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3. Measuring DNA modifications with the comet assay: a compendium of protocols

Author

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

Published
2023
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5. A pooled analysis of molecular epidemiological studies on modulation of DNA repair by host factors

Author

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.

Published
2022
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9. Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations for describing comet assay procedures and results

Author

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.

Published
2020
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10. Gestational Weight Gain by Maternal Pre-pregnancy BMI and Childhood Problem Behaviours in School-Age Years: A Pooled Analysis of Two European Birth Cohorts

Author

Tore, Elena C., Antoniou, Evangelia E., de Groot, Renate H. M., Gielen, Marij, Godschalk, Roger W. L., Roumeliotaki, Theano, and Smits, Luc

Subjects
Obesity -- Complications and side effects, Weight gain -- Health aspects, Child psychopathology -- Risk factors, Child development -- Health aspects, Pregnant women -- Physiological aspects -- Health aspects, Prenatal influences -- Health aspects, Health care industry
Abstract

Objectives Maternal pre-pregnancy weight is known to affect foetal development. However, it has not yet been clarified if gestational weight gain is associated with childhood behavioural development. Methods We performed a pooled analysis of two prospective birth cohorts to investigate the association between gestational weight gain and childhood problem behaviours, and the effect modification of maternal pre-pregnancy BMI. In total, 378 mother-child pairs from the Maastricht Essential Fatty Acids Birth cohort (MEFAB) and 414 pairs from the Rhea Mother-Child cohort were followed up from early pregnancy to 6-7 years post-partum. At follow up, parents assessed their children's behaviour, measured as total problems, internalizing and externalizing behaviours, with the Child Behaviour Checklist. We computed cohort- and subject-specific gestational weight gain trajectories using mixed-effect linear regression models. Fractional polynomial regressions, stratified by maternal pre-pregnancy BMI status, were then used to examine the association between gestational weight gain and childhood problem behaviours. Results In the pre-pregnancy overweight/obese group, greater gestational weight gain was associated with higher problem behaviours. On average, children of women with overweight/obesity who gained 0.5 kg/week scored 25 points higher (on a 0-100 scale) in total problems and internalizing behaviours, and about 18 points higher in externalizing behaviours than children whose mothers gained 0.2 kg/week. Inconsistent results were found in the pre-pregnancy normal weight group. Conclusions for Practice Excessive gestational weight gain in women with pre-pregnancy overweight/obesity might increase problem behaviours in school-age children. Particular attention should be granted to avoid excessive weight gain in women with a pre-pregnancy overweight or obesity., Author(s): Elena C. Tore [sup.1] [sup.2] , Evangelia E. Antoniou [sup.1] , Renate H. M. de Groot [sup.3] [sup.4] , Marij Gielen [sup.4] , Roger W. L. Godschalk [sup.5] , [...]

Published
2020
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17. Optimizing the Comet Assay-Based In Vitro DNA Repair Assay for Placental Tissue: A Pilot Study with Pre-Eclamptic Patients.

Author

Mircheva, Anastasiya, Vangrieken, Philippe, Al-Nasiry, Salwan, van Schooten, Frederik-Jan, Godschalk, Roger W. L., and Langie, Sabine A. S.

Subjects
DNA repair, PREGNANCY proteins, PLACENTA, COMETS, PILOT projects
Abstract

The comet assay-based in vitro DNA repair assay has become a common tool for quantifying base excision repair (BER) activity in human lymphocytes or cultured cells. Here, we optimized the protocol for studying BER in human placental tissue because the placenta is a non-invasive tissue for biomonitoring of early-life exposures, and it can be used to investigate molecular mechanisms associated with prenatal disorders. The optimal protein concentration of placental protein extracts for optimal damage recognition and incision was 2 mg protein/mL. The addition of aphidicolin did not lead to reduced non-specific incisions and was, therefore, not included in the optimized protocol. The interval between sample collection and analysis did not affect BER activity up to 70 min. Finally, this optimized protocol was tested on pre-eclamptic (PE) placental tissues (n = 11) and significantly lower BER activity in PE placentas compared to controls (n = 9) was observed. This was paralleled by a significant reduction in the expression of BER-related genes and increased DNA oxidation in PE placentas. Our study indicates that BER activity can be determined in placentas, and lower activity is present in PE compared with healthy. These findings should be followed up in prospective clinical investigations to examine BER's role in the advancement of PE. [ABSTRACT FROM AUTHOR]

Published
2024
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19. Measuring DNA modifications with the comet assay:a compendium of protocols

Author

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. Short- and long-term reproducibility of the COMET assay for measuring DNA damage biomarkers in frozen blood samples of the EPIC-Heidelberg cohort

Author

Bankoglu, Ezgi Eyluel, primary, Mukama, Trasias, additional, Katzke, Verena, additional, Stipp, Franzisca, additional, Johnson, Theron, additional, Kühn, Tilman, additional, Seyfried, Florian, additional, Godschalk, Roger, additional, Collins, Andrew, additional, Kaaks, Rudolf, additional, and Stopper, Helga, additional

Published
2022
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41. Author Correction: DNA damage in circulating leukocytes measured with the comet assay may predict the risk of death (Scientific Reports, (2021), 11, 1, (16793), 10.1038/s41598-021-95976-7)

Author

Bonassi, Stefano, Ceppi,Marcello, Møller, Peter, Azqueta, Amaya, Milić, Mirta, Neri, Monica, Brunborg, Gunnar, Godschalk, Roger, Koppen, Gudrun, Langie, Sabine A. S., Teixeira, João Paulo, Bruzzone, Marco, Da Silva, Juliana, Benedetti, Danieli, Cavallo, Delia, Ursini, Cinzia Lucia, Giovannelli, Lisa, Moretti, Silvia, Riso, Patrizia, Del Bo’, Cristian, Russo, Patrizia, Dobrzyńska, Malgorzata, Goroshinskaya, Irina A., Surikova, Ekaterina I., Staruchova, Marta, Barančokova, Magdalena, Volkovova, Katarina, Kažimirova, Alena, Smolkova, Bozena, Laffon, Blanca, Valdiglesias, Vanessa, Pastor, Susana, Marcos, Ricard, Hernández, Alba, Gajski, Goran, Spremo-Potparević, Biljana, Živković, Lada, Boutet-Robinet, Elisa, Perdry, Hervé, Lebailly, Pierre, Perez, Carlos L., Basaran, Nursen, Nemeth, Zsuzsanna, Safar, Anna, Dusinska, Maria, Collins, Andrew, Anderson, Diana, Andrade, Vanessa, Pereira, Cristiana Costa, Costa, Solange, Gutzkow, Kristine B., Ladeira, Carina, Moretti, Massimo, Costa, Carla, Orlow, Irene, Rojas, Emilio, Pourrut, Bertrand, Kruszewski, Marcin, Knasmueller, Siegfried, Shaposhnikov, Sergey, Žegura, Bojana, and Stopper, Helga

Abstract

Link to the corrected article: [https://farfar.pharmacy.bg.ac.rs/handle/123456789/3944]

Published
2021

42. DNA damage in circulating leukocytes measured with the comet assay may predict the risk of death

Author

Bonassi, Stefano, Ceppi, Marcello, Møller, Peter, Azqueta, Amaya, Milić, Mirta, Monica, Neri, Brunborg, Gunnar, Godschalk, Roger, Koppen, Gudrun, Langie, Sabine A. S., Teixeira, João Paulo, Bruzzone, Marco, Da Silva, Juliana, Benedetti, Danieli, Cavallo, Delia, Ursini, Cinzia Lucia, Giovannelli, Lisa, Moretti, Silvia, Riso, Patrizia, Del Bo’, Cristian, Russo, Patrizia, Dobrzyńska, Malgorzata, Goroshinskaya, Irina A., Surikova, Ekaterina I., Staruchova, Marta, Barančokova, Magdalena, Volkovova, Katarina, Kažimirova, Alena, Smolkova, Bozena, Laffon, Blanca, Valdiglesias, Vanessa, Pastor, Susana, Marcos, Ricard, Hernández, Alba, Gajski, Goran, Spremo-Potparević, Biljana, Živković, Lada, Boutet-Robinet, Elisa, Perdry, Hervé, Lebailly, Pierre, Perez, Carlos L., Basaran, Nursen, Nemeth, Zsuzsanna, Safar, Anna, Dusinska, Maria, Collins, Andrew, Anderson, Diana, Andrade, Vanessa, Pereira, Cristiana Costa, Costa, Solange, Gutzkow, Kristine B., Ladeira, Carina, Moretti, Massimo, Costa, Carla, Orlow, Irene, Rojas, Emilio, Pourrut, Bertrand, Kruszewski, Marcin, Knasmueller, Siegfried, Shaposhnikov, Sergey, Žegura, Bojana, Stopper, Helga, LESUR, Hélène, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS San Raffaele Pisana), Ospedale Policlinico San Martino [Genoa], University of Copenhagen = Københavns Universitet (UCPH), Universidad de Navarra [Pamplona] (UNAV), Instituto de Investigación Sanitaria de Navarra [Pamplona, Spain] (IdiSNA), Institute for Medical Research and Occupational Health, Norwegian Institute of Public Health [Oslo] (NIPH), School of Nutrition and Translational Research in Metabolism [Maastricht] (NUTRIM), Maastricht University [Maastricht], Flemish Institute for Technological Research (VITO), Instituto Nacional de Saùde Dr Ricardo Jorge [Portugal] (INSA), Universidade Luterana do Brasil - ULBRA (BRAZIL), Universidade Luterana do Brasil (ULBRA), Istituto Nazionale per l’Assicurazione contro gli Infortuni sul Lavoro [Italian Workers Compensation Authority] (INAIL), NEUROFARBA Department [Firenze, Italy], Università degli Studi di Firenze = University of Florence (UniFI), Université de Florence, Università degli Studi di Milano = University of Milan (UNIMI), IRCCS San Raffaele Scientific Institute [Milan, Italie], National Institute of Public Health - National Institute of Hygiene [Poland], N.N. Blokhin National Medical Research Center of Oncology, Slovak Medical University of Bratislava (SMU), Slovak Academy of Science [Bratislava] (SAS), University of A Coruña (UDC), Instituto de Investigación Biomédica de A Coruña [La Corogne, Espagne] (INIBIC), A Coruña University Hospital [La Corogne, Espagne], Universitat Autònoma de Barcelona (UAB), CIBER de Epidemiología y Salud Pública (CIBERESP), School of Medecine [Belgrade], University of Belgrade [Belgrade], Contaminants & Stress Cellulaire (ToxAlim-COMICS), ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre de recherche en épidémiologie et santé des populations (CESP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Unité de recherche interdisciplinaire pour la prévention et le traitement des cancers (ANTICIPE), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Centre Régional de Lutte contre le Cancer François Baclesse [Caen] (UNICANCER/CRLC), Normandie Université (NU)-UNICANCER-Tumorothèque de Caen Basse-Normandie (TCBN)-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM), Instituto de Ciencias Básicas y Preclínicas Victoria de Girón, Hacettepe University = Hacettepe Üniversitesi, National center for public health [Hungary], Norwegian Institute for Air Research (NILU), University of Oslo (UiO), This article is based upon work from COST Action CA15132 (hCOMET), supported by COST (European Cooperation in Science and Technology). The work of SB and PR was supported by funds granted by the Italian Ministry of Health for Institutional Research (Ricerca Corrente)., Farmacologie en Toxicologie, RS: NUTRIM - R3 - Respiratory & Age-related Health, University of Copenhagen = Københavns Universitet (KU), Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), Università degli Studi di Milano [Milano] (UNIMI), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Régional de Lutte contre le Cancer François Baclesse [Caen] (UNICANCER/CRLC), UNICANCER-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-UNICANCER, and Instituto de Saúde Pública da Universidade do Porto

Subjects
Oncology, [SDV]Life Sciences [q-bio], Chromosomal aberrations, Human cancer, Lymphocytes, Predicts, Frequency, Validation, Biomarkers, Hallmarks, Adducts, Repair, Cohort, Kaplan-Meier Estimate, 0302 clinical medicine, Neoplasms, Leukocytes, Medicine, Gel electrophoresis, 0303 health sciences, Multidisciplinary, Hazard ratio, 3. Good health, [SDV] Life Sciences [q-bio], 030220 oncology & carcinogenesis, Biomarker (medicine), LYMPHOCYTES PREDICTS, Comet Assay, Cell-Free Nucleic Acids, medicine.medical_specialty, DNA damage, Science, FREQUENCY, Article, VALIDATION, 03 medical and health sciences, ADDUCTS, Internal medicine, Humans, COHORT, Risk factor, Author Correction, 030304 developmental biology, Proportional Hazards Models, HALLMARKS, REPAIR, HUMAN CANCER, business.industry, Proportional hazards model, Comet assay, biomarker, leukocytes, epidemiology, Comet assay, CHROMOSOMAL-ABERRATIONS, Risk factors, Genotoxicidade Ambiental, business, DNA Damage
Abstract

Author Correction: https://doi.org/10.1038/s41598-021-98620-6. The original version of this Article contained errors: The spelling of the author Monica Neri was incorrectly given as Neri Monica; Additionally, Affiliation 39 was incorrectly given as ‘National Institute of Health, Lisbon, Portugal’, the correct affiliation is listed below: National Institute of Health Doutor Ricardo Jorge, Porto, Portugal. The original Article has been corrected. [Abstract] The comet assay or single cell gel electrophoresis, is the most common method used to measure strand breaks and a variety of other DNA lesions in human populations. To estimate the risk of overall mortality, mortality by cause, and cancer incidence associated to DNA damage, a cohort of 2,403 healthy individuals (25,978 person-years) screened in 16 laboratories using the comet assay between 1996 and 2016 was followed-up. Kaplan–Meier analysis indicated a worse overall survival in the medium and high tertile of DNA damage (p

Published
2021

43. 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

Author

Milić, Mirta, Ceppi, Marcello, Bruzzone, Marco, Azqueta, A., Brunborg, Gunnar, Godschalk, Roger W.L., Koppen, Gudrun F., Sardas, Semra, İstinye Üniversitesi, Eczacılık Fakültesi, Eczacılık Meslek Bilimleri Bölümü, Semra Şardaş / 0000-0001-5456-8636, and Sardas, Semra

Subjects
Human Biomonitoring, Pooled Analysis, Comet Assay, Biomarkers, DNA Damage
Abstract

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. WOS:000658539600011 34083035 Q1

Published
2021

44. 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

Author

Mili, Mirta, Ceppi, Marcello, Bruzzone, Marco, Azqueta, Amaya, Brunborg, Gunnar, Godschalk, Roger, Koppen, Gudrun, Langie, Sabine, Møller, Peter, Teixeira, Joao Paulo, Alija, Avdulla, Anderson, Diana, Andrade, Vanessa, Andreoli, Cristina, Asllani, Fisnik, Bangkoglu, Ezgi Eyluel, Barancokova, Magdalena, Basaran, Nursen, Boutet-Robinet, Elisa, Buschini, Annamaria, Cavallo, Delia, Pereira, Cristiana Costa, Costa, Carla, Costa, Solange, Silva, Juliana Da, Bo, Cristian Del, Sreckovic, Vesna Dimitrijevic, Djelic, Ninoslav, Durackova, Zdenka, Dvorakova, Monika, Gajski, Goran, Galati, Serena, Lima, Omar Garcia, Giovannelli, Lisa, Goroshinskaya, Irina A., Grindel, Annemarie, Gutzkow, Kristine B., Hernandez, Alba, Hernandez, Carlos, Holven, Kirsten B., Ibero-Baraibar, Idoia, Ottestad, Inger, Kadioglu, Ela, Kazimirova, Alena, Kuznetsova, Elena, Ladeira, Carina, Laffon, Blanca, Lamonaca, Palma, Lebailly, Pierre, Louro, Henriqueta, Cardoso, Tania Mandina, Marcon, Francesca, Marcos, Ricard, Moretti, Massimo, Moretti, Silvia, Najafzadeh, Mojgan, Nemeth, Zsuzsanna, Neri, Monica, Novotna, Bozena, Orlow, Irene, Paduchova, Zuzana, Pastor, Susana, Perdry, Herve, Spremo-Potparevic, Biljana, Ramadhani, Dwi, Riso, Patrizia, Rohr, Paula, Rojas, Emilio, Rossner, Pavel, Safar, Anna, Sardas, Semra, Silva, Maria Joao, Sirota, Nikolay, Smolkova, Bozena, Staruchova, Marta, Stetina, Rudolf, Stopper, Helga, Surikova, Ekaterina I., Ulven, Stine M., Ursini, Cinzia Lucia, Valdiglesias, Vanessa, Valverde, Mahara, Vodicka, Pavel, Volkovova, Katarina, Wagner, Karl-Heinz, Zivkovic, Lada, Dusinska, Maria, Collins, Andrew R., Bonassi, Stefano, Mili, Mirta, Ceppi, Marcello, Bruzzone, Marco, Azqueta, Amaya, Brunborg, Gunnar, Godschalk, Roger, Koppen, Gudrun, Langie, Sabine, Møller, Peter, Teixeira, Joao Paulo, Alija, Avdulla, Anderson, Diana, Andrade, Vanessa, Andreoli, Cristina, Asllani, Fisnik, Bangkoglu, Ezgi Eyluel, Barancokova, Magdalena, Basaran, Nursen, Boutet-Robinet, Elisa, Buschini, Annamaria, Cavallo, Delia, Pereira, Cristiana Costa, Costa, Carla, Costa, Solange, Silva, Juliana Da, Bo, Cristian Del, Sreckovic, Vesna Dimitrijevic, Djelic, Ninoslav, Durackova, Zdenka, Dvorakova, Monika, Gajski, Goran, Galati, Serena, Lima, Omar Garcia, Giovannelli, Lisa, Goroshinskaya, Irina A., Grindel, Annemarie, Gutzkow, Kristine B., Hernandez, Alba, Hernandez, Carlos, Holven, Kirsten B., Ibero-Baraibar, Idoia, Ottestad, Inger, Kadioglu, Ela, Kazimirova, Alena, Kuznetsova, Elena, Ladeira, Carina, Laffon, Blanca, Lamonaca, Palma, Lebailly, Pierre, Louro, Henriqueta, Cardoso, Tania Mandina, Marcon, Francesca, Marcos, Ricard, Moretti, Massimo, Moretti, Silvia, Najafzadeh, Mojgan, Nemeth, Zsuzsanna, Neri, Monica, Novotna, Bozena, Orlow, Irene, Paduchova, Zuzana, Pastor, Susana, Perdry, Herve, Spremo-Potparevic, Biljana, Ramadhani, Dwi, Riso, Patrizia, Rohr, Paula, Rojas, Emilio, Rossner, Pavel, Safar, Anna, Sardas, Semra, Silva, Maria Joao, Sirota, Nikolay, Smolkova, Bozena, Staruchova, Marta, Stetina, Rudolf, Stopper, Helga, Surikova, Ekaterina I., Ulven, Stine M., Ursini, Cinzia Lucia, Valdiglesias, Vanessa, Valverde, Mahara, Vodicka, Pavel, Volkovova, Katarina, Wagner, Karl-Heinz, Zivkovic, Lada, Dusinska, Maria, Collins, Andrew R., and Bonassi, Stefano

Abstract

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.(c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creative

Published
2021

48. Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories

Author

Godschalk, Roger W. L., Ersson, Clara, Stępnik, Maciej, Ferlińska, Magdalena, Palus, Jadwiga, Teixeira, João Paulo, Costa, Solange, Jones, George D. D., Higgins, Jennifer A., Kain, Johanna, Möller, Lennart, Forchhammer, Lykke, Loft, Steffen, Lorenzo, Yolanda, Collins, Andrew R., van Schooten, Frederik-Jan, Laffon, Blanca, Valdiglesias, Vanessa, Cooke, Marcus, Mistry, Vilas, Karbaschi, Mahsa, Phillips, David H., Sozeri, Osman, Routledge, Michael N., Nelson-Smith, Kirsty, Riso, Patrizia, Porrini, Marisa, López de Cerain, Adela, Azqueta, Amaya, Matullo, Giuseppe, Allione, Alessandra, and Møller, Peter

Published
2014
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49. Minimum Information for Reporting on the Comet Assay (MIRCA):recommendations for describing comet assay procedures and results

Author

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, Langie, Sabine A.S., 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.

Abstract

The comet assay is a widely used test for the detection of DNA damage and repair activity. However, there are interlaboratory differences in reported levels of baseline and induced damage in the same experimental systems. These differences may be attributed to protocol differences, although it is difficult to identify the relevant conditions because detailed comet assay procedures are not always published. Here, we present a Consensus Statement for the Minimum Information for Reporting Comet Assay (MIRCA) providing recommendations for describing comet assay conditions and results. These recommendations differentiate between ‘desirable’ and ‘essential’ information: ‘essential’ information refers to the precise details that are necessary to assess the quality of the experimental work, whereas ‘desirable’ information relates to technical issues that might be encountered when repeating the experiments. Adherence to MIRCA recommendations should ensure that comet assay results can be easily interpreted and independently verified by other researchers.

Published
2020

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