600 results on '"Laffon, Blanca"'
Search Results
2. Toxicity of zinc oxide nanoparticles: Cellular and behavioural effects
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Fernández-Bertólez, Natalia, Alba-González, Anabel, Touzani, Assia, Ramos-Pan, Lucía, Méndez, Josefina, Reis, Ana Teresa, Quelle-Regaldie, Ana, Sánchez, Laura, Folgueira, Mónica, Laffon, Blanca, and Valdiglesias, Vanessa
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- 2024
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3. Impact of gold nanoparticle exposure on genetic material
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Ramos-Pan, Lucía, Touzani, Assia, Fernández-Bertólez, Natalia, Fraga, Sónia, Laffon, Blanca, and Valdiglesias, Vanessa
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- 2024
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4. Measuring DNA modifications with the comet assay: a compendium of protocols
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Collins, Andrew, Møller, Peter, Gajski, Goran, Vodenková, Soňa, Abdulwahed, Abdulhadi, Anderson, Diana, Bankoglu, Ezgi Eyluel, Bonassi, Stefano, Boutet-Robinet, Elisa, Brunborg, Gunnar, Chao, Christy, Cooke, Marcus S., Costa, Carla, Costa, Solange, Dhawan, Alok, de Lapuente, Joaquin, Bo’, Cristian Del, Dubus, Julien, Dusinska, Maria, Duthie, Susan J., Yamani, Naouale El, Engelward, Bevin, Gaivão, Isabel, Giovannelli, Lisa, Godschalk, Roger, Guilherme, Sofia, Gutzkow, Kristine B., Habas, Khaled, Hernández, Alba, Herrero, Oscar, Isidori, Marina, Jha, Awadhesh N., Knasmüller, Siegfried, Kooter, Ingeborg M., Koppen, Gudrun, Kruszewski, Marcin, Ladeira, Carina, Laffon, Blanca, Larramendy, Marcelo, Hégarat, Ludovic Le, Lewies, Angélique, Lewinska, Anna, Liwszyc, Guillermo E., de Cerain, Adela López, Manjanatha, Mugimane, Marcos, Ricard, Milić, Mirta, de Andrade, Vanessa Moraes, Moretti, Massimo, Muruzabal, Damian, Novak, Matjaž, Oliveira, Rui, Olsen, Ann-Karin, Owiti, Norah, Pacheco, Mário, Pandey, Alok K., Pfuhler, Stefan, Pourrut, Bertrand, Reisinger, Kerstin, Rojas, Emilio, Rundén-Pran, Elise, Sanz-Serrano, Julen, Shaposhnikov, Sergey, Sipinen, Ville, Smeets, Karen, Stopper, Helga, Teixeira, João Paulo, Valdiglesias, Vanessa, Valverde, Mahara, van Acker, Frederique, van Schooten, Frederik-Jan, Vasquez, Marie, Wentzel, Johannes F., Wnuk, Maciej, Wouters, Annelies, Žegura, Bojana, Zikmund, Tomas, Langie, Sabine A. S., and Azqueta, Amaya
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- 2023
- Full Text
- View/download PDF
5. Toxicological Aspects of Iron Oxide Nanoparticles
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Fernández-Bertólez, Natalia, Costa, Carla, Brandão, Fátima, Teixeira, João Paulo, Pásaro, Eduardo, Valdiglesias, Vanessa, Laffon, Blanca, Crusio, Wim E., Series Editor, Dong, Haidong, Series Editor, Radeke, Heinfried H., Series Editor, Rezaei, Nima, Series Editor, Steinlein, Ortrud, Series Editor, Xiao, Junjie, Series Editor, Louro, Henriqueta, editor, and Silva, Maria João, editor
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- 2022
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6. Relationship between DNA damage measured by the comet-assay and cognitive function
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Lorenzo-López, Laura, Lema-Arranz, Carlota, Fernández-Bertólez, Natalia, Costa, Solange, Costa, Carla, Teixeira, João Paulo, Pásaro, Eduardo, Valdiglesias, Vanessa, and Laffon, Blanca
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- 2022
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7. A pooled analysis of molecular epidemiological studies on modulation of DNA repair by host factors
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Opattova, Alena, Langie, Sabine A.S., Milic, Mirta, Collins, Andrew, Brevik, Asgeir, Coskun, Erdem, Dusinska, Maria, Gaivão, Isabel, Kadioglu, Ela, Laffon, Blanca, Marcos, Ricard, Pastor, Susana, Slyskova, Jana, Smolkova, Bozena, Szilágyi, Zsófia, Valdiglesias, Vanessa, Vodicka, Pavel, Volkovova, Katarina, Bonassi, Stefano, and Godschalk, Roger W.L.
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- 2022
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8. Salivary leucocytes as suitable biomatrix for the comet assay in human biomonitoring studies
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Fernández-Bertólez, Natalia, Azqueta, Amaya, Pásaro, Eduardo, Laffon, Blanca, and Valdiglesias, Vanessa
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- 2021
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9. Applicability of flow cytometry γH2AX assay in population studies: suitability of fresh and frozen whole blood samples
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Laffon, Blanca, Sánchez-Flores, María, Fernández-Bertólez, Natalia, Pásaro, Eduardo, and Valdiglesias, Vanessa
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- 2021
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10. Frailty syndrome, biomarkers and environmental factors – A pilot study
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Teixeira-Gomes, Armanda, Lage, Bruna, Esteves, Filipa, Sousa, Ana Catarina, Pastorinho, M. Ramiro, Valdiglesias, Vanessa, Costa, Solange, Laffon, Blanca, and Teixeira, João Paulo
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- 2020
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11. Toxoplasma gondii IgG Serointensity Is Positively Associated With Frailty
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Mohyuddin, Hira, primary, Laffon, Blanca, additional, Teixeira, João P, additional, Costa, Solange, additional, Teixeira-Gomes, Armanda, additional, Pásaro, Eduardo, additional, Constantine, Niel, additional, Dagdag, Aline, additional, Ortmeyer, Heidi K, additional, Tizenberg, Boris, additional, Afram, Liubov, additional, Yen, Poyu, additional, Marano, Christopher, additional, Lowry, Christopher A, additional, Hoisington, Andrew J, additional, RachBeisel, Jill A, additional, Valdiglesias, Vanessa, additional, Lema-Arranz, Carlota, additional, Fernández-Bertólez, Natalia, additional, Maseda, Ana, additional, Millán-Calenti, José C, additional, Kovacs, Elizabeth J, additional, Gostner, Johanna M, additional, Fuchs, Dietmar, additional, Brenner, Lisa A, additional, Lorenzo-López, Laura, additional, and Postolache, Teodor T, additional
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- 2023
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12. Association of inflammatory mediators with frailty status in older adults: results from a systematic review and meta-analysis
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Marcos-Pérez, Diego, Sánchez-Flores, María, Proietti, Stefania, Bonassi, Stefano, Costa, Solange, Teixeira, Joao Paulo, Fernández-Tajes, Juan, Pásaro, Eduardo, Laffon, Blanca, and Valdiglesias, Vanessa
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- 2020
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13. Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations for describing comet assay procedures and results
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Møller, Peter, Azqueta, Amaya, Boutet-Robinet, Elisa, Koppen, Gudrun, Bonassi, Stefano, Milić, Mirta, Gajski, Goran, Costa, Solange, Teixeira, João Paulo, Costa Pereira, Cristiana, Dusinska, Maria, Godschalk, Roger, Brunborg, Gunnar, Gutzkow, Kristine B., Giovannelli, Lisa, Cooke, Marcus S., Richling, Elke, Laffon, Blanca, Valdiglesias, Vanessa, Basaran, Nursen, Del Bo’, Cristian, Zegura, Bojana, Novak, Matjaz, Stopper, Helga, Vodicka, Pavel, Vodenkova, Sona, de Andrade, Vanessa Moraes, Sramkova, Monika, Gabelova, Alena, Collins, Andrew, and Langie, Sabine A. S.
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- 2020
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14. Expanded usage of the Challenge-Comet assay as a DNA repair biomarker in human populations: protocols for fresh and cryopreserved blood samples, and for different challenge agents
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Valdiglesias, Vanessa, Sánchez-Flores, María, Fernández-Bertólez, Natalia, Au, William, Pásaro, Eduardo, and Laffon, Blanca
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- 2020
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15. Cellular and Molecular Toxicity of Iron Oxide Nanoparticles
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Laffon, Blanca, Fernández-Bertólez, Natalia, Costa, Carla, Brandão, Fátima, Teixeira, João Paulo, Pásaro, Eduardo, Valdiglesias, Vanessa, COHEN, IRUN R., Series editor, LAJTHA, ABEL, Series editor, LAMBRIS, JOHN D., Series editor, PAOLETTI, RODOLFO, Series editor, REZAEI, NIMA, Series editor, Saquib, Quaiser, editor, Faisal, Mohammad, editor, Al-Khedhairy, Abdulaziz A., editor, and Alatar, Abdulrahman A., editor
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- 2018
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16. Toxoplasma gondii IgG Serointensity Is Positively Associated With Frailty.
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Mohyuddin, Hira, Laffon, Blanca, Teixeira, João P, Costa, Solange, Teixeira-Gomes, Armanda, Pásaro, Eduardo, Constantine, Niel, Dagdag, Aline, Ortmeyer, Heidi K, Tizenberg, Boris, Afram, Liubov, Yen, Poyu, Marano, Christopher, Lowry, Christopher A, Hoisington, Andrew J, RachBeisel, Jill A, Valdiglesias, Vanessa, Lema-Arranz, Carlota, Fernández-Bertólez, Natalia, and Maseda, Ana
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TUMOR necrosis factor receptors , *TOXOPLASMA gondii , *FRAILTY , *GERIATRIC Depression Scale , *IMMUNOGLOBULIN G , *NEUROCYSTICERCOSIS - Abstract
Background Persistent inflammation related to aging ("inflammaging") is exacerbated by chronic infections and contributes to frailty in older adults. We hypothesized associations between Toxoplasma gondii (T. gondii), a common parasite causing an oligosymptomatic unremitting infection, and frailty, and secondarily between T. gondii and previously reported markers of immune activation in frailty. Methods We analyzed available demographic, social, and clinical data in Spanish and Portuguese older adults [ N = 601; age: mean (SD) 77.3 (8.0); 61% women]. Plasma T. gondii immunoglobulin G (IgG) serointensity was measured with an enzyme-linked immunosorbent assay. The Fried criteria were used to define frailty status. Validated translations of Mini-Mental State Examination, Geriatric Depression Scale, and the Charlson Comorbidity Index were used to evaluate confounders. Previously analyzed biomarkers that were significantly associated with frailty in both prior reports and the current study, and also related to T. gondii serointensity, were further accounted for in multivariable logistic models with frailty as outcome. Results In T. gondii -seropositives, there was a significant positive association between T. gondii IgG serointensity and frailty, accounting for age (p =.0002), and resisting adjustment for multiple successive confounders. Among biomarkers linked with frailty, kynurenine/tryptophan and soluble tumor necrosis factor receptor II were positively associated with T. gondii serointensity in seropositives (p <.05). Associations with other biomarkers were not significant. Conclusions This first reported association between T. gondii and frailty is limited by a cross-sectional design and warrants replication. While certain biomarkers of inflammaging were associated with both T. gondii IgG serointensity and frailty, they did not fully mediate the T. gondii –frailty association. [ABSTRACT FROM AUTHOR]
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- 2024
- Full Text
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17. Effects of Zinc Oxide Nanoparticle Exposure on Human Glial Cells and Zebrafish Embryos
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Valdiglesias, Vanessa, primary, Alba-González, Anabel, additional, Fernández-Bertólez, Natalia, additional, Touzani, Assia, additional, Ramos-Pan, Lucía, additional, Reis, Ana Teresa, additional, Moreda-Piñeiro, Jorge, additional, Yáñez, Julián, additional, Laffon, Blanca, additional, and Folgueira, Mónica, additional
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- 2023
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18. Neurotoxicity assessment of oleic acid-coated iron oxide nanoparticles in SH-SY5Y cells
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Fernández-Bertólez, Natalia, Costa, Carla, Brandão, Fátima, Kiliç, Gözde, Teixeira, Joao Paulo, Pásaro, Eduardo, Laffon, Blanca, and Valdiglesias, Vanessa
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- 2018
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19. Toxicological assessment of silica-coated iron oxide nanoparticles in human astrocytes
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Fernández-Bertólez, Natalia, Costa, Carla, Brandão, Fátima, Kiliç, Gözde, Duarte, José Alberto, Teixeira, Joao Paulo, Pásaro, Eduardo, Valdiglesias, Vanessa, and Laffon, Blanca
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- 2018
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20. Medication use in older patients and age-blind approach: narrative literature review (insufficient evidence on the efficacy and safety of drugs in older age, frequent use of PIMs and polypharmacy, and underuse of highly beneficial nonpharmacological strategies)
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Fialová, Daniela, Laffon, Blanca, Marinković, Valentina, Tasić, Ljiljana, Doro, Peter, Sόos, Gyӧngyver, Mota, Jorge, Dogan, Soner, Brkić, Jovana, Teixeira, João Paulo, Valdiglesias, Vanessa, Costa, Solange, and for the EUROAGEISM H2020 project and WG1b group “Healthy clinical strategies for healthy aging” of the EU COST Action IS 1402
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- 2019
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21. Frailty Status in Older Adults Is Related to Alterations in Indoleamine 2,3-Dioxygenase 1 and Guanosine Triphosphate Cyclohydrolase I Enzymatic Pathways
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Marcos-Pérez, Diego, Sánchez-Flores, María, Maseda, Ana, Lorenzo-López, Laura, Millán-Calenti, José C., Strasser, Barbara, Gostner, Johanna M., Fuchs, Dietmar, Pásaro, Eduardo, Valdiglesias, Vanessa, and Laffon, Blanca
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- 2017
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22. Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results
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Møller, Peter, primary, Azqueta, Amaya, additional, Sanz-Serrano, Julen, additional, Bakuradze, Tamara, additional, Richling, Elke, additional, Bankoglu, Ezgi Eyluel, additional, Stopper, Helga, additional, Bastos, Victoria Claudino, additional, Langie, Sabine A S, additional, Jensen, Annie, additional, Scavone, Francesca, additional, Giovannelli, Lisa, additional, Wojewódzka, Maria, additional, Kruszewski, Marcin, additional, Valdiglesias, Vanessa, additional, Laffon, Blanca, additional, Costa, Carla Trindade, additional, Costa, Solange, additional, Teixeira, João Paulo, additional, Marino, Mirko, additional, Del Bo’, Cristian, additional, Riso, Patrizia, additional, Zheng, Congying, additional, Shaposhnikov, Sergey, additional, and Collins, Andrew, additional
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- 2023
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23. Inter-laboratory variation in measurement of DNA damage by the alkaline comet assay in the hCOMET ring trial
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Møller, Peter, primary, Azqueta, Amaya, additional, Collia, Miguel, additional, Bakuradze, Tamara, additional, Richling, Elke, additional, Bankoglu, Ezgi Eyluel, additional, Stopper, Helga, additional, Bastos, Victoria Claudino, additional, Langie, Sabine A S, additional, Jensen, Annie, additional, Ristori, Sara, additional, Scavone, Francesca, additional, Giovannelli, Lisa, additional, Wojewódzka, Maria, additional, Kruszewski, Marcin, additional, Valdiglesias, Vanessa, additional, Laffon, Blanca, additional, Costa, Carla, additional, Costa, Solange, additional, Teixeira, João Paulo, additional, Marino, Mirko, additional, Del Bo’, Cristian, additional, Riso, Patrizia, additional, Zhang, Congying, additional, Shaposhnikov, Sergey, additional, and Collins, Andrew, additional
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- 2023
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24. Comparative study of human neuronal and glial cell sensitivity for in vitro neurogenotoxicity testing
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Laffon, Blanca, Fernández-Bertólez, Natalia, Costa, Carla, Pásaro, Eduardo, and Valdiglesias, Vanessa
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- 2017
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25. Are iron oxide nanoparticles safe? Current knowledge and future perspectives
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Valdiglesias, Vanessa, Fernández-Bertólez, Natalia, Kiliç, Gözde, Costa, Carla, Costa, Solange, Fraga, Sonia, Bessa, Maria Joao, Pásaro, Eduardo, Teixeira, João Paulo, and Laffon, Blanca
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- 2016
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26. Effects of Zinc Oxide Nanoparticle Exposure on Human Glial Cells and Zebrafish Embryos
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Valdiglesias, Vanessa, Alba-González, Anabel, Fernández-Bertólez, Natalia, Touzani, Assia, Ramos-Pan, Lucía, Reis, Ana Teresa, Moreda-Piñeiro, Jorge, Yáñez, Julián, Laffon, Blanca, Folgueira, Mónica, Valdiglesias, Vanessa, Alba-González, Anabel, Fernández-Bertólez, Natalia, Touzani, Assia, Ramos-Pan, Lucía, Reis, Ana Teresa, Moreda-Piñeiro, Jorge, Yáñez, Julián, Laffon, Blanca, and Folgueira, Mónica
- Abstract
[Abstract] Zinc oxide nanoparticles (ZnO NPs) are among the most widely used nanomaterials. They have multiple applications in cosmetics, textiles, paints, electronics and, recently, also in biomedicine. This extensive use of ZnO NPs notably increases the probability that both humans and wildlife are subjected to undesirable effects. Despite being among the most studied NPs from a toxicological point of view, much remains unknown about their ecotoxicological effects or how they may affect specific cell types, such as cells of the central nervous system. The main objective of this work was to investigate the effects of ZnO NPs on human glial cells and zebrafish embryo development and to explore the role of the released Zn2+ ions in these effects. The effects on cell viability on human A172 glial cells were assessed with an MTT assay and morphological analysis. The potential acute and developmental toxicity was assessed employing zebrafish (Danio rerio) embryos. To determine the role of Zn2+ ions in the in vitro and in vivo observed effects, we measured their release from ZnO NPs with flame atomic absorption spectrometry. Then, cells and zebrafish embryos were treated with a water-soluble salt (zinc sulfate) at concentrations that equal the number of Zn2+ ions released by the tested concentrations of ZnO NPs. Exposure to ZnO NPs induced morphological alterations and a significant decrease in cell viability depending on the concentration and duration of treatment, even after removing the overestimation due to NP interference. Although there were no signs of acute toxicity in zebrafish embryos, a decrease in hatching was detected after exposure to the highest ZnO NP concentrations tested. The ability of ZnO NPs to release Zn2+ ions into the medium in a concentration-dependent manner was confirmed. Zn2+ ions did not seem entirely responsible for the effects observed in the glial cells, but they were likely responsible for the decrease in zebrafish hatching rate. The resu
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- 2023
27. Toxoplasma gondii IgG serointensity is positively associated with frailty
- Author
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Mohyuddin, Hira, Laffon, Blanca, Teixeira, Joao P., Costa, Solange, Teixeira-Gomes, Armanda, Pásaro, Eduardo, Constantine, Niel, Dagdag, Aline, Ortmeyer, Heidi K., Tizenberg, Boris, Afram, Liubov, Yen, Poyu, Marano, Christopher, Lowry, Christopher A., Hoisington, Andrew J., RachBeisel, Jill A., Valdiglesias, Vanessa, Lema-Arranz, Carlota, Fernández-Bertólez, Natalia, Maseda, Ana, Millán-Calenti, José Carlos, Kovacs, Elizabeth J., Gostner, Johanna M., Fuchs, Dietmar, Brenner, Lisa A., Lorenzo-López, Laura, Postolache, Teodor T., Mohyuddin, Hira, Laffon, Blanca, Teixeira, Joao P., Costa, Solange, Teixeira-Gomes, Armanda, Pásaro, Eduardo, Constantine, Niel, Dagdag, Aline, Ortmeyer, Heidi K., Tizenberg, Boris, Afram, Liubov, Yen, Poyu, Marano, Christopher, Lowry, Christopher A., Hoisington, Andrew J., RachBeisel, Jill A., Valdiglesias, Vanessa, Lema-Arranz, Carlota, Fernández-Bertólez, Natalia, Maseda, Ana, Millán-Calenti, José Carlos, Kovacs, Elizabeth J., Gostner, Johanna M., Fuchs, Dietmar, Brenner, Lisa A., Lorenzo-López, Laura, and Postolache, Teodor T.
- Abstract
[Abstract] Background: Persistent inflammation related to aging ("inflammaging") is exacerbated by chronic infections and contributes to frailty in older adults. We hypothesized associations between Toxoplasma gondii (T. gondii), a common parasite causing an oligosymptomatic unremitting infection, and frailty, and secondarily between T. gondii and previously reported markers of immune activation in frailty. Methods: We analyzed available demographic, social, and clinical data in Spanish and Portuguese older adults [N = 601; age: mean (SD) 77.3 (8.0); 61% women]. Plasma T. gondii immunoglobulin G (IgG) serointensity was measured with an enzyme-linked immunosorbent assay. The Fried criteria were used to define frailty status. Validated translations of Mini-Mental State Examination, Geriatric Depression Scale, and the Charlson Comorbidity Index were used to evaluate confounders. Previously analyzed biomarkers that were significantly associated with frailty in both prior reports and the current study, and also related to T. gondii serointensity, were further accounted for in multivariable logistic models with frailty as outcome. Results: In T. gondii-seropositives, there was a significant positive association between T. gondii IgG serointensity and frailty, accounting for age (p = .0002), and resisting adjustment for multiple successive confounders. Among biomarkers linked with frailty, kynurenine/tryptophan and soluble tumor necrosis factor receptor II were positively associated with T. gondii serointensity in seropositives (p < .05). Associations with other biomarkers were not significant. Conclusions: This first reported association between T. gondii and frailty is limited by a cross-sectional design and warrants replication. While certain biomarkers of inflammaging were associated with both T. gondii IgG serointensity and frailty, they did not fully mediate the T. gondii-frailty association.
- Published
- 2023
28. Association of torquetenovirus viremia with physical frailty and cognitive impairment in three independent european cohorts
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Giacconi, Robertina, Laffon, Blanca, Costa, Solange, Teixeira-Gomes, Armanda, Maggi, Fabrizio, Macera, Lisa, Spezia, Pietro Giogio, Piacenza, Francesco, Bürkle, Alexander, Moreno-Villanueva, María, Bonassi, Stefano, Valdiglesias, Vanessa, Teixeira, Joao Paulo, Dollé, Martijn E.T., Rietman, M. Liset, Jansen, Eugène, Grune, Tilman, Gonos, Efstathios S., Franchesci, Claudio, Capri, Miriam, Weinberger, Birgit, Sikora, Ewa, Stuetz, Wolfgang, Toussaint, Olivier, Debacq-Chainiaux, Florence, Hervonen, Antti, Hurme, Mikko, Slagboom, P. Eline, Schön, Christiane, Bernhardt, Jürgen, Breusing, Nicolle, Pásaro, Eduardo, Maseda, Ana, Lorenzo-López, Laura, Millán-Calenti, José Carlos, Provinciali, Mauro, Malavolta, Marco, Giacconi, Robertina, Laffon, Blanca, Costa, Solange, Teixeira-Gomes, Armanda, Maggi, Fabrizio, Macera, Lisa, Spezia, Pietro Giogio, Piacenza, Francesco, Bürkle, Alexander, Moreno-Villanueva, María, Bonassi, Stefano, Valdiglesias, Vanessa, Teixeira, Joao Paulo, Dollé, Martijn E.T., Rietman, M. Liset, Jansen, Eugène, Grune, Tilman, Gonos, Efstathios S., Franchesci, Claudio, Capri, Miriam, Weinberger, Birgit, Sikora, Ewa, Stuetz, Wolfgang, Toussaint, Olivier, Debacq-Chainiaux, Florence, Hervonen, Antti, Hurme, Mikko, Slagboom, P. Eline, Schön, Christiane, Bernhardt, Jürgen, Breusing, Nicolle, Pásaro, Eduardo, Maseda, Ana, Lorenzo-López, Laura, Millán-Calenti, José Carlos, Provinciali, Mauro, and Malavolta, Marco
- Abstract
[Abstract] Introduction: Immunosenescence and inflammaging have been implicated in the pathophysiology of frailty. Torquetenovirus (TTV), a single-stranded DNA anellovirus, the major component of the human blood virome, shows an increased replication rate with advancing age. An elevated TTV viremia has been associated with an impaired immune function and an increased risk of mortality in the older population. The objective of this study was to analyze the relation between TTV viremia, physical frailty, and cognitive impairment. Methods: TTV viremia was measured in 1,131 nonfrail, 45 physically frail, and 113 cognitively impaired older adults recruited in the MARK-AGE study (overall mean age 64.7 ± 5.9 years), and then the results were checked in two other independent cohorts from Spain and Portugal, including 126 frail, 252 prefrail, and 141 nonfrail individuals (overall mean age: 77.5 ± 8.3 years). Results: TTV viremia ≥4log was associated with physical frailty (OR: 4.69; 95% CI: 2.06-10.67, p < 0.0001) and cognitive impairment (OR: 3.49, 95% CI: 2.14-5.69, p < 0.0001) in the MARK-AGE population. The association between TTV DNA load and frailty status was confirmed in the Spanish cohort, while a slight association with cognitive impairment was observed (OR: 1.33; 95% CI: 1.000-1.773), only in the unadjusted model. No association between TTV load and frailty or cognitive impairment was found in the Portuguese sample, although a negative association between TTV viremia and MMSE score was observed in Spanish and Portuguese females. Conclusions: These findings demonstrate an association between TTV viremia and physical frailty, while the association with cognitive impairment was observed only in the younger population from the MARK-AGE study. Further research is necessary to clarify TTV's clinical relevance in the onset and progression of frailty and cognitive decline in older individuals.
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- 2023
29. Assessing the in Vitro Toxicity of Airborne (Nano)Particles to the Human Respiratory System: From Basic to Advanced Models [Review]
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Bessa, Maria João, Brandão, Fátima, Rosário, Fernanda, Moreira, Luciana Maria Vaz, Reis, Ana Teresa, Valdiglesias, Vanessa, Laffon, Blanca, Fraga, S., Teixeira, Joao, Bessa, Maria João, Brandão, Fátima, Rosário, Fernanda, Moreira, Luciana Maria Vaz, Reis, Ana Teresa, Valdiglesias, Vanessa, Laffon, Blanca, Fraga, S., and Teixeira, Joao
- Abstract
[Abstract] Several studies have been conducted to address the potential adverse health risks attributed to exposure to nanoscale materials. While in vivo studies are fundamental for identifying the relationship between dose and occurrence of adverse effects, in vitro model systems provide important information regarding the mechanism(s) of action at the molecular level. With a special focus on exposure to inhaled (nano)particulate material toxicity assessment, this review provides an overview of the available human respiratory models and exposure systems for in vitro testing, advantages, limitations, and existing investigations using models of different complexity. A brief overview of the human respiratory system, pathway and fate of inhaled (nano)particles is also presented.
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- 2023
30. Measuring DNA modifications with the comet assay:a compendium of protocols
- Author
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Collins, Andrew, Moller, Peter, Gajski, Goran, Vodenkova, Sona, Abdulwahed, Abdulhadi, Anderson, Diana, Bankoglu, Ezgi Eyluel, Bonassi, Stefano, Boutet-Robinet, Elisa, Brunborg, Gunnar, Chao, Christy, Cooke, Marcus S. S., Costa, Carla, Costa, Solange, Dhawan, Alok, de Lapuente, Joaquin, Del Bo, Cristian, Dubus, Julien, Dusinska, Maria, Duthie, Susan J. J., El Yamani, Naouale, Engelward, Bevin, Gaivao, Isabel, Giovannelli, Lisa, Godschalk, Roger, Guilherme, Sofia, Gutzkow, Kristine B. B., Habas, Khaled, Hernandez, Alba, Herrero, Oscar, Isidori, Marina, Jha, Awadhesh N. N., Knasmueller, Siegfried, Kooter, Ingeborg M. M., Koppen, Gudrun, Kruszewski, Marcin, Ladeira, Carina, Laffon, Blanca, Larramendy, Marcelo, Le Hegarat, Ludovic, Lewies, Angelique, Lewinska, Anna, Liwszyc, Guillermo E. E., de Cerain, Adela Lopez, Manjanatha, Mugimane, Marcos, Ricard, Milic, Mirta, de Andrade, Vanessa Moraes, Moretti, Massimo, Muruzabal, Damian, Novak, Matjaz, Oliveira, Rui, Olsen, Ann-Karin, Owiti, Norah, Pacheco, Mario, Pandey, Alok K. K., Pfuhler, Stefan, Pourrut, Bertrand, Reisinger, Kerstin, Rojas, Emilio, Runden-Pran, Elise, Sanz-Serrano, Julen, Shaposhnikov, Sergey, Sipinen, Ville, Smeets, Karen, Stopper, Helga, Teixeira, Joao Paulo, Valdiglesias, Vanessa, Valverde, Mahara, van Acker, Frederique, van Schooten, Frederik-Jan, Vasquez, Marie, Wentzel, Johannes F. F., Wnuk, Maciej, Wouters, Annelies, Zegura, Bojana, Zikmund, Tomas, Langie, Sabine A. S., Azqueta, Amaya, Collins, Andrew, Moller, Peter, Gajski, Goran, Vodenkova, Sona, Abdulwahed, Abdulhadi, Anderson, Diana, Bankoglu, Ezgi Eyluel, Bonassi, Stefano, Boutet-Robinet, Elisa, Brunborg, Gunnar, Chao, Christy, Cooke, Marcus S. S., Costa, Carla, Costa, Solange, Dhawan, Alok, de Lapuente, Joaquin, Del Bo, Cristian, Dubus, Julien, Dusinska, Maria, Duthie, Susan J. J., El Yamani, Naouale, Engelward, Bevin, Gaivao, Isabel, Giovannelli, Lisa, Godschalk, Roger, Guilherme, Sofia, Gutzkow, Kristine B. B., Habas, Khaled, Hernandez, Alba, Herrero, Oscar, Isidori, Marina, Jha, Awadhesh N. N., Knasmueller, Siegfried, Kooter, Ingeborg M. M., Koppen, Gudrun, Kruszewski, Marcin, Ladeira, Carina, Laffon, Blanca, Larramendy, Marcelo, Le Hegarat, Ludovic, Lewies, Angelique, Lewinska, Anna, Liwszyc, Guillermo E. E., de Cerain, Adela Lopez, Manjanatha, Mugimane, Marcos, Ricard, Milic, Mirta, de Andrade, Vanessa Moraes, Moretti, Massimo, Muruzabal, Damian, Novak, Matjaz, Oliveira, Rui, Olsen, Ann-Karin, Owiti, Norah, Pacheco, Mario, Pandey, Alok K. K., Pfuhler, Stefan, Pourrut, Bertrand, Reisinger, Kerstin, Rojas, Emilio, Runden-Pran, Elise, Sanz-Serrano, Julen, Shaposhnikov, Sergey, Sipinen, Ville, Smeets, Karen, Stopper, Helga, Teixeira, Joao Paulo, Valdiglesias, Vanessa, Valverde, Mahara, van Acker, Frederique, van Schooten, Frederik-Jan, Vasquez, Marie, Wentzel, Johannes F. F., Wnuk, Maciej, Wouters, Annelies, Zegura, Bojana, Zikmund, Tomas, Langie, Sabine A. S., and Azqueta, Amaya
- Abstract
The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA–DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility., The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.The comet assay is commonly used to assess DNA damage. This collection of consensus protocols includes adaptations for a wide range of species and sample types, assay formats and detection of different types of DNA lesions.
- Published
- 2023
31. Long-term cryopreservation of potassium bromate positive assay controls for measurement of oxidatively damaged DNA by the Fpg-modified comet assay:results from the hCOMET ring trial
- Author
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Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, Collins, Andrew, Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, and Collins, Andrew
- Abstract
The formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent generation of DNA strand breaks. Eight laboratories used the same procedure for the treatment of monocytic THP-1 cells with potassium bromate (0, 0.5, 1.5, and 4.5 mM) and subsequent cryopreservation in a freezing medium consisting of 50% foetal bovine serum, 40% RPMI-1640 medium, and 10% dimethyl sulphoxide. The samples were analysed by the Fpg-modified comet assay three times over a 3-year period. All laboratories obtained a positive concentration–response relationship in cryopreserved samples (linear regression coefficients ranging from 0.79 to 0.99). However, there was a wide difference in the levels of Fpg-sensitive sites between the laboratory with the lowest (4.2% Tail DNA) and highest (74% Tail DNA) values in THP-1 cells after exposure to 4.5 mM KBrO3. In an attempt to assess sources of inter-laboratory variation in Fpg-sensitive sites, comet images from one experiment in each laboratory were forwarded to a central laboratory for visual scoring. There was high consistency between measurements of %Tail DNA values in each laboratory and the visual score of the same comets done in the central laboratory (r = 0.98, P < 0.001, linear regression). In conclusion, the results show that potassium bromate is a suitable positive comet assay control., The formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with little concurrent generation of DNA strand breaks. Eight laboratories used the same procedure for the treatment of monocytic THP-1 cells with potassium bromate (0, 0.5, 1.5 and 4.5 mM) and subsequent cryopreservation in freezing medium consisting of 50% foetal bovine serum, 40% RPMI-1640 medium and 10% dimethyl sulphoxide. The samples were analysed by the Fpg-modified comet assay three times over a three-year period. All laboratories obtained positive concentration-response relationship in cryopreserved samples (linear regression coefficients ranging from 0.79 to 0.99). However, there was a wide difference in the levels of Fpg-sensitive sites between laboratory with the lowest (4.2% Tail DNA) and highest (74% Tail DNA) values in THP-1 cells after exposure to 4.5 mM KBrO3. In an attempt to assess sources of inter-laboratory variation in Fpg-sensitive sites, comet images from one experiment in each laboratory were forwarded to a central laboratory for visual scoring. There was high consistency between measurements of %Tail DNA values in each laboratory and the visual score of the same comets done in the central laboratory (r = 0.98, P < 0.001, linear regression). In conclusion, the results show that potassium bromate is a suitable positive comet assay control.
- Published
- 2023
32. Visual comet scoring revisited:a guide to scoring comet assay slides and obtaining reliable results
- Author
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Møller, Peter, Azqueta, Amaya, Sanz-Serrano, Julen, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla Trindade, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zheng, Congying, Shaposhnikov, Sergey, Collins, Andrew, Møller, Peter, Azqueta, Amaya, Sanz-Serrano, Julen, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla Trindade, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zheng, Congying, Shaposhnikov, Sergey, and Collins, Andrew
- Abstract
Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20%–25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8%, and 15.2% in training sets I–III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I–III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9%–9.6%) as compared to 1 week (CV = 1.3%–6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18%–20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets., Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8% and 15.2% in training sets I-III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I-III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9-9.6%) as compared to one week (CV = 1.3-6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18-20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets.
- Published
- 2023
33. Inter-laboratory variation in measurement of DNA damage by the alkaline comet assay in the hCOMET ring trial
- Author
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Møller, Peter, Azqueta, Amaya, Collia, Miguel, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, Collins, Andrew, Møller, Peter, Azqueta, Amaya, Collia, Miguel, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, and Collins, Andrew
- Abstract
The comet assay is a simple and versatile method for measurement of DNA damage in eukaryotic cells. More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-laboratory variation in DNA migration levels occurs because there is no agreement on optimal assay conditions or suitable assay controls. The purpose of the hCOMET ring trial was to test potassium bromate (KBrO3) as a positive control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. To this end, participating laboratories used semi-standardized protocols for cell culture (i.e. cell culture, KBrO3 exposure, and cryopreservation of cells) and comet assay procedures, whereas the data acquisition was not standardized (i.e. staining of comets and image analysis). Segregation of the total variation into partial standard deviation (SD) in % Tail DNA units indicates the importance of cell culture procedures (SD = 10.9), comet assay procedures (SD = 12.3), staining (SD = 7.9) and image analysis (SD = 0.5) on the overall inter-laboratory variation of DNA migration (SD = 18.2). Future studies should assess sources of variation in each of these steps. On the positive side, the hCOMET ring trial demonstrates that KBrO3 is a robust positive control for the Fpg-modified comet assay. In conclusion, the hCOMET ring trial has demonstrated a high reproducibility of detecting genotoxic effects by the comet assay, but inter-laboratory variation of DNA migration levels is a concern., The comet assay is a simple and versatile method for measurement of DNA damage in eukaryotic cells. More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-laboratory variation in DNA migration levels occurs because there is no agreement on optimal assay conditions or suitable assay controls. The purpose of the hCOMET ring trial was to test potassium bromate (KBrO3) as a positive control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. To this end, participating laboratories used semi-standardized protocols for cell culture (i.e. cell culture, KBrO3 exposure, and cryopreservation of cells) and comet assay procedures, whereas the data acquisition was not standardized (i.e. staining of comets and image analysis). Segregation of the total variation into partial standard deviation (SD) indicates importance of cell culture procedures (SD = 10.9), comet assay procedures (SD = 12.3), staining (SD = 7.9) and image analysis (SD = 0.5) on the overall inter-laboratory variation of DNA migration (SD = 18.2). Future studies should assess sources of variation in each of these steps. On the positive side, the hCOMET ring trial demonstrates that KBrO3 is a robust positive control for the Fpg-modified comet assay. In conclusion, the hCOMET ring trial has demonstrated a high reproducibility of detecting genotoxic effects by the comet assay, but inter-laboratory variation of DNA migration levels is a concern.
- Published
- 2023
34. DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay:results from the hCOMET ring trial
- Author
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Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, Collins, Andrew, Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, Teixeira, João Paulo, Marino, Mirko, Del Bo', Cristian, Riso, Patrizia, Zhang, Congying, Shaposhnikov, Sergey, and Collins, Andrew
- Abstract
The comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 laboratories used standardized cryopreservation and thawing procedures of monocytic (THP-1) or lymphocytic (TK6) cells. Samples were cryopreserved in small aliquots in 50% foetal bovine serum, 40% cell culture medium, and 10% dimethyl sulphoxide. Subsequently, cryopreserved samples were analysed by the standard comet assay on three occasions over a 3-year period. Levels of DNA strand breaks in THP-1 cells were increased (four laboratories), unaltered (four laboratories), or decreased (two laboratories) by long-term storage. Pooled analysis indicates only a modest positive association between storage time and levels of DNA strand breaks in THP-1 cells (0.37% Tail DNA per year, 95% confidence interval: −0.05, 0.78). In contrast, DNA strand break levels were not increased by cryopreservation in TK6 cells. There was inter-laboratory variation in levels of DNA strand breaks in THP-1 cells (SD = 3.7% Tail DNA) and TK6 reference sample cells (SD = 9.4% Tail DNA), whereas the intra-laboratory residual variation was substantially smaller (i.e. SD = 0.4%–2.2% Tail DNA in laboratories with the smallest and largest variation). In conclusion, the study shows that accumulation of DNA strand breaks in cryopreserved mononuclear blood cell lines is not a matter of concern., The comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, ten laboratories used standardized cryopreservation and thawing procedures of monocytic (THP-1) or lymphocytic (TK6) cells. Samples were cryopreserved in small aliquots in 50% foetal bovine serum, 40% cell culture medium and 10% dimethyl sulphoxide. Subsequently, cryopreserved samples were analysed by the standard comet assay on three occasions over a three-year period. Levels of DNA strand breaks in THP-1 cells were increased (4 laboratories), unaltered (4 laboratories) or decreased (2 laboratories) by long-term storage. Pooled analysis indicates only a modest positive association between storage time and levels of DNA strand breaks in THP-1 cells (0.37% Tail DNA per year, 95% confidence interval: -0.05, 0.78). In contrast, DNA strand break levels were not increased by cryopreservation in TK6 cells. There was inter-laboratory variation in levels of DNA strand breaks in THP-1 cells (SD = 3.7% Tail DNA) and TK6 reference sample cells (SD = 9.4% Tail DNA), whereas the intra-laboratory residual variation was substantially smaller (i.e. SD = 0.4% to 2.2% Tail DNA in laboratories with the smallest and largest variation). In conclusion, the study shows that accumulation of DNA strand breaks in cryopreserved mononuclear blood cell lines is not a matter of concern.
- Published
- 2023
35. Long-term cryopreservation of potassium bromate positive assay controls for measurement of oxidatively damaged DNA by the Fpg-modified comet assay: results from the hCOMET ring trial.
- Author
-
Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, and Teixeira, João Paulo
- Subjects
COMETS ,DIMETHYL sulfoxide ,POTASSIUM ,DNA ,POTASSIUM channels ,DNA damage - Abstract
The formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent generation of DNA strand breaks. Eight laboratories used the same procedure for the treatment of monocytic THP-1 cells with potassium bromate (0, 0.5, 1.5, and 4.5 mM) and subsequent cryopreservation in a freezing medium consisting of 50% foetal bovine serum, 40% RPMI-1640 medium, and 10% dimethyl sulphoxide. The samples were analysed by the Fpg-modified comet assay three times over a 3-year period. All laboratories obtained a positive concentration–response relationship in cryopreserved samples (linear regression coefficients ranging from 0.79 to 0.99). However, there was a wide difference in the levels of Fpg-sensitive sites between the laboratory with the lowest (4.2% Tail DNA) and highest (74% Tail DNA) values in THP-1 cells after exposure to 4.5 mM KBrO
3 . In an attempt to assess sources of inter-laboratory variation in Fpg-sensitive sites, comet images from one experiment in each laboratory were forwarded to a central laboratory for visual scoring. There was high consistency between measurements of %Tail DNA values in each laboratory and the visual score of the same comets done in the central laboratory (r = 0.98, P < 0.001, linear regression). In conclusion, the results show that potassium bromate is a suitable positive comet assay control. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF
36. DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial.
- Author
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Møller, Peter, Azqueta, Amaya, Rodriguez-Garraus, Adriana, Bakuradze, Tamara, Richling, Elke, Bankoglu, Ezgi Eyluel, Stopper, Helga, Bastos, Victoria Claudino, Langie, Sabine A S, Jensen, Annie, Ristori, Sara, Scavone, Francesca, Giovannelli, Lisa, Wojewódzka, Maria, Kruszewski, Marcin, Valdiglesias, Vanessa, Laffon, Blanca, Costa, Carla, Costa, Solange, and Teixeira, João Paulo
- Subjects
DNA damage ,BLOOD cells ,CELL lines ,DNA analysis ,CRYOPRESERVATION of cells ,DIMETHYL sulfoxide ,MONONUCLEAR leukocytes - Abstract
The comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 laboratories used standardized cryopreservation and thawing procedures of monocytic (THP-1) or lymphocytic (TK6) cells. Samples were cryopreserved in small aliquots in 50% foetal bovine serum, 40% cell culture medium, and 10% dimethyl sulphoxide. Subsequently, cryopreserved samples were analysed by the standard comet assay on three occasions over a 3-year period. Levels of DNA strand breaks in THP-1 cells were increased (four laboratories), unaltered (four laboratories), or decreased (two laboratories) by long-term storage. Pooled analysis indicates only a modest positive association between storage time and levels of DNA strand breaks in THP-1 cells (0.37% Tail DNA per year, 95% confidence interval: −0.05, 0.78). In contrast, DNA strand break levels were not increased by cryopreservation in TK6 cells. There was inter-laboratory variation in levels of DNA strand breaks in THP-1 cells (SD = 3.7% Tail DNA) and TK6 reference sample cells (SD = 9.4% Tail DNA), whereas the intra-laboratory residual variation was substantially smaller (i.e. SD = 0.4%–2.2% Tail DNA in laboratories with the smallest and largest variation). In conclusion, the study shows that accumulation of DNA strand breaks in cryopreserved mononuclear blood cell lines is not a matter of concern. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
37. Assessing the in vitro toxicity of airborne (nano)particles to the human respiratory system: from basic to advanced models
- Author
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Bessa, Maria João, primary, Brandão, Fátima, additional, Rosário, Fernanda, additional, Moreira, Luciana, additional, Reis, Ana Teresa, additional, Valdiglesias, Vanessa, additional, Laffon, Blanca, additional, Fraga, Sónia, additional, and Teixeira, João Paulo, additional
- Published
- 2023
- Full Text
- View/download PDF
38. Association of Torquetenovirus viremia with physical frailty and cognitive impairment in three independent European cohorts
- Author
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Giacconi, Robertina, primary, Laffon, Blanca, additional, Costa, Solange, additional, Teixeira-Gomes, Armanda, additional, Maggi, Fabrizio, additional, Macera, Lisa, additional, Spezia, Pietro Giorgio, additional, Piacenza, Francesco, additional, Bürkle, Alexander, additional, Moreno-Villanueva, Maria, additional, Bonassi, Stefano, additional, Valdiglesias, Vanessa, additional, Teixeira, João Paulo, additional, Dollé, Martijn E.T., additional, Rietman, M. Liset, additional, Jansen, Eugène, additional, Grune, Tilman, additional, Gonos, Efstathios S., additional, Franceschi, Claudio, additional, Capri, Miriam, additional, Weinberger, Birgit, additional, Sikora, Ewa, additional, Stuetz, Wolfgang, additional, Toussaint, Olivier, additional, Debacq-Chainiaux, Florence, additional, Hervonen, Antti, additional, Hurme, Mikko, additional, Slagboom, P. Eline, additional, Schön, Christiane, additional, Bernhardt, Juergen, additional, Breusing, Nicolle, additional, Pásaro, Eduardo, additional, Maseda, Ana, additional, Lorenzo-López, Laura, additional, Millán-Calenti, José C., additional, Provinciali, Mauro, additional, and Malavolta, Marco, additional
- Published
- 2022
- Full Text
- View/download PDF
39. Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay
- Author
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Fernández-Bertólez, Natalia, primary, Lema-Arranz, Carlota, additional, Fraga, Sónia, additional, Teixeira, João Paulo, additional, Pásaro, Eduardo, additional, Lorenzo-López, Laura, additional, Valdiglesias, Vanessa, additional, and Laffon, Blanca, additional
- Published
- 2022
- Full Text
- View/download PDF
40. Influence of Surface Charge on Biological Behaviour of Gold Nanoparticles in Human SH-SY5Y Neuronal Cells †.
- Author
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Valdiglesias, Vanessa, Paz, Mónica, Touzani, Assia, Baúlde, Sandra, Mosquera, Jesús, Criado, Alejandro, Pásaro, Eduardo, Méndez, Josefina, Laffon, Blanca, and Fernández-Bertólez, Natalia
- Subjects
SURFACE charges ,GOLD nanoparticles ,CELL-mediated cytotoxicity ,GENETIC toxicology ,CATIONS - Abstract
Gold nanoparticles (AuNPs) are one of the most remarkable nanomaterials. Due to their small size, these NPs can cross the blood–brain barrier making them good candidates for the treatment of diseases related to the central nervous system. The main objective of the present work was to evaluate the influence of surface charge on the biological behaviour of AuNPs by assessing the cytotoxic—viability and morphological alterations—and genotoxic—double strand breaks—effects induced in neuronal cells exposed to AuNPs with different charges: cationic, anionic, and neutral. Different toxicological behaviours were obtained depending on the surface charge of the NPs. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
41. Cytotoxic Effects of Zinc Oxide Nanoparticles on Human Glial Cells †.
- Author
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Valdiglesias, Vanessa, Touzani, Assia, Ramos-Pan, Lucía, Alba-González, Anabel, Folgueira, Mónica, Moreda-Piñeiro, Jorge, Méndez, Josefina, Pásaro, Eduardo, Fernández-Bertólez, Natalia, and Laffon, Blanca
- Subjects
ZINC oxide ,NEUROGLIA ,NANOPARTICLES ,CELL survival ,CELL-mediated cytotoxicity - Abstract
Despite being one of the most studied nanomaterials, much remains unknown about the mechanism of action of zinc oxide (ZnO) nanoparticles (NP). The aim of this work was to evaluate the effects on cell viability caused by the exposure of glial cells to ZnO NP, and the role of Zn
2+ in the observed effects. The impact of ZnO NP or Zn2+ ions on cell viability was assessed by MTT assay. The exposure to ZnO NP induced a significant decrease in cell viability. The presence of Zn2+ ions released from ZnO NP was not entirely responsible for the observed cytotoxic effects. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF
42. Neuron and Glial Cells Exposed to Cerium Dioxide Nanoparticles: Results from MTT and γH2AX Assays †.
- Author
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Fernández-Bertólez, Natalia, Touzani, Assia, Martínez, Luisa, Méndez, Josefina, Reis, Ana Teresa, Costa, Carla, Fraga, Sonia, Teixeira, João Paulo, Pásaro, Eduardo, Laffon, Blanca, and Valdiglesias, Vanessa
- Subjects
CERIUM compounds ,CELL-mediated cytotoxicity ,GENETIC toxicology ,NANOMEDICINE ,BIOCOMPATIBILITY - Abstract
Cerium dioxide nanoparticles (CeO
2 NP) show antioxidant enzyme-like properties and reactive oxygen species (ROS) scavenging activity, making them a promising material for potential therapeutic applications in neurodegenerative diseases. The objective of this work was to assess the biological behavior of CeO2 NP in human SH-SY5Y neuronal and A172 glial cells by means of the MTT assay and the γH2AX assay. Despite the significant dose- and time-dependent NP internalization by both cell lines, nanoceria generally presented scarce cytotoxicity or genotoxicity, essentially restricted to the highest NP doses and longest exposure times. In conclusion, a high biocompatibility of CeO2 NP was observed under the conditions tested. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF
43. Endocrine and immunological parameters in individuals involved in Prestige spill cleanup tasks seven years after the exposure
- Author
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Laffon, Blanca, Aguilera, Francisco, Ríos-Vázquez, Julia, García-Lestón, Julia, Fuchs, Dietmar, Valdiglesias, Vanessa, and Pásaro, Eduardo
- Published
- 2013
- Full Text
- View/download PDF
44. Suitability of Salivary Leucocytes to Assess DNA Repair Ability in Human Biomonitoring Studies by the Challenge-Comet Assay
- Author
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Fernández-Bertólez, Natalia, Lema Arranz, Carlota, Fraga, S., Teixeira, Joao, Pasaro, Eduardo, Lorenzo-López, Laura, Valdiglesias, Vanessa, Laffon, Blanca, Fernández-Bertólez, Natalia, Lema Arranz, Carlota, Fraga, S., Teixeira, Joao, Pasaro, Eduardo, Lorenzo-López, Laura, Valdiglesias, Vanessa, and Laffon, Blanca
- Abstract
[Abstract] The challenge-comet assay is a simple but effective approach that provides a quantitative and functional determination of DNA repair ability, and allows to monitor the kinetics of repair process. Peripheral blood mononuclear cells (PBMC) are the cells most frequently employed in human biomonitoring studies using the challenge-comet assay, but having a validated alternative of non-invasive biomatrix would be highly convenient for certain population groups and circumstances. The objective of this study was to validate the use of salivary leucocytes in the challenge-comet assay. Leucocytes were isolated from saliva samples and challenged (either in fresh or after cryopreservation) with three genotoxic agents acting by different action mechanisms: bleomycin, methyl methanesulfonate, and ultraviolet radiation. Comet assay was performed just after treatment and at other three additional time points, in order to study repair kinetics. The results obtained demonstrated that saliva leucocytes were as suitable as PBMC for assessing DNA damage of different nature that was efficiently repaired over the evaluated time points, even after 5 months of cryopreservation (after a 24 h stimulation with PHA). Furthermore, a new parameter to determine the efficacy of the repair process, independent of the initial amount of damage induced, is proposed, and recommendations to perform the challenge-comet assay with salivary leucocytes depending on the type of DNA repair to be assessed are suggested. Validation studies are needed to verify whether the method is reproducible and results reliable and comparable among laboratories and studies.
- Published
- 2022
45. From trihalomethanes chronic daily intake through multiple exposure routes to cancer and non-cancer health risk assessment: Evidence from public Portuguese indoor swimming pools facilities using a probabilistic approach
- Author
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Costa, Carla, primary, Assunção, Ricardo, additional, Sequeira, Diana, additional, Esteves, Filipa, additional, Valdiglesias, Vanessa, additional, Laffon, Blanca, additional, Teixeira, João Paulo, additional, and Madureira, Joana, additional
- Published
- 2022
- Full Text
- View/download PDF
46. 266. Depression Moderates the Link Between Toxoplasma Gondii and Frailty.
- Author
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Mohyuddin, Hira, Laffon, Blanca, Teixeira, João P., Costa, Solange, Constantine, Niel, Kanwar, Jyoti, Dagdag, Aline, Lowry, Christopher A., Brenner, Lisa A., Volkov, Janna, Hemadeh, Ali, Lema-Arranz, Carlota, Maseda, Ana, Millán-Calenti, José C., Lorenzo-López, Laura, Valdiglesias, Vanessa, and Postolache, Teodor T.
- Subjects
- *
FRAILTY , *TOXOPLASMA gondii - Published
- 2024
- Full Text
- View/download PDF
47. Biomonitoring of a population of Portuguese workers exposed to lead
- Author
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García-Lestón, Julia, Roma-Torres, Joana, Vilares, Maria, Pinto, Rui, Cunha, Luís M., Prista, João, Teixeira, Joao Paulo, Mayan, Olga, Pásaro, Eduardo, Méndez, Josefina, and Laffon, Blanca
- Published
- 2011
- Full Text
- View/download PDF
48. Exploring Early Detection of Frailty Syndrome in Older Adults: Evaluation of Oxi-Immune Markers, Clinical Parameters and Modifiable Risk Factors
- Author
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Teixeira-Gomes, Armanda, primary, Laffon, Blanca, additional, Valdiglesias, Vanessa, additional, Gostner, Johanna M., additional, Felder, Thomas, additional, Costa, Carla, additional, Madureira, Joana, additional, Fuchs, Dietmar, additional, Teixeira, João Paulo, additional, and Costa, Solange, additional
- Published
- 2021
- Full Text
- View/download PDF
49. Immunometabolism as predictor of frailty
- Author
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Gostner, Johanna M., primary, Laffon, Blanca, additional, and Felder, Thomas K., additional
- Published
- 2021
- Full Text
- View/download PDF
50. Genotoxicity assessment of TiO2 nanoparticles in SH-SY5Y cells: suitability of the cytokinesis-block micronucleus test
- Author
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Fernández-Bertólez, Natalia, Brandão, Fátima, Costa, Carla, Lema-Arranz, C., Rodríguez-Fernández, R., Pásaro, E., Teixeira, J.P., Laffon, Blanca, and Valdiglesias, Vanessa
- Subjects
SH-SY5Y Cell ,Nanoparticles ,TiO2 ,Genotoxicity ,Genotoxicidade Ambiental - Abstract
Objective: to determine whether Cyt-B could interfere with micronuclei (MN) induction by TiO2 NP in human SH-SY5Y cells, as assessed by CBMN test. info:eu-repo/semantics/publishedVersion
- Published
- 2021
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