The EU Interreg Project "GEREMIA" on waste management for the improvement of port waters: results on monitoring the health status of fish as bioindicator.

Highly anthropized areas as ports represent complex scenarios that require accurate monitoring plans aimed to address the environmental status. In this context, the activities of the EU Interreg Project "GEstione dei REflui per il MIglioramento delle Acque portuali (GEREMIA)" were focused on comparing sites differently affected by human presence, as the Port of Genoa and the natural area of the S'Ena Arrubia fishpond: a panel of analyses was carried out on Mugilidae fish sampled in these two areas, aimed to address trace metal accumulation in the liver, gills, and muscle, as well as cytochrome P450 (CYP450) induction in liver and biliary polycyclic aromatic hydrocarbon (PAH) metabolites, and histopathological alterations in the liver and gills. Chemical analyses in the liver, gills, and muscle of specimens collected in the port area showed an overall higher degree of trace metal contamination compared to the natural fishpond, and similar results were obtained in terms of CYP450 induction and biliary PAH metabolites, suggesting a higher exposure to organic compounds. In addition, histopathological analyses revealed a significant alteration and then a loss of functionality of liver and gill tissue in individuals from the port. Overall, this study describes the complex environmental pollution scenario in the Port of Genoa, confirming the importance of using multidisciplinary approaches and different types of analyses to address both the presence and the effects of contaminants in marine environments.

System biology mediated assessment of molecular mechanism for sinapic acid against breast cancer: via network pharmacology and molecular dynamic simulation.

Sinapic acid is a hydroxycinnamic acid widespread in the plant kingdom, known to be a potent anti-oxidant used for the treatment of cancer, infections, oxidative stress, and inflammation. However, the mode of action for its chemotherapeutic properties has yet not been unleashed. Hence, we aimed to identify potential targets to propose a possible molecular mechanism for sinapic acid against breast cancer. We utilized multiple system biology tools and databases like DisGeNET, DIGEP-Pred, Cytoscape, STRING, AutoDock 4.2, AutoDock vina, Schrodinger, and gromacs to predict a probable molecular mechanism for sinapic acid against breast cancer. Targets for the disease breast cancer, were identified via DisGeNET database which were further matched with proteins predicted to be modulated by sinapic acid. In addition, KEGG pathway analysis was used to identify pathways; a protein-pathway network was constructed via Cytoscape. Molecular docking was performed using three different algorithms followed by molecular dynamic simulations and MMPBSA analysis. Moreover, cluster analysis and gene ontology (GO) analysis were performed. A total of 6776 targets were identified for breast cancer; 95.38% of genes predicted to be modulated by sinapic acid were common with genes of breast cancer. The 'Pathways in cancer' was predicted to be modulated by most umber of proteins. Further, PRKCA, CASP8, and CTNNB1 were predicted to be the top 3 hub genes. In addition, molecular docking studies revealed CYP3A4, CYP1A1, and SIRT1 to be the lead proteins identified from AutoDock 4.2, AutoDock Vina, and Schrodinger suite Glide respectively. Molecular dynamic simulation and MMPBSA were performed for the complex of sinapic acid with above mentioned proteins which revealed a stable complex throughout simulation. The predictions revealed that the mechanism of sinapic acid in breast cancer may be due to regulation of multiple proteins like CTNNB1, PRKCA, CASP8, SIRT1, and cytochrome enzymes (CYP1A1 & CYP3A4); the majorly regulated pathway was predicted to be 'Pathways in cancer'. This indicates the rationale for sinapic acid to be used in the treatment of breast cancer. However, these are predictions and need to be validated and looked upon in-depth to confirm the exact mechanism of sinapic acid in the treatment of breast cancer; this is future scope as well as a drawback of the current study.

Comparative toxicity of conventional versus compostable plastic consumer products: An in-vitro assessment.

This study investigates the toxicity of methanolic extracts obtained from compostable plastics (BPs) and conventional plastics (both virgin and recycled). Additionally, it explores the potential influence of plastic photodegradation and composting on toxic responses using a battery of in vitro assays conducted in PLHC-1 cells. The extracts of BPs, but not those of conventional plastics, induced a significant decrease in cell viability (<70%) in PLHC-1 cells after 24 h of exposure. Toxicity was enhanced by either photodegradation or composting of BPs. Extracts of conventional plastics, and particularly those of recycled plastics, induced 7-ethoxyresorufin-O-deethylase (EROD) activity and micronucleus formation in exposed cells, indicating the presence of significant amounts of CYP1A inducers and genotoxic compounds in the extracts, which was enhanced by photodegradation. These findings highlight the importance of investigating the effects of degradation mechanisms such as sunlight and composting on the toxicity of BPs. It is also crucial to investigate the composition of newly developed formulations for BPs, as they may be more harmful than conventional ones.

Assessment of cytochrome P450 1A1 gene polymorphism and vitamin A serum level in psoriasis vulgaris.

Psoriasis is characterized by cutaneous hyperproliferation, secondary to immune system dysregulation. Vitamin A regulates the immune response and sustains epithelial tissue hemostasis. The CYP1A1 gene, has many biological actions, including vitamin A metabolism. To evaluate CYP1A1 gene polymorphism and serum vitamin A level in patients with psoriasis vulgaris, a case-control study involving two groups was conducted: group 1 (45 patients with psoriasis vulgaris) served as the cased group and group 2 (45 healthy participants who were sex and age matched) acted as the control group. CYP1A1 (rs1048943) gene polymorphism and vitamin A serum level were assessed by TaqMan allelic discrimination (PCR) and ELISA, respectively. AG genotype was present only in cases (22.2%), while AA genotype was present in all controls (P=.001). Vitamin A levels were lower in cases than in controls (32.0 ± 7.41 vs. 46.2 ± 15.7 µg/ml, respectively) (P<.001). AG genotype was associated with a lower vitamin A level (P=.001). The detected genotype difference between psoriasis patients and controls, which was associated with a lower serum vitamin A level and was also lower in more severe cases, suggests a role of the CYP1A1 gene and vitamin A in disease pathogenesis and prognosis.

Toxicity and ecological risk assessment for two AhR agonistic pesticides mepanipyrim and cyprodinil and their metabolites.

Mepanipyrim and cyprodinil are widely used to control and/or prevent fungal diseases in fruit culture. They are frequently detected in the aquatic environment and some food commodities. Different from TCDD, mepanipyrim and cyprodinil are more easily metabolised in the environments. However, the risk of their metabolites to the ecological environment is unclear and needs to be further confirmed. In this study, we investigated the temporal pattern of mepanipyrim- and cyprodinil-induced CYP1A and AhR2 expression and EROD enzyme activity at different time frames during zebrafish embryonic and larval development. Then, we assessed the ecological risk of mepanipyrim, cyprodinil, and their metabolites to aquatic organisms. Our results showed that mepanipyrim and cyprodinil exposure could increase the expression level of cyp1a and ahr2 genes and EROD activity by a dynamic pattern in different developmental stages of zebrafish. Besides, their several metabolites showed strong AhR agonistic activity. Importantly, these metabolites could cause potential ecological risks to aquatic organisms and should be paid more attention to. Our results would provide an important reference value for environmental pollution control and the use management of mepanipyrim and cyprodinil.

In vitro toxicity assessment of polyethylene terephthalate and polyvinyl chloride microplastics using three cell lines from rainbow trout (Oncorhynchus mykiss).

The RTgill-W1 (gill), RTG-2 (gonad), and RTL-W1 (liver) cell lines derived from a freshwater fish rainbow trout (Oncorhynchus mykiss), were used to assess the toxicity of polyethylene terephthalate (PET) and two forms of polyvinyl chloride (PVC). Two size fractions (25-µm and 90-µm particles) were tested for all materials. The highest tested concentration was 1 mg/ml, corresponding to from 70 000 ± 9000 to 620 000 ± 57 000 particles/ml for 25-µm particles and from 2300 ± 100 to 11 000 ± 1000 particles/ml for 90-µm particles (depending on the material). Toxicity differences between commercial PVC dry blend powder and secondary microplastics created from a processed PVC were newly described. After a 24-h exposure, the cells were analyzed for changes in viability, 7-ethoxyresorufin-O-deethylase (EROD) activity, and reactive oxygen species (ROS) generation. In addition to the microplastic suspensions, leachates and particles remaining after leaching resuspended in fresh exposure medium were tested. The particles were subjected to leaching for 1, 8, and 15 days. The PVC dry blend (25 µm and 90 µm) and processed PVC (25 µm) increased ROS generation, to which leached chemicals appeared to be the major contributor. PVC dry blend caused substantially higher ROS induction than processed PVC, showing that the former is not suitable for toxicity testing, as it can produce different results from those of secondary PVC. The 90-µm PVC dry blend increased ROS generation only after prolonged leaching. PET did not induce any changes in ROS generation, and none of the tested polymers had any effect on viability or EROD activity. The importance of choosing realistic extraction procedures for microplastic toxicity experiments was emphasized. Conducting long-term experiments is crucial to detect possible environmentally relevant effects. In conclusion, the tested materials showed no acute toxicity to the cell lines.

Polluting characteristics, sources, cancer risk, and cellular toxicity of PAHs bound in atmospheric particulates sampled from an economic transformation demonstration area of Dongguan in the Pearl River Delta, China.

The Songshan Lake Science and Technology Industrial Park is a national economic transition demonstration area, which centers at a traditional industrial region, in Dongguan, China. We were interested in the involved atmospheric particulates-bound PAHs regarding their sources, cancer risk, and related cellular toxicity for those in other areas under comparable conditions. In this study, the daily concentrations of TSP, PM10, and PM2.5 were averaged 127.95, 95.91, and 67.62 µg/m3, and the bound PAHs were averaged 1.31, 1.22, and 0.77 ng/m3 in summer and 12.72, 20.51 and 40.27 ng/m3 in winter, respectively. The dominant PAHs were those with 5-6 rings, and 4-6 rings in summer and winter, respectively. The incremental lifetime cancer risk (ILCR) (90th percentile probability) of total PAHs was above 1.00E-06 in each age group, particularly high in adolescents. Sensitivity analysis indicated that slope factor and body weight had greater impact than exposure duration and inhalation rate on the ILCR. Moreover, treatment of human bronchial epithelial BEAS-2B cells with mixed five indicative PAHs increased the formation of ROS, DNA damage (elevation in γ-H2AX), and protein levels of CAR, PXR, CYP1A1, 1A2, 1B1, while reduced the AhR protein, with the winter mixture more potent than summer. For the sources of PAHs, the stable carbon isotope ratio analysis and diagnostic ratios consistently pointed to petroleum and fossil fuel combustion as major sources. In conclusion, our findings suggest that particulates-bound PAHs deserve serious concerns for a cancer risk in such environment, and the development of new power sources for reducing fossil fuel combustion is highly encouraged.

Competitive Metabolism of Polycyclic Aromatic Hydrocarbons (PAHs): An Assessment Using In Vitro Metabolism and Physiologically Based Pharmacokinetic (PBPK) Modeling.

Humans are routinely exposed to complex mixtures such as polycyclic aromatic hydrocarbons (PAHs) rather than to single compounds, as are often assessed for hazards. Cytochrome P450 enzymes (CYPs) metabolize PAHs, and multiple PAHs found in mixtures can compete as substrates for individual CYPs (e.g., CYP1A1, CYP1B1, etc.). The objective of this study was to assess competitive inhibition of metabolism of PAH mixtures in humans and evaluate a key assumption of the Relative Potency Factor approach that common human exposures will not cause interactions among mixture components. To test this objective, we co-incubated binary mixtures of benzo[a]pyrene (BaP) and dibenzo[def,p]chrysene (DBC) in human hepatic microsomes and measured rates of enzymatic BaP and DBC disappearance. We observed competitive inhibition of BaP and DBC metabolism and measured inhibition coefficients (Ki), observing that BaP inhibited DBC metabolism more potently than DBC inhibited BaP metabolism (0.061 vs. 0.44 µM Ki, respectively). We developed a physiologically based pharmacokinetic (PBPK) interaction model by integrating PBPK models of DBC and BaP and incorporating measured metabolism inhibition coefficients. The PBPK model predicts significant increases in BaP and DBC concentrations in blood AUCs following high oral doses of PAHs (≥100 mg), five orders of magnitude higher than typical human exposures. We also measured inhibition coefficients of Supermix-10, a mixture of the most abundant PAHs measured at the Portland Harbor Superfund Site, on BaP and DBC metabolism. We observed similar potencies of inhibition coefficients of Supermix-10 compared to BaP and DBC. Overall, results of this study demonstrate that these PAHs compete for the same enzymes and, at high doses, inhibit metabolism and alter internal dosimetry of exposed PAHs. This approach predicts that BaP and DBC exposures required to observe metabolic interaction are much higher than typical human exposures, consistent with assumptions used when applying the Relative Potency Factor approach for PAH mixture risk assessment.

The Amphibian Short-Term Assay: Evaluation of a New Ecotoxicological Method for Amphibians Using Two Organophosphate Pesticides Commonly Found in Nature-Assessment of Biochemical, Morphological, and Life-History Traits.

Amphibia is the most threatened class among vertebrates, with >40% of the species threatened with extinction. Pollution is thought to alter amphibian population dynamics. With the growing interest in behavioral ecotoxicology, the neurotoxic organophosphate pesticides are of special concern. Understanding how exposure to neurotoxics leads to behavioral alterations is of crucial importance, and mechanistic endpoints should be included in ecotoxicological methods. In the present study, we tested an 8-day assay to evaluate the toxicity of two organophosphates, diazinon and chlorpyrifos, on Xenopus laevis, that is, on biochemical, morphological, and life-history traits related to locomotion capacities. The method involves measuring biomarkers such as glutathione-S-transferase (GST) and ethoxyresorufin-O-deethylase (EROD; two indicators of the detoxifying system) in the 8-day-old larvae as well as acetylcholinesterase (AChE) activity (involved in the nervous system) in 4-day-old embryos and 8-day-old larvae. Snout-to-vent length and snout-to-tail length of 4-day-old embryos and 8-day larvae were recorded as well as the corresponding growth rate. Fin and tail muscle widths were measured as well for testing changes in tail shape. Both tests showed effects of both organophosphates on AChE activity; however, no changes were observed in GST and EROD. Furthermore, exposure to chlorpyrifos demonstrated impacts on morphological and life-history traits, presaging alteration of locomotor traits. In addition, the results suggest a lower sensitivity to chlorpyrifos of 4-day-old embryos compared to 8-day-old larvae. Tests on other organophosphates are needed to test the validity of this method for the whole organophosphate group. Environ Toxicol Chem 2022;41:2688-2699. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Metal exposure and breast cancer among Northern Mexican women: assessment of genetic susceptibility.

This study aims to assess breast cancer (BC) association with metals and whether polymorphisms in CYP1A1, CYP1B1, GSTM1 and GSTT1 act as confounders or as modifiers of those relationships. We performed a secondary analysis of 499 histologically confirmed BC cases and the same number of age-matched population controls. We measured urinary concentrations of 18 metals with mass spectrometry. We determined the genetic variants of interest by allelic discrimination and multiplex PCR. After adjusting for covariates, we found BC negatively associated with arsenic, barium, cobalt, copper, magnesium, molybdenum and vanadium concentrations and positively with those of caesium, manganese, tin and thallium. Most associations remained after stratifying by the genetic variants. We identified that polymorphisms in CYP1B1, CYP1A1 and GSTM1 genes interacted with some metals on BC: interaction p-values CYP1B1 G119T × antimony= 0.036, CYP1B1 G119T × cobalt <0.001, CYP1B1 G119T × tin= 0.032, CYP1A1 A4889G × aluminium= 0.018, CYP1A1 A4889G × arsenic= 0.031, CYP1A1 A4889G × nickel= 0.036, CYP1A1 A4889G × vanadium= 0.031 and GSTM1 deletion × barium= 0.035. Exposure to various individual metals, along with genetic characteristics may contribute to BC development. Further studies are warranted to confirm our results.

Physical Evidence of Oil Uptake and Toxicity Assessment of Amphiphilic Grafted Nanoparticles Used as Oil Dispersants.

Herein, we report the toxicity evaluation of a new prototype dispersant system, silicon dioxide nanoparticles (NPs) functionalized with (3-glycidoxypropyl)triethoxysilane (GPS) and grafted poly(ε-caprolactone)-block-poly[oligo(ethylene glycol)methyl methacrylate mono-methyl ether] (NP-PCL-POEGMA). This serves as a follow up of our previous study where grafted silicon dioxide NPs functionalized with GPS and grafted hyperbranched poly(glycidol) (NP-HPG) were evaluated for reducing the toxicity in embryo, juvenile, and adult fish populations. In this study, the NP-HPG sample is used as a baseline to compare against the new NP-PCL-POEGMA samples. The relative size was established for three NP-PCL-POEGMA samples via cryogenic transmission electron microscopy. A quantitative mortality study determined that these NPs are non-toxic to embryo populations. An ethoxyresorufin-O-deethylase assay was performed on these NP-PCL-POEGMA samples to test for reduced cytochrome P450 1A after the embryos were exposed to the water-accommodated fraction of crude oil. Overall, these NP-PCL-POEGMA NPs better protected the embryo populations than the previous NP-HPG sample (using a protein activity end point), showing a trend in the right direction for prototype dispersants to replace the commercially utilized Corexit.

Impacts of size-fractionation on toxicity of marine microplastics: Enhanced integrated biomarker assessment in the tropical mussels, Perna viridis.

Accumulation of microplastics (MP) in oceanic waters is eroding the health of marine biota. We investigated how size-fractionated MP influence the toxicity risks towards a tropical keystone species, Perna viridis. Tissue-specific bioaccumulation and in vivo toxicity of polystyrene (PS) particles (0.5, 5, and 50 µm) were measured upon continuous exposure for 7 days, followed by 7 days depuration. P. viridis were exposed to equivalent mass (0.6 mg/L), corresponding to 4.0-4.6 particles/mL, 4.6-7.1 × 103 particles/mL, and 1.1-4.8 × 106 particles/mL for 50 µm, 5 µm and 0.5 µm PS particles, respectively. Onset toxicity were quantified through the enhanced integrated multi-biomarker response (EIBR) model, measured by weighting of biological organisation levels of eight biomarkers: (i) molecular (i.e., DNA damage (comet), 7-ethoxy resorufin O-deethylase (EROD), Catalase (CAT), Superoxide dismutase (SOD), Ferric Reducing Antioxidant Power (FRAP)); (ii) cellular (i.e., Neutral red retention (NRR), phagocytosis); and (iii) physiological (i.e., filtration rate). Data showed slightly elevated lysosomal instability (NRR) and antioxidant defences (FRAP, SOD, CAT, EROD) in specimens exposed to nano-PS (0.5 µm) compared to micro-PS (5 and 50 µm). Immunotoxicity (phagocytosis) and genotoxicity (comet) for haemocyte cells were significantly higher in specimens exposed to nano-PS (p < 0.05). EIBR index corroborated increasing toxicity modulated by MP sizes in descending order: 0.5 µm > 5 µm > 50 µm, with nano-PS exerted significantly higher biological effects (EIBR = 19.77 ± 5.89) than the unexposed group (EIBR = 10.97 ± 2.02; p < 0.05). Symptomatic organismal depression was manifested by the depleting filtering proficiency and weakened defence against invasive Zymosan bioparticles in the phagocytosis assay. Although impaired mussels duly recovered during depuration, individuals affected by nano-PS showed immunocompetence deficiency and gill responses that were not readily reversible, which could potentially increase their vulnerability towards further environmental stressors.

Toxicity assessment of urban marine sediments from Western Norway using a battery of stress-activated receptors and cell-based bioassays from fish.

A luciferase reporter gene-based bioassay battery consisting of stress-activated receptors from fish, complemented with traditional fish cell-based bioassays, were used to assess the toxicity of marine sediment samples from the Byfjorden area around the city of Bergen (Norway). The reporter assays covered a wide range of cellular signalling and metabolic pathways, representing different molecular initiating events in the adverse outcome pathway framework. Cytotoxicity, generation of reactive oxygen-species, and induction of 7-ethoxyresorufin-O-deethylase activity were analysed using fish liver and gill cell lines. Chemical analyses of the sediment extracts revealed complex contamination profiles, especially at the innermost stations, which contained a wide array of persistent organic pollutants, polycyclic aromatic hydrocarbons, and metals. Sediment extracts from these sites were more potent in activating the stress-activated receptors than the other extracts, reflecting their toxicant profiles. Importantly, receptor- and cell-based bioassays complemented the chemical analyses and provided important data for future environmental risk assessments of urban marine sediments.

Structure-based virtual screening of CYP1A1 inhibitors: towards rapid tier-one assessment of potential developmental toxicants.

Cytochrome P450 1A1 (CYP1A1) metabolizes estrogens, melatonin, and other key endogenous signaling molecules critical for embryonic/fetal development. The enzyme has increasing expression during pregnancy, and its inhibition or knockout increases embryonic/fetal lethality and/or developmental problems. Here, we present a virtual screening model for CYP1A1 inhibitors based on the orthosteric and predicted allosteric sites of the enzyme. Using 1001 reference compounds with CYP1A1 activity data, we optimized the decision thresholds of our model and classified the training compounds with 68.3% balanced accuracy (91.0% sensitivity and 45.7% specificity). We applied our final model to 11 known CYP1A1 orthosteric binders and related compounds, and found that our ranking of the known orthosteric binders generally agrees with the relative activity of CYP1A1 in metabolizing these compounds. We also applied the model to 22 new test compounds with unknown/unclear CYP1A1 inhibitory activity, and predicted 16 of them are CYP1A1 inhibitors. The CYP1A1 potency and modes of inhibition of these 22 compounds were experimentally determined. We confirmed that most predicted inhibitors, including drugs contraindicated during pregnancy (amiodarone, bicalutamide, cyproterone acetate, ketoconazole, and tamoxifen) and environmental agents suspected to be endocrine disruptors (bisphenol A, diethyl and dibutyl phthalates, and zearalenone), are indeed potent inhibitors of CYP1A1. Our results suggest that virtual screening may be used as a rapid tier-one method to screen for potential CYP1A1 inhibitors, and flag them out for further experimental evaluations.

Toxicity Assessment of Transfluthrin, Benzyl Butyl Phthalate, and 17ß-Estradiol on the Primary Fibroblast of the Striped Field Mouse, Apodemus agrarius.

Environmental pollution (EP) is a well-known threat to wild animals, but its toxicological impact is poorly understood. In vitro toxicity evaluation using cells of lower predators could be a promising way to assess and monitor the effects of EPs on whole wildlife populations that are related in the food web. Here, we describe EPs' toxic effect and mechanism in the primary fibroblast derived from the embryo of the striped field mouse, Apodemus agrarius. Characterization of the primary fibroblast was via morphology, genetics, immunocytochemistry, and stable culture conditions for optimal toxicity screening. Cell viability assays-MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and lactate dehydrogenase (LDH)-were performed to observe cytotoxicity, and quantitative PCR was conducted to confirm gene alteration by EP exposure. MTT and LDH assays confirmed the cytotoxicity of transfluthrin (TF), benzyl butyl phthalate (BBP), and 17ß-estradiol (E2) with IC50 values of 10.56 µM, 10.82 µM, and 24.08 µM, respectively, following 48-h exposures. mRNA expression of androgen-binding protein, growth hormone receptor, cytochrome C oxidase, and cytochrome P450-1A1 was induced after exposure to TF, BBP, and E2. We unveiled new EP mechanisms at the mammalian cellular level and discovered potential biomarker genes for monitoring of EPs. Based on our findings, we propose the primary fibroblast of A. agrarius as a valuable model to assess the toxicological effects of EP on wildlife.

First assessment of POPs and cytochrome P450 expression in Cuvier's beaked whales (Ziphius cavirostris) skin biopsies from the Mediterranean Sea.

The Cuvier's beaked whale (Ziphius cavirostris) is one of the least known cetacean species worldwide. The decreasing population trend and associated threats has led to the IUCN categorising the Mediterranean subpopulation as Vulnerable on the Red List of Threatened Species. This study aimed to investigate for the first time the ecotoxicological status of Cuvier's beaked whale in the NW Mediterranean Sea. The study sampled around the 20% of the individuals belonging to the Ligurian subpopulation, collecting skin biopsies from free-ranging specimens. The levels of polychlorinated biphenyl (PCBs), polybrominated diphenyl ethers (PBDEs) and induction of cytochrome's P450 (CYP1A1 and CYP2B isoforms) were evaluated. Results highlighted that the pattern of concentration for the target contaminants was PCBs > PBDEs and the accumulation values were linked to age and sex, with adult males showing significantly higher levels than juvenile. Concerns raised by the fact that 80% of the individuals had PCB levels above the toxicity threshold for negative physiological effects in marine mammals. Therefore, these findings shed light on this silent and serious threat never assessed in the Mediterranean Cuvier's beaked whale population, indicating that anthropogenic pressures, including chemical pollution, may represent menaces for the conservation of this species in the Mediterranean Sea.

Toxicity assessment of a novel oil dispersant based on silica nanoparticles using Fathead minnow.

Oil spill accidents are a major concern for aquatic organisms. In recent history, the Deepwater Horizon blowout spilled 500 million liters of crude oil into the Gulf of Mexico. Corexit 9500A was used to disperse the oil since it was the method approved at that time, despite safety concerns about its use. A better solution is necessary for dispersing oil from spills that reduces the toxicity to exposed aquatic organisms. To address this challenge, novel engineered nanoparticles were designed using silica cores grafted with hyperbranched poly(glycidol) branches. Because the silica core and polymers are known to be biocompatible, we hypothesized that these particles are nontoxic to fathead minnows (Pimephales promelas) and would decrease their exposure to oil polyaromatic hydrocarbons. Fathead minnow embryos, juveniles and adult stages were exposed to the particles alone or in combination with a water-accommodated fraction of oil. Acute toxicity of nanoparticles to fish was tested by measuring mortality. Sub-lethal effects were also measured including gene expression of cytochrome P450 1a (cyp1a) mRNA and heart rate in embryos. In addition, a mixture of particles plus the water-accommodated fraction was directly introduced to adult female fathead minnows by gavage. Three different nanoparticle concentrations were used (2, 10, and 50 mg/L) in either artificial fresh water or the water-accommodated fraction of the oil. In addition, nanoparticle-free controls were carried out in the two solutions. No significant mortality was observed for any age group or nanoparticle concentration, suggesting the safety of the nanoparticles. In the presence of the water-accommodated fraction alone, juvenile and adult fathead minnows responded by increasing expression of cyp1a. The addition of nanoparticles to the water-accommodated fraction reduced cyp1a gene expression in treatments. Heart rate was also restored to normal parameters in embryos co-exposed to nanoparticles and to the water-accommodated fraction. Measurement of polyaromatic hydrocarbons confirmed their presence in the tested solutions and the reduction of available PAH in WAF treated with the nanoparticles. Our findings suggest the engineered nanoparticles may be protecting the fish by sequestering polyaromatic hydrocarbons from oil, measured indirectly by the induction of cypa1 mRNAs. Furthermore, chemical analysis showed a reduction in PAH content in the water accommodated fraction with the presence of nanoparticles.

Risk assessment of Arctic aquatic species using ecotoxicological biomarkers and Bayesian network.

The risk to Arctic aquatic species due to accidental oil spills is not well studied. One of the key reasons for this limitation is the lack of understanding of the dose-response relations for the species in the Arctic region. The present study addresses this knowledge gap. It proposes a new approach to develop dose-response curves for Arctic aquatic species. The application of the approach is demonstrated using the estimation of mortality risk in Boreogadus saida (polar cod) due to exposure from polycyclic aromatic hydrocarbons (PAH). The proposed approach considers the toxicity mechanism in Arctic species (i.e. polar cod) and regional environmental factors, and models these as a belief-based Bayesian Network (BN). The BN model integrates diverse ecotoxicology biomarker data types and predicts the cell death probability due to exposure to a toxicant (PAH in crude oil). The input data and results from the model were verified using data available in the literature. Seasonal sea ice played a major role in containing PAH exposure and subsequent risk to polar cod. However, the physiological factors, such as presence of higher Phase II activity, and higher oxyradical scavenging ability, had greater impact on PAH risk mitigation.

Assessment of safety and efficacy of pine bark extract in normal and ovariectomized mice.

We evaluated the influence of pine bark extract (PBE) on organs, the cytochrome-P450 (CYP) activities in liver and estrogenic effects in normal and ovariectomized (OVX) female mice. The PBE did not affect organ weights and liver-function indexes (activities of alkaline phosphatase, aspartate amino transferase, and alanine amino transferase) at doses; 0.04%, 0.4%, and 2.0% PBE in the diet, in normal and OVX female mice. In the OVX mice, CYP1A1 activity was significantly higher in the 0.4% and 2.0% PBE groups than in the OVX control group, and in the 0.4% and 2.0% PBE groups were significantly higher than in the 0.04% PBE group. CYP1A2 and 3A4 activities were significantly higher in the 2.0% PBE group than in all other groups. The PBE did not affect uterine weight and femoral bone mineral density at all PBE doses. These results showed that the dose of PBE at the recommended human intake, had no toxic and estrogenic effects in normal female and OVX mice, however, it may need attention to use the excess intake of PBE with some drugs in postmenopausal women.

Toxicity assessment of pollutants sorbed on environmental sample microplastics collected on beaches: Part I-adverse effects on fish cell line.

Microplastics (MPs), are tiny plastic fragments from 1 µm to 5 mm generally found in the aquatic environment which can be easily ingested by organisms and may cause chronic physical but also toxicological effects. Toxicological assays on fish cell lines are commonly used as an alternative tool to provide fast and reliable assessment of the toxic and ecotoxic properties of chemicals or mixtures. Rainbow trout liver cell line (RTLW-1) was used to evaluate the toxicity of pollutants sorbed to MPs sampled in sandy beaches from different islands around the world during the first Race for Water Odyssey in 2015. The collected MPs were analyzed for polymer composition and associated persistent organic pollutants: polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT). In addition, DMSO-extracts from virgin MPs, MPs artificially coated with B[a]P and environmental MPs were analyzed with different bioassays: MTT reduction assay (MTT), ethoxyresorufin-O-deethylase (EROD) assay and comet assay. Microplastics from sand beaches were dominated by polyethylene, followed by polypropylene fragments with variable proportions. Organic pollutants found on plastic from beach sampling was PAHs (2-71 ng g-1). Samples from Bermuda (Somerset Long Bay) and Hawaii (Makapu'u) showed the highest concentration of PAHs and DDT respectively. No toxicity was observed for virgin microplastics. No cytotoxicity was observed on cells exposed to MP extract. However, EROD activity was induced and differently modulated depending on the MPs locations suggesting presence of different pollutants or additives in extract. DNA damage was observed after exposure to four microplastics samples on the six tested. Modification of EROD activity level and DNA damage rate highlight MPs extract toxicity on fish cell line.