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Prof. Aurelia Tubaro
Department of Life Science, University of Trieste, Via A. Valerio, 6, 34127 Trieste, Italy

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0 Nanotoxicology
0 Natural toxins
0 Toxicological studies
0 Mechanism of toxicity
0 Dermotoxicity

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Paper
Published: 21 October 2020 in Environmental Science: Nano
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The environmental impact of graphene oxide was evaluated on the model organism Artemia franciscana for ecotoxicological studies considering different biological parameters.

ACS Style

Federica Cavion; Laura Fusco; Silvio Sosa; Chiara Manfrin; Beatriz Alonso; Amaia Zurutuza; Roberto Della Loggia; Aurelia Tubaro; Maurizio Prato; Marco Pelin. Ecotoxicological impact of graphene oxide: toxic effects on the model organism Artemia franciscana. Environmental Science: Nano 2020, 7, 3605 -3615.

AMA Style

Federica Cavion, Laura Fusco, Silvio Sosa, Chiara Manfrin, Beatriz Alonso, Amaia Zurutuza, Roberto Della Loggia, Aurelia Tubaro, Maurizio Prato, Marco Pelin. Ecotoxicological impact of graphene oxide: toxic effects on the model organism Artemia franciscana. Environmental Science: Nano. 2020; 7 (11):3605-3615.

Chicago/Turabian Style

Federica Cavion; Laura Fusco; Silvio Sosa; Chiara Manfrin; Beatriz Alonso; Amaia Zurutuza; Roberto Della Loggia; Aurelia Tubaro; Maurizio Prato; Marco Pelin. 2020. "Ecotoxicological impact of graphene oxide: toxic effects on the model organism Artemia franciscana." Environmental Science: Nano 7, no. 11: 3605-3615.

Journal article
Published: 15 August 2020 in Nanomaterials
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In the frame of graphene-based material (GBM) hazard characterization, particular attention should be given to the cutaneous effects. Hence, this study investigates if HaCaT skin keratinocytes exposed to high concentrations of few-layer graphene (FLG) or partially dehydrated graphene oxide (d-GO) for a short time can recover from the cytotoxic insult, measured by means of cell viability, mitochondrial damage and oxidative stress, after GBM removal from the cell medium. When compared to 24 or 72 h continuous exposure, recovery experiments suggest that the cytotoxicity induced by 24 h exposure to GBM is only partially recovered after 48 h culture in GBM-free medium. This partial recovery, higher for FLG as compared to GO, is not mediated by autophagy and could be the consequence of GBM internalization into cells. The ability of GBMs to be internalized inside keratinocytes together with the partial reversibility of the cellular damage is important in assessing the risk associated with skin exposure to GBM-containing devices.

ACS Style

Marco Pelin; Hazel Lin; Arianna Gazzi; Silvio Sosa; Cristina Ponti; Amaya Ortega; Amaia Zurutuza; Ester Vázquez; Maurizio Prato; Aurelia Tubaro; Alberto Bianco. Partial Reversibility of the Cytotoxic Effect Induced by Graphene-Based Materials in Skin Keratinocytes. Nanomaterials 2020, 10, 1602 .

AMA Style

Marco Pelin, Hazel Lin, Arianna Gazzi, Silvio Sosa, Cristina Ponti, Amaya Ortega, Amaia Zurutuza, Ester Vázquez, Maurizio Prato, Aurelia Tubaro, Alberto Bianco. Partial Reversibility of the Cytotoxic Effect Induced by Graphene-Based Materials in Skin Keratinocytes. Nanomaterials. 2020; 10 (8):1602.

Chicago/Turabian Style

Marco Pelin; Hazel Lin; Arianna Gazzi; Silvio Sosa; Cristina Ponti; Amaya Ortega; Amaia Zurutuza; Ester Vázquez; Maurizio Prato; Aurelia Tubaro; Alberto Bianco. 2020. "Partial Reversibility of the Cytotoxic Effect Induced by Graphene-Based Materials in Skin Keratinocytes." Nanomaterials 10, no. 8: 1602.

Journal article
Published: 14 August 2020 in International Journal of Molecular Sciences
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The marine polyether palytoxin (PLTX) is one of the most toxic natural compounds, and is involved in human poisonings after oral, inhalation, skin and/or ocular exposure. Epidemiological and molecular evidence suggest different inter-individual sensitivities to its toxic effects, possibly related to genetic-dependent differences in the expression of Na+/K+-ATPase, its molecular target. To identify Na+/K+-ATPase subunits, isoforms correlated with in vitro PLTX cytotoxic potency, sensitivity parameters (EC50: PLTX concentration reducing cell viability by 50%; Emax: maximum effect induced by the highest toxin concentration; 10−7 M) were assessed in 60 healthy donors’ monocytes by the MTT (methylthiazolyl tetrazolium) assay. Sensitivity parameters, not correlated with donors’ demographic variables (gender, age and blood group), demonstrated a high inter-individual variability (median EC50 = 2.7 × 10−10 M, interquartile range: 0.4–13.2 × 10−10 M; median Emax = 92.0%, interquartile range: 87.5–94.4%). Spearman’s analysis showed significant positive correlations between the β2-encoding ATP1B2 gene expression and Emax values (rho = 0.30; p = 0.025) and between Emax and the ATP1B2/ATP1B3 expression ratio (rho = 0.38; p = 0.004), as well as a significant negative correlation between Emax and the ATP1B1/ATP1B2 expression ratio (rho = −0.30; p = 0.026). This toxicogenetic study represents the first approach to define genetic risk factors that may influence the onset of adverse effects in human PLTX poisonings, suggesting that individuals with high gene expression pattern of the Na+/K+-ATPase β2 subunit (alone or as β2/β1 and/or β2/β3 ratio) could be highly sensitive to PLTX toxic effects.

ACS Style

Marco Pelin; Gabriele Stocco; Chiara Florio; Silvio Sosa; Aurelia Tubaro. In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform. International Journal of Molecular Sciences 2020, 21, 5833 .

AMA Style

Marco Pelin, Gabriele Stocco, Chiara Florio, Silvio Sosa, Aurelia Tubaro. In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform. International Journal of Molecular Sciences. 2020; 21 (16):5833.

Chicago/Turabian Style

Marco Pelin; Gabriele Stocco; Chiara Florio; Silvio Sosa; Aurelia Tubaro. 2020. "In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform." International Journal of Molecular Sciences 21, no. 16: 5833.

Journal article
Published: 28 January 2020 in Toxins
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Pinnatoxin G (PnTx-G) is a marine cyclic imine toxin produced by the dinoflagellate Vulcanodinium rugosum, frequently detected in edible shellfish from Ingril Lagoon (France). As other pinnatoxins, to date, no human poisonings ascribed to consumption of PnTx-G contaminated seafood have been reported, despite its potent antagonism at nicotinic acetylcholine receptors and its high and fast-acting toxicity after intraperitoneal or oral administration in mice. The hazard characterization of PnTx-G by oral exposure is limited to a single acute toxicity study recording lethality and clinical signs in non-fasted mice treated by gavage or through voluntary food ingestion, which showed differences in PnTx-G toxic potency. Thus, an acute toxicity study was carried out using 3 h-fasted CD-1 female mice, administered by gavage with PnTx-G (8–450 µg kg−1). At the dose of 220 µg kg−1 and above, the toxin induced a rapid onset of clinical signs (piloerection, prostration, hypothermia, abdominal breathing, paralysis of the hind limbs, and cyanosis), leading to the death of mice within 30 min. Except for moderate mucosal degeneration in the small intestine recorded at doses of 300 µg kg−1, the toxin did not induce significant morphological changes in the other main organs and tissues, or alterations in blood chemistry parameters. This acute oral toxicity study allowed to calculate an oral LD50 for PnTx-G equal to 208 μg kg−1 (95% confidence limits: 155–281 µg kg−1) and to estimate a provisional NOEL of 120 µg kg−1.

ACS Style

Silvio Sosa; Marco Pelin; Federica Cavion; Fabienne Hervé; Philipp Hess; Aurelia Tubaro. Acute Oral Toxicity of Pinnatoxin G in Mice. Toxins 2020, 12, 87 .

AMA Style

Silvio Sosa, Marco Pelin, Federica Cavion, Fabienne Hervé, Philipp Hess, Aurelia Tubaro. Acute Oral Toxicity of Pinnatoxin G in Mice. Toxins. 2020; 12 (2):87.

Chicago/Turabian Style

Silvio Sosa; Marco Pelin; Federica Cavion; Fabienne Hervé; Philipp Hess; Aurelia Tubaro. 2020. "Acute Oral Toxicity of Pinnatoxin G in Mice." Toxins 12, no. 2: 87.

Journal article
Published: 24 December 2019 in Carbon
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Skin provides the first interface between body and environment, representing one of the most feasible exposure routes to graphene-based materials (GBMs). However, interactions of GBMs with the skin are poorly understood. In particular, low-concentration effects have not been investigated. Here we explored the ability of endotoxin-free, few-layer graphene (FLG) and dehydrated graphene oxide (d-GO) to initiate an inflammatory response at the cutaneous level by using human HaCaT keratinocytes. HaCaT cell exposure to low concentrations (0.01–1.0 μg/mL) of FLG or d-GO did not affect cell viability. FLG triggered the secretion of pro-inflammatory tumor necrosis factor-α (TNF-α), interleukin (IL)-1α, and IL-6, while d-GO, and to a lesser extent FLG, prompted IL-8 (CXCL8) production. However, conditioned medium from HaCaT cells exposed to FLG or d-GO had no effect on THP-1 monocyte activation. Moreover, co-culture experiments did not show any effect of FLG- or d-GO-treated HaCaT cells on THP-1 cell migration. These results suggest that while GBMs are able to initiate an inflammatory response in keratinocytes, this does not necessarily lead to activation of monocytes. The present findings are relevant for potential dermal exposures to GBMs in occupational settings as well as the use of GBMs for cutaneous applications such as in wearable sensors.

ACS Style

Laura Fusco; Marco Pelin; Sourav Mukherjee; Sandeep Keshavan; Silvio Sosa; Cristina Martín; Viviana González; Ester Vázquez; Maurizio Prato; Bengt Fadeel; Aurelia Tubaro. Keratinocytes are capable of selectively sensing low amounts of graphene-based materials: Implications for cutaneous applications. Carbon 2019, 159, 598 -610.

AMA Style

Laura Fusco, Marco Pelin, Sourav Mukherjee, Sandeep Keshavan, Silvio Sosa, Cristina Martín, Viviana González, Ester Vázquez, Maurizio Prato, Bengt Fadeel, Aurelia Tubaro. Keratinocytes are capable of selectively sensing low amounts of graphene-based materials: Implications for cutaneous applications. Carbon. 2019; 159 ():598-610.

Chicago/Turabian Style

Laura Fusco; Marco Pelin; Sourav Mukherjee; Sandeep Keshavan; Silvio Sosa; Cristina Martín; Viviana González; Ester Vázquez; Maurizio Prato; Bengt Fadeel; Aurelia Tubaro. 2019. "Keratinocytes are capable of selectively sensing low amounts of graphene-based materials: Implications for cutaneous applications." Carbon 159, no. : 598-610.

Journals
Published: 27 November 2019 in Nanoscale
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Graphene related materials, if prepared with non-irritant exfoliation agents, do not induce skin irritation on a 3D model of human epidermis, following the OECD guideline 439.

ACS Style

Laura Fusco; Marina Garrido; Cristina Martín; Silvio Sosa; Cristina Ponti; Alba Centeno; Beatriz Alonso; Amaia Zurutuza; Ester Vázquez; Aurelia Tubaro; Maurizio Prato; Marco Pelin. Skin irritation potential of graphene-based materials using a non-animal test. Nanoscale 2019, 12, 610 -622.

AMA Style

Laura Fusco, Marina Garrido, Cristina Martín, Silvio Sosa, Cristina Ponti, Alba Centeno, Beatriz Alonso, Amaia Zurutuza, Ester Vázquez, Aurelia Tubaro, Maurizio Prato, Marco Pelin. Skin irritation potential of graphene-based materials using a non-animal test. Nanoscale. 2019; 12 (2):610-622.

Chicago/Turabian Style

Laura Fusco; Marina Garrido; Cristina Martín; Silvio Sosa; Cristina Ponti; Alba Centeno; Beatriz Alonso; Amaia Zurutuza; Ester Vázquez; Aurelia Tubaro; Maurizio Prato; Marco Pelin. 2019. "Skin irritation potential of graphene-based materials using a non-animal test." Nanoscale 12, no. 2: 610-622.

Journal article
Published: 25 May 2019 in Toxins
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In September 2015, a massive occurrence of the Ostreopsis species was recorded in central Adriatic Kaštela Bay. In order to taxonomically identify the Ostreopsis species responsible for this event and determine their toxin profile, cells collected in seawater and from benthic macroalgae were analyzed. Conservative taxonomic methods (light microscopy and SEM) and molecular methods (PCR-based assay) allowed the identification of the species Ostreopsis cf. ovata associated with Coolia monotis. The abundance of O. cf. ovata reached 2.9 × 104 cells L−1 in seawater, while on macroalgae, it was estimated to be up to 2.67 × 106 cells g−1 of macroalgae fresh weight and 14.4 × 106 cells g−1 of macroalgae dry weight. An indirect sandwich immunoenzymatic assay (ELISA) and liquid chromatography–high-resolution mass spectrometry (LC-HRMS) were used to determine the toxin profile. The ELISA assay revealed the presence of 5.6 pg palytoxin (PLTX) equivalents per O. cf. ovata cell. LC-HRMS was used for further characterization of the toxin profile, which showed that there were 6.3 pg of the sum of ovatoxins (OVTXs) and isobaric PLTX per O. cf. ovata cell, with a prevalence of OVTXs (6.2 pg cell−1), while the isobaric PLTX concentration was very low (0.1 pg cell−1). Among OVTXs, the highest concentration was recorded for OVTX-a (3.6 pg cell−1), followed by OVTX-b (1.3 pg cell−1), OVTX-d (1.1 pg cell−1), and OVTX-c (0.2 pg cell−1).

ACS Style

Živana Ninčević Gladan; Jasna Arapov; Silvia Casabianca; Antonella Penna; Giorgio Honsell; Valentina Brovedani; Marco Pelin; Luciana Tartaglione; Silvio Sosa; Carmela Dell’Aversano; Aurelia Tubaro; Ante Žuljević; Branka Grbec; Matea Čavar; Mia Bužančić; Ana Bakrač; Sanda Skejić. Massive Occurrence of the Harmful Benthic Dinoflagellate Ostreopsis cf. ovata in the Eastern Adriatic Sea. Toxins 2019, 11, 300 .

AMA Style

Živana Ninčević Gladan, Jasna Arapov, Silvia Casabianca, Antonella Penna, Giorgio Honsell, Valentina Brovedani, Marco Pelin, Luciana Tartaglione, Silvio Sosa, Carmela Dell’Aversano, Aurelia Tubaro, Ante Žuljević, Branka Grbec, Matea Čavar, Mia Bužančić, Ana Bakrač, Sanda Skejić. Massive Occurrence of the Harmful Benthic Dinoflagellate Ostreopsis cf. ovata in the Eastern Adriatic Sea. Toxins. 2019; 11 (5):300.

Chicago/Turabian Style

Živana Ninčević Gladan; Jasna Arapov; Silvia Casabianca; Antonella Penna; Giorgio Honsell; Valentina Brovedani; Marco Pelin; Luciana Tartaglione; Silvio Sosa; Carmela Dell’Aversano; Aurelia Tubaro; Ante Žuljević; Branka Grbec; Matea Čavar; Mia Bužančić; Ana Bakrač; Sanda Skejić. 2019. "Massive Occurrence of the Harmful Benthic Dinoflagellate Ostreopsis cf. ovata in the Eastern Adriatic Sea." Toxins 11, no. 5: 300.

Journal article
Published: 08 May 2019 in Marine Drugs
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Azaspiracids (AZAs) are marine toxins that are produced by Azadinium and Amphidoma dinoflagellates that can contaminate edible shellfish inducing a foodborne poisoning in humans, which is characterized by gastrointestinal symptoms. Among these, AZA1, -2, and -3 are regulated in the European Union, being the most important in terms of occurrence and toxicity. In vivo studies in mice showed that, in addition to gastrointestinal effects, AZA1 induces liver alterations that are visible as a swollen organ, with the presence of hepatocellular fat droplets and vacuoles. Hence, an in vitro study was carried out to investigate the effects of AZA1, -2, and -3 on liver cells, using human non-tumor IHH hepatocytes. The exposure of IHH cells to AZA1, -2, or -3 (5 × 10-12-1 × 10-7 M) for 24 h did not affect the cell viability and proliferation (Sulforhodamine B assay and 3H-Thymidine incorporation assay), but they induced a significant concentration-dependent increase of mitochondrial dehydrogenases activity (MTT reduction assay). This effect depends on the activity of mitochondrial electron transport chain complex I and II, being counteracted by rotenone and tenoyl trifluoroacetone, respectively. Furthermore, AZAs-increased mitochondrial dehydrogenase activity was almost totally suppressed in the K+-, Cl--, and Na+-free media and sensitive to the specific inhibitors of KATP and hERG potassium channels, Na+/K+, ATPase, and cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels. These results suggest that AZA mitochondrial effects in hepatocytes derive from an imbalance of intracellular levels of K+ and, in particular, Cl- ions, as demonstrated by the selective reduction of toxin effects by CFTR chloride channel inhibition.

ACS Style

Marco Pelin; Jane Kilcoyne; Chiara Florio; Philipp Hess; Aurelia Tubaro; Silvio Sosa. Azaspiracids Increase Mitochondrial Dehydrogenases Activity in Hepatocytes: Involvement of Potassium and Chloride Ions. Marine Drugs 2019, 17, 276 .

AMA Style

Marco Pelin, Jane Kilcoyne, Chiara Florio, Philipp Hess, Aurelia Tubaro, Silvio Sosa. Azaspiracids Increase Mitochondrial Dehydrogenases Activity in Hepatocytes: Involvement of Potassium and Chloride Ions. Marine Drugs. 2019; 17 (5):276.

Chicago/Turabian Style

Marco Pelin; Jane Kilcoyne; Chiara Florio; Philipp Hess; Aurelia Tubaro; Silvio Sosa. 2019. "Azaspiracids Increase Mitochondrial Dehydrogenases Activity in Hepatocytes: Involvement of Potassium and Chloride Ions." Marine Drugs 17, no. 5: 276.

Journal article
Published: 22 August 2018 in Nanoscale
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The most significant routes associated with occupational exposure to graphene-based materials (GBMs) are the inhalation, cutaneous, ocular and oral ones. The manuscript presents a review of the in vivo toxicity data of GBMs after these exposure routes.

ACS Style

Marco Pelin; Silvio Sosa; Maurizio Prato; Aurelia Tubaro. Occupational exposure to graphene based nanomaterials: risk assessment. Nanoscale 2018, 10, 15894 -15903.

AMA Style

Marco Pelin, Silvio Sosa, Maurizio Prato, Aurelia Tubaro. Occupational exposure to graphene based nanomaterials: risk assessment. Nanoscale. 2018; 10 (34):15894-15903.

Chicago/Turabian Style

Marco Pelin; Silvio Sosa; Maurizio Prato; Aurelia Tubaro. 2018. "Occupational exposure to graphene based nanomaterials: risk assessment." Nanoscale 10, no. 34: 15894-15903.

Journal article
Published: 14 August 2018 in Toxins
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The marine algal toxin palytoxin (PLTX) and its analogues are some of the most toxic marine compounds. Their accumulation in edible marine organisms and entrance into the food chain represent their main concerns for human health. Indeed, several fatal human poisonings attributed to these compounds have been recorded in tropical and subtropical areas. Due to the increasing occurrence of PLTX in temperate areas such as the Mediterranean Sea, the European Food Safety Authority (EFSA) has suggested a maximum limit of 30 µg PLTX/kg in shellfish meat, and has recommended the development of rapid, specific, and sensitive methods for detection and quantitation of PLTX in seafood. Thus, a novel, sensitive cell-based ELISA was developed and characterized for PLTX quantitation in mussels. The estimated limits of detection (LOD) and quantitation (LOQ) were 1.2 × 10−11 M (32.2 pg/mL) and 2.8 × 10−11 M (75.0 pg/mL), respectively, with good accuracy (bias = 2.5%) and repeatability (15% and 9% interday and intraday relative standard deviation of repeatability (RSDr), respectively). Minimal interference of 80% aqueous methanol extract allows PLTX quantitation in mussels at concentrations lower than the maximum limit suggested by EFSA, with an LOQ of 9.1 µg PLTX equivalent/kg mussel meat. Given its high sensitivity and specificity, the cell-based ELISA should be considered a suitable method for PLTX quantitation.

ACS Style

Marco Pelin; Silvio Sosa; Valentina Brovedani; Laura Fusco; Mark Poli; Aurelia Tubaro. A Novel Sensitive Cell-Based Immunoenzymatic Assay for Palytoxin Quantitation in Mussels. Toxins 2018, 10, 329 .

AMA Style

Marco Pelin, Silvio Sosa, Valentina Brovedani, Laura Fusco, Mark Poli, Aurelia Tubaro. A Novel Sensitive Cell-Based Immunoenzymatic Assay for Palytoxin Quantitation in Mussels. Toxins. 2018; 10 (8):329.

Chicago/Turabian Style

Marco Pelin; Silvio Sosa; Valentina Brovedani; Laura Fusco; Mark Poli; Aurelia Tubaro. 2018. "A Novel Sensitive Cell-Based Immunoenzymatic Assay for Palytoxin Quantitation in Mussels." Toxins 10, no. 8: 329.

Journals
Published: 11 June 2018 in Nanoscale
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Graphene based nanomaterials induce a reactive oxygen species-mediated mitochondrial depolarization, caused by the activation of NADH dehydrogenase and xanthine oxidase.

ACS Style

Marco Pelin; Laura Fusco; Cristina Martín; Silvio Sosa; Javier Frontiñán-Rubio; Jose Miguel González-Domínguez; Mario Durán-Prado; Ester Vázquez; Maurizio Prato; Aurelia Tubaro. Graphene and graphene oxide induce ROS production in human HaCaT skin keratinocytes: the role of xanthine oxidase and NADH dehydrogenase. Nanoscale 2018, 10, 11820 -11830.

AMA Style

Marco Pelin, Laura Fusco, Cristina Martín, Silvio Sosa, Javier Frontiñán-Rubio, Jose Miguel González-Domínguez, Mario Durán-Prado, Ester Vázquez, Maurizio Prato, Aurelia Tubaro. Graphene and graphene oxide induce ROS production in human HaCaT skin keratinocytes: the role of xanthine oxidase and NADH dehydrogenase. Nanoscale. 2018; 10 (25):11820-11830.

Chicago/Turabian Style

Marco Pelin; Laura Fusco; Cristina Martín; Silvio Sosa; Javier Frontiñán-Rubio; Jose Miguel González-Domínguez; Mario Durán-Prado; Ester Vázquez; Maurizio Prato; Aurelia Tubaro. 2018. "Graphene and graphene oxide induce ROS production in human HaCaT skin keratinocytes: the role of xanthine oxidase and NADH dehydrogenase." Nanoscale 10, no. 25: 11820-11830.

Conference abstract
Published: 30 May 2018 in Toxicon
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ACS Style

Marco Pelin; Silvio Sosa; Cristina Ponti; Davide Gibellini; Chiara Florio; Aurelia Tubaro. Pro-inflammatory effects of palytoxin on skin keratinocytes and indirect modulation of inflammatory cells. Toxicon 2018, 149, 97 .

AMA Style

Marco Pelin, Silvio Sosa, Cristina Ponti, Davide Gibellini, Chiara Florio, Aurelia Tubaro. Pro-inflammatory effects of palytoxin on skin keratinocytes and indirect modulation of inflammatory cells. Toxicon. 2018; 149 ():97.

Chicago/Turabian Style

Marco Pelin; Silvio Sosa; Cristina Ponti; Davide Gibellini; Chiara Florio; Aurelia Tubaro. 2018. "Pro-inflammatory effects of palytoxin on skin keratinocytes and indirect modulation of inflammatory cells." Toxicon 149, no. : 97.

Journal article
Published: 12 January 2017 in Scientific Reports
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Impressive properties make graphene-based materials (GBMs) promising tools for nanoelectronics and biomedicine. However, safety concerns need to be cleared before mass production of GBMs starts. As skin, together with lungs, displays the highest exposure to GBMs, it is of fundamental importance to understand what happens when GBMs get in contact with skin cells. The present study was carried out on HaCaT keratinocytes, an in vitro model of skin toxicity, on which the effects of four GBMs were evaluated: a few layer graphene, prepared by ball-milling treatment (FLG), and three samples of graphene oxide (GOs, a research-grade GO1, and two commercial GOs, GO2 and GO3). Even though no significant effects were observed after 24 h, after 72 h the less oxidized compound (FLG) was the less cytotoxic, inducing mitochondrial and plasma-membrane damages with EC50s of 62.8 μg/mL (WST-8 assay) and 45.5 μg/mL (propidium iodide uptake), respectively. By contrast, the largest and most oxidized compound, GO3, was the most cytotoxic, inducing mitochondrial and plasma-membrane damages with EC50s of 5.4 and 2.9 μg/mL, respectively. These results suggest that only high concentrations and long exposure times to FLG and GOs could impair mitochondrial activity associated with plasma membrane damage, suggesting low cytotoxic effects at the skin level.

ACS Style

Marco Pelin; Laura Fusco; Verónica León; Cristina Martín; Alejandro Criado; Silvio Sosa; Ester Vázquez; Aurelia Tubaro; Maurizio Prato. Differential cytotoxic effects of graphene and graphene oxide on skin keratinocytes. Scientific Reports 2017, 7, 40572 .

AMA Style

Marco Pelin, Laura Fusco, Verónica León, Cristina Martín, Alejandro Criado, Silvio Sosa, Ester Vázquez, Aurelia Tubaro, Maurizio Prato. Differential cytotoxic effects of graphene and graphene oxide on skin keratinocytes. Scientific Reports. 2017; 7 (1):40572.

Chicago/Turabian Style

Marco Pelin; Laura Fusco; Verónica León; Cristina Martín; Alejandro Criado; Silvio Sosa; Ester Vázquez; Aurelia Tubaro; Maurizio Prato. 2017. "Differential cytotoxic effects of graphene and graphene oxide on skin keratinocytes." Scientific Reports 7, no. 1: 40572.

Case reports
Published: 19 August 2016 in Toxicon
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Palytoxin (PLTX) is a lethal natural toxin often found in Palythoa zoantharians that, together with its congeners, may induce adverse effects in humans after inhalation of toxic aerosols both in open-air and domestic environments, namely in the vicinity of public and private aquaria. In this study, we describe a poisoning of an aquarium hobbyist who was hospitalized after handling a PLTXs-containing zoantharian hexacoral. Furthermore, we provide evidence for water detoxification. The zoantharian was morphologically and genetically identified as Palythoa cf. toxica (Cnidaria: Anthozoa). Palytoxin itself and two new PLTX congeners, a hydroxyPLTX and a deoxyPLTX, were detected and structurally identified by liquid chromatography high resolution multiple stage mass spectrometry (LC-HRMSn, n = 1, 2). Total and individual toxins were quantified by LC-HRMS and sandwich ELISA both in the zoantharian (93.4 and 96.80 μg/g, respectively) and in the transport water (48.3 and 42.56 μg/mL, respectively), with an excellent mean bias of 1.3% between the techniques. Activated carbon adsorbed 99.7% of PLTXs contained in the seawater and this represents a good strategy for preventing aquarium hobbyist poisonings.

ACS Style

Luciana Tartaglione; Marco Pelin; Massimo Morpurgo; Carmela Dell'Aversano; Javier Montenegro; Giuseppe Sacco; Silvio Sosa; James Davis Reimer; Patrizia Ciminiello; Aurelia Tubaro. An aquarium hobbyist poisoning: Identification of new palytoxins in Palythoa cf. toxica and complete detoxification of the aquarium water by activated carbon. Toxicon 2016, 121, 41 -50.

AMA Style

Luciana Tartaglione, Marco Pelin, Massimo Morpurgo, Carmela Dell'Aversano, Javier Montenegro, Giuseppe Sacco, Silvio Sosa, James Davis Reimer, Patrizia Ciminiello, Aurelia Tubaro. An aquarium hobbyist poisoning: Identification of new palytoxins in Palythoa cf. toxica and complete detoxification of the aquarium water by activated carbon. Toxicon. 2016; 121 ():41-50.

Chicago/Turabian Style

Luciana Tartaglione; Marco Pelin; Massimo Morpurgo; Carmela Dell'Aversano; Javier Montenegro; Giuseppe Sacco; Silvio Sosa; James Davis Reimer; Patrizia Ciminiello; Aurelia Tubaro. 2016. "An aquarium hobbyist poisoning: Identification of new palytoxins in Palythoa cf. toxica and complete detoxification of the aquarium water by activated carbon." Toxicon 121, no. : 41-50.

Journal article
Published: 23 June 2016 in Toxicon
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Palytoxin (PLTX) and its analogues have been detected as seafood contaminants associated with a series of human foodborne poisonings. Due to a number of fatalities ascribed to the ingestion of PLTX-contaminated marine organisms, the development of methods for its detection in seafood has been recommended by the European Food Safety Authority (EFSA). Due to its feasibility, the spectrophotometric hemolytic assay is widely used to detect PLTX in different matrices, even though a standardized protocol is still lacking. Thus, on the basis of available assay procedures, a new standardized protocol was set up using purified human erythrocytes exposed to PLTX (working range: 3.9 × 10−10–2.5 × 10−8 M) in a K+-free phosphate buffered saline solution, employing a 5 h incubation at 41 °C. An intra-laboratory characterization demonstrated its sensitivity (limit of detection, LOD = 1.4 × 10−10 M and quantitation, LOQ = 3.4 × 10−10 M), accuracy (bias = −0.8%), repeatability (RSDr = 15% and 6% for intra- and inter-day repeatability, respectively) and specificity. However, the standardized method seems not to be suitable for PLTX quantitation in complex matrices, such as mussel (Mytilus galloprovincialis) extracts, at least below the limit suggested by EFSA (30 μg PLTXs/Kg shellfish meat). Thus, the hemolytic assay for PLTX quantitation in seafood should be used only after a careful evaluation of the specific matrix effects.

ACS Style

Valentina Brovedani; Silvio Sosa; Mark Poli; Martino Forino; Katia Varello; Aurelia Tubaro; Marco Pelin. A revisited hemolytic assay for palytoxin detection: Limitations for its quantitation in mussels. Toxicon 2016, 119, 225 -233.

AMA Style

Valentina Brovedani, Silvio Sosa, Mark Poli, Martino Forino, Katia Varello, Aurelia Tubaro, Marco Pelin. A revisited hemolytic assay for palytoxin detection: Limitations for its quantitation in mussels. Toxicon. 2016; 119 ():225-233.

Chicago/Turabian Style

Valentina Brovedani; Silvio Sosa; Mark Poli; Martino Forino; Katia Varello; Aurelia Tubaro; Marco Pelin. 2016. "A revisited hemolytic assay for palytoxin detection: Limitations for its quantitation in mussels." Toxicon 119, no. : 225-233.

Journals
Published: 17 May 2016 in Toxicology Research
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Keratinocytes are actively involved in the recruitment of inflammatory cells in response to cutaneous contact with palytoxin.

ACS Style

Marco Pelin; Chiara Florio; Cristina Ponti; Marianna Lucafò; Davide Gibellini; Aurelia Tubaro; Silvio Sosa. Pro-inflammatory effects of palytoxin: an in vitro study on human keratinocytes and inflammatory cells. Toxicology Research 2016, 5, 1172 -1181.

AMA Style

Marco Pelin, Chiara Florio, Cristina Ponti, Marianna Lucafò, Davide Gibellini, Aurelia Tubaro, Silvio Sosa. Pro-inflammatory effects of palytoxin: an in vitro study on human keratinocytes and inflammatory cells. Toxicology Research. 2016; 5 (4):1172-1181.

Chicago/Turabian Style

Marco Pelin; Chiara Florio; Cristina Ponti; Marianna Lucafò; Davide Gibellini; Aurelia Tubaro; Silvio Sosa. 2016. "Pro-inflammatory effects of palytoxin: an in vitro study on human keratinocytes and inflammatory cells." Toxicology Research 5, no. 4: 1172-1181.

Review
Published: 04 February 2016 in Marine Drugs
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Palytoxin (PLTX), one the most potent marine toxins, and/or its analogs, have been identified in different marine organisms, such as Palythoa soft corals, Ostreopsis dinoflagellates, and Trichodesmium cyanobacteria. Although the main concern for human health is PLTXs entrance in the human food chain, there is growing evidence of adverse effects associated with inhalational, cutaneous, and/or ocular exposure to aquarium soft corals contaminated by PLTXs or aquaria waters. Indeed, the number of case reports describing human poisonings after handling these cnidarians is continuously increasing. In general, the signs and symptoms involve mainly the respiratory (rhinorrhea and coughing), skeletomuscular (myalgia, weakness, spasms), cardiovascular (electrocardiogram alterations), gastrointestinal (nausea), and nervous (paresthesia, ataxia, tremors) systems or apparates. The widespread phenomenon, the entity of the signs and symptoms of poisoning and the lack of control in the trade of corals as aquaria decorative elements led to consider these poisonings an emerging sanitary problem. This review summarizes literature data on human poisonings due to, or ascribed to, PLTX-containing soft corals, focusing on the different PLTX congeners identified in these organisms and their toxic potential.

ACS Style

Marco Pelin; Valentina Brovedani; Silvio Sosa; Aurelia Tubaro. Palytoxin-Containing Aquarium Soft Corals as an Emerging Sanitary Problem. Marine Drugs 2016, 14, 33 .

AMA Style

Marco Pelin, Valentina Brovedani, Silvio Sosa, Aurelia Tubaro. Palytoxin-Containing Aquarium Soft Corals as an Emerging Sanitary Problem. Marine Drugs. 2016; 14 (2):33.

Chicago/Turabian Style

Marco Pelin; Valentina Brovedani; Silvio Sosa; Aurelia Tubaro. 2016. "Palytoxin-Containing Aquarium Soft Corals as an Emerging Sanitary Problem." Marine Drugs 14, no. 2: 33.

Research article
Published: 12 January 2016 in Environmental Science & Technology
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This study provides the first evaluation of the cytotoxic effects of the recently identified palytoxin (PLTX) analog, ovatoxin-a (OVTX-a), the major toxin produced by Ostreopsis cf. ovata in the Mediterranean Sea. Its increasing detection during Ostreopsis blooms and in seafood highlights the need to characterize its toxic effects and to set up appropriate detection methods. OVTX-a is about 100 fold less potent than PLTX in reducing HaCaT cells viability (EC50 = 1.1 × 10–9 M vs 1.8 × 10–11 M, MTT test) in agreement with a reduced binding affinity (Kd = 1.2 × 10–9 vs 2.7 × 10–11 M, saturation experiments on intact cells). Similarly, OVTX-a hemolytic effect is lower than that of the reference PLTX compound. Ost-D shows the lowest cytotoxicity toward HaCaT keratinocytes, suggesting the lack of a hydroxyl group at C44 as a critical feature for PLTXs cytotoxic effects. A sandwich ELISA developed for PLTX detects also OVTX-a in a sensitive (LOD = 4.2 and LOQ = 5.6 ng/mL) and accurate manner (Bias = 0.3%), also in O. cf. ovata extracts and contaminated mussels. Although in vitro OVTX-a appears less toxic than PLTX, its cytotoxicity at nanomolar concentrations after short exposure time rises some concern for human health. The sandwich ELISA can be a viable screening method for OVTXs detection in monitoring program.

ACS Style

Marco Pelin; Martino Forino; Valentina Brovedani; Luciana Tartaglione; Carmela Dell’Aversano; Rossella Pistocchi; Mark Poli; Silvio Sosa; Chiara Florio; Patrizia Ciminiello; Aurelia Tubaro. Ovatoxin-a, A Palytoxin Analogue Isolated from Ostreopsis cf. ovata Fukuyo: Cytotoxic Activity and ELISA Detection. Environmental Science & Technology 2016, 50, 1544 -1551.

AMA Style

Marco Pelin, Martino Forino, Valentina Brovedani, Luciana Tartaglione, Carmela Dell’Aversano, Rossella Pistocchi, Mark Poli, Silvio Sosa, Chiara Florio, Patrizia Ciminiello, Aurelia Tubaro. Ovatoxin-a, A Palytoxin Analogue Isolated from Ostreopsis cf. ovata Fukuyo: Cytotoxic Activity and ELISA Detection. Environmental Science & Technology. 2016; 50 (3):1544-1551.

Chicago/Turabian Style

Marco Pelin; Martino Forino; Valentina Brovedani; Luciana Tartaglione; Carmela Dell’Aversano; Rossella Pistocchi; Mark Poli; Silvio Sosa; Chiara Florio; Patrizia Ciminiello; Aurelia Tubaro. 2016. "Ovatoxin-a, A Palytoxin Analogue Isolated from Ostreopsis cf. ovata Fukuyo: Cytotoxic Activity and ELISA Detection." Environmental Science & Technology 50, no. 3: 1544-1551.

Reference work
Published: 09 June 2015 in Marine and Freshwater Toxins
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Okadaic acid (OA) and its derivatives are toxic lipophilic polyethers produced by marine dinoflagellates and accumulated in filter-feeding organisms, where they can undergo biotransformation. The ingestion of marine shellfish contaminated by these toxins causes a gastrointestinal illness in humans known as diarrhetic shellfish poisoning (DSP). OA and its analogues are weak tumor promoters and specific inhibitors of serine/threonine protein phosphatases. Inhibition of these enzymes affects various cellular processes, including the events at the basis of diarrhea and tumor promotion, although other mechanisms of action cannot be excluded. Studies in rodents indicate that these diarrheic toxins are absorbed through the gastrointestinal tract and distributed in the whole body, inducing toxic effects. Their lethal oral doses after acute administration in mice are 2–5 times higher than intraperitoneal ones, with the small intestine and liver as main target organs, while repeated oral OA exposure affects mainly the forestomach, lymphoid organs, pancreas, and, at ultrastructural level, myocardium. The increasing number of toxicological studies on OA, including cytotoxicity, genotoxicity, neurotoxicity, and tumor promotion, suggests that the adverse effects of this compound and/or its analogues could not be limited to acute hazards related to diarrhea induction but can also pose a significant risk after sub(chronic) exposure. Thus, further studies are needed to assess the adverse effects of long-term exposure to low doses of diarrheic toxins. Moreover, periodic monitoring of seafood and harmful algal species in the seawater is recommended to protect public health from foodborne poisonings and to minimize negative economic impacts to the shellfish industry.

ACS Style

Silvio Sosa; Aurelia Tubaro. Okadaic Acid and Other Diarrheic Toxins: Toxicological Profile. Marine and Freshwater Toxins 2015, 1 -17.

AMA Style

Silvio Sosa, Aurelia Tubaro. Okadaic Acid and Other Diarrheic Toxins: Toxicological Profile. Marine and Freshwater Toxins. 2015; ():1-17.

Chicago/Turabian Style

Silvio Sosa; Aurelia Tubaro. 2015. "Okadaic Acid and Other Diarrheic Toxins: Toxicological Profile." Marine and Freshwater Toxins , no. : 1-17.

Journal article
Published: 24 July 2014 in Toxicology Letters
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Palytoxin (PLTX) is one of the most toxic algal biotoxin known so far. It transforms the Na+/K+-ATPase into a cationic channel inducing a massive intracellular Na+ influx. However, from a mechanistic point of view, the features and the intracellular pathways leading to PLTX-induced cell death are still not completely characterized. This study on skin HaCaT keratinocytes demonstrates that PLTX induces necrosis since propidium iodide uptake was observed already after 1 h toxin exposure, an effect that was not lowered by toxin removal. Furthermore, necrotic-like morphological alterations were evidenced by confocal microscopy. Apoptosis occurrence was excluded since no caspases 3/7, caspase 8, and caspase 9 activation as well as no apoptotic bodies formation were recorded. Necrosis was preceded by a very early mitochondrial damage as indicated by JC-1 fluorescence shift, recorded already after 5 min toxin exposure. This shift was totally abolished when Na+ and Ca2+ ions were withdrawn from culture medium, whereas cyclosporine-A was ineffective, excluding the occurrence of a controlled biochemical response. These results clearly establish necrosis as the primary mechanism for PLTX-induced cell death in HaCaT cells. The rapidity of mitochondrial damage and the consequent irreversible necrosis rise serious concerns about the very fast onset of PLTX toxic effects.

ACS Style

Marco Pelin; Silvio Sosa; Sabrina Pacor; Aurelia Tubaro; Chiara Florio. The marine toxin palytoxin induces necrotic death in HaCaT cells through a rapid mitochondrial damage. Toxicology Letters 2014, 229, 440 -450.

AMA Style

Marco Pelin, Silvio Sosa, Sabrina Pacor, Aurelia Tubaro, Chiara Florio. The marine toxin palytoxin induces necrotic death in HaCaT cells through a rapid mitochondrial damage. Toxicology Letters. 2014; 229 (3):440-450.

Chicago/Turabian Style

Marco Pelin; Silvio Sosa; Sabrina Pacor; Aurelia Tubaro; Chiara Florio. 2014. "The marine toxin palytoxin induces necrotic death in HaCaT cells through a rapid mitochondrial damage." Toxicology Letters 229, no. 3: 440-450.