Chagas Disease Megaesophagus Patients Carrying Variant MRPS18B P260A Display Nitro-Oxidative Stress and Mitochondrial Dysfunction in Response to IFN-γ Stimulus

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Chagas Disease Megaesophagus Patients Carrying Variant MRPS18B P260A Display Nitro-Oxidative Stress and Mitochondrial Dysfunction in Response to IFN-γ Stimulus
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Study links genetic variant to digestive disturbances in patients with Chagasdisease frontiersin

viable PBMCs were incubated in 400 µL of R20 medium ) supplemented with 20% inactivated FBS , 10 U/mL of penicillin, 100 µg/mL of streptomycin and 0.25 µg/mL of amphotericin B , with 200 µL of the EBV-enriched supernatant and 2.4 µL of cyclosporin A 1.0 mg/mL in a 1.9 cmfor seven days without disturbance.

The MitoSOX signal was measured using the software Columbus. First, we identified the total number of cells by using the nucleus probe Hoechst 33342. Then, dead cells were excluded by using a second probe that stains the nucleus green in dead cells. By excluding dead cells, we measured MitoSOX fluorescence intensity inside live cells at approximately 580 nm .

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