A novel ruthenium complex with 5-fluorouracil suppresses colorectal cancer stem cells by inhibiting Akt/mTOR signaling.

[Ru(5-FU)(PPh3)2(bipy)]PF6 (Ru/5-FU) is a novel ruthenium complex with 5-fluorouracil with promising potential against colorectal cancer (CRC). In the present study, we investigated the molecular mechanism of Ru/5-FU action in HCT116 CRC cells. Ru/5-FU exhibited potent cytotoxicity on a panel of cancer cell lines and on primary cancer cells and induced apoptosis in HCT116 CRC cells. Ru/5-FU reduced AKT1 gene transcripts, as well as the expression of Akt1 and Akt (pS473) and downstream Akt proteins mTOR (pS2448), S6 (pS235/pS236), 4EBP1 (pT36/pT45), GSK-3ß (pS9) and NF-κB p65 (pS529), but not Akt upstream proteins Hsp90 and PI3K p85/p55 (pT458/pT199), indicating an inhibitory action of Akt/mTOR signaling. Ru/5-FU increased LC3B expression and reduced p62/SQSTM1 levels, indicating autophagy induction. Curiously, the autophagy inhibitors 3-methyladenine and chloroquine increased Ru/5-FU-induced cell death, indicating an induction of cytoprotective autophagy by this compound. Ru/5-FU also reduced clonogenic survival, as well as the percentage of CD133+ cells and colonosphere formation, indicating that Ru/5-FU can suppress stem cells in HCT116 cells. Ru/5-FU inhibited cell migration and invasion in wound healing assays and Transwell cell invasion assays, along with a reduction in vimentin expression and an increase in E-cadherin levels, indicating that Ru/5-FU can interfere with epithelial-mesenchymal transition. Ru/5-FU also inhibited in vivo HCT116 cell development and experimental lung metastases in mouse xenograft models. Altogether, these results indicate that Ru/5-FU is an anti-CRC chemotherapy drug candidate with the ability to suppress stemness in CRC cells by inhibiting Akt/mTOR signaling.

New ruthenium-xanthoxylin complex eliminates colorectal cancer stem cells by targeting the heat shock protein 90 chaperone.

In this work, we describe a novel ruthenium-xanthoxylin complex, [Ru(phen)2(xant)](PF6) (RXC), that can eliminate colorectal cancer (CRC) stem cells by targeting the chaperone Hsp90. RXC exhibits potent cytotoxicity in cancer cell lines and primary cancer cells, causing apoptosis in HCT116 CRC cells, as observed by cell morphology, YO-PRO-1/PI staining, internucleosomal DNA fragmentation, mitochondrial depolarization, and PARP cleavage (Asp214). Additionally, RXC can downregulate the HSP90AA1 and HSP90B1 genes and the expression of HSP90 protein, as well as the expression levels of its downstream/client elements Akt1, Akt (pS473), mTOR (pS2448), 4EBP1 (pT36/pT45), GSK-3ß (pS9), and NF-κB p65 (pS529), implying that these molecular chaperones can be molecular targets for RXC. Moreover, this compound inhibited clonogenic survival, the percentage of the CRC stem cell subpopulation, and colonosphere formation, indicating that RXC can eliminate CRC stem cells. RXC reduced cell migration and invasion, decreased vimentin and increased E-cadherin expression, and induced an autophagic process that appeared to be cytoprotective, as autophagy inhibitors enhanced RXC-induced cell death. In vivo studies showed that RXC inhibits tumor progression and experimental metastasis in mice with CRC HCT116 cell xenografts. Taken together, these results highlight the potential of the ruthenium complex RXC in CRC therapy with the ability to eliminate CRC stem cells by targeting the chaperone Hsp90.

Assessing the cytotoxic potential of glycoalkaloidic extract in nanoparticles against bladder cancer cells.

OBJECTIVE: This study proposed to use the nanotechnology to deliver glycoalkaloidic extract (AE) to bladder cancer cells, evaluating their activity in 2D and 3D models and the biological mechanism of cell death. METHODS: NPs were prepared by nanoprecipitation method using polylactic acid (PLA) and characterized considering their size, charge, particle concentration and stability. The cytotoxicity was evaluated in 2D and 3D model, and the apoptosis and cell cycle were investigated using flow cytometry. KEY FINDINGS: NPs loading AE (NP-AE) had diameter around 125 ± 6 nm (PdI <0.1) and negative charge. The encapsulation efficiency of SM and SS was higher than 85% for both compounds. The obtained formulation showed a significant in-vitro cytotoxic effect against RT4 cells in a dose-dependent manner with IC50 two fold lower than the free AE. The cytotoxic effect of NP-AE was mediated by apoptosis and cell cycle arrested in the S phase. RT4 cells cultured under 3D conditions exhibited a higher resistance to the treatments (IC50 ~ three fold higher than in 2D cell culture). CONCLUSION: The NP-AE might be a promising nanocarrier to load and deliver glycoalkaloids against bladder cancer.

Oral health complications in Brazilian and French diabetic older people: A comparative study.

INTRODUCTION: Diabetes mellitus (DM) is a risk factor for periodontitis for over 40 years and novel evidence suggests that periodontitis has an impact on glycemic control in patients with diabetes. This study aimed to compare oral health complications in diabetic older patients from Brazil and France. METHODS: This cross-sectional study included 120 patients aged 65 and over diagnosed with type 2 diabetes. Sixty patients were admitted to a center for diabetes and hypertension care in Brazil and 60 patients were admitted to the Rouen University hospital. Dental conditions were assessed through the decayed, missing and filled teeth index and periodontal condition was assessed using the Community Periodontal Index. The significance threshold was p < 0.05. RESULTS: Decayed teeth differed statistically between the groups (p = 0.001). The French group presented more tooth mobility, gingival recession and furcation involvement (p < 0.001). Tooth brushing frequency differed significantly between the groups (p < 0.030). The main cause of missing teeth was periodontitis in the French group and caries and periodontitis in the Brazilian group (p < 0.001). Statistical significance was found for use of fixed upper (p = 0.013) and lower (p = 0.013) dentures in the French group. The French group needed upper denture rehabilitation (p = 0.010) while the Brazilian group needed lower denture rehabilitation (p = 0.003). CONCLUSION: Edentulism was prevalent in diabetic older people in both countries. However, the French participants presented with better oral health.

Multigenerational exposure of Folsomia candida to silver: Effect of different contamination scenarios (continuous versus pulsed and recovery).

Effects of pollutants are mostly assessed using standard testing procedures, which cover a fraction of the animals' life cycle. Although, in nature species are exposed during multiple generations of sub-lethal doses of persistent chemicals. In the present study, we focused on the multigenerational (MG) effects of silver in Folsomia candida during 6 generations using the EC50 for reproduction as exposure concentration. We tested 9 different exposure scenarios, going from continuous 6 generations Ag exposure over pulse exposure (i.e. one generation clean, next contaminated, next clean etc.) to gradually increasing the number of exposure generations, with a final transfer to clean media. The biological endpoints assessed included survival, reproduction and size, with reproduction being the most sensitive. The biological response depended on the specific MG scenario, e.g. the 6 Ag MG caused a decreased number of juveniles from F4, whereas the pulse exposure experienced an increase in reproductive output when in clean soil. It is uncertain whether Ag causes transgenerational effects, but the reproduction levels in both pulse exposures are lower than in continuous control over the 6 generations which could be due to transference of Ag by the maternal generation. Overall, population size distribution seemed to indicate a delay in time for egg laying, with close relationship between adult survival, organisms size and reproduction output. Size monitoring allowed significant added interpretation possibilities and we strongly recommend the addition of this endpoint to the standard guideline. The smaller observed size range can have implications in terms of adaptation potential, carrying associated increased risk.

Standardization and Prevalence of the Booster Phenomenon: Evaluation Using a Two-Step Skin Test with 43 kDa Glycoprotein in Individuals from an Endemic Region of Paracoccidioidomycosis.

BACKGROUND: We estimated the occurrence rate of the booster phenomenon by using an intradermal test with 43 kDa glycoprotein in an endemic area of paracoccidioidomycosis in the central-west region of Brazil. METHODS: Individuals who had a negative result on a survey performed by using an intradermal test with 43 kDa glycoprotein in an endemic area of paracoccidioidomycosis underwent a second intradermal test after 10-15 days to determine the presence or absence of the booster phenomenon. Statistical analyses were performed using the Chi-square test, Chi-square for linear trend test, Student's t test, and binomial test; p < 0.05 was considered significant. RESULTS: For the first time, we reported the occurrence of the booster phenomenon to an intradermal reaction caused by 43 kDa glycoprotein at a rate of 5.8-8.4%, depending on the test's cutoff point. This suggests that a cutoff point should be considered for the booster phenomenon in intradermal tests with 43 kDa glycoprotein: a difference of 6-7 mm between readings according to the first and second tests, depending on the purpose of the evaluation. CONCLUSION: The results indicate that the prevalence of paracoccidioidal infection in endemic areas is underestimated, as the booster phenomenon has not been considered in epidemiological surveys for this infection.

Transporte inter-hospitalar de recém-nascido em estado crítico: revisão integrativa da literatura / Inter-hospital transport of newborns in critical condition:integrative literature review

Modelo do estudo: revisão integrativa. Objetivo: identificar condutas organizacionais descritas na literatura sobre transporte inter-hospitalar de recém-nascido em estado crítico de saúde realizado por meio de ambulância. Metodologia: revisão integrativa com consulta nas bases de dados: LILACS, CINAHL, MEDLINE, Web of Science e SCOPUS, no período de setembro a novembro de 2013. Para realizar a pesquisa, foram utilizados os descritores não controlados newborn transport, newborn infants, protocol e os controlados transporte de pacientes, recém-nascido e ambulâncias. A busca limitou-se a artigos publicados em português, inglês e espanhol, sem período estabelecido, que versassem sobre transporte inter-hospitalar de recém-nascido crítico por ambulância. Resultados: selecionou-se 12 artigos. As condutas identificadas foram: comunicação pré-transporte; composição da equipe que realizará o transporte; referência e contra referência; seleção do veículo para o transporte; equipamentos e medicações que poderão ser utilizados durante o transporte. Conclusões: Os estudos encontrados versam sobre importantes condutas a serem aplicadas em todas as fases do transporte inter-hospitalar do recém nascido por ambulância, importantes para uma assistência de qualidade e efetiva...

Study model: integrative review. Objective: to identify organizational behaviors described in the literature about inter-hospital transport of newborns in critical health condition performed by means of ambulance. Methodology: integrative review with query in the databases: LILACS, CINAHL, MEDLINE, Web of Science and Scopus, during the period from September to November 2013. In order to perform the research, the non-controlled descriptors newborn transport’, ‘newborn infants’ and ‘protocol’ were used, aswell as the controlled descriptors transport of patients’, ‘newborn’ and ‘ambulances’. The search was limited to papers published in Portuguese, English and Spanish, without an established period, which addressed the inter-hospital transport of critically ill newborn by ambulance. Results: 12 papers were selected. The identified behaviors were: pre-transport communication; composition of the team that will conduct the transport; reference and counter-reference; selection of vehicle for transportation; equipment and medications that may be used during transportation. Conclusions: The selected studies address the important measures to be applied at all stages of the inter-hospital transport of newborns by ambulance, which are important for quality and effective care...

Optimization of the production of mycorrhizal inoculum on substrate with organic fertilizer.

The system for production of inoculum of arbuscular mycorrhizal fungi (AMF) using sand and vermiculite irrigated with nutrient solution is promising. However, organic amendments added to the substrate can stimulate sporulation of AMF and replace the nutrient solution. The aim of this study was to maximize the production of AMF (Acaulospora longula, Claroideoglomus etunicatum, Dentiscutata heterogama and Gigaspora albida) using selected organic substrates (vermicompost, coir dust and Tropstrato) together with sand and vermiculite. The production of spores varied among the tested AMF and according to the organic source added to the substrate. The vermicompost promoted higher sporulation of A. longula in relation to the other AMF and substrates. The Tropstrato inhibited the sporulation of D. heterogama while the reproduction of C. etunicatum was not affected by the organic compounds. The inoculum of A. longula also showed a high number of infective propagules and promoted biomass accumulation in maize plants. The system of inoculum production using sand and vermiculite + 10% vermicompost favors the production of infective inoculum of A. longula with the fungus benefiting growth of corn plants.

Impact of benzo(a)pyrene, Cu and their mixture on the proteomic response of Mytilus galloprovincialis.

In natural waters, chemical interactions between mixtures of contaminants can result in potential synergistic and/or antagonic effects in aquatic animals. Benzo(a)pyrene (BaP) and copper (Cu) are two widespread environmental contaminants with known toxicity towards mussels Mytilus spp. The effects of the individual and the interaction of BaP and Cu exposures were assessed in mussels Mytilus galloprovincialis using proteomic analysis. Mussels were exposed to BaP [10 µg L(-1) (0.396 µM)], and Cu [10 µg L(-1) (0.16 µM)], as well as to their binary mixture (mixture) for a period of 7 days. Proteomic analysis showed different protein expression profiles associated to each selected contaminant condition. A non-additive combined effect was observed in mixture in terms of new and suppressed proteins. Proteins more drastically altered (new, suppressed and 2-fold differentially expressed) were excised and analyzed by mass spectrometry, and eighteen putatively identified. Protein identification demonstrated the different accumulation, metabolism and chemical interactions of BaP, Cu and their mixture, resulting in different modes of action. Proteins associated with adhesion and motility (catchin, twitchin and twitchin-like protein), cytoskeleton and cell structure (α-tubulin and actin), stress response (heat shock cognate 71, heat shock protein 70, putative C1q domain containing protein), transcription regulation (zinc-finger BED domain-containing and nuclear receptor subfamily 1G) and energy metabolism (ATP synthase F0 subunit 6 protein and mannose-6-phosphate isomerase) were assigned to all three conditions. Cu exposure alone altered proteins associated with oxidative stress (glutathione-S-transferase) and digestion, growth and remodelling processes (chitin synthase), while the mixture affected only one protein (major vault protein) possibly related to multi drug resistance. Overall, new candidate biomarkers, namely zinc-finger BED domain-containing protein, chitin synthase and major vault protein, were also identified for BaP, Cu and mixture, respectively.

Improved in vitro and in vivo cutaneous delivery of protoporphyrin IX from PLGA-based nanoparticles.

We report the development of D, L lactic co-glycolic acid) (PLGA)-based nanoparticles (NPs) for topical delivery of protoporphyrin IX (PpIX), a photosensitizer (PS), in treatments like photodynamic therapy (PDT) of skin cancers. PpIX-NPs were obtained in ~75.0% yield, encapsulation efficiency of 67.7%, drug content of 50.3 µg mg(-1), average diameter of 290 nm maintained up to 30 days and a zeta potential of 32.3 mV. Sustained in vitro release of PpIX through artificial membranes following Higuchi kinetics was kept up to 10 days. In vitro retentions of PpIX both in stratum corneum (SC) and epidermis + dermis ([EP + D]) were higher from NPs (23.0 and 10.0 times, respectively) compared to control solutions at all times. Quantification of PpIX by extraction, after in vivo skin application of NPs-PpIX on hairless mice, showed higher retention of the PS both in SC and in [EP + D] (3.0 and 2.0 times, respectively) compared to control solutions. Taken together, the results indicate that NPs are suitable for PpIX encapsulation showing minimal permeation through the skin and a localized effect, characteristics of a potential and promising delivery system for PDT-associated treatments of skin cancers, photodiagnosis and their off-label uses.

Evaluation of Paracoccidioides brasiliensis infection by gp 43 intradermal test in rural settlements in Central-West Brazil.

Epidemiological studies of paracoccidioidomycosis have been based on surveys achieved with intradermal tests, and paracoccidioidin is the most common antigen used in most cases. The glycoprotein of 43-kDa (gp43) has been used in intradermal tests. It is the most antigenic component of Paracoccidioides brasiliensis, and it provides greater specificity to evaluate infection for this fungus. In this study, the prevalence of P. brasiliensis infection was estimated with intradermal tests involving gp43 for 695 people in rural Central-West Brazil. The infection rate was 45.8 % (95 % CI = 42.1-49.5), and the average age of those infected was 45.8 ± 18.2 years. The prevalence did not show gender-based differences but increased with age. The results demonstrate the importance of P. brasiliensis infection in rural settlements and the early exposure of children in the region to the fungus. Despite the high antigenicity and specificity of gp43, its usage must be standardized, so that epidemiological surveys will be comparable and more accurately reflect P. brasiliensis infection in endemic areas.

Mycorrhizal technology and phosphorus in the production of primary and secondary metabolites in cebil (Anadenanthera colubrina (Vell.) Brenan) seedlings.

BACKGROUND: The application of arbuscular mycorrhizal fungi (AMF) can increase the growth and concentration of primary and secondary metabolites in several plant species. Cebil (Anadenanthera colubrina), a medicinal plant, benefits from mycorrhizal association, but the influence of the symbiosis on the production of its bioactive compounds is unknown. In this study the effect of mycorrhizal inoculation and phosphorus (P) supply on the production of primary and secondary metabolites in cebil seedlings was determined. RESULTS: The production of proteins and carbohydrates in terms of both concentration and content was enhanced by inoculation with AMF, but this benefit was mitigated at higher levels of P (30 and 50 mg dm(-3) soil). The concentration of phenols, flavonoids and total tannins was favoured by mycorrhizal inoculation even at the highest levels of P (30 and 50 mg dm(-3) soil). CONCLUSION: The production of primary and secondary metabolites in leaves of A. colubrina can be maximised by mycorrhization, with the benefit depending on supplementation of soil phosphate.

Oral lesions in elderly patients of a community in Brazilian Northeast.

OBJECTIVE: To investigate the oral soft-tissue injuries in people aged 60 or older. METHODS: Cross-sectional research conducted with 262 elders aged 60-93 years (mean 69.84, SD ± 6.212) from the Dende community in Northeast, Brazil. Data were collected through an identification questionnaire and the Community Indicator in Oral Health. Data were computed by the Software SPSS, version 15. RESULTS: There was a predominance of female gender (64.1%), retired, low schooling and income of up to one minimum wage. It was detected a prevalence of soft-tissue injuries in 38.4% (n = 101) of the elders, distributed as follows: red spots 60(57.7%), blisters 20(19.2%), lesions and/or wound 16(15.4%), and white spot 8(7.7%). Among the elders, 40(15.3%) smoked and 20(7.6%) used alcohol. It was verified a statistical association between injuries and using prosthesis (p = 0.039), medicine (p = 0.023) and calculus (p = 0.016). CONCLUSION: The oral health of this population is precarious, and there is a high prevalence of oral lesions.

Biological evaluation of the bone healing process after application of two potentially osteogenic proteins: an animal experimental model.

OBJECTIVE: The aim of this work was to analyse qualitatively and quantitatively the newly formed bone after insertion of rhBMP-2 and protein extracted from Hevea brasiliensis (P-1), associated or not with a carrier in critical bone defects created in Wistar rat calvarial bone, using histological and histomorphometrical analyses. MATERIALS AND METHODS: Eighty-four male Wistar rats were used, divided into two groups, according to the period of time until the sacrifice (2 and 6 weeks). Each one of these groups was subdivided into six groups with seven animals each, according to the treatments: (1) 5 µg of pure rhBMP-2, (2) 5 µg of rhBMP-2/monoolein gel, (3) pure monoolein gel, (4) 5 µg of pure P-1, (5) 5 µg of P-1/monoolein gel and (6) critical bone defect controls. The animals were euthanised and the calvarial bone tissue removed for histological and histomorphometrical analyses. RESULT AND CONCLUSION: The results showed an improvement in the bone healing process using the rhBMP-2 protein, associated or not with a material carrier in relation to the other groups, and this process demonstrated to be time dependent.

Antioxidant and lipid peroxidation responses in Mytilus galloprovincialis exposed to mixtures of benzo(a)pyrene and copper.

This study aimed to assess the antioxidant system potential and lipid peroxidative effects, in the gill and digestive gland of Mytilus galloprovincialis exposed to individual and binary mixtures of benzo(a)pyrene (BaP) and Cu for 7 days. Data demonstrated that in mussels exposed to BaP antioxidant enzymes (catalase--CAT, total glutathione peroxidase--tGPx, glutathione S-transferase--GST and glutathione reductase--GR) and lipid peroxidation (LPO) increased in the gill. On the contrary, in the digestive gland inhibitory antioxidant effects (superoxide dismutase-SOD, GR, metallothioneins-MT) and no changes in LPO levels were detected. Cu was also a potent oxidant agent since MT and LPO levels increased in mussel gill, despite no LPO effect in the digestive gland. For both single contaminants the organ specificity and distinct physiologic/metabolism roles were evident in terms of antioxidant capacity. Gill SOD inhibition, MT and GST unchanged was a result of "simple independent action" of exposure to BaP and Cu. "Interactions" in the binary mixtures, led to absence of changes in LPO effects. In the digestive gland, BaP and Cu interactions were also responsible for the GST and LPO enhancement (antagonistic effects). The current findings demonstrate the differences in antioxidant responses where the organ dependency highlights each contaminant particular mode of action. Generally, in the gill "non-interactive" effects occurred with the lowest Cu concentration while "interactions" exist for the mixture with the highest Cu concentrations. In the digestive gland, "interactions" and "no interaction" effects occurred in all the binary mixtures. Complex contaminant mixtures interact differently based on target tissue which may lead to an imbalance in the mussels health status.

Comparison of metal accumulation between 'Artificial Mussel' and natural mussels (Mytilus galloprovincialis) in marine environments.

The passive sampler called 'Artificial Mussel' (AM) developed by Wu et al. (2007) can provide a time-integrated estimate of metal concentrations in the marine environment, and offers a potential device to assess and compare metal concentration in different marine environments worldwide. The aim of this study was to compare metal accumulation on AM and natural mussel Mytilus galloprovincialis at three sites with different metal loads along the Portuguese coast for four months. M. galloprovincialis were placed in cages alongside AMs at each site. Samples were collected monthly and Cd, Cr, Cu, Pb, and Zn concentrations in whole soft tissues and AMs compared. For both Cu and Cd, the results were similar between AMs and natural mussels. Higher concentrations of Zn were observed in natural mussels, whereas the inverse was shown for Pb (about 10-fold higher). Our results showed that AMs are promising tools for assessing metal concentrations in marine environments.

Golden grey mullet and sea bass oxidative DNA damage and clastogenic/aneugenic responses in a contaminated coastal lagoon.

Several xenobiotics or their metabolites have redox-cycling properties and potential to induce oxidative stress and DNA damage. The current work aimed to study, under environmental conditions, oxidative DNA damage (8-hydroxy-2'-deoxyguanosine (8-OHdG)) and its association with chromosomal damage measured as erythrocytic nuclear abnormalities (ENAs), in Liza aurata and Dicentrarchus labrax, caught at a costal lagoon (Ria de Aveiro, Portugal) having sites with different contamination profiles. The quantified parameters were also used to assess the lagoon's environmental status. Five critical sites were assessed comparing to a reference site. L. aurata displayed higher 8-OHdG levels and ENAs frequency, respectively, at Laranjo and Vagos. D. labrax 8-OHdG levels were higher at the sites where quantification was possible whereas no differences were found in terms of ENAs. No correlation was found in both species between 8-OHdG and ENAs. Despite no direct linkage between the two biomarkers was found, this study demonstrates species and site dependent genotoxic responses.

Evaluation of oxidative DNA lesions in plasma and nuclear abnormalities in erythrocytes of wild fish (Liza aurata) as an integrated approach to genotoxicity assessment.

Genetic lesions (8-hydroxy-2'-deoxyguanosine (8-OHdG) and erythrocytic nuclear abnormalities (ENA) were seasonally quantified in the blood of Liza aurata caught at Ria de Aveiro (Portugal), a multi-contaminated aquatic system. Thus, five critical sites were assessed and compared with a reference site (Torreira). Oxidative DNA damage was found in Gafanha (harbour-water area), Laranjo (metal-contaminated) and Vagos (contaminated with PAHs) in the spring; Rio Novo do Principe (near a former paper-mill effluent) in the autumn; Rio Novo do Principe and Vagos in the winter. ENA were higher than Torreira at VAG (spring and winter). Torreira did not display seasonal variation neither in terms of 8-OHdG or total ENA. A positive correlation between 8-OHdG and ENA was found, suggesting oxidative stress as a mechanism involved in the formation of ENA. This study clearly demonstrates the presence of DNA-damaging substances in Ria de Aveiro and recommends the use of 8-OHdG and ENA as biomarkers of environmental contamination.

Seasonal Liza aurata tissue-specific DNA integrity in a multi-contaminated coastal lagoon (Ria de Aveiro, Portugal).

In this study, the DNA integrity of golden grey mullet (Liza aurata) collected in differently contaminated sites of a coastal lagoon, Ria de Aveiro (Portugal), was assessed, over the period of 1 year, using the DNA alkaline unwinding assay, in four different tissues (gill, kidney, liver and blood) and compared to a reference site. The four tissues displayed different DNA integrity basal levels, clearly affected by seasonal factors. Gill and kidney were, respectively, the most and least sensitive tissues. All sites demonstrated the capacity to interfere with DNA integrity. The sites displaying the highest and lowest DNA damage capability were, respectively, Barra (subject to naval traffic) and Vagos (contaminated with polycyclic aromatic hydrocarbons). In terms of seasonal variability, autumn seems to be the more critical season (more DNA damage) unlike summer when no DNA damage was found in any tissue. Data recommend the continued monitoring of this aquatic system.

Antioxidant responses versus DNA damage and lipid peroxidation in golden grey mullet liver: a field study at Ria de Aveiro (Portugal).

The present work aimed to investigate golden grey mullet (Liza aurata) liver protection versus damage responses at a polluted coastal lagoon, Ria de Aveiro (Portugal), as a tool to evaluate the human impacts on environmental health at five critical sites in Ria de Aveiro (Portugal) in comparison to a reference site (Torreira; TOR). Protection was evaluated by measuring non-enzymatic [total glutathione (GSHt) and non-protein thiols (NPT)] and enzymatic [catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and glutathione reductase (GR)] antioxidant defenses. Damage was assessed as DNA integrity loss and lipid peroxidation (LPO). No significant differences were found between sites in terms of non-enzymatic defenses (GSHt and NPT). CAT did not display significant differences among sites. However, GPx at Barra (BAR, associated with naval traffic), Gafanha (GAF, harbor and dry-dock activities area), Laranjo (LAR, metal contaminated associated with chlor-alkali plant), and Vagos (VAG, contaminated by polycyclic aromatic hydrocarbons) was significantly lower than the reference site. GST was lower at GAF, Rio Novo do Príncipe (RIO, pulp mill effluent area), LAR, and VAG, whereas GR was lower at RIO. The loss of antioxidant defenses was paralleled by higher LPO levels only at GAF and VAG. However, no DNA integrity loss was found. Results highlight the importance of the adopted multibiomarkers as applied in the liver of L. aurata in coastal water pollution monitoring. The integration of liver antioxidant defense and damage responses can improve the aquatic contamination assessment.