RESUMO Na região Amazônica, o óleo de andiroba (Carapa guianensis) é um dos produtos mais utilizados pela população local devido às suas propriedades medicinais. Anteriormente, nosso grupo descreveu a atividade leishmanicida de três frações do óleo de andiroba ricas em limonóides. Este trabalho objetivou avaliar a atividade antioxidante e alterações ultraestruturais em Leishmania amazonensis tratadas com essas frações ricas em limonóides do óleo de andiroba: 6α,11β-diacetoxigedunina; 11β-hidroxigedunina; e uma fração com ambas 6α,11β-diacetoxigedunina + 11β-hidroxigedunina. A atividade antioxidante foi avaliada pelo método DPPH, e alterações ultraestruturais foram avaliadas por microscopia eletrônica de transmissão. A fração com ambas 6α,11β-diacetoxigedunina + 11β-hidroxigedunina exibiu a melhor atividade antioxidante (IC50 = 330,35 ± 8,60 µg mL-1; p = 0.0022) comparado com a fração 6α,11β-diacetoxigedunina. As três frações induziram alterações estruturais nas mitocôndrias e cinetoplasto de L. amazonensis, além de corpúsculos lipídicos, vacuolizações e vesículas na bolsa flagelar, indicando que as frações ricas em limonóides do óleo de andiroba podem induzir dano estrutural a Leishmania. Amazônica Carapa guianensis medicinais Anteriormente 6α,11βdiacetoxigedunina 6α11βdiacetoxigedunina αβdiacetoxigedunina 6α,11β diacetoxigedunina 6α 11β α β 11βhidroxigedunina βhidroxigedunina hidroxigedunina 11βhidroxigedunina. hidroxigedunina. DPPH transmissão IC50 IC (IC5 33035 330 35 330,3 860 8 60 8,6 mL1 mL 1 mL-1 0.0022 00022 0 0022 6α,11βdiacetoxigedunina. diacetoxigedunina. L lipídicos flagelar 11βdiacetoxigedunina βdiacetoxigedunina 6α11β αβ IC5 (IC 3303 33 3 330, 86 6 8, mL- 0.002 0002 002 0.00 000 00 0.0 0.

ABSTRACT In the Amazon region, andiroba (Carapa guianensis) oil is among the most used products by the local population due to its medicinal properties. Previously, our group described the leishmanicidal activity of three limonoid-rich fractions from andiroba oil. This work aimed to evaluate the antioxidant activity and ultrastructural changes in Leishmania amazonensis treated with these limonoid-rich fractions from andiroba oil: 6α,11β-diacetoxygedunin; 11β-hydroxygedunin; and a fraction with both 6α,11β-diacetoxygedunin +11β-hydroxygedunin. The antioxidant activity was evaluated by the DPPH method and ultrastructural changes were evaluated by transmission electron microscopy. The fraction with both 6α,11β-diacetoxygedunin + 11β-hydroxygedunin displayed the best antioxidant activity (IC50 330.35 ± 8.60 µg mL-1; p=0.0022) compared to the 6α,11β-diacetoxygedunin fraction. The three fractions induced structural changes to mitochondria and kinetoplasts of L. amazonensis, in addition to lipid bodies, vacuolization, and vesicles in the flagellar pocket, indicating that the limonoid-rich fractions from andiroba oil can induce structural damage to Leishmania. region Carapa guianensis properties Previously limonoidrich limonoid rich 6α,11βdiacetoxygedunin 6α11βdiacetoxygedunin αβdiacetoxygedunin 6α,11β diacetoxygedunin 6α 11β α β 11βhydroxygedunin βhydroxygedunin hydroxygedunin +11βhydroxygedunin. +11β hydroxygedunin. +11β-hydroxygedunin microscopy IC50 IC (IC5 33035 330 35 330.3 860 8 60 8.6 mL1 mL 1 mL-1 p=0.0022 p00022 p 0 0022 L bodies vacuolization pocket 11βdiacetoxygedunin βdiacetoxygedunin 6α11β αβ +11βhydroxygedunin IC5 (IC 3303 33 3 330. 86 6 8. mL- p=0.002 p0002 002 p=0.00 p000 00 p=0.0 p00 p=0. p0 p=0 p=

ABSTRACT A study with transmission electron microscopy of mycoplasma-contaminated HeLa cells using five cell donors referred to as donors A, B, C, D and E, observations are herein presented. Experiments performed with cells from donors B, C and D, revealed the presence of Mycoplasma hyorhinis after PCR and sequencing experiments. Bacteria probably originated from a cytoplasm with compacted tiny granular particles replacing the normal cytosol territories, or from the contact with the cytoplasm through a clear semi-solid material. The compact granularity (CG) of the cytoplasm was crossed by stripes of smooth and rough endoplasmic reticulum cisternae. Among apparently normal mitochondria, it was noted, in variable proportions, mitochondria with crista-delimited lucent central regions that expand to and occupied the interior of a crista-less organelle, which can undergo fission. Other components of the scenarios of mycoplasma-induced cell demolition are villus-like structures with associated 80-200 nm vesicles and a clear, flexible semi-solid, process-sensitive substance that we named jam-like material. This material coated the cytoplasmic surface, its recesses, irregular protrusions and detached cytoplasmic fragments. It also cushioned forming bacteria. Cyst-like structures were often present in the cytoplasm. Cells, mainly apoptotic, exhibiting ample cytoplasmic sectors with characteristic net-like profile due to adjoined vacuoles, as well as ovoid or elongated profiles, consistently appeared in all cells from the last four cell donors. These cells were named “modified host cells” because bacteria arose in the vacuoles. The possibility that, in some samples, there was infection and/or coinfection of the host cell by another organism(s) cannot be ruled out.

A microscopia confocal por varredura a laser vem se tornando extremamente útil na biologia celular e patologia. Com o uso de anticorpos monoclonais, pode ser uma poderosa ferramenta de diagnóstico assim como para estudos detalhados das diferentes formas do Trypanosoma cruzi em vários tecidos infectados.

Confocal scanning fluorescence microscopy has become widely used in cell biology and pathology. In conjunction with monoclonal antibodies it may turn out to be a powerful diagnostic tool that also enables detailed studies of tissue forms of Trypanosoma cruzi.