Axenic culture of Giardia lamblia in TYI-S-33 Medium Supplemented with Bile
Transactions of the Royal Society of Tropical Medicine and Hygiene (Impact Factor: 1.84). 02/1983; 77(4):487-8. DOI: 10.1016/0035-9203(83)90120-7
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- "G. lamblia trophozoites assemblages A (human-derived) was offered by Prof. Zhaorong Lun from Southern China Research Center of Parasitic Biology, Sun Yat-Sen University, China, and the trophozoites were cultivated axenically with modified TYI-S-33 medium (Keister, 1983). Genomic DNAs were extracted from G. lamblia trophozoite assemblage A (human-derived) using DNA extraction kit (Promega, WI, USA). "
ABSTRACT: To study the genetic variation and prokaryotic expression of α18 giardin gene of Giardia lamblia zoonotic assemblage A and host-specific assemblage F, the α18 genes were amplified from G. lamblia assemblages A and F by PCR and sequenced. The PCR product was cloned into the prokaryotic expression vector pET-28a(+) and the positive recombinant plasmid was transformed into Escherichia coli Rosetta (DE3) strain for the expression. The expressed α18 giardin fusion protein was validated by SDS-PAGE and Western blot analysis, and purified by Ni-Agarose resin. The putative sequence of α18 giardin amino acid was analyzed by bioinformatics software. Results showed that the α18 giardin gene was 861 bp in length, encoding 286 amino acids; it was 100% homologous between human-derived and dog-derived G. lamblia assemblage A, but it was 86.8% homologous with G. lamblia assemblage F (cat-derived). Giardin α18 was about 36 kDa in molecular weight, with good reactivity. Prediction based on in silico analyses: it had hydrophobicity, without signal peptide and transmembrane domain, and contained 11 alpha regions, 13 beta sheets, 1 beta turn and 7 random coils in secondary structure. The above information would lay the foundation for research about the subcellular localization and biological function of α18 giardin in G. lamblia.
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- "Trophozoite Cultures, Growth of ABZ-Resistant Clones and Obtention of H 2 O 2 -Resistant Trophozoites Giardia duodenalis trophozoites of the WB strain (ATCC#30957) and ABZ-resistant clones were maintained in TYI-S-33 medium supplemented with 10% adult bovine serum (HyClone) and antibiotic/antimycotic solution (Thermo, USA) at 37 • C (Keister, 1983) in 4.5 mL screw-capped vials. ABZ-resistant trophozoites were selected by continuous subculture under increasing sublethal concentrations of ABZ (Sigma cat. "
ABSTRACT: The control of Giardia duodenalis infections is carried out mainly by drugs, among these albendazole (ABZ) is commonly used. Although the cytotoxic effect of ABZ usually involves binding to β-tubulin, it has been suggested that oxidative stress may also play a role in its parasiticidal mechanism. In this work the effect of ABZ in Giardia clones that are susceptible or resistant to different concentrations (1.35, 8, and 250 μM) of this drug was analyzed. Reactive oxygen species (ROS) were induced by ABZ in susceptible clones and this was associated with a decrease in growth that was alleviated by cysteine supplementation. Remarkably, ABZ-resistant clones exhibited partial cross-resistance to H 2 O 2, whereas a Giardia H 2 O 2-resistant strain can grow in the presence of ABZ. Lipid oxidation and protein carbonylation in ABZ-treated parasites did not show significant differences as compared to untreated parasites; however, ABZ induced the formation of 8OHdG adducts and DNA degradation, indicating nucleic acid oxidative damage. This was supported by observations of histone H2AX phosphorylation in ABZ-susceptible trophozoites treated with 250 μM ABZ. Flow cytometry analysis showed that ABZ partially arrested cell cycle in drug-susceptible clones at G2/M phase at the expense of cells in G1 phase. Also, ABZ treatment resulted in phosphatidylserine exposure on the parasite surface, an event related to apoptosis. All together these data suggest that ROS induced by ABZ affect Giardia genetic material through oxidative stress mechanisms and subsequent induction of apoptotic-like events.
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- "E. histolytica strain HM-1:IMSS, T. vaginalis strain GT-13, and G. lamblia strain 0989:IMSS were used in this study. E. histolytica and T. vaginalis were grown in PEHPS medium (Said-Fernández et al., 1988), and G. lamblia in TYI-S-33 supplemented with bile (Keister, 1983). All three species were subcultured three times each week. "
ABSTRACT: Background: Crataegus mexicana, Hyptis albida, Larrea tridentata, Ocimum baislicum, Prunus serotina, and Smilax spp. are used in Mexican traditional medicine to treat respiratory and gastrointestinal diseases such as flu, cough, diarrhea, dysentery, and other parasitic or microbial infections. Therefore this study was aimed at the pharmacological prospection of these plants against eleven bacterial species and three amitochondrial protist pathogens. Material and methods: The fruits or aerial parts of C. mexicana, H. albida, L. tridentata, O. baislicum, P. serotina, and Smilax spp. were extracted with different solvents. The antibacterial properties of organic and aqueous extracts of these plants were determined by the microdilution method and the microplate alamar blue assay against Stenotrophomonas maltophilia, Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae, Staphylococcus aureus, methicillin-resistant S. aureus, Listeria monocytogenes, Enterococcus faecalis, and Mycobacterium tuberculosis, whereas anti-protozoal activities of extracts were evaluated by a vial micro-assay against strains of Entamoeba histolytica, Trichomonas vaginalis, and Giardia lamblia. Results: H. albida, Smilax spp, and C. mexicana showed good activity against the Gram-positive strains, S. aureus, methicillin-resistant S. aureus, and E. faecalis. Four extracts (C. mexicana, H. albida, O. basilicum, and L. tridentata) showed good activity against E. histolytica, T. vaginalis, and G. lamblia. Conclusion: The extracts of these six medicinal plants could be a source for new antibacterial and antiprotozoal drugs. For this reason they are currently under investigation to isolate and characterize their active compounds.
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