Antioxidant mechanisms to counteract TiO2-nanoparticles toxicity in wheat leaves and roots are organ dependent

Nanoparticles (NP) bioactivity is under deep scrutiny. In this work, the antioxidant response to TiO2-NP in wheat (Triticum aestivum) was determined. For that, enzymatic and the non-enzymatic antioxidants were evaluated in plants exposed to the P25 anatase:rutile material composed of TiO2-NP and under environmentally realistic doses (0; 5; 50; 150 mg/L for 20 days). Shoot but not root growth was reduced. In leaves, thiol metabolism and ascorbate accumulation were the preferred route whereas in roots the pre-existing antioxidant capacity was preferentially utilized. Both leaves and roots showed increased glutathione reductase and dehydroascorbate reductase activities and decreased ascorbate peroxidase activity. Roots, nevertheless, presented higher enzymatic basal levels than leaves. On the other hand, when examining non-enzymatic antioxidants, the ratio of reduced-to-oxidized glutathione (GSH/GSSG) increased in leaves and decreased in roots. Exposed leaves also presented higher total ascorbate accumulation compared to roots. TiO2-NP exposure down regulated, with more prominence in roots, antioxidant enzyme genes encoding catalase, ascorbate peroxidase, monodehydroascorbate reductase and dehydroascorbate reductase. In leaves, superoxide dismutase gene expression was increased. All data pinpoint to TiO2-NP toxicity above 5 mg/L, with aerial parts being more susceptible, which draws concerns on the safety doses for the use of these NPs in agricultural practices.publishe

Inorganic Hg toxicity in plants: a comparison of different genotoxic parameters

Inorganic Mercury (Hg) contamination persists an environmental problem, but its cyto- and genotoxicity in plants remains yet unquantified. To determine the extent of Hg-induced cyto- and genotoxicity, and assess most sensitive endpoints in plants, Pisum sativum L. seedlings were exposed for 14 days to different HgCl2 concentrations up to 100 μM. Shoots and roots from hydroponic exposure presented growth impairment and/or morphological disorders for doses >1 μM, being the roots more sensitive. Plant growth, ploidy changes, clastogenicity (HPCV), cell cycle dynamics (G1-S-G2), Comet-tail moment (TM), Comet-TD, Mitotic-index (MI) and cell proliferation index (CPI) were used to evaluate Hg-induced cyto/genotoxicity. Both leaf and root DNA-ploidy levels, assessed by flow cytometry (FCM), remained unaltered after exposure. Root cell cycle impairment occurred at lower doses (≥1 μM) than structural DNA damages (≥10 μM). Cytostatic effects depended on the Hg concentration, with delays during S-phase at lower doses, and arrests at G1 at higher ones. This arrest was paralleled with decreases of both mitotic index (MI) and cell proliferation index (CPI). DNA fragmentation, assessed by the Comet assay parameters of TD and TM, could be visualized for conditions ≥10 μM, while FCM-clastogenic parameter (FPCV) and micronuclei (MNC) were only altered in roots exposed to 100 μM. We demonstrate that inorganic-Hg induced cytostaticity is detectable even at 1 μM (a value found in contaminated sites), while structural DNA breaks/damage are only visualized in plants at concentrations ≥10 μM. We also demonstrate that among the different techniques tested for cyto- and genotoxicity, TD and TM Comet endpoints were more sensitive than FPCV or MNC. Regarding cytostatic effects, cell cycle analysis by FCM, including the difference in % cell cycle phases and CPI were more sensitive than MI or MNC frequency. Our data contribute to better understand Hg cyto- and genotoxicity in plants and to understand the information and sensitivity provided by each of the genotoxic techniques used.publishe

The Effect of Lycopene Preexposure on UV-B-Irradiated Human Keratinocytes

Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and RT-PCR: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer

“Quem ensina também aprende” : a formação pela prática de professores primários na província do Paraná

Resumo Segundo a historiografia da educação brasileira, muitas foram as ações relacionadas aos modos de formar professores primários durante o período imperial. Desses estudos, a maioria se centra na formação de professores atrelada à instituição das escolas normais, entretanto, há uma parcela menor de trabalhos que se propõem a discutir outro aspecto da formação de professores ao longo do século XIX, mais especificamente, a forma como sujeitos que não frequentaram esse espaço institucional (a escola normal), constituíram-se docentes primários. O artigo que aqui se apresenta partilha dessa perspectiva, e volta o olhar para os modos de formação pela prática de professores primários no Paraná na segunda metade do século XIX, por compreender que esse tipo de formação marcou um período em que a instrução pública estava se consolidando em meio a ações, deliberações, dificuldades e tensões, na tentativa de melhorias de sua condição. A pesquisa valeu-se da consulta da legislação educacional do período e de documentos advindos dos sujeitos envolvidos com a instrução pública, naquele momento, disponíveis no acervo do Arquivo Público do Paraná. No cotejamento e análise das fontes, é possível afirmar, que a formação pela prática dos professores primários na província do Paraná se deu no decorrer do desenvolvimento do processo de constituição do magistério primário

Proteomic Analysis of the Secretory Response of Aspergillus niger to D-Maltose and D-Xylose

Fungi utilize polysaccharide substrates through extracellular digestion catalyzed by secreted enzymes. Thus far, protein secretion by the filamentous fungus Aspergillus niger has mainly been studied at the level of individual proteins and by genome and transcriptome analyses. To extend these studies, a complementary proteomics approach was applied with the aim to investigate the changes in secretome and microsomal protein composition resulting from a shift to a high level secretion condition. During growth of A. niger on d-sorbitol, small amounts of d-maltose or d-xylose were used as inducers of the extracellular amylolytic and xylanolytic enzymes. Upon induction, protein compositions in the extracellular broth as well as in enriched secretory organelle (microsomal) fractions were analyzed using a shotgun proteomics approach. In total 102 secreted proteins and 1,126 microsomal proteins were identified in this study. Induction by d-maltose or d-xylose resulted in the increase in specific extracellular enzymes, such as glucoamylase A on d-maltose and β-xylosidase D on d-xylose, as well as of microsomal proteins. This reflects the differential expression of selected genes coding for dedicated extracellular enzymes. As expected, the addition of extra d-sorbitol had no effect on the expression of carbohydrate-active enzymes, compared to addition of d-xylose or d-maltose. Furthermore, d-maltose induction caused an increase in microsomal proteins related to translation (e.g., Rpl15) and vesicular transport (e.g., the endosomal-cargo receptor Erv14). Millimolar amounts of the inducers d-maltose and d-xylose are sufficient to cause a direct response in specific protein expression levels. Also, after induction by d-maltose or d-xylose, the induced enzymes were found in microsomes and extracellular. In agreement with our previous findings for d-xylose induction, d-maltose induction leads to recruitment of proteins involved in proteasome-mediated degradation

Proteomics of industrial fungi: trends and insights for biotechnology

Filamentous fungi are widely known for their industrial applications, namely, the production of food-processing enzymes and metabolites such as antibiotics and organic acids. In the past decade, the full genome sequencing of filamentous fungi increased the potential to predict encoded proteins enormously, namely, hydrolytic enzymes or proteins involved in the biosynthesis of metabolites of interest. The integration of genome sequence information with possible phenotypes requires, however, the knowledge of all the proteins in the cell in a system-wise manner, given by proteomics. This review summarises the progress of proteomics and its importance for the study of biotechnological processes in filamentous fungi. A major step forward in proteomics was to couple protein separation with high-resolution mass spectrometry, allowing accurate protein quantification. Despite the fact that most fungal proteomic studies have been focused on proteins from mycelial extracts, many proteins are related to processes which are compartmentalised in the fungal cell, e.g. β-lactam antibiotic production in the microbody. For the study of such processes, a targeted approach is required, e.g. by organelle proteomics. Typical workflows for sample preparation in fungal organelle proteomics are discussed, including homogenisation and sub-cellular fractionation. Finally, examples are presented of fungal organelle proteomic studies, which have enlarged the knowledge on areas of interest to biotechnology, such as protein secretion, energy production or antibiotic biosynthesis

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio