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Article ; Online: In memory of Dr. Ir. Gudrun Koppen (1969-2024).

Collins, Andrew R / Azqueta, Amaya / Schoeters, Greet / Slingers, Gitte / Dusinska, Maria / Langie, Sabine A S

Mutation research. Genetic toxicology and environmental mutagenesis

2024 Volume 895, Page(s) 503751

Language	English
Publishing date	2024-03-19
Publishing country	Netherlands
Document type	Editorial
ISSN	1879-3592
ISSN (online)	1879-3592
DOI	10.1016/j.mrgentox.2024.503751
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Article ; Online: Further development of CometChip technology to measure DNA damage in vitro and in vivo: Comparison with the 2 gels/slide format of the standard and enzyme-modified comet assay.

Collia, Miguel / Møller, Peter / Langie, Sabine A S / Vettorazzi, Ariane / Azqueta, Amaya

Toxicology

2023 Volume 501, Page(s) 153690

Abstract: DNA damage plays a pivotal role in carcinogenesis and other diseases. The comet assay has been used for more than three decades to measure DNA damages. The 1-2 gels/slide format is the most used version of the assay. In 2010, a high throughput 96 ... ...

Abstract	DNA damage plays a pivotal role in carcinogenesis and other diseases. The comet assay has been used for more than three decades to measure DNA damages. The 1-2 gels/slide format is the most used version of the assay. In 2010, a high throughput 96 macrowell format with a spatially encoded array of microwells patterned in agarose was developed, called the CometChip. The commercial version (CometChip®) has been used for the in vitro standard version of the comet assay (following the manufacturer's protocol), although it has not been compared directly with the 2 gels/slide format. The aim of this work is to developed new protocols to allow use of DNA repair enzymes as well as the analysis of in vivo frozen tissue samples in the CometChip®, to increase the throughput, and to compare its performance with the classic 2 gels/slide format. We adapted the manufacturer's protocol to allow the use of snap frozen tissue samples, using male Wistar rats orally dosed with methyl methanesulfonate (MMS, 200 mg/kg b.w.), and to detect altered nucleobases using DNA repair enzymes, with TK6 cells treated with potassium bromate (KBrO
MeSH term(s)	Male ; Animals ; Rats ; Comet Assay/methods ; Rats, Wistar ; DNA Damage ; DNA Repair Enzymes ; Gels
Chemical Substances	DNA Repair Enzymes (EC 6.5.1.-) ; Gels
Language	English
Publishing date	2023-11-29
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	184557-3
ISSN	1879-3185 ; 0300-483X
ISSN (online)	1879-3185
ISSN	0300-483X
DOI	10.1016/j.tox.2023.153690
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Zs.A 888: Show issues

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Article ; Online: Optimizing the Comet Assay-Based In Vitro DNA Repair Assay for Placental Tissue: A Pilot Study with Pre-Eclamptic Patients.

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

International journal of molecular sciences

2023 Volume 25, Issue 1

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 ... ...

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 (
MeSH term(s)	Pregnancy ; Humans ; Female ; Pilot Projects ; Comet Assay ; Prospective Studies ; Placenta ; DNA Repair ; Pre-Eclampsia/genetics
Language	English
Publishing date	2023-12-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms25010187
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Article ; Online: Upregulation of mNEIL3 in Ogg1-null cells is a potential backup mechanism for 8-oxoG repair.

Higgs, Ellen B / Godschalk, Roger / Langie, Sabine A S / van Schooten, Frederik-Jan / Hodges, Nikolas J

Mutagenesis

2021 Volume 36, Issue 6, Page(s) 437–444

Abstract: Reactive oxygen species formation and resultant oxidative damage to DNA are ubiquitous events in cells, the homeostasis of which can be dysregulated in a range of pathological conditions. Base excision repair (BER) is the primary repair mechanism for ... ...

Abstract	Reactive oxygen species formation and resultant oxidative damage to DNA are ubiquitous events in cells, the homeostasis of which can be dysregulated in a range of pathological conditions. Base excision repair (BER) is the primary repair mechanism for oxidative genomic DNA damage. One prevalent oxidised base modification, 8-oxoguanine (8-oxoG), is recognised by 8-oxoguanine glycosylase-1 (OGG1) initiating removal and repair via BER. Surprisingly, Ogg1 null mouse embryonic fibroblasts (mOgg1-/- MEFs) do not accumulate 8-oxoG in the genome to the extent expected. This suggests that there are backup repair mechanisms capable of repairing 8-oxoG in the absence of OGG1. In the current study, we identified components of NER (Ercc1, Ercc4, Ercc5), BER (Lig1, Tdg, Nthl1, Mpg, Mgmt, NEIL3), MMR (Mlh1, Msh2, Msh6) and DSB (Brip1, Rad51d, Prkdc) pathways that are transcriptionally elevated in mOgg1-/- MEFs. Interestingly, all three nucleotide excision repair genes identified: Ercc1 (2.5 ± 0.2-fold), Ercc4 (1.5 ± 0.1-fold) and Ercc5 (1.7 ± 0.2-fold) have incision activity. There was also a significant functional increase in NER activity (42.0 ± 7.9%) compared to WT MEFs. We also observed upregulation of both Neil3 mRNA (37.9 ± 1.6-fold) and protein in mOgg1-/- MEFs. This was associated with a 3.4 ± 0.4-fold increase in NEIL3 substrate sites in genomic DNA of cells treated with BSO, consistent with the ability of NEIL3 to remove 8-oxoG oxidation products from genomic DNA. In conclusion, we suggest that in Ogg1-null cells, upregulation of multiple DNA repair proteins including incision components of the NER pathway and Neil3 are important compensatory responses to prevent the accumulation of genomic 8-oxoG.
MeSH term(s)	Animals ; Cells, Cultured ; Comet Assay/methods ; DNA Damage ; DNA Glycosylases/genetics ; DNA Glycosylases/metabolism ; DNA Repair ; DNA-Binding Proteins/metabolism ; Embryo, Mammalian/metabolism ; Endodeoxyribonucleases/genetics ; Endodeoxyribonucleases/metabolism ; Endonucleases/metabolism ; Fibroblasts/metabolism ; Gene Expression Regulation ; Guanine/analogs & derivatives ; Guanine/metabolism ; Lymphocytes, Null/metabolism ; Mice ; Nuclear Proteins/metabolism ; Oxidative Stress ; Reactive Oxygen Species/metabolism ; Transcription Factors/metabolism
Chemical Substances	DNA excision repair protein ERCC-5 ; DNA-Binding Proteins ; Nuclear Proteins ; Reactive Oxygen Species ; Transcription Factors ; xeroderma pigmentosum group F protein ; 8-hydroxyguanine (5614-64-2) ; Guanine (5Z93L87A1R) ; Endodeoxyribonucleases (EC 3.1.-) ; Endonucleases (EC 3.1.-) ; Ercc1 protein, mouse (EC 3.1.-) ; NEIL3 protein, mouse (EC 3.1.-) ; DNA Glycosylases (EC 3.2.2.-) ; Ogg1 protein, mouse (EC 3.2.2.-)
Language	English
Publishing date	2021-10-13
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632903-2
ISSN	1464-3804 ; 0267-8357
ISSN (online)	1464-3804
ISSN	0267-8357
DOI	10.1093/mutage/geab038
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Article ; Online: Epigenome-wide analysis of maternal exposure to green space during gestation and cord blood DNA methylation in the ENVIRONAGE cohort.

Alfano, Rossella / Bijnens, Esmée / Langie, Sabine A S / Nawrot, Tim S / Reimann, Brigitte / Vanbrabant, Kenneth / Wang, Congrong / Plusquin, Michelle

Environmental research

2022 Volume 216, Issue Pt 4, Page(s) 114828

Abstract: Background: DNA methylation programming is sensitive to prenatal life environmental influences, but the impact of maternal exposure to green space on newborns DNA methylation has not been studied yet.: Methods: We conducted a meta-epigenome-wide ... ...

Abstract	Background: DNA methylation programming is sensitive to prenatal life environmental influences, but the impact of maternal exposure to green space on newborns DNA methylation has not been studied yet. Methods: We conducted a meta-epigenome-wide association study (EWAS) of maternal exposure to green space during gestation with cord blood DNA methylation in two subsets of the ENVIRONAGE cohort (N = 538). Cord blood DNA methylation was measured by Illumina HumanMethylation 450K in one subset (N = 189) and EPICarray in another (N = 349). High (vegetation height>3 m (m)), low (vegetation height<3 m) and total (including both) high-resolution green space exposures during pregnancy were estimated within 100 m and 1000 m distance around maternal residence. In each subset, we sought cytosine-phosphate-guanine (CpG) sites via linear mixed models adjusted on newborns' sex, ethnicity, gestational age, season at delivery, sampling day, maternal parity, age, smoking, education, and estimated blood cell proportions. EWASs results were meta-analysed via fixed-effects meta-analyses. Differentially methylated regions (DMRs) were identified via ENmix-combp and DMRcate algorithms. Sensitivity analyses were additionally adjusted on PM Results: 147 DMRs were identified, 85 of which were still significant upon adjustment for PM Conclusions: Our results demonstrate that maternal exposure to green space during pregnancy is associated with cord blood DNA methylation, mainly at loci organized in regions, in genes playing important roles in neurological development (e.g., HTR2A).
MeSH term(s)	Pregnancy ; Female ; Humans ; Infant, Newborn ; Maternal Exposure ; Epigenome ; DNA Methylation ; Fetal Blood/metabolism ; Parks, Recreational ; Prenatal Exposure Delayed Effects/genetics ; Particulate Matter/metabolism ; Kruppel-Like Transcription Factors/genetics ; Kruppel-Like Transcription Factors/metabolism ; Receptors, Progesterone/metabolism
Chemical Substances	Particulate Matter ; ZNF274 protein, human ; Kruppel-Like Transcription Factors ; PAQR9 protein, human ; Receptors, Progesterone
Language	English
Publishing date	2022-11-15
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	205699-9
ISSN	1096-0953 ; 0013-9351
ISSN (online)	1096-0953
ISSN	0013-9351
DOI	10.1016/j.envres.2022.114828
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Zs.A 726: Show issues

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Article ; Online: Regenerative responses following DNA damage - β-catenin mediates head regrowth in the planarian

Wouters, Annelies / Ploem, Jan-Pieter / Langie, Sabine A S / Artois, Tom / Aboobaker, Aziz / Smeets, Karen

Journal of cell science

2020 Volume 133, Issue 8

Abstract: Pluripotent stem cells hold great potential for regenerative medicine. Increased replication and division, such is the case during regeneration, concomitantly increases the risk of adverse outcomes through the acquisition of mutations. Seeking for ... ...

Abstract	Pluripotent stem cells hold great potential for regenerative medicine. Increased replication and division, such is the case during regeneration, concomitantly increases the risk of adverse outcomes through the acquisition of mutations. Seeking for driving mechanisms of such outcomes, we challenged a pluripotent stem cell system during the tightly controlled regeneration process in the planarian
MeSH term(s)	Animals ; DNA Damage/genetics ; Head ; Mediterranea ; Planarians/genetics ; Planarians/metabolism ; Wnt Signaling Pathway ; beta Catenin/genetics ; beta Catenin/metabolism
Chemical Substances	beta Catenin
Language	English
Publishing date	2020-04-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.237545
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Zs.A 1200: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Maternal exercise during pregnancy modulates genotoxicity caused by high fructose consumption in mice offspring.

Magenis, Marina Lummertz / Damiani, Adriani Paganini / Monteiro, Isadora de Oliveira / Dagostin, Ligia Salvan / Silva, Nicollas Dos Santos / Scussel, Rahisa / Nagashima, Seigo / Langie, Sabine A S / Pinho, Ricardo Aurino / de Andrade, Vanessa Moraes

Mutagenesis

2023 Volume 39, Issue 2, Page(s) 119–140

Abstract: Pregnancy is a period that is characterized by several metabolic and physiological changes and requires special attention, especially with regard to the relationship between feeding and foetal development. Therefore, the objective of this study was to ... ...

Abstract	Pregnancy is a period that is characterized by several metabolic and physiological changes and requires special attention, especially with regard to the relationship between feeding and foetal development. Therefore, the objective of this study was to evaluate whether the practice of voluntary physical exercise (VPE) in combination with chronic consumption of fructose (FRU) from the beginning of life and/or until the gestational period causes genotoxic changes in pregnant females and in their offspring. Seventy Swiss female mice received FRU in the hydration bottle and/or practiced VPE for 8 weeks (prepregnancy/pregnancy). After the lactation period, the offspring groups were separated by sex. It was observed that the consumption of FRU affected the food consumption, serum concentration of FRU, and glycemic profile in the mothers and that the VPE decreases these parameters. In addition, FRU was genotoxic in the mothers' peripheral tissues and VPE had a preventive effect on these parameters. The offspring showed changes in food consumption, serum FRU concentration, and body weight, in addition to an increase in the adiposity index in male offspring in the FRU (FRU) group and a decrease in the FRU + VPE group. FRU leads to hepatic steatosis in the offspring and VPE was able to decrease the area of steatosis. In addition, FRU led to genotoxicity in the offspring and VPE was able to modulate this effect, reducing damages. In conclusion, we observed that all interventions with VPE had nutritional, genetic, and biochemical benefits of the mother and her offspring.
MeSH term(s)	Pregnancy ; Mice ; Male ; Female ; Animals ; Humans ; Fructose/adverse effects ; Obesity ; Body Weight ; Adiposity ; Lactation ; Prenatal Exposure Delayed Effects/metabolism
Chemical Substances	Fructose (30237-26-4)
Language	English
Publishing date	2023-11-29
Publishing country	England
Document type	Journal Article
ZDB-ID	632903-2
ISSN	1464-3804 ; 0267-8357
ISSN (online)	1464-3804
ISSN	0267-8357
DOI	10.1093/mutage/gead035
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Article ; Online: The enzyme-modified comet assay: Enzyme incubation step in 2 vs 12-gels/slide systems.

Muruzabal, Damian / Langie, Sabine A S / Pourrut, Bertrand / Azqueta, Amaya

Mutation research

2018 Volume 845, Page(s) 402981

Abstract: The enzyme-modified comet assay is a commonly used method to detect specific DNA lesions. However, still a lot of errors are made by many users, leading to dubious results and even misinterpretations. This technical note describes some critical points in ...

Abstract	The enzyme-modified comet assay is a commonly used method to detect specific DNA lesions. However, still a lot of errors are made by many users, leading to dubious results and even misinterpretations. This technical note describes some critical points in the use of the enzyme-modified comet assay, such as the enzyme concentration, the time of incubation, the format used and the equipment. To illustrate the importance of these conditions/parameters, titration experiments of formamidopyrimidine DNA glycosylase (Fpg) were performed using the 2 gels/slide and the 12 minigels/slide formats (plus the 12-Gel Comet Assay Unit™). Incubation times of 15 and 30 min, and 1 h were used. Results showed that the 12 minigels/slide system requires a lower volume and concentration of Fpg. A longer time of incubation has a bigger impact when using such format. Moreover, the paper describes how to perform and interpret a titration experiment when using the enzyme-modified comet assay.
MeSH term(s)	8-Hydroxy-2'-Deoxyguanosine/analysis ; Alkylating Agents/toxicity ; Cell Line ; Comet Assay/instrumentation ; Comet Assay/methods ; DNA Damage ; DNA-Formamidopyrimidine Glycosylase/pharmacology ; Dose-Response Relationship, Drug ; Electrophoresis, Agar Gel ; Gels ; Humans ; Lymphocytes/drug effects ; Reproducibility of Results ; Time Factors ; Titrimetry/methods
Chemical Substances	Alkylating Agents ; Gels ; 8-Hydroxy-2'-Deoxyguanosine (88847-89-6) ; DNA-Formamidopyrimidine Glycosylase (EC 3.2.2.23)
Language	English
Publishing date	2018-11-22
Publishing country	Netherlands
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	206607-5
ISSN	1873-135X ; 1383-5718 ; 0027-5107 ; 0165-1110 ; 0165-1161 ; 0165-7992 ; 0921-8777 ; 0165-1218 ; 1383-5726 ; 0167-8817 ; 0921-8734 ; 1383-5742
ISSN (online)	1873-135X
ISSN	1383-5718 ; 0027-5107 ; 0165-1110 ; 0165-1161 ; 0165-7992 ; 0921-8777 ; 0165-1218 ; 1383-5726 ; 0167-8817 ; 0921-8734 ; 1383-5742
DOI	10.1016/j.mrgentox.2018.11.005
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Article ; Online: The telomere-mitochondrial axis of aging in newborns.

Van Der Stukken, Charlotte / Nawrot, Tim S / Alfano, Rossella / Wang, Congrong / Langie, Sabine A S / Plusquin, Michelle / Janssen, Bram G / Martens, Dries S

Aging

2022 Volume 14, Issue 4, Page(s) 1627–1650

Abstract: Aging starts at the beginning of life as evidenced by high variability in telomere length (TL) and mitochondrial DNA content (mtDNAc) at birth. Whether p53 and PGC-1α are connected to these age-related markers in early life is unclear. In this study, we ... ...

Abstract	Aging starts at the beginning of life as evidenced by high variability in telomere length (TL) and mitochondrial DNA content (mtDNAc) at birth. Whether p53 and PGC-1α are connected to these age-related markers in early life is unclear. In this study, we hypothesized that these hallmarks of aging are associated at birth. In 613 newborns from the ENVIRONAGE birth cohort, p53 and PGC-1α protein levels were measured in cord plasma, while TL and mtDNAc were measured in both cord blood and placental tissue. Cord blood methylation data of genes corresponding to the measured protein levels were available from the Human MethylationEPIC 850K BeadChip array. Pearson correlations and linear regression models were applied while accounting for selected covariates. In cord, a 10% increase in TL was associated with 5.22% (95% CI: 3.26 to 7.22;
MeSH term(s)	Aging/genetics ; DNA, Mitochondrial/genetics ; Female ; Fetal Blood ; Humans ; Placenta ; Pregnancy ; Telomere/genetics ; Tumor Suppressor Protein p53/genetics
Chemical Substances	DNA, Mitochondrial ; Tumor Suppressor Protein p53
Language	English
Publishing date	2022-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1945-4589
ISSN (online)	1945-4589
DOI	10.18632/aging.203897
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Article: The comet assay: past, present, and future.

Langie, Sabine A S / Azqueta, Amaya / Collins, Andrew R

Frontiers in genetics

2015 Volume 6, Page(s) 266

Language	English
Publishing date	2015-08-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2015.00266
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