The role of glucose in the pathology of EHEC O157: H7
The pathogen enterohemorrhagic Escherichia coli (EHEC) O157: H7 is responsible for hemorrhagic colitis and hemolytic uremic syndrome in humans [1]. During the colonization process in the gastrointestinal tract, EHEC needs to adapt to changes in nutrient availability [2]. The objective of this stu...
| Autores principales: | , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Cambridge University Press
2021
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/10224 https://www.cambridge.org/core/journals/microscopy-and-microanalysis/article/role-of-glucose-in-the-pathology-of-ehec-o157-h7/4F5327F46D17EE9933438979E0134B28 https://doi.org/10.1017/S1431927620001129 |
| Sumario: | The pathogen enterohemorrhagic Escherichia coli (EHEC) O157: H7 is responsible for hemorrhagic
colitis and hemolytic uremic syndrome in humans [1]. During the colonization process in the
gastrointestinal tract, EHEC needs to adapt to changes in nutrient availability [2]. The objective of this
study was to evaluate the influence of glucose on physiology and processes involved in the pathogenesis
of EHEC O157: H7 in order to improve our understanding of the mechanisms controlling EHEC growth
and survival in the bovine gut.
In this study we first analyzed the growth rate of EHEC O157: H7 Rafaela II clade 8, a strain isolated
from a bovine in Argentina, grown in the medium DMEM supplemented with either 4.5% glucose (Highglucose - DHG) or 1% glucose (Low-glucose - DLG). In addition, we assessed the bacterial adhesion
capacity and actin pedestal formation induced by EHEC [3] by performing infection assays. For this
purpose, Caco-2 epithelial cells were exposed for 5 h with Rafaela II grown with the different
concentrations of glucose. Subsequently, the samples were fixed (paraformaldehyde 4%) and
permeabilized (triton); actin and nucleic acids (DNA) were stained with rhodamine-phalloidin and TOPRO-3, respectively. Bacterial adhesion capacity and pedestal formation of cells were evaluated using a
Leica TCS SP5 laser scanning confocal microscope (MC). Each Image was acquired by monitoring a
single focal plane over time (xyt scanning mode) using a 40X/1.25 oil objective lens and 543nm HeNe
and 633 nm HeNe lasers. The frequency and resolution for acquiring images were set at 200 Hz and 1,024
x 1,024 pixels, while maintaining the same settings for laser powers, gain, and offset. |
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