Microorganisms use specialized systems to export virulence factors into host cells. Secretion of effector proteins into the extracellular environment has been described in Trypanosoma cruzi; however, a comprehensive proteomic analysis of the secretome and the secretion mechanisms involved remain elusive. Here, we present evidence that T. cruzi releases proteins associated with vesicles that are formed by at least two different mechanisms. Transmission electron microscopy showed larger vesicles budding from the plasma membrane of noninfective epimastigotes and infective metacyclic trypomastigotes, as well as smaller vesicles within the flagellar pocket of both forms. Parasite conditioned culture supernatant was fractionated and characterized by morphological, immunochemical, and proteomic analyses. Three fractions were obtained by differential ultracentrifugation: the first enriched in larger vesicles resembling ectosomes, the second enriched in smaller vesicles resembling exosomes, and a third fraction enriched in soluble proteins not associated with extracellular vesicles. Label-free quantitative proteomic analysis revealed a rich collection of proteins involved in metabolism, signaling, nucleic acid binding, and parasite survival and virulence. These findings support the notion that T. cruzi uses different secretion pathways to excrete/secrete proteins. Moreover, our results suggest that metacyclic forms may use extracellular vesicles to deliver cargo into host cells.

Little is known about the fate, transport, and bioavailability of CeO2 nanoparticles (NPs) in soil. Moreover, there are no reports on the effect of surface coating upon NPs uptake by plants. In this study, Zea mays plants were grown for one month in unenriched and organic soils treated with coated and uncoated CeO2 NPs. In addition, plants were exposed to fluorescein isothiocyanate (FITC)-stained CeO2 NPs and analyzed in a confocal microscope. In organic soil, roots from uncoated and coated NPs at 100, 200, 400, and 800 mg kg−1 had 40, 80, 130, and 260% and 10, 70, 90, and 40% more Ce, respectively, compared to roots from unenriched soil. Conversely, shoots of plants from unenriched soil had significantly more Ce compared with shoots from organic soil. Confocal fluorescence images showed FITC-stained CeO2 NP aggregates in cell walls of epidermis and cortex, suggesting apoplastic pathway. The μXRF results revealed the presence of CeO2 NP aggregates within vascular tissues. To the authors knowledge this is the first report on the effects of surface coating and organic matter on Ce uptake from CeO2 NPs and upon the mechanisms of CeO2 NPs uptake by higher plants

The rapid development of nanotechnology will inevitably release nanoparticles (NPs) into the environment with unidentified consequences. In addition, the potential toxicity of CeO2 NPs to plants, and the possible transfer into the food chain, are still unknown. Corn plants (Zea mays) were germinated and grown in soil treated with CeO2 NPs at 400 or 800 mg/kg. Stress related parameters, such as: H2O2, catalase (CAT) and ascorbate peroxidase (APX) activity, heat shock protein 70 (HSP 70), lipid peroxidation, cell death and leaf gas exchange were analyzed at 10, 15, and 20 days post germination. Confocal laser scanning microscopy was used to image H2O2 distribution in corn leaves. Results showed that the CeO2 NP treatments increased accumulation of H2O2, up to day 15, in phloem, xylem, bundle sheath cells, and epidermal cells of shoots. The CAT and APX activities were also increased in the corn shoot, concomitant with the H2O2 levels. Both 400 and 800 mg/kg CeO2 NPs triggered the up regulation of the HSP 70 in roots, indicating a systemic stress response. None of the CeO2 NPs increased the level of thiobarbituric acid reacting substances, indicating that no lipid peroxidation occurred. CeO2 NPs, at both concentrations, did not induce ion leakage in either roots or shoots, suggesting membrane integrity was not compromised. Leaf net photosynthetic rate, transpiration, and stomatal conductance were not affected by CeO2 NPs. Our results suggest that the CAT, APX and HSP 70 might help the plants defend against CeO2 NPs induced oxidative injury and survive NP exposure.

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Cerium oxide nanoparticles (nCeO2) have been shown to have significant interactions in plants; however, there are limited reports on their impacts in rice (Oryza sativa). Given the widespread environmental dispersal of nCeO2, it is paramount to understand its biochemical and molecular impacts on a globally important agricultural crop, such as rice. This study was carried out to determine the impact of nCeO2 on the oxidative stress, membrane damage, antioxidant enzymes' activities, and macromolecular changes in the roots of rice seedlings. Rice seeds (medium amylose) were grown for 10 days in nCeO2 suspensions (0-500 mg L(-1)). Results showed that Ce in root seedlings increased as the external nCeO2 increased without visible signs of toxicity. Relative to the control, the 62.5 mg nCeO2 L(-1) reduced the H2O2 generation in the roots by 75%. At 125 mg nCeO2 L(-1), the roots showed enhanced lipid peroxidation and electrolyte leakage, while at 500 mg L(-1), the nCeO2 increased the H2O2 generation in roots and reduced the fatty acid content. The lignin content decreased by 20% at 500 mg nCeO2 L(-1), despite the parallel increase in H2O2 content and peroxidase activities. Synchrotron μ-XRF confirmed the presence of Ce in the vascular tissues of the roots.