Effect of MG-132 on gene expression profile in <i>Schistosoma mansoni</i> adult worms.

<p>Adult worm pairs were treated for 24 h with 50 μM MG-132. Microarrays were used to measure gene expression on a large scale. The figure shows a group of 1,919 genes with a statistically significant (<i>q-value</i> ≤ 0.025) differential expression in adult worms treated with MG-132 <i>versus</i> controls. Each horizontal line represents a gene and each column represents an experimental replicate. There are two technical replicates for each one of four biological replicates. Genes with transcription induced by treatment are shown in red, genes with repressed transcription are in green, and the color intensity is proportional to the log2 ratio (treated/control), as indicated by the color scale at the bottom.</p

Microarray results validation by real-time PCR.

<p>Validation is shown for a group of selected differentially expressed genes in <i>S</i>. <i>mansoni</i> adult worms treated with MG-132 compared with control parasites. Real time PCR data, expressed as Fold Change (normalized to the control group) are displayed as a bar graph while the corresponding data from the microarray (fold change) are shown below in numbers. The asterisk (*) indicates a statistically significant change (p < 0.05, t-test) when comparing treated with control samples.</p

Scanning electron microscopy of the tegument of <i>S</i>. <i>mansoni</i> adult worms.

<p>(A, x600) Normal morphology of adult worm tegument. (B, x1200) and (C, x3000) magnification of normal morphology of <i>S</i>. <i>mansoni</i> tegument showing the large number of tubercles (tu) and spines (sp). (D, x5000) Magnification of normal morphology of <i>S</i>. <i>mansoni</i> tegument showing spines (sp). (E, x600) Morphology of <i>S</i>. <i>mansoni</i> tegument after treatment with 50 μM MG-132 for 24 h. (F, x1200) Magnification showing tegumental changes in treated male adult worms: peeling (p), swelling (s), outbreak (o). (G, x3000) and (H, x5000) Magnification showing peeling (p) and bubbles (b) in the tegument promoted by MG-132.</p

Scanning electron microscopy of the tegument of <i>S</i>. <i>mansoni</i> adult worms.

<p>(A, x600) Normal morphology of adult worm tegument. (B, x1200) and (C, x3000) magnification of normal morphology of <i>S</i>. <i>mansoni</i> tegument showing the large number of tubercles (tu) and spines (sp). (D, x5000) Magnification of normal morphology of <i>S</i>. <i>mansoni</i> tegument showing spines (sp). (E, x600) Morphology of <i>S</i>. <i>mansoni</i> tegument after treatment with 50 μM MG-132 for 24 h. (F, x1200) Magnification showing tegumental changes in treated male adult worms: peeling (p), swelling (s), outbreak (o). (G, x3000) and (H, x5000) Magnification showing peeling (p) and bubbles (b) in the tegument promoted by MG-132.</p

Representative list of 30 induced genes in <i>S</i>. <i>mansoni</i> adult worms in response to MG-132 treatment.

<p>Representative list of 30 induced genes in <i>S</i>. <i>mansoni</i> adult worms in response to MG-132 treatment.</p

Effect of MG-132 on gene expression profile in <i>Schistosoma mansoni</i> adult worms.

<p>Adult worm pairs were treated for 24 h with 50 μM MG-132. Microarrays were used to measure gene expression on a large scale. The figure shows a group of 1,919 genes with a statistically significant (<i>q-value</i> ≤ 0.025) differential expression in adult worms treated with MG-132 <i>versus</i> controls. Each horizontal line represents a gene and each column represents an experimental replicate. There are two technical replicates for each one of four biological replicates. Genes with transcription induced by treatment are shown in red, genes with repressed transcription are in green, and the color intensity is proportional to the log2 ratio (treated/control), as indicated by the color scale at the bottom.</p

Number of genes with expression detected in each category generated by the analysis of microarray experiments.

<p>Number of genes with expression detected in each category generated by the analysis of microarray experiments.</p

Chemical composition and <i>in vitro</i> antileishmanial and cytotoxic activities of the essential oils of <i>Ocotea dispersa</i> (Nees) Mez and <i>Ocotea odorifera</i> (Vell) Rohwer (Lauraceae)

<p>We investigate the chemical composition and the <i>in vitro</i> antileishmanial and cytotoxic activities of the essential oils extracted from the leaves of <i>Ocotea dispersa</i> (Nees) Mez (OD-EO) and <i>Ocotea odorifera</i> (Vell) Rohwer (OO-EO). On the basis of GC-FID and GC-MS, α-eudesmol (20.9%), valencene (10.2%), δ-elemene (9.3%) and isospathulenol (7.3%) are the major constituents of OD-EO, whereas safrole (36.3%), γ-cadinene (6.6%), camphor (6.5%) and α-copaene (6.0%) are the main constituents of OO-EO. Both OD-EO and OO-EO display significant activity against the promastigote forms of <i>Leishmania amazonensis</i>, with IC₅₀ values of 4.67 ± 0.95 and 11.67 ± 2.16 μg/mL, respectively. The 50% cytotoxic concentration (CC₅₀) of OD-EO and OO-EO to mouse peritoneal macrophages is 26.77 ± 4.06 and 49.52 ± 1.04 μg/mL, respectively. This is the first report on the chemical composition of the essential oil extracted from the leaves of <i>O. dispersa</i>. Our results suggest that OD-EO and OO-EO are a promising source of new antileishmanial agents.</p

<i>In vitro</i> schistosomicidal activity of the lignan (−)-6,6′-dinitrohinokinin (DNHK) loaded into poly(lactic-co-glycolic acid) nanoparticles against <i>Schistosoma mansoni</i>

<p><b>Context:</b> (−)-6,6′-Dinitrohinokinin (DNHK) display remarkable antiparasitic activity and was, therefore, incorporated into a nanoparticle formulation.</p> <p><b>Objective:</b> Incorporation of DNHK in poly lactic-co-glycolic acid (PLGA) nanoparticles aiming to improve its biological activities.</p> <p><b>Materials and methods:</b> Synthesis, characterization and incorporation of DNHK into glycolic acid (PLGA) nanoparticles by nanoprecipitation method. The nanoparticles were characterized by ultraviolet-visible spectroscopy, X-ray diffraction, field emission electron microscopic scanning <i>mansoni</i> (FESEM), and dynamic light scattering (DLS). For the <i>in vitro</i> test with <i>Schistosoma mansoni</i>, the DNHK-loaded PLGA was diluted into the medium, and added at concentrations 10–200 µM to the culture medium containing one adult worm pair. The parasites were kept for 120 h and monitored every 24 h to evaluate their general condition, including: pairing, alterations in motor activity and mortality.</p> <p><b>Results:</b> The loaded PLGA nanoparticles gave an encapsulation efficiency of 42.2% and showed spherical characteristics in monodisperse polymeric matrix. The adult worm pairs were separated after 120 h of incubation for concentrations higher than 50 µM of DNHK-loaded PLGA. The groups incubated with 150 and 200 µM of DNHK-loaded PLGA for 24 and 120 h killed 100% of adult worms, afforded LC₅₀ values of 137.0 ± 2.12 µM and 79.01 ± 1.90 µM, respectively, which was similar to the effect displayed by 10 µM of praziquantel.</p> <p><b>Discussion and conclusions:</b> The incorporation of DNHK-loaded showed schistosomicidal activity and allowed its sustained release. The loaded PLGA system can be administered intravenously, as well as it may be internalized by endocytosis by the target organisms.</p

<i>In vitro</i> schistosomicidal activity of the lignan (−)-6,6′-dinitrohinokinin (DNHK) loaded into poly(lactic-co-glycolic acid) nanoparticles against <i>Schistosoma mansoni</i>

<p><b>Context:</b> (−)-6,6′-Dinitrohinokinin (DNHK) display remarkable antiparasitic activity and was, therefore, incorporated into a nanoparticle formulation.</p> <p><b>Objective:</b> Incorporation of DNHK in poly lactic-co-glycolic acid (PLGA) nanoparticles aiming to improve its biological activities.</p> <p><b>Materials and methods:</b> Synthesis, characterization and incorporation of DNHK into glycolic acid (PLGA) nanoparticles by nanoprecipitation method. The nanoparticles were characterized by ultraviolet-visible spectroscopy, X-ray diffraction, field emission electron microscopic scanning <i>mansoni</i> (FESEM), and dynamic light scattering (DLS). For the <i>in vitro</i> test with <i>Schistosoma mansoni</i>, the DNHK-loaded PLGA was diluted into the medium, and added at concentrations 10–200 µM to the culture medium containing one adult worm pair. The parasites were kept for 120 h and monitored every 24 h to evaluate their general condition, including: pairing, alterations in motor activity and mortality.</p> <p><b>Results:</b> The loaded PLGA nanoparticles gave an encapsulation efficiency of 42.2% and showed spherical characteristics in monodisperse polymeric matrix. The adult worm pairs were separated after 120 h of incubation for concentrations higher than 50 µM of DNHK-loaded PLGA. The groups incubated with 150 and 200 µM of DNHK-loaded PLGA for 24 and 120 h killed 100% of adult worms, afforded LC₅₀ values of 137.0 ± 2.12 µM and 79.01 ± 1.90 µM, respectively, which was similar to the effect displayed by 10 µM of praziquantel.</p> <p><b>Discussion and conclusions:</b> The incorporation of DNHK-loaded showed schistosomicidal activity and allowed its sustained release. The loaded PLGA system can be administered intravenously, as well as it may be internalized by endocytosis by the target organisms.</p