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Entamoeba histolytica HM-1:IMSS

ABSTRACT: RNA-Seq expression analysis of Entamoeba histolytica trophozoites adapted to grow in the presence of toxic amount of the Nitric Oxide donor S-Nitrosoglutathione (GSNO)

PROVIDER: PRJNA309317 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR3110550.fastq.gz	Fastqsanger.gz
	SRR3110551.fastq.gz	Fastqsanger.gz
	SRR3110552.fastq.gz	Fastqsanger.gz
	SRR3110553.fastq.gz	Fastqsanger.gz
	SRR3110554.fastq.gz	Fastqsanger.gz

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RNA-Seq expression analysis of Entamoeba histolytica trophozoites adapted to grow in the presence of toxic amount of the Nitric Oxide donor S-Nitrosoglutathione (GSNO)

Project description:Entamoeba histolytica is a protozoan parasite responsible for amebiasis, a disease which is characterized by acute inflammation of the colon. Adaptation of the parasite to toxic level of nitric oxide (NO) produced by phagocytes may be essential for the establishment of chronic amebiasis and for the survival of parasite within the host. To obtain insight into the mechanism of adaptation to NO, E. histolytica trophozoites were selected in vitro by stepwise exposures to increasing amounts of NO donor S-Nitrosoglutathione (GSNO) up to a concentration of 110 µM. These NO adapted trophozoites (NAT) were more resistance to acute exposure of GSNO (350μM), to activated macrophages and have a better capability to invade porcine colon explants compared to wild-type trophozoites. The transcriptome of NAT was investigated by RNA-sequencing (RNA-seq) and it showed only a weak overlapping with the transcriptome of trophozoites exposed to acute nitrosative stress (TEANS).

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Project description:The ability of Entamoeba histolytica to phagocytose host cells correlates to observed virulence in vivo. To better understand the mechanism of phagocytosis we used paramagnetic beads coated with host ligand and sorted trophozoites based on phagocytic ability. Gene expression was then measured in both the sorted phagocytic and non-phagocytic populations using a custom Affymetrix chip for E. histolytica. Feed forward regulation of phagocytosis by Entamoeba histolytica. Infection and Immunity. PMID 23045476 M280 Streptavidin Dynabeads (Invitrogen) were labeled with 20ug/mL biotinylated Human C1q (Quidel). Entamoeba histolytica (strain HM1) were washed twice with PBS then resuspended with the labeled beads at a 10:1 ratio of beads to trophozoites. Samples were incubated at 37Â°C for 45 minutes, washed twice with agitation to remove adherent beads, then resuspended in MACS buffer. Samples were loaded into magnetic columns (Miltenyi Biotec) and trophozoites were seperated according to manufacturer's protocols. phagocytic vs. non-phagocytic Entamoeba histolytica populations

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