Paula C Hernández

National University of Colombia, Μπογκοτά, Bogota D.C., Colombia

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Publications (5)7.37 Total impact

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    ABSTRACT: We examined the effect of aphidicolin, colchicine, demecolcine, fluorouracil, hydroxyurea, and nocodazole, as well as nutrient deprivation on the Giardia intestinalis cell cycle. Aphidicolin was the only drug that was able to block the cell cycle at a specific stage (G1/S), and permit cells to resume growth at a high rate upon its removal. Nutrient deprivation resulted in a portion of G2/M cells completing mitosis and cytokinesis in synchrony during the recovery period, but this synchrony was shortly lost and a sample containing a predominance of G1 cells could not be obtained. Flow cytometry analysis of normal and untreated Giardia cultures showed the occasional appearance of a small percentage of cells with a DNA content of 16C, which is twice the DNA content of G2 cells. However, this 16C peak is larger and more frequently observed in drug-treated Giardia. These 16C are likely produced from endoreplication of 8C/G2 cells, and we propose that they represent a pre-encystation stage that is induced by drug treatments and other stressors.
    Molecular and Biochemical Parasitology 12/2012; 187(1). DOI:10.1016/j.molbiopara.2012.11.005 · 2.24 Impact Factor
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    ABSTRACT: The reproductive mechanism of Giardia intestinalis, considered one of the earliest divergent eukaryotes, has not been fully defined yet. Some evidence supports the hypothesis that Giardia is an exclusively asexual organism with a clonal population structure. However, the high genetic variability, the variation in ploidy during its life cycle, the low heterozygosity and the existence of genes involved in the meiotic-like recombination pathway in the parasite's genome cast doubt on exclusively asexual nature of Giardia. In this work, semiquantitative RT-PCR analysis was used to assess the transcription pattern of three meiosis-like-specific genes involved in homologues recombination: dmc1, hop1 and spo11. The mRNAs were amplified during the parasite's differentiation processes, encystation and excystation, and expression was found at each stage of its life cycle. A semiquantitative assessment also suggests that expression of some of the genes is regulated during encystation process.
    Memórias do Instituto Oswaldo Cruz 06/2008; 103(4):347-50. DOI:10.1590/S0074-02762008000400006 · 1.57 Impact Factor
  • Paula C Hernandez · Moises Wasserman
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    ABSTRACT: Giardia intestinalis undergo biochemical and morphological changes to survive under extreme environmental conditions. One of these changes is encystation. The trophozoites colonizing the upper part of the small intestine differentiate into a cyst, the infective form of the parasite. In in vitro cultures, the formation of cysts is induced by the depletion of cholesterol. It was reported that Giardia cannot synthesize cholesterol de novo. However, through bioinformatic studies, we found the genes that codify for the enzymes in the cholesterol biosynthesis pathway. We were able to verify the existence and define the transcription of four genes in the trophozoite and in parasites subjected to the encystation and excystation processes.
    Parasitology Research 03/2006; 98(3):194-9. DOI:10.1007/s00436-005-0039-1 · 2.33 Impact Factor
  • Paula C Hernández · María Leonor Caldas · Moisés Wasserman
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    ABSTRACT: The reconstruction of Giardia lamblia life cycle in vitro is an excellent tool for the study of the parasite's molecular biology. The present work describes techniques developed that better define parasite differentiation. An encystation protocol is presented along with a method for isolation and purification of the produced cysts. The cyst morphology at the light microscopy level is identical to that of in vivo cysts. A two-dimension protein map obtained by high-resolution electrophoresis indicated that most of the parasite's proteins are acid. Based on this result, the two dimension gel electrophoresis used a pH 4-7 gradient in the first, isoelectric focusing dimension. Differences in protein expression during the stages of encystation were clearly discerned, as well as images of the parasite obtained by light and by transmission electron microscopy that describe the morphological and the ultrastructural changes that occur as the cysts are produced in vitro.
    Biomédica: revista del Instituto Nacional de Salud 10/2002; 22(3):253-62. · 0.62 Impact Factor
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    Paula C Hernández · María Leonor Caldas · Moisés Wasserman
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    ABSTRACT: La reconstrucción in vitro del ciclo de vida de Giardia lamblia es un excelente instrumento para el estudio de la biología molecular del parásito. El presente trabajo pretende contribuir al desarrollo de algunas técnicas por las cuales se puede definir mejor un modelo de diferenciación celular del parásito. El estudio presenta un protocolo de enquistación in vitro y establece un método para el aislamiento y la purificación de los quistes producidos, cuyas características morfológicas, por microscopía de luz, coinciden con las de los quistes obtenidos in vivo. Se estudió el mapa de las proteínas de G. lamblia por medio de electroforesis bidimensional de alta resolución y de electroforesis en una dimensión. Estos estudios mostraron que la mayoría de las proteínas del parásito son de carácter ácido. Se amplió, con base en ese resultado, la resolución de la región ácida por medio de un isoelectroenfoque de pH 4-7 en la primera dimensión. Se encontraron diferencias en la expresión de las proteínas durante el proceso de enquistación. Se obtuvieron, además, imágenes por microscopía de luz y electrónica de transmisión que permitieron observar morfológica y ultraestructuralmente las células producidas durante el proceso de la enquistación. Palabras clave: Giardia lamblia, enquistación, microscopía electrónica, electroforesis bidimensional, diferenciación, parásito, protista.
    Biomédica: revista del Instituto Nacional de Salud 09/2002; 22(3). DOI:10.7705/biomedica.v22i3.1162 · 0.62 Impact Factor