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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 24 (1977), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS A controlled-environment culture system was used to show that both physical and biologic parameters can influence the penetration of vertebrate cells by Toxoplasma gondii. The optimum bicarbonate ion concentration for the penetration of bovine embryo skeletal muscle (BESM) cells is 36.25 mM. Higher or lower bicarbonate ion concentrations are increasingly inhibitory to penetration. As CO2 increases in the range from 0.5–3.7 mM, penetration is progressively inhibited. No relationship was found between penetration and pH in the pH range of 6.949–7.765. The culture age of the BESM cells directly influenced the ability of the parasites to penetrate the cells. Older BESM cells were more refractory to penetration than younger cells.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 34 (1987), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Utilizing the previously reported inter-clonal differences in total DNA/organism, flow cytometry was used to analyze the population dynamics of Trypanosoma cruzi clone mixtures growing in liquid medium or vertebrate cells. The growth of clone mixtures in liquid medium can be described by unique parameters reflecting exponential growth rate (r), stationary phase population density (1/k), and the interaction between the clones (h). The relative numbers of each clone in the population change rapidly with time and the results arc in quantitative agreement with mathematical models of competitive population growth. The relationship between the parameters for T. cruzi is such that, in general, there is no dynamic equilibrium with coexistence of clones with different growth rates; under all culture protocols, the faster growing clone will prevail. A computer simulation of the vertebrate cell cycle of T. cruzi suggests that clone mixtures grow relatively independently; the basic attributes of the model were substantiated experimentally. Although wide fluctuations in the proportion of each clone released occurred, the faster growing clone again predominated. Finally, these results underline the importance of working with well-defined clones in the laboratory to avoid inconsistencies and paradoxical results and stress the importance of the rapid isolation of single cell clones from clinical specimens when studying the relationship of the parasite to human disease.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 31 (1984), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The infection of bovine embryo skin and muscle cells by trypomastigotes of four Trypanosoma cruzi clones (CA-I/71, /72, Miranda/76, /80) was quantified. Stable and reproducible intra-isolate differences were observed; an almost 70-fold difference in infectivity occurred between clones. The CA-I/71 clone was not susceptible to N-acetyl-D-glucosamine at a concentration that inhibits the infection of vertebrate cells by Ernestina and Y-strain parasites. Eight other monosaccharides that are common constitutents of vertebrate cell surface glycoproteins also failed to inhibit the infection of vertebrate cells by the CA-I/71 clone.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 32 (1985), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The complete intracellular cycle of five cloned stocks of Trypanosoma cruzi was quantified. Marked but stable interclonal differences were found in the length of the pre-replicative lag period (18.2-34.2 h), amastigote doubling lime (8.6-21.5 h), and duration of the complete intracellular cycle (96-215 h). Strong correlations were demonstrated between these characteristics as well as to the growth rate of the epimastigote stage of the same clones grown in liquid medium. These data demonstrate that the marked heterogeneity of the natural population of T. cruzi extends to the intracellular cycle of the parasite and has important implications for our understanding of Chagas’ disease.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 27 (1980), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS. the pattern of RNA synthesis of intracellular Trypanosoma cruzi amastigotes, immediately following infection of Lesch-Nyhan human fibroblasts, was studied by autoradiography. Amastigote RNA synthesis, determined by [3H]guanine incorporation, was not detected until 2 h after infection. At 8 h postinfection more than 90% of intracellular amastigotes were labeled. It was verified that extracellular trypomastigotes also synthesized RNA. Therefore it was concluded that, if RNA is required for trypomastigote-to-amastigote transformation, this nucleic acid is already present in the trypomastigotes before infection of the vertebrate cell. It is probable that the RNA synthesized by amastigotes during the prereplicative lag period (the period between initial infection and the onset of DNA synthesis) is required for intracellular growth and reproduction.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 23 (1976), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS. A video technic is described that permits a quantification of the degree of attraction of Trypanosoma cruzi trypomastigotes to vertebrate cells in vitro. Bovine embryo skeletal muscle cells (BESM), HeLa cells and Vero cells all attract a myotropic strain of T. cruzi trypomastigotes. BESM cells, however, are 2-fold more attractive to trypomastigotes than HeLa cells and 10-fold more attractive than Vero cells. Heat-inactivation of BESM cells abolishes their ability to respire and also to attract T. cruzi trypomastigotes. As there is no difference in the endogenous oxygen consumption between BESM, HeLa, and Vero cells, it is unlikely that differences in the attraction of trypomastigotes to the 3 cell types are due to variations in the magnitude of pO2 or pCO2 gradients in the milieu around the cells.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 42 (1995), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: . We developed a method to study the DNA synthetic cycles of Entamoeba histolytica and Entamoeba invadens by flow cytometry (FCM) based on a preparative procedure to reduce both high levels of natural fluorescence and non-specific adsorption of fluorochromes. We modeled G1, S, and G2 phases as a series of overlapping Gaussian curves. Both E. histolytica and E. invadens displayed G1, S, and G2 proportions that are consistent with eukaryotic cell populations in exponential or stationary growth phase. Exponential phase E. histolytica populations contained a hypodiploid subset with a mass of about 20% less than the diploid value which we estimate by FCM to be 24 × 10-14 g DNA/cell. Exponential phase E. invadens populations contained a hypodiploid subset with a mass of about 6% less than the diploid value which we estimate by FCM to be 30 × 10-14 g DNA/cell.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 38 (1991), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Flow cytometry and DNA binding-specific fluorescent reagents were used to compare the total DNA, G-C, and A-T content of the epimastigote and trypomastigote stages of Trypanosoma cruzi stocks. Significant total DNA differences of 2–12% between epimastigotes and trypomastigotes were found in three of six stocks studied. The epimastigote G-C content of five of six stocks was 4–8% higher than trypomastigotes, whereas the trypomastigote A-T content was 2.5–13% higher than the epimastigote A-T content. Although no obvious developmental stage association between total DNA and base composition was found, intrastage associations do exist. These observations were unaffected by nucleoprotein extraction implying that the observed differences between trypomastigotes and epimastigotes are not a consequence of nucleoprotein interference with DNA-binding fluorochromes. The nuclei and kinetoplasts of four T. cruzi stocks were isolated and analyzed. Developmental stage differences in nuclear and kinetoplast DNA are stock-dependent and base composition-dependent; both organelles contribute to the observed differences in DNA of intact cells. We found a nearly linear association between the percentage of total kinetoplast DNA, G-C, and A-T content. During metacyclogenesis, the G-C content decreases by approximately 7% as epimastigotes transform into metacyclic trypomastigotes. The decrease in G-C content precedes changes in morphology or in complement resistance. If the DNA changes are causally connected to developmental stage transformations in T. cruzi remains to be determined. However, our results could facilitate studies of the molecular genetic processes the parasite uses to successfully complete various phases of its life cycle and, consequently, the disease process it evokes.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 32 (1985), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: C57B1/6 mice were infected with Brasil strain Trypanosoma cruzi trypomastigotes. The leg muscles of the mice were serial-sectioned with a cryostat, and individual fibers were classified histochemically as type I or type II on the basis of succinic dehydrogenase or adenosine triphosphatase activity. Although markedly more type II fibers were present in the leg muscles, the percentage of infected type I fibers was nearly five-fold higher than type II. This is the first demonstration of a preferential in vivo distribution of T. cruzi in muscle fibers based upon muscle type.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Nineteen clones of Trypanosoma cruzi were obtained as single-cell isolates from Triatoma infestans. Ten of the clones were isolates from a patient with chronic Chagas' disease; nine clones were isolates from a dog infected with T. cruzi strain CA-I isolated originally from a chronic chagasic patient. The growth kinetics and peak modal Coulter volume of these clones were characterized. Significant inter- and intra-group differences between growth rates and peak modal volumes were found. These data indicate that subpopulations and, consequently, genetic heterogeneity of T. cruzi exist in chronic chagasic patients. All of the clones infected vertebrate cells in vitro.
    Type of Medium: Electronic Resource
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