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DNA damage and repair activity are often assessed in blood samples from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyse the samples on the day of collection without any type of storage. For instance, certain studies use repeated sampling of cells from the same subject or samples from different subjects collected at different time-points, and it is desirable to analyse all these samples in the same comet assay experiment. In addition, flawless comet assay analyses on frozen samples open up the possibility of using this technique on biobank material. In this article we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC) and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors’ experiences indicate that various types of blood samples can be cryopreserved with only a minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen samples of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs and WB samples.
Peter Møller; Ezgi Eyluel Bankoglu; Helga Stopper; Lisa Giovannelli; Carina Ladeira; Gudrun Koppen; Goran Gajski; Andrew Collins; Vanessa Valdiglesias; Blanca Laffon; Elisa Boutet-Robinet; Hervé Perdry; Cristian Del Bo’; Sabine A S Langie; Maria Dusinska; Amaya Azqueta. Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies. Mutagenesis 2021, 36, 193 -212.
AMA StylePeter Møller, Ezgi Eyluel Bankoglu, Helga Stopper, Lisa Giovannelli, Carina Ladeira, Gudrun Koppen, Goran Gajski, Andrew Collins, Vanessa Valdiglesias, Blanca Laffon, Elisa Boutet-Robinet, Hervé Perdry, Cristian Del Bo’, Sabine A S Langie, Maria Dusinska, Amaya Azqueta. Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies. Mutagenesis. 2021; 36 (3):193-212.
Chicago/Turabian StylePeter Møller; Ezgi Eyluel Bankoglu; Helga Stopper; Lisa Giovannelli; Carina Ladeira; Gudrun Koppen; Goran Gajski; Andrew Collins; Vanessa Valdiglesias; Blanca Laffon; Elisa Boutet-Robinet; Hervé Perdry; Cristian Del Bo’; Sabine A S Langie; Maria Dusinska; Amaya Azqueta. 2021. "Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies." Mutagenesis 36, no. 3: 193-212.
The alkaline comet assay, or single cell gel electrophoresis, is one of the most popular methods for assessing DNA damage in human population. One of the open issues concerning this assay is the identification of those factors that can explain the large inter-individual and inter-laboratory variation. International collaborative initiatives such as the hCOMET project - a COST Action launched in 2016 - represent a valuable tool to meet this challenge. The aims of hCOMET were to establish reference values for the level of DNA damage in humans, to investigate the effect of host factors, lifestyle and exposure to genotoxic agents, and to compare different sources of assay variability. A database of 19,320 subjects was generated, pooling data from 105 studies run by 44 laboratories in 26 countries between 1999 and 2019. A mixed random effect log-linear model, in parallel with a classic meta-analysis, was applied to take into account the extensive heterogeneity of data, due to descriptor, specimen and protocol variability. As a result of this analysis interquartile intervals of DNA strand breaks (which includes alkali-labile sites) were reported for tail intensity, tail length, and tail moment (comet assay descriptors). A small variation by age was reported in some datasets, suggesting higher DNA damage in oldest age-classes, while no effect could be shown for sex or smoking habit, although the lack of data on heavy smokers has still to be considered. Finally, highly significant differences in DNA damage were found for most exposures investigated in specific studies. In conclusion, these data, which confirm that DNA damage measured by the comet assay is an excellent biomarker of exposure in several conditions, may contribute to improving the quality of study design and to the standardization of results of the comet assay in human populations.
Mirta Milić; Marcello Ceppi; Marco Bruzzone; Amaya Azqueta; Gunnar Brunborg; Roger Godschalk; Gudrun Koppen; Sabine Langie; Peter Møller; João Paulo Teixeira; Avdulla Alija; Diana Anderson; Vanessa Andrade; Cristina Andreoli; Fisnik Asllani; Ezgi Eyluel Bangkoglu; Magdalena Barančoková; Nursen Basaran; Elisa Boutet-Robinet; Annamaria Buschini; Delia Cavallo; Cristiana Costa Pereira; Carla Costa; Solange Costa; Juliana Da Silva; Cristian Del Boˊ; Vesna Dimitrijević Srećković; Ninoslav Djelić; Malgorzata Dobrzyńska; Zdenka Duračková; Monika Dvořáková; Goran Gajski; Serena Galati; Omar García Lima; Lisa Giovannelli; Irina A. Goroshinskaya; Annemarie Grindel; Kristine B. Gutzkow; Alba Hernández; Carlos Hernández; Kirsten B. Holven; Idoia Ibero-Baraibar; Inger Ottestad; Ela Kadioglu; Alena Kažimirová; Elena Kuznetsova; Carina Ladeira; Blanca Laffon; Palma Lamonaca; Pierre Lebailly; Henriqueta Louro; Tania Mandina Cardoso; Francesca Marcon; Ricard Marcos; Massimo Moretti; Silvia Moretti; Mojgan Najafzadeh; Zsuzsanna Nemeth; Monica Neri; Bozena Novotna; Irene Orlow; Zuzana Paduchova; Susana Pastor; Hervé Perdry; Biljana Spremo-Potparević; Dwi Ramadhani; Patrizia Riso; Paula Rohr; Emilio Rojas; Pavel Rossner; Anna Safar; Semra Sardas; Maria João Silva; Nikolay Sirota; Bozena Smolkova; Marta Staruchova; Rudolf Stetina; Helga Stopper; Ekaterina I. Surikova; Stine M. Ulven; Cinzia Lucia Ursini; Vanessa Valdiglesias; Mahara Valverde; Pavel Vodicka; Katarina Volkovova; Karl-Heinz Wagner; Lada Živković; Maria Dušinská; Andrew R. Collins; Stefano Bonassi. The hCOMET project: International database comparison of results with the comet assay in human biomonitoring. Baseline frequency of DNA damage and effect of main confounders. Mutation Research/Reviews in Mutation Research 2021, 787, 108371 .
AMA StyleMirta Milić, Marcello Ceppi, Marco Bruzzone, Amaya Azqueta, Gunnar Brunborg, Roger Godschalk, Gudrun Koppen, Sabine Langie, Peter Møller, João Paulo Teixeira, Avdulla Alija, Diana Anderson, Vanessa Andrade, Cristina Andreoli, Fisnik Asllani, Ezgi Eyluel Bangkoglu, Magdalena Barančoková, Nursen Basaran, Elisa Boutet-Robinet, Annamaria Buschini, Delia Cavallo, Cristiana Costa Pereira, Carla Costa, Solange Costa, Juliana Da Silva, Cristian Del Boˊ, Vesna Dimitrijević Srećković, Ninoslav Djelić, Malgorzata Dobrzyńska, Zdenka Duračková, Monika Dvořáková, Goran Gajski, Serena Galati, Omar García Lima, Lisa Giovannelli, Irina A. Goroshinskaya, Annemarie Grindel, Kristine B. Gutzkow, Alba Hernández, Carlos Hernández, Kirsten B. Holven, Idoia Ibero-Baraibar, Inger Ottestad, Ela Kadioglu, Alena Kažimirová, Elena Kuznetsova, Carina Ladeira, Blanca Laffon, Palma Lamonaca, Pierre Lebailly, Henriqueta Louro, Tania Mandina Cardoso, Francesca Marcon, Ricard Marcos, Massimo Moretti, Silvia Moretti, Mojgan Najafzadeh, Zsuzsanna Nemeth, Monica Neri, Bozena Novotna, Irene Orlow, Zuzana Paduchova, Susana Pastor, Hervé Perdry, Biljana Spremo-Potparević, Dwi Ramadhani, Patrizia Riso, Paula Rohr, Emilio Rojas, Pavel Rossner, Anna Safar, Semra Sardas, Maria João Silva, Nikolay Sirota, Bozena Smolkova, Marta Staruchova, Rudolf Stetina, Helga Stopper, Ekaterina I. Surikova, Stine M. Ulven, Cinzia Lucia Ursini, Vanessa Valdiglesias, Mahara Valverde, Pavel Vodicka, Katarina Volkovova, Karl-Heinz Wagner, Lada Živković, Maria Dušinská, Andrew R. Collins, Stefano Bonassi. The hCOMET project: International database comparison of results with the comet assay in human biomonitoring. Baseline frequency of DNA damage and effect of main confounders. Mutation Research/Reviews in Mutation Research. 2021; 787 ():108371.
Chicago/Turabian StyleMirta Milić; Marcello Ceppi; Marco Bruzzone; Amaya Azqueta; Gunnar Brunborg; Roger Godschalk; Gudrun Koppen; Sabine Langie; Peter Møller; João Paulo Teixeira; Avdulla Alija; Diana Anderson; Vanessa Andrade; Cristina Andreoli; Fisnik Asllani; Ezgi Eyluel Bangkoglu; Magdalena Barančoková; Nursen Basaran; Elisa Boutet-Robinet; Annamaria Buschini; Delia Cavallo; Cristiana Costa Pereira; Carla Costa; Solange Costa; Juliana Da Silva; Cristian Del Boˊ; Vesna Dimitrijević Srećković; Ninoslav Djelić; Malgorzata Dobrzyńska; Zdenka Duračková; Monika Dvořáková; Goran Gajski; Serena Galati; Omar García Lima; Lisa Giovannelli; Irina A. Goroshinskaya; Annemarie Grindel; Kristine B. Gutzkow; Alba Hernández; Carlos Hernández; Kirsten B. Holven; Idoia Ibero-Baraibar; Inger Ottestad; Ela Kadioglu; Alena Kažimirová; Elena Kuznetsova; Carina Ladeira; Blanca Laffon; Palma Lamonaca; Pierre Lebailly; Henriqueta Louro; Tania Mandina Cardoso; Francesca Marcon; Ricard Marcos; Massimo Moretti; Silvia Moretti; Mojgan Najafzadeh; Zsuzsanna Nemeth; Monica Neri; Bozena Novotna; Irene Orlow; Zuzana Paduchova; Susana Pastor; Hervé Perdry; Biljana Spremo-Potparević; Dwi Ramadhani; Patrizia Riso; Paula Rohr; Emilio Rojas; Pavel Rossner; Anna Safar; Semra Sardas; Maria João Silva; Nikolay Sirota; Bozena Smolkova; Marta Staruchova; Rudolf Stetina; Helga Stopper; Ekaterina I. Surikova; Stine M. Ulven; Cinzia Lucia Ursini; Vanessa Valdiglesias; Mahara Valverde; Pavel Vodicka; Katarina Volkovova; Karl-Heinz Wagner; Lada Živković; Maria Dušinská; Andrew R. Collins; Stefano Bonassi. 2021. "The hCOMET project: International database comparison of results with the comet assay in human biomonitoring. Baseline frequency of DNA damage and effect of main confounders." Mutation Research/Reviews in Mutation Research 787, no. : 108371.
This study aimed to evaluate occupational exposure to a styrene and xylene mixture through environmental exposure assessment and identify the potential genotoxic effects through biological monitoring. Secondly, we also exposed human peripheral blood cells in vitro to both xylene and styrene either alone or in mixture at concentrations found in occupational settings in order to understand their mechanism of action. The results obtained by air monitoring were below the occupational exposure limits for both substances. All biomarkers of effect, except for nucleoplasmic bridges, had higher mean values in workers (N = 17) compared to the corresponding controls (N = 17). There were statistically significant associations between exposed individuals and the presence of nuclear buds and oxidative damage. As for in vitro results, there was no significant influence on primary DNA damage in blood cells as evaluated by the comet assay. On the contrary, we did observe a significant increase of micronuclei and nuclear buds, but not nucleoplasmic bridges upon in vitro exposure. Taken together, both styrene and xylene have the potential to induce genomic instability either alone or in combination, showing higher effects when combined. The obtained data suggested that thresholds for individual chemicals might be insufficient for ensuring the protection of human health.
Carina Ladeira; Goran Gajski; Márcia Meneses; Marko Gerić; Susana Viegas. The genotoxicity of an organic solvent mixture: A human biomonitoring study and translation of a real-scenario exposure to in vitro. Regulatory Toxicology and Pharmacology 2020, 116, 104726 .
AMA StyleCarina Ladeira, Goran Gajski, Márcia Meneses, Marko Gerić, Susana Viegas. The genotoxicity of an organic solvent mixture: A human biomonitoring study and translation of a real-scenario exposure to in vitro. Regulatory Toxicology and Pharmacology. 2020; 116 ():104726.
Chicago/Turabian StyleCarina Ladeira; Goran Gajski; Márcia Meneses; Marko Gerić; Susana Viegas. 2020. "The genotoxicity of an organic solvent mixture: A human biomonitoring study and translation of a real-scenario exposure to in vitro." Regulatory Toxicology and Pharmacology 116, no. : 104726.
The comet assay is a well-accepted biomonitoring tool to examine the effect of dietary, lifestyle, environmental and occupational exposure on levels of DNA damage in human cells. With such a wide range of determinants for DNA damage levels, it becomes challenging to deal with confounding and certain factors are inter-related (e.g. poor nutritional intake may correlate with smoking status). This review describes the effect of intrinsic (i.e. sex, age, tobacco smoking, occupational exposure and obesity) and extrinsic (season, environmental exposures, diet, physical activity and alcohol consumption) factors on the level of DNA damage measured by the standard or enzyme-modified comet assay. Although each factor influences at least one comet assay endpoint, the collective evidence does not indicate single factors have a large impact. Thus, controlling for confounding may be necessary in a biomonitoring study, but none of the factors is strong enough to be regarded a priori as a confounder. Controlling for confounding in the comet assay requires a case-by-case approach. Inter-laboratory variation in levels of DNA damage and to some extent also reproducibility in biomonitoring studies are issues that have haunted the users of the comet assay for years. Procedures to collect specimens, and their storage, are not standardized. Likewise, statistical issues related to both sample-size calculation (before sampling of specimens) and statistical analysis of the results vary between studies. This review gives guidance to statistical analysis of the typically complex exposure, co-variate, and effect relationships in human biomonitoring studies.
Amaya Azqueta; Carina Ladeira; Lisa Giovannelli; Elisa Boutet-Robinet; Stefano Bonassi; Monica Neri; Goran Gajski; Susan Duthie; Cristian Del Bo’; Patrizia Riso; Gudrun Koppen; Nursen Basaran; Andrew Collins; Peter Møller. Application of the comet assay in human biomonitoring: An hCOMET perspective. Mutation Research/Reviews in Mutation Research 2019, 783, 108288 .
AMA StyleAmaya Azqueta, Carina Ladeira, Lisa Giovannelli, Elisa Boutet-Robinet, Stefano Bonassi, Monica Neri, Goran Gajski, Susan Duthie, Cristian Del Bo’, Patrizia Riso, Gudrun Koppen, Nursen Basaran, Andrew Collins, Peter Møller. Application of the comet assay in human biomonitoring: An hCOMET perspective. Mutation Research/Reviews in Mutation Research. 2019; 783 ():108288.
Chicago/Turabian StyleAmaya Azqueta; Carina Ladeira; Lisa Giovannelli; Elisa Boutet-Robinet; Stefano Bonassi; Monica Neri; Goran Gajski; Susan Duthie; Cristian Del Bo’; Patrizia Riso; Gudrun Koppen; Nursen Basaran; Andrew Collins; Peter Møller. 2019. "Application of the comet assay in human biomonitoring: An hCOMET perspective." Mutation Research/Reviews in Mutation Research 783, no. : 108288.
The comet assay has become one of the methods of choice for the evaluation and measurement of DNA damage. It is sensitive, quick to perform and relatively affordable for the evaluation of DNA damage and repair at the level of individual cells. The comet assay can be applied to virtually any cell type derived from different organs and tissues. Even though the comet assay is predominantly used on human cells, the application of the assay for the evaluation of DNA damage in yeast, plant and animal cells is also quite high, especially in terms of biomonitoring. The present extensive overview on the usage of the comet assay in animal models will cover both terrestrial and water environments. The first part of the review was focused on studies describing the comet assay applied in invertebrates. The second part of the review, (Part 2) will discuss the application of the comet assay in vertebrates covering cyclostomata, fishes, amphibians, reptiles, birds and mammals, in addition to chordates that are regarded as a transitional form towards vertebrates. Besides numerous vertebrate species, the assay is also performed on a range of cells, which includes blood, liver, kidney, brain, gill, bone marrow and sperm cells. These cells are readily used for the evaluation of a wide spectrum of genotoxic agents both in vitro and in vivo. Moreover, the use of vertebrate models and their role in environmental biomonitoring will also be discussed as well as the comparison of the use of the comet assay in vertebrate and human models in line with ethical principles. Although the comet assay in vertebrates is most commonly used in laboratory animals such as mice, rats and lately zebrafish, this paper will only briefly review its use regarding laboratory animal models and rather give special emphasis to the increasing usage of the assay in domestic and wildlife animals as well as in various ecotoxicological studies.
Goran Gajski; Bojana Žegura; Carina Ladeira; Matjaž Novak; Monika Sramkova; Bertrand Pourrut; Cristian Del Bo’; Mirta Milić; Kristine Bjerve Gutzkow; Solange Costa; Maria Dusinska; Gunnar Brunborg; Andrew Collins. The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates). Mutation Research/Reviews in Mutation Research 2019, 781, 130 -164.
AMA StyleGoran Gajski, Bojana Žegura, Carina Ladeira, Matjaž Novak, Monika Sramkova, Bertrand Pourrut, Cristian Del Bo’, Mirta Milić, Kristine Bjerve Gutzkow, Solange Costa, Maria Dusinska, Gunnar Brunborg, Andrew Collins. The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates). Mutation Research/Reviews in Mutation Research. 2019; 781 ():130-164.
Chicago/Turabian StyleGoran Gajski; Bojana Žegura; Carina Ladeira; Matjaž Novak; Monika Sramkova; Bertrand Pourrut; Cristian Del Bo’; Mirta Milić; Kristine Bjerve Gutzkow; Solange Costa; Maria Dusinska; Gunnar Brunborg; Andrew Collins. 2019. "The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)." Mutation Research/Reviews in Mutation Research 781, no. : 130-164.
The comet assay offers the opportunity to measure both DNA damage and repair. Various comet assay based methods are available to measure DNA repair activity, but some requirements should be met for their effective use in human biomonitoring studies. These conditions include i) robustness of the assay, ii) sources of inter- and intra-individual variability must be known, iii) DNA repair kinetics should be assessed to optimize sampling timing; and iv) DNA repair in accessible surrogate tissues should reflect repair activity in target tissues prone to carcinogenic effects. DNA repair phenotyping can be performed on frozen and fresh samples, and is a more direct measurement than genomic or transcriptomic approaches. There are mixed reports concerning the regulation of DNA repair by environmental and dietary factors. In general, exposure to genotoxic agents did not change base excision repair (BER) activity, whereas some studies reported that dietary interventions affected BER activity. On the other hand, in vitro and in vivo studies indicated that nucleotide excision repair (NER) can be altered by exposure to genotoxic agents, but studies on other life style related factors, such as diet, are rare. Thus, crucial questions concerning the factors regulating DNA repair and inter-individual variation remain unanswered. Intra-individual variation over a period of days to weeks seems limited, which is favourable for DNA repair phenotyping in biomonitoring studies. Despite this reported low intra-individual variation, timing of sampling remains an issue that needs further investigation. A correlation was reported between the repair activity in easily accessible peripheral blood mononuclear cells (PBMCs) and internal organs for both NER and BER. However, no correlation was found between tumour tissue and blood cells. In conclusion, although comet assay based approaches to measure BER/NER phenotypes are feasible and promising, more work is needed to further optimize their application in human biomonitoring and intervention studies.
Amaya Azqueta; Sabine A.S. Langie; Elisa Boutet-Robinet; Susan Duthie; Carina Ladeira; Peter Møller; Andrew R. Collins; Roger W.L. Godschalk. DNA repair as a human biomonitoring tool: Comet assay approaches. Mutation Research/Reviews in Mutation Research 2019, 781, 71 -87.
AMA StyleAmaya Azqueta, Sabine A.S. Langie, Elisa Boutet-Robinet, Susan Duthie, Carina Ladeira, Peter Møller, Andrew R. Collins, Roger W.L. Godschalk. DNA repair as a human biomonitoring tool: Comet assay approaches. Mutation Research/Reviews in Mutation Research. 2019; 781 ():71-87.
Chicago/Turabian StyleAmaya Azqueta; Sabine A.S. Langie; Elisa Boutet-Robinet; Susan Duthie; Carina Ladeira; Peter Møller; Andrew R. Collins; Roger W.L. Godschalk. 2019. "DNA repair as a human biomonitoring tool: Comet assay approaches." Mutation Research/Reviews in Mutation Research 781, no. : 71-87.
This study was designed within the frame of the COST Action hCOMET 15132 (Working Group 6), with the aim of comparing different peripheral blood cell preparations for their feasibility in human biomonitoring studies, using the comet assay for the evaluation of DNA damage. Basal levels of strand breaks/ALS and formamidopyrimidine DNA glycosylase (Fpg) - sites, and H2O2 (500 μM)-induced strand breaks, were measured in whole blood, peripheral blood mononuclear cells - lymphocytes and monocytes - and buffy coat; in fresh and 1, 4 and 12 weeks-frozen samples. The comparison among the fresh preparations showed that the basal levels of DNA damage were all very low and similar in the three samples. Frozen whole blood samples stored in cryostraws without cryoprotection showed similar basal levels of DNA damage as fresh samples, indicating that this preparation, often chosen for biobanks, resists efficiently freezing/thawing artifacts. However, long-term storage of frozen buffy coat samples in cryostraws and with no cryopreservative did not appear feasible. Storage up to 3 months of frozen cryoprotected peripheral blood mononuclear cells induced small increases in basal strand breaks and no other statistically significant modification. Altogether, this study suggests that whole blood could be the most suitable sample to be used to perform comet assay in human epidemiological biomonitoring for genotoxicity assessment in frozen samples, such as those stored in biobanks.
Carina Ladeira; Gudrun Koppen; Francesca Scavone; Lisa Giovannelli. The comet assay for human biomonitoring: Effect of cryopreservation on DNA damage in different blood cell preparations. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2019, 843, 11 -17.
AMA StyleCarina Ladeira, Gudrun Koppen, Francesca Scavone, Lisa Giovannelli. The comet assay for human biomonitoring: Effect of cryopreservation on DNA damage in different blood cell preparations. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2019; 843 ():11-17.
Chicago/Turabian StyleCarina Ladeira; Gudrun Koppen; Francesca Scavone; Lisa Giovannelli. 2019. "The comet assay for human biomonitoring: Effect of cryopreservation on DNA damage in different blood cell preparations." Mutation Research/Genetic Toxicology and Environmental Mutagenesis 843, no. : 11-17.
It is relevant to develop new monitoring techniques of carcinogenic risk associated to environmental exposition to genotoxic chemicals. The conventional biomonitoring techniques are based on laborious, expensive methods as the ones requiring isolation of lymphocytes from peripheral blood, in vitro cell culture, followed by e.g. cytokinesis-block assay and microscope observation of chromosomal abnormalities. The present work evaluated an infrared spectroscopy method, based on a simple, more economic and high-throughput procedure of analysis of whole blood processed with methanol. It was possible to identify ratios of spectral bands that are statistically different between hospital professionals occupationally exposed to antineoplastic drugs, such as 5-fluorouracil and non-hospital professionals without this exposure. It was also identified ratios of spectral bands which are statistically different between participants presenting lymphocytes with chromosomal abnormalities (as micronucleus, nuclear buds and nucleoplasmatic bridges) and participants not presenting these abnormalities. The infrared spectroscopy-based method presents therefore appealing characteristics to be applied in more intensive and/or large-scale studies of monitoring genotoxic risks.
Helder Paz Teixeira; Luís Ramalhete; Carina Ladeira; Cecília Calado. Spectral Biomarkers of Genotoxicity from Methanol Extracts of Blood. 2019 IEEE 6th Portuguese Meeting on Bioengineering (ENBENG) 2019, 1 -4.
AMA StyleHelder Paz Teixeira, Luís Ramalhete, Carina Ladeira, Cecília Calado. Spectral Biomarkers of Genotoxicity from Methanol Extracts of Blood. 2019 IEEE 6th Portuguese Meeting on Bioengineering (ENBENG). 2019; ():1-4.
Chicago/Turabian StyleHelder Paz Teixeira; Luís Ramalhete; Carina Ladeira; Cecília Calado. 2019. "Spectral Biomarkers of Genotoxicity from Methanol Extracts of Blood." 2019 IEEE 6th Portuguese Meeting on Bioengineering (ENBENG) , no. : 1-4.
The comet assay, also called single cell gel electrophoresis, is a sensitive, rapid and low-cost technique for quantifying and analysing DNA damage and repair at the level of individual cells. The assay itself can be applied on virtually any cell type derived from different organs and tissues of eukaryotic organisms. Although it is mainly used on human cells, the assay has applications also in the evaluation of DNA damage in yeast, plant and animal cells. Therefore, the purpose of this review is to give an extensive overview on the usage of the comet assay in animal models from invertebrates to vertebrates, covering both terrestrial and water biota. The comet assay is used in a variety of invertebrate species since they are regarded as interesting subjects in ecotoxicological research due to their significance in ecosystems. Hence, the first part of the review (Part 1) will discuss the application of the comet assay in invertebrates covering protozoans, platyhelminthes, planarians, cnidarians, molluscs, annelids, arthropods and echinoderms. Besides a large number of animal species, the assay is also performed on a variety of cells, which includes haemolymph, gills, digestive gland, sperm and embryo cells. The mentioned cells have been used for the evaluation of a broad spectrum of genotoxic agents both in vitro and in vivo. Moreover, the use of invertebrate models and their role from an ecotoxicological point of view will also be discussed as well as the comparison of the use of the comet assay in invertebrate and human models. Since the comet assay is still developing, its increasing potential in assessing DNA damage in animal models is crucial especially in the field of ecotoxicology and biomonitoring at the level of different species, not only humans.
Goran Gajski; Bojana Žegura; Carina Ladeira; Bertrand Pourrut; Cristian Del Bo’; Matjaž Novak; Monika Sramkova; Mirta Milic; Kristine Bjerve Gutzkow; Solange Costa; Maria Dusinska; Gunnar Brunborg; Andrew Collins. The comet assay in animal models: From bugs to whales – (Part 1 Invertebrates). Mutation Research/Reviews in Mutation Research 2019, 779, 82 -113.
AMA StyleGoran Gajski, Bojana Žegura, Carina Ladeira, Bertrand Pourrut, Cristian Del Bo’, Matjaž Novak, Monika Sramkova, Mirta Milic, Kristine Bjerve Gutzkow, Solange Costa, Maria Dusinska, Gunnar Brunborg, Andrew Collins. The comet assay in animal models: From bugs to whales – (Part 1 Invertebrates). Mutation Research/Reviews in Mutation Research. 2019; 779 ():82-113.
Chicago/Turabian StyleGoran Gajski; Bojana Žegura; Carina Ladeira; Bertrand Pourrut; Cristian Del Bo’; Matjaž Novak; Monika Sramkova; Mirta Milic; Kristine Bjerve Gutzkow; Solange Costa; Maria Dusinska; Gunnar Brunborg; Andrew Collins. 2019. "The comet assay in animal models: From bugs to whales – (Part 1 Invertebrates)." Mutation Research/Reviews in Mutation Research 779, no. : 82-113.
Bisphenol A (BPA) is one of the most widely utilized endocrine disruptors to which humans are exposed, particularity through ingestion. BPA is an aneugenic compound with a putative association to tumorigenesis. Although extensively studied in estrogen responsive cells, information regarding its effects on cells from the upper gastrointestinal tract exposed to free/active forms of BPA is still scarce. Similarly, BPA interactions with other drugs have been neglected, although it has been suggested to have a potential role in doxorubicin (DOX) chemoresistance. This study is intended to assess potential cytotoxic and genotoxic effects of BPA, as well as its interactions with DOX, in Human epithelial type 2 cells (Hep-2) originated from a human laryngeal carcinoma and in a DNA damage responsive cell line, the human lung fibroblasts (MRC-5). Cell viability was analyzed through the resazurin assay. The G protein-coupled estrogen receptor 1 (GPER) expression was visualized by immunodetection. Genotoxicity, namely DNA damage and oxidative DNA damage, were assessed by comet assay and micronuclei induction, and mitotic disruption was evaluated cytologically by fluorescent microscopy with DAPI staining. Cytotoxicity analysis showed that exposure to BPA per se does not affect cellular viability. Nevertheless, the genotoxic analysis showed that BPA induced an increase of DNA damage in the Hep-2 cell line and in oxidative damage in the MRC-5 cell line. An increase of micronuclei was also observed in both cell lines following BPA exposure. BPA and DOX co-exposures suggested that BPA acts as an antagonist of DOX effects in both cell lines. The interaction with DOX appears to be cell type dependent, exhibiting a non-monotonic response curve in MRC-5 cells, a GPER expressing cell line. Our study emphasizes the need for a deeper knowledge of BPA interactions, particularly with chemotherapeutic agents, in the context of risk assessment and public health.
Carina Ramos; Carina Ladeira; Sofia Zeferino; Ana Dias; Isabel Faria; Elisabete Cristovam; Manuel Gomes; Edna Ribeiro. Cytotoxic and genotoxic effects of environmental relevant concentrations of bisphenol A and interactions with doxorubicin. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2018, 838, 28 -36.
AMA StyleCarina Ramos, Carina Ladeira, Sofia Zeferino, Ana Dias, Isabel Faria, Elisabete Cristovam, Manuel Gomes, Edna Ribeiro. Cytotoxic and genotoxic effects of environmental relevant concentrations of bisphenol A and interactions with doxorubicin. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2018; 838 ():28-36.
Chicago/Turabian StyleCarina Ramos; Carina Ladeira; Sofia Zeferino; Ana Dias; Isabel Faria; Elisabete Cristovam; Manuel Gomes; Edna Ribeiro. 2018. "Cytotoxic and genotoxic effects of environmental relevant concentrations of bisphenol A and interactions with doxorubicin." Mutation Research/Genetic Toxicology and Environmental Mutagenesis 838, no. : 28-36.
Cytostatic drugs are highly cytotoxic agents used in cancer treatment and although their benefit is unquestionable, they have been recognized as hazardous to healthcare professionals in occupational settings. In a working environment, simultaneous exposure to cytostatics may occur creating a higher risk than that of a single substance. Hence, the present study evaluated the combined cyto/genotoxicity of a mixture of selected cytostatics with different mechanisms of action (MoA; 5-fluorouracil, cyclophosphamide and paclitaxel) towards human lymphocytes in vitro at a concentration range relevant for occupational as well as environmental exposure. The results suggest that the selected cytostatic drug mixture is potentially cyto/genotoxic and that it can induce cell and genome damage even at low concentrations. This indicates not only that such mixture may pose a risk to cell and genome integrity, but also that single compound toxicity data are not sufficient for the prediction of toxicity in a complex working environment. The presence of drugs in different amounts and with different MoA suggests the need to study the relationship between the presence of genotoxic components in the mixture and the resulting effects, taking into account the MoA of each component by itself. Therefore, this study provides new data sets necessary for scientifically-based risk assessments of cytostatic drug mixtures in occupational as well as environmental settings.
Goran Gajski; Carina Ladeira; Marko Gerić; Vera Garaj-Vrhovac; Susana Viegas. Genotoxicity assessment of a selected cytostatic drug mixture in human lymphocytes: A study based on concentrations relevant for occupational exposure. Environmental Research 2018, 161, 26 -34.
AMA StyleGoran Gajski, Carina Ladeira, Marko Gerić, Vera Garaj-Vrhovac, Susana Viegas. Genotoxicity assessment of a selected cytostatic drug mixture in human lymphocytes: A study based on concentrations relevant for occupational exposure. Environmental Research. 2018; 161 ():26-34.
Chicago/Turabian StyleGoran Gajski; Carina Ladeira; Marko Gerić; Vera Garaj-Vrhovac; Susana Viegas. 2018. "Genotoxicity assessment of a selected cytostatic drug mixture in human lymphocytes: A study based on concentrations relevant for occupational exposure." Environmental Research 161, no. : 26-34.
Cancer is one of the diseases of greatest concern in developed countries and much effort has been invested in discovering and developing therapeutics for curing cancer. Despite the improvements in antineoplastic therapeutics in the last decades, cancer is still one of the most harmful diseases worldwide. The global burden of cancer also implies financial costs: these can be direct costs, such as those related to treatment, care, and rehabilitation and indirect, which include the loss of economic output due to missed work (morbidity costs) and premature death (mortality costs). There are also hidden costs such as health insurance premiums and nonmedical expenses that are worth noting. This paper intends to present an overview of the generally forgotten impacts that the increasing number of cancer cases can have on the environment, workers who handle antineoplastic drugs, and health services. The knowledge available of each of the impacts will be addressed and discussed regarding the expected development. Overall, lessons learnt reflect on the impact of cancer through aspects not commonly evidenced in the literature or even considered in socio-economic analysis, in part due to the fact that these are difficult to contemplate in direct and indirect cancer costs already defined. Attention may be drawn to the need of continuous investment in prevention to reduce the negative impact on the environment, and in the health of workers who handle antineoplastic drugs for patients’ treatment.
Susana Viegas; Carina Ladeira; Ana Costa-Veiga; Julian Perelman; Goran Gajski. Forgotten public health impacts of cancer – an overview. Archives of Industrial Hygiene and Toxicology 2017, 68, 287 -297.
AMA StyleSusana Viegas, Carina Ladeira, Ana Costa-Veiga, Julian Perelman, Goran Gajski. Forgotten public health impacts of cancer – an overview. Archives of Industrial Hygiene and Toxicology. 2017; 68 (4):287-297.
Chicago/Turabian StyleSusana Viegas; Carina Ladeira; Ana Costa-Veiga; Julian Perelman; Goran Gajski. 2017. "Forgotten public health impacts of cancer – an overview." Archives of Industrial Hygiene and Toxicology 68, no. 4: 287-297.
The role of mycotoxins –e.g. aflatoxins, ochratoxins, trichothecenes, zearalenone, fumonisins, tremorgenic toxins, and ergot alkaloids - has been recognized in the etiology of a number of diseases. In many African countries the public health impact of chronic (indoor) and/or repeated (dietary) mycotoxin exposure is largely ignored hitherto, with impact on human health, food security, and export of African agricultural food products. Notwithstanding, African scientific research reached milestones that, when linked to findings gained by the international scientific community, make the design and implementation of science-driven governance schemes feasible. Starting from Nigeria as leading African Country, the paper i) overviews available data on mycotoxins exposure in Africa, ii) discusses new food safety issues, such as the environment-feed-food chain and toxic exposures of food producing animals in risk assessment and management; iii) identifies milestones for mycotoxins risk management already reached in West Africa; iv) points out preliminary operationalization aspects for shielding communities from direct (on health) and indirect (on trade, economies and livelihoods) effects of mycotoxins. An African science-driven engaging of scientific knowledge by development actors is expected therefore. In particular, One health/One prevention is suggested, as it proved to be a strategic and sustainable development framework.
Carina Ladeira; Chiara Frazzoli; Orish Ebere Orisakwe. Engaging One Health for Non-Communicable Diseases in Africa: Perspective for Mycotoxins. Frontiers in Public Health 2017, 5, 266 .
AMA StyleCarina Ladeira, Chiara Frazzoli, Orish Ebere Orisakwe. Engaging One Health for Non-Communicable Diseases in Africa: Perspective for Mycotoxins. Frontiers in Public Health. 2017; 5 ():266.
Chicago/Turabian StyleCarina Ladeira; Chiara Frazzoli; Orish Ebere Orisakwe. 2017. "Engaging One Health for Non-Communicable Diseases in Africa: Perspective for Mycotoxins." Frontiers in Public Health 5, no. : 266.
Molecular epidemiology is an approach increasingly used in the establishment of associations between exposure to hazardous substances and development of disease, including the possible modulation by genetic susceptibility factors. Environmental chemicals and contaminants from anthropogenic pollution of air, water and soil, but also originating specifically in occupational contexts, are potential sources of risk of development of disease. Also, diet presents an important role in this process, with some well characterized associations existing between nutrition and some types of cancer. Genotoxicity biomarkers allow the detection of early effects that result from the interaction between the individual and the environment; they are therefore important tools in cancer epidemiology and are extensively used in human biomonitoring studies. This work intends to give an overview of the potential for genotoxic effects assessment, specifically with the cytokinesis blocked micronucleus assay and comet assay in environmental and occupational scenarios, including diet. The plasticity of these techniques allows their inclusion in human biomonitoring studies, adding important information with the ultimate aim of disease prevention, in particular cancer, and so it is important that they be included as genotoxicity assays in molecular epidemiology.
Carina Ladeira; Lenka Smajdova. The use of genotoxicity biomarkers in molecular epidemiology: applications in environmental, occupational and dietary studies. AIMS Genetics 2017, 04, 166 -191.
AMA StyleCarina Ladeira, Lenka Smajdova. The use of genotoxicity biomarkers in molecular epidemiology: applications in environmental, occupational and dietary studies. AIMS Genetics. 2017; 04 (03):166-191.
Chicago/Turabian StyleCarina Ladeira; Lenka Smajdova. 2017. "The use of genotoxicity biomarkers in molecular epidemiology: applications in environmental, occupational and dietary studies." AIMS Genetics 04, no. 03: 166-191.
F.M. De Oliveira; Ana Carmona; C. Ladeira. Is mobile phone radiation genotoxic? An analysis of micronucleus frequency in exfoliated buccal cells. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2017, 822, 41 -46.
AMA StyleF.M. De Oliveira, Ana Carmona, C. Ladeira. Is mobile phone radiation genotoxic? An analysis of micronucleus frequency in exfoliated buccal cells. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2017; 822 ():41-46.
Chicago/Turabian StyleF.M. De Oliveira; Ana Carmona; C. Ladeira. 2017. "Is mobile phone radiation genotoxic? An analysis of micronucleus frequency in exfoliated buccal cells." Mutation Research/Genetic Toxicology and Environmental Mutagenesis 822, no. : 41-46.
Healthy mobile phone users aged 18-30 y.o. provided exfoliated buccal cells samples from the right and left inner cheeks. A total of 2000 cells per subject were screened for the presence of micronuclei as a sign of genotoxic damage, according to the mobile phone use profile of each user.
F.M. De Oliveira; Ana Carmona; C. Ladeira. Genotoxicity assessment data for exfoliated buccal cells exposed to mobile phone radiation. Data in Brief 2017, 15, 344 -347.
AMA StyleF.M. De Oliveira, Ana Carmona, C. Ladeira. Genotoxicity assessment data for exfoliated buccal cells exposed to mobile phone radiation. Data in Brief. 2017; 15 ():344-347.
Chicago/Turabian StyleF.M. De Oliveira; Ana Carmona; C. Ladeira. 2017. "Genotoxicity assessment data for exfoliated buccal cells exposed to mobile phone radiation." Data in Brief 15, no. : 344-347.
Bisphenol A (BPA), 2,2-bis(4-hydroxyphenyl) propane, is one of the most utilized industrial chemicals worldwide, with the ability to interfere with/or mimic estrogenic hormones with associated biological responses. Environmental human exposure to this endocrine disruptor, mostly through oral intake, is considered a generalized phenomenon, particularly in developed countries. However, in the context of occupational exposure, non-dietary exposure sources (e.g., air and contact) cannot be underestimated. Here, we performed a review of the literature on BPA occupational exposure and associated health effects. Relevantly, the authors only identified 19 studies from 2009 to 2017 that demonstrate that occupationally exposed individuals have significantly higher detected BPA levels than environmentally exposed populations and that the detection rate of serum BPA increases in relation to the time of exposure. However, only 12 studies performed in China have correlated potential health effects with detected BPA levels, and shown that BPA-exposed male workers are at greater risk of male sexual dysfunction across all domains of sexual function; also, endocrine disruption, alterations to epigenetic marks (DNA methylation) and epidemiological evidence have shown significant effects on the offspring of parents exposed to BPA during pregnancy. This overview raises awareness of the dramatic and consistent increase in the production and exposure of BPA and creates urgency to assess the actual exposure of workers to this xenoestrogen and to evaluate potential associated adverse health effects.
Edna Ribeiro; Carina Ladeira; Susana Viegas. Occupational Exposure to Bisphenol A (BPA): A Reality That Still Needs to Be Unveiled. Toxics 2017, 5, 22 .
AMA StyleEdna Ribeiro, Carina Ladeira, Susana Viegas. Occupational Exposure to Bisphenol A (BPA): A Reality That Still Needs to Be Unveiled. Toxics. 2017; 5 (3):22.
Chicago/Turabian StyleEdna Ribeiro; Carina Ladeira; Susana Viegas. 2017. "Occupational Exposure to Bisphenol A (BPA): A Reality That Still Needs to Be Unveiled." Toxics 5, no. 3: 22.
Endocrine disrupting chemicals (EDCs) are exogenous chemicals that may occur naturally (e.g., phytoestrogens), while others are industrial substances and plasticizers commonly utilized worldwide to which human exposure, particularly at low-doses, is omnipresent, persistent and occurs in complex mixtures. EDCs can interfere with/or mimic estrogenic hormones and, consequently, can simultaneously trigger diverse signaling pathways which result in diverse and divergent biological responses. Additionally, EDCs can also bioaccumulate in lipid compartments of the organism forming a mixed “body burden” of contaminants. Although the independent action of chemicals has been considered the main principle in EDCs mixture toxicity, recent studies have demonstrated that numerous effects cannot be predicted when analyzing single compounds independently. Co-exposure to these agents, particularly in critical windows of exposure, may induce hazardous health effects potentially associated with a complex “body burden” of different origins. Here, we performed an exhaustive review of the available literature regarding EDCs mixtures exposure, toxicity mechanisms and effects, particularly at the most vulnerable human life stages. Although the assessment of potential risks to human health due to exposure to EDCs mixtures is a major topic for consumer safety, information regarding effective mixtures effects is still scarce.
Edna Ribeiro; Carina Ladeira; Susana Viegas. EDCs Mixtures: A Stealthy Hazard for Human Health? Toxics 2017, 5, 5 .
AMA StyleEdna Ribeiro, Carina Ladeira, Susana Viegas. EDCs Mixtures: A Stealthy Hazard for Human Health? Toxics. 2017; 5 (1):5.
Chicago/Turabian StyleEdna Ribeiro; Carina Ladeira; Susana Viegas. 2017. "EDCs Mixtures: A Stealthy Hazard for Human Health?" Toxics 5, no. 1: 5.
The links between diet and genomic instability have been under investigation for several decades, and evidence suggests a significant causal or preventive role for various dietary factors. This study investigates the influence of macronutrients (calories, protein, and glucides) and micronutrients, such as vitamins and minerals, as assessed by a food frequency questionnaire, on genotoxicity biomarkers measured by cytokinesis-blocked micronucleus assay and comet assay. The results found significant positive and negative correlations. Micronucleus frequency tends to increase with higher intake of caffeine, calcium, magnesium, zinc, and protein ( P < .05, Spearman correlation). Calorie and omega-6 intakes are negatively correlated with DNA damage measured by the comet assay. These results are somewhat controversial because some of the correlations found are contrary to dominant views in the literature; however, we suggest that unraveling the association between diet and genetic instability requires a much better understanding of the modulating role of macronutrients and micronutrients.
Carina Ladeira; Elisabete Carolino; Manuel C Gomes; Miguel Brito. Role of Macronutrients and Micronutrients in DNA Damage: Results From a Food Frequency Questionnaire. Nutrition and Metabolic Insights 2017, 10, 1 .
AMA StyleCarina Ladeira, Elisabete Carolino, Manuel C Gomes, Miguel Brito. Role of Macronutrients and Micronutrients in DNA Damage: Results From a Food Frequency Questionnaire. Nutrition and Metabolic Insights. 2017; 10 ():1.
Chicago/Turabian StyleCarina Ladeira; Elisabete Carolino; Manuel C Gomes; Miguel Brito. 2017. "Role of Macronutrients and Micronutrients in DNA Damage: Results From a Food Frequency Questionnaire." Nutrition and Metabolic Insights 10, no. : 1.
E. Ribeiro; C. Ladeira; S. Viegas. EDCs mixture effects in human cell lines. Toxicology Letters 2016, 258, S320 .
AMA StyleE. Ribeiro, C. Ladeira, S. Viegas. EDCs mixture effects in human cell lines. Toxicology Letters. 2016; 258 ():S320.
Chicago/Turabian StyleE. Ribeiro; C. Ladeira; S. Viegas. 2016. "EDCs mixture effects in human cell lines." Toxicology Letters 258, no. : S320.