta densidad de parásitos por área, aceleran el proceso de diferenciación.

          Translated abstract

          Transformation of blood Trypanosoma cruzi tripomastigotes into epimastigotes occurs naturally at the insect gut and is identified as epimastigogenesis. Here we report that during in vitro epimastigogenesis, tripomastigotes transform into rounded forms, biologically and antigenically equivalent to the amastigote stadium, with a persistance time which depends on the height of the culture medium. Tripomastigote forms from cultures of infected tissues were incubated in flasks with different LITB medium heights (3 and 83 mm), and kinetics of transformation to epimastigotes were compared. Masses of parasites at different differentiation times were collected for antigen production. Morphological changes, inoculum increase and resistance to complement were studied by phase contrast microscopy and Giemsa stained smears. Antigenic changes were analyzed through Western Blot using an IgY anti-amastigote antibody. Results showed that rounded forms resist lysis by complement, express amastigote-specific antigens, and that the speed of transformation to epimastigotes depends on the height of the supernatant medium in the culture. Evidences suggest that apart from a temperature fall, a low dissolved oxygen tension and a high parasite per area density accelerate the differentiation process.

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          Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

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            The life cycle of Trypanosoma cruzi revisited.

            The basic features of the life cycle of Trypanosoma cruzi have been known for nearly a century. Various aspects of the life cycle, however, have been elucidated only recently, whilst others remain either controversial or unstudied. Here, we present a revised life cycle influenced by recent findings and specific questions that remain unresolved.
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              Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications

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                Author and article information

                Journal
                rsvm
                Revista de la Sociedad Venezolana de Microbiología
                Rev. Soc. Ven. Microbiol.
                Organo Oficial de la Sociedad Venezolana de Microbiología. (Caracas, DF, Venezuela )
                1315-2556
                June 2013
                : 33
                : 1
                : 72-79
                Affiliations
                [01] Carabobo orgnameUniversidad de Carabobo orgdiv1Facultad de Ciencias de la Salud orgdiv2Instituto de Biología Molecular de Parásitos (InBioMolP) Venezuela
                Article
                S1315-25562013000100014 S1315-2556(13)03300114
                076ceca3-24aa-4d83-bbbc-c78a46f427e6

                http://creativecommons.org/licenses/by/4.0/

                History
                : 12 March 2012
                : 12 December 2012
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 28, Pages: 8
                Product

                SciELO Venezuela

                Categories
                Artículos Originales

                Trypanosoma cruzi,in vitro epimastigogenesis,amastigote-specific antigens,epimastigogénesis in vitro,antígenos amastigota específicos

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