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  • 1
    Electronic Resource
    Electronic Resource
    Serial Subcultivation of Giardia lamblia in Keister's Modified TYI-S-33 Medium Containing Ultroser G (1992)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 39 (1992), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: . The ability of Giardia strains of the duodenalis type to grow in Keister's modified TYI-S-33 medium varies with serum lot. Recently, strains of Giardia including MR4, WB, and Human-1-Portland, have been cultivated in Keister's modified TYI-S-33 medium containing the serum substitute Ultroser G and have been cultured serially at least 40 times. An optimal concentration of 8% Ultroser G promotes maximal growth in Keister's modified TYI-S-33 medium for all three strains. This concentration of Ultroser G will produce a two-log increase in the number of trophozoites in approximately three days post-inoculation. Generation times for the trophozoites ranging from 6 to 11 h have been achieved in Keister's modified TYI-S-33 containing 10% adult bovine serum and from 8 to 13 h in Keister's modified TYI-S-33 with 8% Ultroser G. Despite the excellent growth of Giardia strains in medium containing Ultroser G, the maximum trophozoite density is only about half of that achieved in Keister's modified TYI-S-33 medium supplemented with 10% adult bovine serum. Comparisons of trophozoites grown with serum or the serum substitute reveal no discernable differences in morphology and motility. Additionally, these strains have been successfully cryopreserved and revived in Keister's modified TYI-S-33 medium supplemented with Ultroser G. Because Ultroser G is a characterized mixture of six main groups of ingredients (growth factors, adhesion factors, mineral trace elements, hormones, binding proteins, and vitamins), the variability in cell proliferation that may occur when changing serum lots should be minimized when using this product.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1992.tb01304.x
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  • 2
    Electronic Resource
    Electronic Resource
    Enzymes of Carbohydrate Metabolism in Four Human Species of Leishmania: A Comparative Survey (1976)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 23 (1976), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS. The occurrence and levels of activity of various enzymes of carbohydrate catabolism in culture forms (promastigotes) of 4 human species of Leishmania (L. brasiliensis, L. donovani, L. mexicana, and L. tropica) were compared. These organisms possess enzymes of the Embden-Meyerhof pathway but lack lactate dehydrogenase. No evidence could be found for the production of lactic acid by growing cultures and lactic acid could not be detected either in cell-free preparations or after incubation of cell-free extracts with pyruvate and NADH under appropriate conditions. All 4 species possess α-glycerophosphate dehydrogenase and α-glycerophosphate phosphatase which together could regenerate NAD, thus compensating for the absence of lactate dehydrogenase. The oxidative and nonoxidative reactions of the hexose monophosphate pathway are present in all 4 species. Cell-free extracts have pyruvate dehydrogenase activity which allows the entry of pyruvate into and its subsequent oxidation through the tricarboxylic acid cycle. All enzymes of this cycle, including a thiamine pyrophosphate dependent α-ketoglutarate dehydrogenase are present. Both NAD and NADP-linked malate dehydrogenase activities are present. The isocitrate dehydrogenase is NADP specific. There is an active glutamate dehydrogenase which could compete with α-ketoglutarate dehydrogenase for the common substrate (α-ketoglutarate). Replenishment of C4 acids is accomplished by heterotrophic CO2 fixation catalyzed by pyruvate carboxylase. All 4 species have high levels of NADH oxidase activity. Several enzymes thus far not found in any species of Leishmania have been demonstrated. These are: phosphoglucose isomerase, triose phosphate isomerase, fructose-1, 6-diphosphatase, 3-phosphoglycerate kinase, enolase, α-glycerophosphate dehydrogenase, α-glycerophosphate phosphatase, pyruvate dehydrogenase complex, citrate synthase, aconitase, α-ketoglutarate dehydrogenase, glutamate dehydrogenase, and NADH oxidase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1976.tb03850.x
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  • 3
    Electronic Resource
    Electronic Resource
    Specificity of the Glucose Transport System in Leishmania tropica Promastigotes (1976)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 23 (1976), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS. The glucose transport system in Leishmania tropica promastigotes was characterized by the use of labeled 2-deoxy-D-glucose (2-DOG), a nonmetabolizable glucose analog. The uptake system has a Q10 of 2 and a heat of activation of 10.2 kcal/mole. The glucose transport system is subject to competitive inhibition by 2-DOG, glucosamine, N-acetyl glucosamine, mannose, galactose, and fructose which suggests that substitutions in the hexose chain at carbons 2 and 4 do not affect carrier specificity. In contrast, changes at carbon 1 (α-methyl-D-glucoside, 1,5-anhydroglucitol) and carbon 3 (3–0-methyl glucose) lead to loss of carrier affinity since these sugars do not compete for the glucose carrier. Sugars that compete with the glucose carrier have one common feature—they all exist in the pyranose form in solution. The carrier for D-glucose does not interact with L-glucose or any of the pentose sugars tested. Uptake of 2-DOG is inhibited by glycerol. This inhibition, however, is noncompetitive; it is evident, therefore, that glucose and glycerol do not compete for the same carrier. Glycerol does not repress the glucose carrier since cells grown in presence of glycerol transport the sugar normally.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1976.tb03810.x
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  • 4
    Electronic Resource
    Electronic Resource
    Delayed in vitro Utilization of Glucose by Leishmania tropica Promastigotes (1974)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 21 (1974), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Leishmania tropica promastigotes do not utilize glucose provided in the medium until late log phase. Rapid depletion of glucose from the medium, however, occurs during late log and stationary phases. At about the same time, the cells show maximal rates of glucose uptake as well as peak levels of phosphofructokinase and pyruvate kinase activities. The glucose analog, 2-deoxy-D-glucose inhibits glucose transport. Incorporation of this analog in the growth medium results in inhibition of growth. The hexokinase of L. tropica phosphorylates 2-deoxy-D-glucose. Pyruvate kinase is activated by fructose-1, 6-diphosphate and adenosine monophosphate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1974.tb03676.x
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  • 5
    Electronic Resource
    Electronic Resource
    Effects of Ethidium Bromide and Several Acridine Dyes on the Kinetoplast DNA of Leishmania tropica (1972)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 19 (1972), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Whole cell DNA from Leishmania tropica has 2 peaks when banded by CsCl equilibrium density centrifugation. The main band has a buoyant density of 1.721 and the satellite band a buoyant density of 1.705, with Clostridium perfringens DNA (ρ= 1.6915) used as a reference. The satellite band has been identified as the kinetoplast DNA by purifying DNA from isolated kinetoplasts. L. tropica has the highest G + C content of both nuclear and kinetoplastic DNA thus far reported for trypanosomatids. The effects of ethidium bromide, acriflavin, proflavin, and 5-aminoacridine on the kinetoplast of L. tropica have been compared. Ethidium bromide and acriflavin, but not proflavin or 5-aminoacridine, induce dyskinetoplasty. L. tropica is one of the most sensitive trypanosomatids to ethidium bromide and acriflavin. Examination of the DNA from drug-treated cells in CsCl gradients revealed a loss of the satellite band after ethidium bromide or acriflavin treatment, but not after proflavin or 5-aminoacridine treatment. Cell division was required to produce these effects on the kinetoplast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1972.tb03557.x
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  • 6
    Electronic Resource
    Electronic Resource
    The Glucose Transport System in Leishmania tropica Promastigotes (1974)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 21 (1974), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Transport of glucose by Leishmania tropica promastigotes was measured by the uptake of the nonutilizable glucose analog, 2-deoxy-D-glucose (2-DOG), using the rapid filtration method. Both D-glucose and 2-DOG show identical rates of initial uptake. Intracellular 2-DOG readily exchanges with extracellular D-glucose and 2-DOG uptake is competitively inhibited by D-glucose. These observations suggest that both sugars are taken up by the same system. Neither the glucose analog α-methyl-D-glucoside (α-MG) nor 3-0-methyl glucose (3-0-MG) is taken up to any appreciable extent. Transport of 2-DOG shows saturation kinetics with a Vmax of 3.2 nmoles/mg cells/min and a Km of 0.16 mM. There is thus a stereospecific, carrier-mediated transport system for glucose uptake in L. tropica. About 2/3 of the intracellular pool following transport consists of 2-deoxy-D-glucose phosphate (2-DOG-P) and the remainder is free, unaltered 2-DOG.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.1974.tb03708.x
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  • 7
    Electronic Resource
    Electronic Resource
    Fluorescent in situ Detection of Encephalitozoon hellem Spores with a 6-Carboxyfluorescein-Labeled Ribosomal RNA-Targeted Oligonucleotide Probe (2000)
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 47 (2000), S. 0 
    Details
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: . A fluorescent in situ hybridization assay has been developed for the detection of the human-pathogenic microsporidian, Encephalitozoon hellem in water samples using epifluorescence microscopy. The assay employs a 19-nucleotide species-specific 6-carboxyfluorescein-labeled oligonucleotide probe, HEL878F, designed to be complementary to the nucleic acid sequence 878–896, a highly variable segment of the 16S ribosomal RNA of E. hellem spores. The specificity of this probe for its ribosomal RNA target site was confirmed using RNA degradation, ribosomal RNA target site competition, and nucleotide base mismatch control probe assays. Furthermore, the specificity of the HEL878F oligonucleotide probe for E, hellem spores was established when it was evaluated on spores from all three species of the genus Encephalitozoon that had been seeded in reagent water and environmental water concentrates. The specificity of the HEL878F oligonucleotide probe was further corroborated when tested on algae, bacteria, and protozoa commonly found in environmental water. The study demonstrates the applicability of a fluorescent in situ hybridization assay using a species-specific fluorescent-labeled oligonucleotide probe for the detection of E. hellem spores in water samples.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1550-7408.2000.tb00051.x
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  • 8
    Electronic Resource
    Electronic Resource
    Harvesting of leukocytes from intestinal lumen in murine giardiasis and preliminary characterization of these cells (1985)
    Springer
    Digestive diseases and sciences 30 (1985), S. 149-153 
    Details
    ISSN: 1573-2568
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aims of this study were to develop a method for harvesting leukocytes from the mouse small intestinal lumen and to identify leukocytes which enter the intestinal lumen of mice infected withGiardia muris. Giardia-infected and uninfectedBALB/c mice were anesthetized, and the small intestine was removed. The intestinal lumen was then flushed with nutrient medium, and the luminal washings were found to contain substantial numbers of mononuclear leukocytes. The number of these cells harvested from the intestinal lumen of 24 mice infected withGiardia muris was 22±3×104 (mean value ± standard error). Most of the cells were lymphocytes, although small numbers of macrophages were also obtained. By staining with monoclonal antibodies, approximately 50% of the intraluminal leukocytes were shown to be T lymphocytes. Similar numbers of mononuclear leukocytes were obtained from the intestinal lumen ofGiardia-infected and uninfected mice. However, T lymphocytes and other mononuclear cells from infected animals were frequently observed in contact withGiardia trophozoites. This observation is consistent with the hypothesis that intraluminal leukocytes play a part in gastrointestinal immune defense.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01308202
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