Journal: ACS Omega

Article Title: Isoxazole Derivative Induces Apoptosis-like Death and Autophagy through Oxidative Stress in Leishmania amazonensis

doi: 10.1021/acsomega.5c11341

Figure Lengend Snippet: Optical microscopy of J774A.1 macrophages infected with L. amazonensis amastigotes (arrows) and treated with isoxazole 4 for 48 h. Untreated cells (A). Cells treated with 1 μM (B) and 0.1 μM (C) of isoxazole 4 . Scale bars: 10 μm.

Article Snippet: J774A.1 murine macrophages (Rio de Janeiro Cell Bank, Rio de Janeiro, Brazil) and L929 fibroblasts (clone NCTC 929, L cell, L-929; ATCC CCL1, Manassas, VA, USA) were cultured in supplemented RPMI 1640 medium (10% FBS, 5000 U mL –1 penicillin, and 5 mg mL –1 streptomycin) and supplemented DMEM (2 mM l -glutamine, 10% FBS, 5000 U mL –1 penicillin, and 5 mg mL –1 streptomycin), respectively, at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ).

Techniques: Microscopy, Infection

Journal: ACS Omega

Article Title: Isoxazole Derivative Induces Apoptosis-like Death and Autophagy through Oxidative Stress in Leishmania amazonensis

doi: 10.1021/acsomega.5c11341

Figure Lengend Snippet: Morphological and ultrastructural alterations in intracellular amastigotes of L. amazonensis treated with IC 50 (0.96 μM) and 2 × IC 50 (1.92 μM) of isoxazole 4 for 48 h, analyzed by scanning electron microscopy (SEM) (A–F) and transmission electron microscopy (TEM) (A′–F′). SEM images show untreated amastigotes within the parasitophorous vacuole of macrophages with normal morphology (a) (A, B). A reduced number of amastigotes is observed following treatment with IC 50 (C-E) and 2 × IC 50 (F) of isoxazole 4 (white arrows). TEM images show intracellular amastigotes inside macrophages, with untreated parasites displaying typical cellular structures, including the endoplasmic reticulum (er), Golgi complex (Gc), nucleus (n), mitochondrion (m), and the macrophage nucleus (nM) (A′, B′). Treated amastigotes show accumulation of lipid droplets (*) and an increase in autophagic vacuoles (▶) at IC 50 (C′–E′) and 2 × IC 50 (F′). Scale bars: 5 μm (A–F), 1 μm (A′–C′), 0.5 μm (D′–F′).

Article Snippet: J774A.1 murine macrophages (Rio de Janeiro Cell Bank, Rio de Janeiro, Brazil) and L929 fibroblasts (clone NCTC 929, L cell, L-929; ATCC CCL1, Manassas, VA, USA) were cultured in supplemented RPMI 1640 medium (10% FBS, 5000 U mL –1 penicillin, and 5 mg mL –1 streptomycin) and supplemented DMEM (2 mM l -glutamine, 10% FBS, 5000 U mL –1 penicillin, and 5 mg mL –1 streptomycin), respectively, at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ).

Techniques: Electron Microscopy, Transmission Assay