Uso de biomarcadores de contaminação ambiental na avaliação do impacto do vazamento de óleo derivado de petróleo em ecossistemas de água doce /

Orientador : Ciro Alberto de Oliveira RibeiroDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Ecologia e Conservaçao. Defesa: Curitiba, 2006Inclui bibliografiaÁrea de concentração : Ecotoxicologi

Polystyrene nanoplastics and microplastics can act as Trojan horse carriers of benzo(a)pyrene to mussel hemocytes in vitro

[EN]In this work we studied the ability of polystyrene (PS) nanoplastics (NPs) and microplastics (MPs) to transfer benzo(a)pyrene (BaP) to mussel hemocytes and to produce toxic effects in vitro. For this, intracellular fate and toxicity of PS NPs (0.05 μm) and MPs (0.5 and 4.5 μm) alone or with BaP and of BaP alone were assessed. Particles of 0.05 and 0.5 µm largely aggregated in the exposure medium whereas presence of BaP reduced particle aggregation. Cells internalized PS NPs and MPs alone or with BaP and these were found inside and outside lysosomes, depending on their size. PS particles alone or with BaP were cytotoxic to hemocytes only at the highest concentrations tested. The same was true for most sublethal endpoints except for increased phagocytic activity provoked by NPs and 0.5 μm MPs at lower concentrations. Plastic particles appeared to be the main drivers for reduced plasma membrane integrity and increased phagocytic and lysosomal activities whereas BaP appeared to contribute more to reduced cell viability and phagocytosis and increased ROS production and genotoxicity. Overall, PS NPs and MPs can act as carriers of BaP to mussel hemocytes, rising concerns about risks plastics associated to pollutants may pose to aquatic organisms.This work was carried out within the EU Project PLASTOX (JPI Oceans 005/2015) and was funded by the Spanish MINECO (NACE Project, CTM2016-81130-R) and Basque Government (Consolidated Research Groups Grant IT1302-19)

Functionalized Fluorescent Silica Nanoparticles for Bioimaging of Cancer Cells

Functionalized fluorescent silica nanoparticles were designed and synthesized to selectively target cancer cells for bioimaging analysis. The synthesis method and characterization of functionalized fluorescent silica nanoparticles (50–60 nm), as well as internalization and subcellular localization in HeLa cells is reported here. The dye, rhodamine 101 (R101) was physically embedded during the sol–gel synthesis. The dye loading was optimized by varying the synthesis conditions (temperature and dye concentration added to the gel) and by the use of different organotriethoxysilanes as a second silica precursor. Additionally, R101, was also covalently bound to the functionalized external surface of the silica nanoparticles. The quantum yields of the dye-doped silica nanoparticles range from 0.25 to 0.50 and demonstrated an enhanced brightness of 230–260 fold respect to the free dye in solution. The shell of the nanoparticles was further decorated with PEG of 2000 Da and folic acid (FA) to ensure good stability in water and to enhance selectivity to cancer cells, respectively. In vitro assays with HeLa cells showed that fluorescent nanoparticles were internalized by cells accumulating exclusively into lysosomes. Quantitative analysis showed a significantly higher accumulation of FA functionalized fluorescent silica nanoparticles compared to nanoparticles without FA, proving that the former may represent good candidates for targeting cancer cells.This research was funded by the Basque Government, grant numbers IT912-16 and IT-1302-19; Ministry of Economy and Competitiveness (MINECO), grant numbers MAT2017-83856-C3-3-P and CTM2016-81130-R; and the University of the Basque Country (UPV/EHU), grant number COLAB19/01

Cell and tissue level responses in mussels Mytilus galloprovincialis dietarily exposed to PVP/PEI coated Ag nanoparticles at two seasons

Silver nanoparticles (Ag NPs) are present in numerous consumer products due to their antimicrobial and other unique properties, thus concerns about their potential input into aquatic ecosystems are increasing. Toxicity of Ag NPs in waterborne exposed aquatic organisms has been widely investigated, but studies assessing the potential toxic effects caused after ingestion through the food web, especially at low realistic concentrations, remain scarce. Moreover, it is not well known whether season may influence toxic effects of Ag NPs. The main objective of this study was to determine cell and tissue level responses in mussels Mytilus galloprovincialis dietarily exposed to poly-N-vinyl-2-pirrolidone/polyethyleneimine (PVP/PEI) coated 5 nm Ag NPs for 1, 7 and 21 days both in autumn and spring. Mussels were fed every day with microalgae Isochrysis galbana exposed for 24 h to a low dose (1 mu g Ag/L Ag NPs) in spring and to a higher dose (10 mu g Ag/L Ag NPs) in spring and autumn. Mussels fed with microalgae exposed to the high dose accumulated Ag significantly after 21 days in both seasons, higher levels being measured in autumn compared to spring. Intralysosomal metal accumulation measured in mussel digestive gland and time- and dose-dependent reduction of mussels health status was similar in both seasons. DNA strand breaks increased significantly in hemocytes at both exposure doses along the 21 days in spring and micronuclei frequency showed an increasing trend after 1 and 7 days of exposure to 1 mu g Ag/L Ag NPs in spring and to 10 mu g Ag/L in both seasons. Values decreased after 21 days of exposure in all the cases. In conclusion, PVP/PEI coated 5 nm Ag NPs ingested through the food web were significantly accumulated in mussel tissues and caused adverse cell and tissue level effects both in autumn and in spring.This work has been funded by the Spanish Ministry of Economy and Competitiveness (Nano Silver Omics project MAT2012-39372), Basque Government (SAIOTEK project S-PE13UN142 and Consolidated Research Group GIC IT810-13 and IT1302-19), the University of the Basque Country UPV/EHU (UFI 11/37 and PhD fellowship to N.D.) and French Ministry of Higher Education and Research (PhD fellowship to M.M.)

Biomarkers responses in fish (Atherinella brasiliensis) of paranaguá bay, southern Brazil, for assessment of pollutant effects

Paranaguá bay is a complex estuary located in southern Brazil containing three protected areas listed by UNESCO. Historically, the estuary has been affected by urban, industrial, agricultural and harbor activities, and occasional accidents. Specifically, the explosion of the Chilean ship Vicuña in December 2004 spilled methanol and crude and fuel oils which affected both protected and non-protected areas. The present study sought to investigate the pollution threat to aquatic organisms in order to evaluate the potential effects of pollutants. One hundred and twenty adult fish Atherinella brasiliensis were collected from different sites within Paranaguá estuary, including the harbor and open ocean, during summer, autumn and winter of 2005. Among the biomarkers, the somatic index, chemical analysis of bile, biochemical, genetic and morphological parameters were considered. Chemical analysis of bile showed a continuous bioavailability of polycyclic aromatic hydrocarbons (PAHs) according to proximity to the harbor site. The histopathological findings have demonstrated aconsiderable incidence of severe pathologies in the liver and gills, corroborated by biochemical disturbances and genetic damage. These findings indicate that more studies are necessary to evaluate both water quality and fish health so as to permit a better analysis of the impact of pollution in Paranaguá estuary.A Baia de Paranaguá é um complexo estuarino localizado no sul do Brasil constituído de três áreas de proteção ambiental listadas pela UNESCO. Historicamente, o estuário tem sido afetado por atividade urbana, industrial, agricultura e portuária, e eventualmente por acidentes. Particularmente a explosão do navio Chileno Vicuña em dezembro de 2004 derramou metanol, óleo cru e combustível atingindo áreas protegidas e não protegidas. O presente estudo tem por objetivo investigar a poluição em organismos aquáticos. Cento e vinte indivíduos adultos do peixe Atherinella brasiliensis foram coletados em quatro diferentes pontos de coleta no estuário de Paranaguá, partindo do porto até o oceano aberto nos períodos de verão, inverno e primavera de 2005. Os índices somáticos, parâmetros químicos, enzimáticos, genéticos e morfológicos foram considerados. As análises histopatológicas demonstraram expressiva incidência de patologia no fígado e nas brânquias algumas vezes corroboradas pelas alterações bioquímicas. Danos genéticos e anormalidades genéticas também foram observados. As análises químicas na bile mostraram uma contínua biodisponibilidade de hidrocarbonetos policíclicos aromáticos para os organismos aquáticos. Os dados obtidos indicam que a qualidade da água e a saúde dos peixes encontram-se bastante comprometidos no estuário de Paranaguá

A comparison of dermal toxicity models; assessing suitability for safe(r)-by-design decision-making and for screening nanomaterial hazards

The objective of Safe-by-Design (SbD) is to support the development of safer products and production processes, and enable safe use throughout a materials' life cycle; an intervention at an early stage of innovation can greatly benefit industry by reducing costs associated with the development of products later found to elicit harmful effects. Early hazard screening can support this process, and is needed for all of the expected nanomaterial exposure routes, including inhalation, ingestion and dermal. In this study, we compare in vitro and ex vivo cell models that represent dermal exposures (including HaCaT cells, primary keratinocytes, and reconstructed human epidermis (RhE)), and when possible consider these in the context of regulatory accepted OECD TG for in vitro dermal irritation. Various benchmark nanomaterials were used to assess markers of cell stress in each cell model. In addition, we evaluated different dosing strategies that have been used when applying the OECD TG for dermal irritation in assessment of nanomaterials, and how inconsistencies in the approach used can have considerable impact of the conclusions made. Although we could not demonstrate alignment of all models used, there was an indication that the simpler in vitro cell model aligned more closely with RhE tissue than ex vivo primary keratinocytes, supporting the use of HaCaT cells for screening of dermal toxicity of nanomaterials and in early-stage SbD decision-making

Red haloBODIPYs as theragnostic agents: The role of the substitution at meso position

Three different molecular designs based on BODIPY dye have been proposed as photosensitizers (PSs) for photodynamic therapy (PDT) by the inclusion of halogen atoms (Iodine) at 2,6-positions and with extended conjugation at 3, 5-positions and varying the substitution at meso position. The synthesis is described and their main photophysical features including singlet oxygen production and triplet states were characterized by absorption and fluorescence spectroscopy (steady-state and time-correlated) and nanosecond transient absorption spectroscopy. The results were compared with the commercial Chlorin e6. The three new red-halogen-BODIPYs showed a great balance between singlet oxygen generation (Phi(Delta)>= 0.40) and fluorescence (Phi(fl)>= 0.22) for potential application on PDT, and particularly in theragnosis. In vitro experiments in HeLa cells were done to study their performance and to elucidate the best potential candidate for PDT.This research was funded by the Basque Government, grant numbers IT912-16, IT-1302-19 and IT1639-22. This work is supported by Min-isterio de Ciencia, Innovacion y Universidades-Agencia Estatal de Investigacion (MCI/AEI) , grant numbers MAT2017-83856-C3-2-P and 3-P, PID2020-114347RB-C32, PID2020-114755 GB-C32 and the Uni-versity of the Basque Country (UPV/EHU) , grant number COLAB19/01. R.P.M. thanks UPV/EHU for postdoctoral felowship (DOCREC 20/55) . Open Access funding is provided by University of Basque Country

Functionalization of photosensitized silica nanoparticles for advanced photodynamic therapy of cancer

BODIPY dyes have recently attracted attention as potential photosensitizers. In this work, commercial and novel photosensitizers (PSs) based on BODIPY chromophores (haloBODIPYs and orthogonal dimers strategically designed with intense bands in the blue, green or red region of the visible spectra and high singlet oxygen production) were covalently linked to mesoporous silica nanoparticles (MSNs) further functionalized with PEG and folic acid (FA). MSNs approximately 50 nm in size with different functional groups were synthesized to allow multiple alternatives of PS-PEG-FA decoration of their external surface. Different combinations varying the type of PS (commercial Rose Bengal, Thionine and Chlorine e6 or custom-made BODIPY-based), the linkage design, and the length of PEG are detailed. All the nanosystems were physicochemically characterized (morphology, diameter, size distribution and PS loaded amount) and photophysically studied (absorption capacity, fluorescence efficiency, and singlet oxygen production) in suspension. For the most promising PS-PEG-FA silica nanoplatforms, the biocompatibility in dark conditions and the phototoxicity under suitable irradiation wavelengths (blue, green, or red) at regulated light doses (10–15 J/cm2) were compared with PSs free in solution in HeLa cells in vitro

Red haloBODIPYs as theragnostic agents: The role of the substitution at meso position

Three different molecular designs based on BODIPY dye have been proposed as photosensitizers (PSs) for photodynamic therapy (PDT) by the inclusion of halogen atoms (Iodine) at 2,6-positions and with extended conjugation at 3, 5-positions and varying the substitution at meso position. The synthesis is described and their main photophysical features including singlet oxygen production and triplet states were characterized by absorption and fluorescence spectroscopy (steady-state and time-correlated) and nanosecond transient absorption spectroscopy. The results were compared with the commercial Chlorin e6. The three new red-halogen-BODIPYs showed a great balance between singlet oxygen generation (ΦΔ≥0.40) and fluorescence (Φfl≥0.22) for potential application on PDT, and particularly in theragnosis. In vitro experiments in HeLa cells were done to study their performance and to elucidate the best potential candidate for PDT

Evidence of contamination by oil and oil products in the Santos-Sao Vicente estuary, Sao Paulo, Brazil

Different components of the mixed function oxidase (MFO) system and the levels of fluorescent aromatic compounds in bile (FACs) were measured in Cathorops spixii in order to assess the impact of polycyclic aromatic hydrocarbons (PAHs). Fish were sampled in an estuary (Santos/Sao Vicente) with a history of contamination by PAHs, mainly due to the presence of the industrial complex of Cubatao city and of another of low anthropogenic influence (Cananeia) on the Brazilian coast. FACs were higher in fish from the polluted site, and the PAH 5 and 6-ring metabolites were the most frequent - with 14% and 15%, respectively. Levels of the different components of the MFO system showed the same variation profile as the FACs for both estuaries. Therefore, the values found for somatic indexes and biomarkers with data of bile PAH metabolites indicate the presence of organic contaminants, especially in the area subject to the influence of the industrial complex on the Santos/Sao Vicente estuary.CAPES (Brazilian Agency for Science and Technology); Oceanographic Institute of Sao Paulo University; Laboratory of Ecotoxicology and Environmental Chemistry of the University of Algarve; CAPES-PDEE [BEX 2176/07-6]info:eu-repo/semantics/publishedVersio