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  • 1990-1994  (2)
  • 1
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Analysis of the initial interaction betweenEntamoeba histolytica trophozoites and the large intestine is impossible in humans and very difficult in experimental animals. To circumvent this obstacle we treated the luminal side of full-thickness rabbit colon segments mounted in Ussing-type chambers with trophozoite lysates of theE. histolytica HM1 virulent strain. Exposure to lysates for up to 90 min produced dose- and time-dependent effects on the colon, consisting of (a) increased decay rates for potential difference, short-circuit current, and transmural resistance and (b) mucosal damage ranging from vacuolation at the bases and shortening of epithelial cells to the loss of intercellular junctions, destruction of microvilli, and necrosis of interglandular epithelial zones. This acute model of intestinal amebiasis is sensitive, fast, and reliable.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0749-503X
    Keywords: Firefly luciferase ; yeast expression ; gene reporter ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The LUC gene coding for Photinus pyralis firefly luciferase was cloned in different yeast episomal plasmids in order to assess its possibilities as an in vivo reporter gene. Activity of the enzyme in transformed cells in vivo was measured by following light emission and assay conditions optimized in intact cells, with regard to oxygen concentration, temperature, cell concentration in assay mixtures and external ATP concentration. Among the factors tested, light emission was drastically influenced by the external pH in the assay (which resulted in a ten-fold amplification signal) and by substrate permeability. The growth phase of the cells was also important for the level of activity detected. Cloning of firefly luciferase gene under the control of different yeast-regulated promoters (ADH1, GAL1-10) enabled us to measure their strength which correlated well with previously described data. We conclude that firefly luciferase is an adequate gene reporter for the in vivo sensitive determination of gene expression and promoter strength in yeast.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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