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5. Selective Estrogen Receptor Modulators' (SERMs) Influence on TET3 Expression in Breast Cancer Cell Lines with Distinct Biological Subtypes.

Author

Linowiecka, Kinga, Szpotan, Justyna, Godlewska, Marlena, Gaweł, Damian, Zarakowska, Ewelina, Gackowski, Daniel, Brożyna, Anna A., and Foksiński, Marek

Subjects
LIQUID chromatography-mass spectrometry, SELECTIVE estrogen receptor modulators, DNA demethylation, GENE expression, HIGH performance liquid chromatography, ESTROGEN receptors
Abstract

Tamoxifen, a selective estrogen receptor modulator (SERM), exhibits dual agonist or antagonist effects contingent upon its binding to either G-protein-coupled estrogen receptor (GPER) or estrogen nuclear receptor (ESR). Estrogen signaling plays a pivotal role in initiating epigenetic alterations and regulating estrogen-responsive genes in breast cancer. Employing three distinct breast cancer cell lines—MCF-7 (ESR+; GPER+), MDA-MB-231 (ESR−; GPER−), and SkBr3 (ESR−; GPER+)—this study subjected them to treatment with two tamoxifen derivatives: 4-hydroxytamoxifen (4-HT) and endoxifen (Endox). Through 2D high-performance liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS), varying levels of 5-methylcytosine (5-mC) were found, with MCF-7 displaying the highest levels. Furthermore, TET3 mRNA expression levels varied among the cell lines, with MCF-7 exhibiting the lowest expression. Notably, treatment with 4-HT induced significant changes in TET3 expression across all cell lines, with the most pronounced increase seen in MCF-7 and the least in MDA-MB-231. These findings underscore the influence of tamoxifen derivatives on DNA methylation patterns, particularly through modulating TET3 expression, which appears to be contingent on the presence of estrogen receptors. This study highlights the potential of targeting epigenetic modifications for personalized anti-cancer therapy, offering a novel avenue to improve treatment outcomes. [ABSTRACT FROM AUTHOR]

Published
2024
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6. Exploring the crop epigenome: a comparison of DNA methylation profiling techniques

Author

European Cooperation in Science and Technology, Agius, Dolores Rita [0000-0003-1819-2883], Kapazoglou, Aliki [0000-0002-7584-5994], Avramidou, Evangelia V. [0000-0002-5932-1189], Baránek, Miroslav [0000-0002-1583-3588], Carneros, Elena [0000-0003-2066-6320], Caro, Elena [0000-0002-1034-1621], Castiglione, Stefano [0000-0002-0632-4677], Cicatelli, Angela [0000-0001-6996-3495], Radanovic, Aleksandra [0000-0002-7467-9728], Ebejer, Jean Paul [0000-0003-0888-2637], Gackowski, Daniel [0000-0002-7259-1511], Guarino, Francesco [0000-0001-8536-4257], Gulyás, Andrea [0000-0001-6978-1341], Hidvégi, Norbert [0000-0003-3665-2724], Hoenicka, Hans [0000-0001-7226-3772], Inácio, Vera [0000-0002-5652-2540], Johannes, Frank [0000-0002-7962-2907], Karalija, Erna [0000-0001-7262-0645], Lieberman-Lazarovich, Michal [0000-0002-1776-8257], Martinelli, Federico [0000-0002-8502-767X], Maury, Stéphane [0000-0003-0481-0847], Mladenov, Velimir [0000-0002-2182-6579], Morais-Cecílio, Leonor [0000-0001-9313-2253], Pecinka, Ales [0000-0001-9277-1766], Tani, Eleni [0000-0001-6178-0459], Testillano, P. S. [0000-0003-4509-7646], Valledor, Luis [0000-0002-0636-365X], Vassileva, Valya [0000-0002-9055-8002], Agius, Dolores Rita, Kapazoglou, Aliki, Avramidou, Evangelia V., Baránek, Miroslav, Carneros, Elena, Caro, Elena, Castiglione, Stefano, Cicatelli, Angela, Radanovic, Aleksandra, Ebejer, Jean Paul, Gackowski, Daniel, Guarino, Francesco, Gulyás, Andrea, Hidvégi, Norbert, Hoenicka, Hans, Inácio, Vera, Johannes, Frank, Karalija, Erna, Lieberman-Lazarovich, Michal, Martinelli, Federico, Maury, Stéphane, Mladenov, Velimir, Morais-Cecílio, Leonor, Pecinka, Ales, Tani, Eleni, Testillano, Pilar S., Todorov, Dimitar, Valledor, Luis, Vassileva, Valya, European Cooperation in Science and Technology, Agius, Dolores Rita [0000-0003-1819-2883], Kapazoglou, Aliki [0000-0002-7584-5994], Avramidou, Evangelia V. [0000-0002-5932-1189], Baránek, Miroslav [0000-0002-1583-3588], Carneros, Elena [0000-0003-2066-6320], Caro, Elena [0000-0002-1034-1621], Castiglione, Stefano [0000-0002-0632-4677], Cicatelli, Angela [0000-0001-6996-3495], Radanovic, Aleksandra [0000-0002-7467-9728], Ebejer, Jean Paul [0000-0003-0888-2637], Gackowski, Daniel [0000-0002-7259-1511], Guarino, Francesco [0000-0001-8536-4257], Gulyás, Andrea [0000-0001-6978-1341], Hidvégi, Norbert [0000-0003-3665-2724], Hoenicka, Hans [0000-0001-7226-3772], Inácio, Vera [0000-0002-5652-2540], Johannes, Frank [0000-0002-7962-2907], Karalija, Erna [0000-0001-7262-0645], Lieberman-Lazarovich, Michal [0000-0002-1776-8257], Martinelli, Federico [0000-0002-8502-767X], Maury, Stéphane [0000-0003-0481-0847], Mladenov, Velimir [0000-0002-2182-6579], Morais-Cecílio, Leonor [0000-0001-9313-2253], Pecinka, Ales [0000-0001-9277-1766], Tani, Eleni [0000-0001-6178-0459], Testillano, P. S. [0000-0003-4509-7646], Valledor, Luis [0000-0002-0636-365X], Vassileva, Valya [0000-0002-9055-8002], Agius, Dolores Rita, Kapazoglou, Aliki, Avramidou, Evangelia V., Baránek, Miroslav, Carneros, Elena, Caro, Elena, Castiglione, Stefano, Cicatelli, Angela, Radanovic, Aleksandra, Ebejer, Jean Paul, Gackowski, Daniel, Guarino, Francesco, Gulyás, Andrea, Hidvégi, Norbert, Hoenicka, Hans, Inácio, Vera, Johannes, Frank, Karalija, Erna, Lieberman-Lazarovich, Michal, Martinelli, Federico, Maury, Stéphane, Mladenov, Velimir, Morais-Cecílio, Leonor, Pecinka, Ales, Tani, Eleni, Testillano, Pilar S., Todorov, Dimitar, Valledor, Luis, and Vassileva, Valya

Abstract

Epigenetic modifications play a vital role in the preservation of genome integrity and in the regulation of gene expression. DNA methylation, one of the key mechanisms of epigenetic control, impacts growth, development, stress response and adaptability of all organisms, including plants. The detection of DNA methylation marks is crucial for understanding the mechanisms underlying these processes and for developing strategies to improve productivity and stress resistance of crop plants. There are different methods for detecting plant DNA methylation, such as bisulfite sequencing, methylation-sensitive amplified polymorphism, genome-wide DNA methylation analysis, methylated DNA immunoprecipitation sequencing, reduced representation bisulfite sequencing, MS and immuno-based techniques. These profiling approaches vary in many aspects, including DNA input, resolution, genomic region coverage, and bioinformatics analysis. Selecting an appropriate methylation screening approach requires an understanding of all these techniques. This review provides an overview of DNA methylation profiling methods in crop plants, along with comparisons of the efficacy of these techniques between model and crop plants. The strengths and limitations of each methodological approach are outlined, and the importance of considering both technical and biological factors are highlighted. Additionally, methods for modulating DNA methylation in model and crop species are presented. Overall, this review will assist scientists in making informed decisions when selecting an appropriate DNA methylation profiling method.

Published
2023

7. The urinary excretion of epigenetically modified DNA as a marker of pediatric ALL status and chemotherapy response

Author

Rozalski, Rafal, Gackowski, Daniel, Skalska-Bugala, Aleksandra, Starczak, Marta, Siomek-Gorecka, Agnieszka, Zarakowska, Ewelina, Modrzejewska, Martyna, Dziaman, Tomasz, Szpila, Anna, Linowiecka, Kinga, Guz, Jolanta, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Gackowska, Lidia, Foksinski, Marek, Olinska, Elwira, Wasilow, Aleksandra, Koltan, Andrzej, Styczynski, Jan, and Olinski, Ryszard

Published
2021
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8. LINE-1 transcription in round spermatids is associated with accretion of 5-carboxylcytosine in their open reading frames

Author

Blythe, Martin J., Kocer, Ayhan, Rubio-Roldan, Alejandro, Giles, Tom, Abakir, Abdulkadir, Ialy-Radio, Côme, Wheldon, Lee M., Bereshchenko, Oxana, Bruscoli, Stefano, Kondrashov, Alexander, Drevet, Joël R., Emes, Richard D., Johnson, Andrew D., McCarrey, John R., Gackowski, Daniel, Olinski, Ryszard, Cocquet, Julie, Garcia-Perez, Jose L., and Ruzov, Alexey

Published
2021
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9. PB1914: UNPRECEDENTED ACCUMULATION OF 5-HYDROXYMETHYLURACIL IN CHRONIC LYMPHOCYTIC LEUKEMIA –POTENTIAL SOURCES AND CLINICAL IMPLICATIONS

Author

Gackowski, Daniel, primary, Czyz, Jaroslaw, additional, Starczak, Marta, additional, Gawronski, Maciej, additional, Wasilow, Aleksandra, additional, Mijewski, Pawel, additional, Baginska, Patrycja, additional, Rozalski, Rafal, additional, Siomek-Gorecka, Agnieszka, additional, Skalska-Bugala, Aleksandra, additional, Wołowiec, Anna, additional, Gackowska, Lidia, additional, and Olinski, Ryszard, additional

Published
2023
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10. Context dependent effects of ascorbic acid treatment in TET2 mutant myeloid neoplasia

Author

Guan, Yihong, Greenberg, Edward F., Hasipek, Metis, Chen, Shi, Liu, Xiaochen, Kerr, Cassandra M., Gackowski, Daniel, Zarakowska, Ewelina, Radivoyevitch, Tomas, Gu, Xiaorong, Willard, Belinda, Visconte, Valeria, Makishima, Hideki, Nazha, Aziz, Mukherji, Mridul, Sekeres, Mikkael A., Saunthararajah, Yogen, Oliński, Ryszard, Xu, Mingjiang, Maciejewski, Jaroslaw P., and Jha, Babal K.

Published
2020
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11. Exploring the crop epigenome: a comparison of DNA methylation profiling techniques

Author

Agius, Dolores Rita, primary, Kapazoglou, Aliki, additional, Avramidou, Evangelia, additional, Baranek, Miroslav, additional, Carneros, Elena, additional, Caro, Elena, additional, Castiglione, Stefano, additional, Cicatelli, Angela, additional, Radanovic, Aleksandra, additional, Ebejer, Jean-Paul, additional, Gackowski, Daniel, additional, Guarino, Francesco, additional, Gulyás, Andrea, additional, Hidvégi, Norbert, additional, Hoenicka, Hans, additional, Inácio, Vera, additional, Johannes, Frank, additional, Karalija, Erna, additional, Lieberman-Lazarovich, Michal, additional, Martinelli, Federico, additional, Maury, Stéphane, additional, Mladenov, Velimir, additional, Morais-Cecílio, Leonor, additional, Pecinka, Ales, additional, Tani, Eleni, additional, Testillano, Pilar S., additional, Todorov, Dimitar, additional, Valledor, Luis, additional, and Vassileva, Valya, additional

Published
2023
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12. Exploring the crop epigenome: a comparison of DNA methylation profiling techniques

Author

Agius, Dolores Rita, Agius, Dolores Rita, Kapazoglou, Aliki, Avramidou, Evangelia, Baranek, Miroslav, Carneros, Elena, Caro, Elena, Castiglione, Stefano, Cicatelli, Angela, Radanović, Aleksandra, Ebejer, Jean-Paul, Gackowski, Daniel, Guarino, Francesco, Gulya, Andrea, Hidvegi, Norbert, Hoenicka, Hans, Inacio, Vera, Johannes, Frank, Karalija, Erna, Lieberman-Lazarovich, Michal, Martinelli, Federico, Maury, Stephane, Mladenov, Velimir, Morais-Cecılio, Leonor, Pecinka, Ales, Tani, Eleni, Testillano, Pilar S., Todorov, Dimitar, Valledor, Luis, Vassileva, Valya, Agius, Dolores Rita, Agius, Dolores Rita, Kapazoglou, Aliki, Avramidou, Evangelia, Baranek, Miroslav, Carneros, Elena, Caro, Elena, Castiglione, Stefano, Cicatelli, Angela, Radanović, Aleksandra, Ebejer, Jean-Paul, Gackowski, Daniel, Guarino, Francesco, Gulya, Andrea, Hidvegi, Norbert, Hoenicka, Hans, Inacio, Vera, Johannes, Frank, Karalija, Erna, Lieberman-Lazarovich, Michal, Martinelli, Federico, Maury, Stephane, Mladenov, Velimir, Morais-Cecılio, Leonor, Pecinka, Ales, Tani, Eleni, Testillano, Pilar S., Todorov, Dimitar, Valledor, Luis, and Vassileva, Valya

Abstract

Epigenetic modifications play a vital role in the preservation of genome integrity and in the regulation of gene expression. DNA methylation, one of the key mechanisms of epigenetic control, impacts growth, development, stress response and adaptability of all organisms, including plants. The detection of DNA methylation marks is crucial for understanding the mechanisms underlying these processes and for developing strategies to improve productivity and stress resistance of crop plants. There are different methods for detecting plant DNA methylation, such as bisulfite sequencing, methylation-sensitive amplified polymorphism, genome-wide DNA methylation analysis, methylated DNA immunoprecipitation sequencing, reduced representation bisulfite sequencing, MS and immuno-based techniques. These profiling approaches vary in many aspects, including DNA input, resolution, genomic region coverage, and bioinformatics analysis. Selecting an appropriate methylation screening approach requires an understanding of all these techniques. This review provides an overview of DNA methylation profiling methods in crop plants, along with comparisons of the efficacy of these techniques between model and crop plants. The strengths and limitations of each methodological approach are outlined, and the importance of considering both technical and biological factors are highlighted. Additionally, methods for modulating DNA methylation in model and crop species are presented. Overall, this review will assist scientists in making informed decisions when selecting an appropriate DNA methylation profiling method.

Published
2023

15. Unprecedentedly High Level of Intracellular Vitamin C and DNA Epigenetic Marks in Prostate: Relevant for Male Fertility?

Author

Guz, Jolanta, Zarakowska, Ewelina, Mijewski, Pawel, Wasilow, Aleksandra, Szpotan, Justyna, Foksinski, Marek, Brzoszczyk, Bartosz, Gackowski, Daniel, Jarzemski, Piotr, and Olinski, Ryszard

Subjects
VITAMIN C, LIQUID chromatography-mass spectrometry, SEMEN, PROSTATE, BENIGN prostatic hyperplasia, FREE radical scavengers, DNA demethylation, FERTILITY
Abstract

Background/Aims: Seminal plasma composition is affected by the physiological state of the prostate, the major male reproductive gland. Semen components, like vitamin C, can modulate sperm function. Vitamin C is an effective scavenger of free radicals and is an essential component of enzymes such as TET proteins involved in the DNA demethylation process. In the present study, a broad range of parameters which may influence the metabolic state of the prostate gland were analysed including blood and prostate tissue vitamin C, epigenetic DNA modifications and 8-oxo-7,8-dihydro-2'-deoxyguanosine in DNA of leukocytes and prostate tissues. Methods: The experimental material were tissue samples from patients with benign prostatic hyperplasia (BPH), normal/marginal prostate tissues from prostate cancer patients, leukocytes from healthy donors, and blood plasma from BPH patients and healthy donors. We applied ultra-performance liquid chromatography methods with mass spectrometry and/or UV detection. Results: We found an unprecedentedly high level of intracellular vitamin C in all analysed prostatic tissues (benign prostatic hyperplasia and normal, marginal ones), a value much higher than in leukocytes and most human tissues. DNA epigenetic patterns in prostate cells are similar to other soft tissues like the colon, however, its uniqueness is the unprecedentedly high level of 5-(hydroxymethyl)-2'-deoxyuridine and a significant increase in 5-formyl-2'-deoxycytidine value compared to aforementioned tissues. Moreover, the level of 8-oxo-7,8-dihydro-2'-deoxyguanosine, an established marker of oxidative stress, is significantly higher in prostate tissues than in leukocytes and many previously studied soft tissues. Conclusion: Our results pointed out that prostatic vitamin C (regarded as the main supplier of the vitamin C to seminal plasma) and the DNA modifications (which may be linked to the regeneration of prostate epithelium) may play important role to maintain the prostate health. [ABSTRACT FROM AUTHOR]

Published
2023
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16. In vivo evidence of ascorbate involvement in the generation of epigenetic DNA modifications in leukocytes from patients with colorectal carcinoma, benign adenoma and inflammatory bowel disease

Author

Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Dziaman, Tomasz, Szpila, Anna, Linowiecka, Kinga, Guz, Jolanta, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Gackowski, Daniel, and Olinski, Ryszard

Published
2018
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17. Characteristic profiles of DNA epigenetic modifications in colon cancer and its predisposing conditions—benign adenomas and inflammatory bowel disease

Author

Dziaman, Tomasz, Gackowski, Daniel, Guz, Jolanta, Linowiecka, Kinga, Bodnar, Magdalena, Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Szpila, Anna, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Marszalek, Andrzej, and Olinski, Ryszard

Published
2018
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18. Plasma micronutrients, trace elements, and breast cancer in BRCA1 mutation carriers: an exploratory study

Author

Kotsopoulos, Joanne, Sukiennicki, Grzegorz, Muszyńska, Magdalena, Gackowski, Daniel, Kąklewski, Krzysztof, Durda, Katarzyna, Jaworska, Katarzyna, Huzarski, Tomasz, Gronwald, Jacek, Byrski, Tomasz, Ashuryk, Oleg, Dębniak, Tadeusz, Tołoczko-Grabarek, Aleksandra, Stawicka, Małgorzata, Godlewski, Dariusz, Oliński, Ryszard, Jakubowska, Anna, Narod, Steven A., and Lubinski, Jan

Published
2012

19. Diagnostic and Prognostic Power of Active DNA Demethylation Pathway Intermediates in Acute Myelogenous Leukemia and Myelodysplastic Syndromes

Author

Skalska-Bugala, Aleksandra, primary, Starczak, Marta, additional, Szukalski, Łukasz, additional, Gawronski, Maciej, additional, Siomek-Gorecka, Agnieszka, additional, Szpotan, Justyna, additional, Labejszo, Anna, additional, Zarakowska, Ewelina, additional, Szpila, Anna, additional, Jachalska, Anna, additional, Szukalska, Adriana, additional, Kruszewski, Marcin, additional, Sadowska, Anna, additional, Wasilow, Aleksandra, additional, Baginska, Patrycja, additional, Czyz, Jaroslaw, additional, Olinski, Ryszard, additional, Rozalski, Rafal, additional, and Gackowski, Daniel, additional

Published
2022
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22. An IDH-independent mechanism of DNA hypermethylation upon VHL inactivation in cancer.

Author

Artemov, Artem V., Zhenilo, Svetlana, Kaplun, Daria, Starshin, Alexey, Sokolov, Alexey, Mazur, Alexander M., Szpotan, Justyna, Gawronski, Maciej, Modrzejewska, Martyna, Gackowski, Daniel, and Prokhortchouk, Egor B.

Subjects
TUMOR suppressor genes, DNA demethylation, DNA, DNA methylation, RENAL cancer, DNA methyltransferases
Abstract

Hypermethylation of tumour suppressors and other aberrations of DNA methylation in tumours play a significant role in cancer progression. DNA methylation can be affected by various environmental conditions, including hypoxia. The response to hypoxia is mainly achieved through activation of the transcriptional program associated with HIF1A transcription factor. Inactivation of Von Hippel-Lindau Tumour Suppressor gene (VHL) by genetic or epigenetic events, which also induces aberrant activation of HIF1A, is the most common driver event for renal cancer. With whole-genome bisulphite sequencing and LC-MS, we demonstrated that VHL inactivation induced global genome hypermethylation in human kidney cancer cells under normoxic conditions. This effect was reverted by exogenous expression of wild-type VHL. We showed that global genome hypermethylation in VHL mutants can be explained by transcriptional changes in MDH and L2HGDH genes that cause the accumulation of 2-hydroxyglutarate – a metabolite that inhibits DNA demethylation by TET enzymes. Unlike the known cases of DNA hypermethylation in cancer, 2-hydroxyglutarate was accumulated in the cells with the wild-type isocitrate dehydrogenases. [ABSTRACT FROM AUTHOR]

Published
2022
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23. Targeted DNA oxidation by LSD1–SMAD2/3 primes TGF-β1/ EMT genes for activation or repression

Author

Pezone, Antonio, primary, Taddei, Maria Letizia, additional, Tramontano, Alfonso, additional, Dolcini, Jacopo, additional, Boffo, Francesca Ludovica, additional, De Rosa, Mariarosaria, additional, Parri, Matteo, additional, Stinziani, Stefano, additional, Comito, Giuseppina, additional, Porcellini, Antonio, additional, Raugei, Giovanni, additional, Gackowski, Daniel, additional, Zarakowska, Ewelina, additional, Olinski, Ryszard, additional, Gabrielli, Armando, additional, Chiarugi, Paola, additional, and Avvedimento, Enrico Vittorio, additional

Published
2020
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25. Mass spectrometry reveals the presence of specific set of epigenetic DNA modifications in the Norway spruce genome

Author

Yakovlev, Igor, Gackowski, Daniel, Abakir, Abdulkadir, Viejo, Marcos, Ruzov, Alexey, Olinski, Ryszard, Starczak, Marta, Fossdal, Carl Gunnar, and Krutovsky, Konstantin V.

Subjects
Epigenomics, DNA methylation, Norway, lcsh:R, fungi, Mass spectrometry, epigenetic DNA, Norway spruce genome, lcsh:Medicine, Mass Spectrometry, Article, Epigenesis, Genetic, Cytosine, 5-Methylcytosine, lcsh:Q, Picea, lcsh:Science, Chromatography, High Pressure Liquid, Genome, Plant
Abstract

5-Methylcytosine (5mC) is an epigenetic modification involved in regulation of gene expression in metazoans and plants. Iron-(II)/α-ketoglutarate-dependent dioxygenases can oxidize 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). Although these oxidized forms of 5mC may serve as demethylation intermediates or contribute to transcriptional regulation in animals and fungi, experimental evidence for their presence in plant genomes is ambiguous. Here, employing reversed-phase HPLC coupled with sensitive mass spectrometry, we demonstrated that, unlike 5caC, both 5hmC and 5fC are detectable in non-negligible quantities in the DNA of a conifer, Norway spruce. Remarkably, whereas 5hmC content of spruce DNA is approximately 100-fold lower relative to human colorectal carcinoma cells, the levels of both - 5fC and a thymine base modification, 5-hydroxymethyluracil, are comparable in these systems. We confirmed the presence of modified DNA bases by immunohistochemistry in Norway spruce buds based on peroxidase-conjugated antibodies and tyramide signal amplification. Our results reveal the presence of specific range of noncanonical DNA bases in conifer genomes implying potential roles for these modifications in plant development and homeostasis. Open-Access-Publikationsfonds 2019 peerReviewed

Published
2019

28. MOESM1 of In vivo evidence of ascorbate involvement in the generation of epigenetic DNA modifications in leukocytes from patients with colorectal carcinoma, benign adenoma and inflammatory bowel disease

Author

Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Dziaman, Tomasz, Szpila, Anna, Linowiecka, Kinga, Guz, Jolanta, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Gackowski, Daniel, and Olinski, Ryszard

Abstract

Additional file 1. Tables S1â S3, Figures S1â S10, additional method.

Published
2018
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29. Additional file 3: of Characteristic profiles of DNA epigenetic modifications in colon cancer and its predisposing conditionsâ benign adenomas and inflammatory bowel disease

Author

Dziaman, Tomasz, Gackowski, Daniel, Guz, Jolanta, Linowiecka, Kinga, Bodnar, Magdalena, Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Szpila, Anna, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Marszalek, Andrzej, and Olinski, Ryszard

Subjects
body regions, nervous system, fungi, biochemical phenomena, metabolism, and nutrition
Abstract

Table S3. Primers and short hydrolysis probes used for TETs and AID mRNA expression analysis. (PDF 442Â kb)

Published
2018
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30. Additional file 2: of Characteristic profiles of DNA epigenetic modifications in colon cancer and its predisposing conditionsâ benign adenomas and inflammatory bowel disease

Author

Dziaman, Tomasz, Gackowski, Daniel, Guz, Jolanta, Linowiecka, Kinga, Bodnar, Magdalena, Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Szpila, Anna, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Marszalek, Andrzej, and Olinski, Ryszard

Abstract

Table S2. Transition patterns specific detector settings and sources of standards for analyzed compounds (relative response ratio = area under the peak of qualifierion/area under the peak of quantifier ion). (PDF 82Â kb)

Published
2018
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31. Additional file 1: of Characteristic profiles of DNA epigenetic modifications in colon cancer and its predisposing conditionsâ benign adenomas and inflammatory bowel disease

Author

Dziaman, Tomasz, Gackowski, Daniel, Guz, Jolanta, Linowiecka, Kinga, Bodnar, Magdalena, Starczak, Marta, Zarakowska, Ewelina, Modrzejewska, Martyna, Szpila, Anna, Szpotan, Justyna, Gawronski, Maciej, Labejszo, Anna, Liebert, Ariel, Banaszkiewicz, Zbigniew, Klopocka, Maria, Foksinski, Marek, Marszalek, Andrzej, and Olinski, Ryszard

Subjects
body regions, nervous system, fungi
Abstract

Table S1. Characteristics of antibodies used in immunohistochemical staining. (PDF 499Â kb)

Published
2018
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32. TET2 Loss Accelerates Leukemogenesis By Disrupting Mismatch Repair Proteins

Author

Kishtagari, Ashwin, primary, Guan, Yihong, additional, Kerr, Cassandra M, additional, Awada, Hassan, additional, Durrani, Jibran, additional, Greenberg, Edward F, additional, Gu, Xiaorong, additional, Pan, Feng, additional, Visconte, Valeria, additional, You, Dewen, additional, Qu, Guangbo, additional, Graham, Amy C, additional, Przychodzen, Bartlomiej P., additional, Laframboise, Thomas, additional, Makishima, Hideki, additional, Gackowski, Daniel, additional, Ólinski, Ryszard, additional, Cheng, Yu-Wei, additional, Sekeres, Mikkael A., additional, Xu, Mingjiang, additional, Jha, Babal K., additional, and Maciejewski, Jaroslaw P., additional

Published
2019
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33. 5-formylcytosine and 5-hydroxymethyluracil as surrogate markers of TET2 and SF3B1 mutations in myelodysplastic syndrome, respectively

Author

Gackowski, Daniel, primary, Gawronski, Maciej, additional, Kerr, Cassandra, additional, Radivoyevitch, Tomas, additional, Zarakowska, Ewelina, additional, Starczak, Marta, additional, Abakir, Abdulkadir, additional, Ruzov, Alexey, additional, Maciejewski, Jaroslaw P., additional, and Olinski, Ryszard, additional

Published
2019
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34. Modulation of TET2 Activity By Ascorbic Acid and Factors Affecting Lysine Acetylation

Author

Guan, Yihong, primary, Greenberg, Edward F, additional, Hasipek, Metis, additional, Hirsch, Cassandra M., additional, Graham, Amy C, additional, Radivoyevitch, Tomas, additional, Chen, Shi, additional, Liu, Xiaochen, additional, Visconte, Valeria, additional, You, Dewen, additional, Negoro, Eiju, additional, Przychodzen, Bartlomiej P., additional, Makishima, Hideki, additional, Nazha, Aziz, additional, Gackowski, Daniel, additional, Zarakowska, Ewelina, additional, Mukherji, Mridul, additional, Sekeres, Mikkael A., additional, Xu, Mingjiang, additional, Ólinski, Ryszard, additional, Maciejewski, Jaroslaw P., additional, and Jha, Babal K., additional

Published
2018
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36. Correction: Profiles of a broad spectrum of epigenetic DNA modifications in normal and malignant human cell lines: Proliferation rate is not the major factor responsible for the 5-hydroxymethyl-2′-deoxycytidine level in cultured cancerous cell lines

Author

Foksinski, Marek, primary, Zarakowska, Ewelina, additional, Gackowski, Daniel, additional, Skonieczna, Magdalena, additional, Gajda, Karolina, additional, Hudy, Dorota, additional, Szpila, Anna, additional, Bialkowski, Karol, additional, Starczak, Marta, additional, Labejszo, Anna, additional, Czyz, Jaroslaw, additional, Rzeszowska-Wolny, Joanna, additional, and Olinski, Ryszard, additional

Published
2018
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37. Profiles of a broad spectrum of epigenetic DNA modifications in normal and malignant human cell lines: Proliferation rate is not the major factor responsible for the 5-hydroxymethyl-2′-deoxycytidine level in cultured cancerous cell lines

Author

Foksinski, Marek, primary, Zarakowska, Ewelina, additional, Gackowski, Daniel, additional, Skonieczna, Magdalena, additional, Gajda, Karolina, additional, Hudy, Dorota, additional, Szpila, Anna, additional, Bialkowski, Karol, additional, Starczak, Marta, additional, Labejszo, Anna, additional, Czyz, Jaroslaw, additional, Rzeszowska-Wolny, Joanna, additional, and Olinski, Ryszard, additional

Published
2017
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40. 8-Oxo-7,8-dihydroguanine and uric acid as efficient predictors of survival in colon cancer patients

Author

Dziaman, Tomasz, Banaszkiewicz, Zbigniew, Roszkowski, Krzysztof, Gackowski, Daniel, Wisniewska, Ewa, Rozalski, Rafał, Foksinski, Marek, Siomek, Agnieszka, Speina, Elzbieta, Winczura, Alicja, Marszalek, Andrzej, Tudek, Barbara, and Olinski, Ryszard

Subjects
Biological markers, Survival, Oxidative stress, DNA damage, DNA repair, 8-oxoGua, Uric acid, Colon cancer
Abstract

The aim of this work was to answer the question whether the broad range of parameters which describe oxidative stress and oxidatively damaged DNA and repair are appropriate prognosis factors of colon cancer (CRC) patients survival? The following parameters were analyzed for 89 CRC patients: concentration of uric acid and vitamins A, E, C in plasma; levels of 8-oxodGuo (8-oxo-7,8-dihydro-2'-deoxyguanosine) in DNA of leukocyte and colon tissues; urinary excretion rates of 8-oxodGuo and 8-oxoGua (8-oxo-7,8-dihydroguanine); the activity and mRNA or protein level of repair enzymes OGG1, APE1, ANPG, TDG and PARP1. All DNA modifications and plasma antioxidants were analyzed using high performance liquid chromatography (HPLC) or HPLC/gas chromatography-mass spectrometry techniques. Expression of repair proteins was analyzed by QPCR, Western or immunohistochemistry methods. Longer survival coincided with low levels of 8-oxodGuo/8oxoGua in urine and 8-oxodGuo in DNA as well as with high concentration of uric acid plasma level. In contrast to expectations, longer survival coincided with lower mRNA level in normal colon tissue of the main 8-oxoGua DNA glycosylase, OGG1, but no association was found for PARP-1 expression. When analyzing simultaneously two parameters the discriminating power increased significantly. Combination of low level of urinary 8-oxoGua together with low level of 8-oxodGuo in leukocyte (both below median value) or high concentration of plasma uric acid (above median value) have the best prediction power. Since prediction value of these parameters seems to be comparable to conventional staging procedure, they could possibly be used as markers to predict clinical success in CRC treatment. The authors of this article (R.O., T.D., R.R., D.G., A.S., M.F.) are partners of ECNIS and ECNIS2 (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a Network of Excellence operating within the European Union 6th and 7th Framework Program (contract numbers: 513943 and 266198, respectively).

Published
2013

41. Systemic oxidoreductive balance in patients without clinical manifestation of atherosclerosis.

Author

Majer, Marcin, Gackowski, Daniel, Różalski, Rafał, Siomek-Górecka, Agnieszka, Oliński, Ryszard, and Budzyński, Jacek

Subjects
*ATHEROSCLEROSIS, *OXIDOREDUCTASES, *OXIDATIVE stress, *ANTIOXIDANTS, *VITAMIN E
Abstract

Introduction. Oxidative stress plays an important role in atherosclerosis, but numerous clinical trials have not confirmed a favourable effect of antioxidant supplementation. We aimed to determine the oxidative stress parameters in patients without clinical manifestation of vascular disease. Material and methods. Forty-eight patients were divided into two groups in relation to the presence or absence of clinically silent signs of atherosclerosis (ankle-brachial index < 0.9, intima-media thickness = 0.9 mm, the presence of carotid atherosclerotic plaques, silent ischaemia in a treadmill stress test or focal myocardial contractility found in echocardiography). Plasma concentrations of: retinol, ascorbic acid, alpha-tocopherol and uric acid, as well as the products of oxidative DNA damage repair: 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in blood leucocytes and urine, and 8-oxo-7,8-dihydroguanine (8-oxo-Gua) in urine. Results. Patients with signs of subclinical atherosclerosis had lower blood concentration of alpha-tocopherol, and a non-significantly greater urine concentration of 8-oxoGua. Women had significantly greater blood concentration of ascorbic acid and alpha-tocopherol, as well as lower level of retinol and uric acid. They also had greater leucocyte concentration of 8-oxodG. Plasma concentration of alpha-tocopherol 30.34 µM distinguished patients with and without signs of subclinical atherosclerosis. Conclusions. Oxidative stress has clinical importance in the early stages of atherosclerosis and may be helpful in predicting its subclinical stage. Women had higher level of antioxidant defence, which explains their natural protection against early atherosclerosis development. Further studies are needed to determine the usefulness of tocopherol determination as a biomarker for atherosclerosis risk evaluation. [ABSTRACT FROM AUTHOR]

Published
2018
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45. Establishing the background level of base oxidation in human lymphocyte DNA: results of an interlaboratory validation study

Author

Gedik, Catherine, Collins, Andrew, Dubois, Jacques, Duez, Pierre, Kouegnigan, Léonard, Rees, Jean-François, Loft, Steffen, Möller, Peter, Jensen, Annie, Poulsen, Henrik, Riis, Bente, Weimann, Allan, Cadet, Jean, Douki, Thierry, Ravanat, Jean-Luc, Sauvaigo, Sylvie, Faure, Henri, Morel, Isabelle, Morin, Bénédicte, Epe, Bernd, Eckert, Inge, Hartwig, Andrea, Schwerdtle, Tanja, Dolara, Piero, Giovannelli, Lisa, Lodovici, Maura, Guglielmi, Francesco, Olinski, Ryszard, Bialkowski, Karol, Foksinski, Marek, Gackowski, Daniel, Duračková, Zdena, Muchová, Jana, Korytar, Peter, Sivonová, Monika, Dusinska, Maria, Mislanova, Csilla, Petrovska, Helena, Smolkova, Bozena, Vina, José, Lloret, Ana, Saez, Guillermo, Hofer, Tim, Möller, Lennart, Eriksson, Hanna, Gremaud, Eric, Herbert, Karl, Wild, Chris, Kelly, Frank, Dunster, Christina, White, Ann, Wood, Sharon, Vaughan, Nicolas, Department of Computer Science [Liverpool], University of Liverpool, France, Amériques, Espagne – Sociétés, pouvoirs, acteurs (FRAMESPA), Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université libre de Bruxelles (ULB), Université Catholique de Louvain = Catholic University of Louvain (UCL), Universität Ulm - Ulm University [Ulm, Allemagne], Laboratoire Lésions des Acides Nucléiques (LAN), Service de Chimie Inorganique et Biologique (SCIB - UMR E3), Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS), Chimie Interface Biologie pour l’Environnement, la Santé et la Toxicologie (CIBEST ), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), CHU Grenoble, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble, Université de Rennes (UR), Université de Bordeaux (UB), Department of Preclinical and Clinical Pharmacology, Università degli Studi di Firenze = University of Florence (UniFI), Norwegian Institute for Air Research (NILU), Interface Physique et Chimie pour le Vivant (IPCV), Centre d'Etudes Nucléaires de Bordeaux Gradignan (CENBG), Université Sciences et Technologies - Bordeaux 1 (UB)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1 (UB)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse - Jean Jaurès (UT2J)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes (UNIV-RENNES), Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), and Université Sciences et Technologies - Bordeaux 1-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Guanine, Analytical chemistry, Biology, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Cell Line, Tumor, Genetics, Humans, AP site, Lymphocytes, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Chromatography, 8-Hydroxy-2'-deoxyguanosine, Deoxyguanosine, DNA oxidation, Formamidopyrimidine DNA glycosylase, DNA, Orders of magnitude (mass), Comet assay, chemistry, 8-Hydroxy-2'-Deoxyguanosine, 030220 oncology & carcinogenesis, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Laboratories, Oxidation-Reduction, Biotechnology, HeLa Cells
Abstract

Accurate measurement of low levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in DNA is hampered by the ease with which guanine is oxidized during preparation of DNA for analysis. ESCODD, a consortium of mainly European laboratories, has attempted to minimize this artifact and to provide standard, reliable protocols for sample preparation and analysis. ESCODD has now analyzed 8-oxodGuo in the DNA of lymphocytes isolated from venous blood from healthy young male volunteers in several European countries. Two approaches were used. Analysis of 8-oxodGuo by HPLC with electrochemical detection was performed on lymphocytes from 10 groups of volunteers, in eight countries. The alternative enzymic approach was based on digestion of DNA with formamidopyrimidine DNA glycosylase (FPG) to convert 8-oxo-7,8-dihydroguanine (8-oxoGua) to apurinic sites, subsequently measured as DNA breaks using the comet assay (7 groups of volunteers, in six countries). The median concentration of 8-oxodGuo in lymphocyte DNA, calculated from the mean values of each group of subjects as determined by HPLC, was 4.24 per 10(6) guanines. The median concentration of FPG-sensitive sites, measured with the comet assay, was 0.34 per 10(6) guanines. Identical samples of HeLa cells were supplied to all participants as a reference standard. The median values for 8-oxodGuo in HeLa cells were 2.78 per 10(6) guanines (by HPLC) and 0.50 per 10(6) guanines (by enzymic methods). The discrepancy between chromatographic and FPG-based approaches may reflect overestimation by HPLC (if spurious oxidation is still not completely controlled) or underestimation by the enzymic method. Meanwhile, it is clear that the true background level of base oxidation in DNA is orders of magnitude lower than has often been claimed in the past.

Published
2004
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47. Human and methodological sources of variability in the measurement of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine

Author

Barregard, Lars, Møller, Peter, Henriksen, Trine, Mistry, Vilas, Koppen, Gudrun, Rossner, Pavel, Sram, Radim J, Weimann, Allan, Poulsen, Henrik E, Nataf, Robert, Andreoli, Roberta, Manini, Paola, Marczylo, Tim, Lam, Patricia, Evans, Mark D, Kasai, Hiroshi, Kawai, Kazuaki, Li, Yun-Shan, Sakai, Kazuo, Singh, Rajinder, Teichert, Friederike, Farmer, Peter B, Rozalski, Rafal, Gackowski, Daniel, Siomek, Agnieszka, Saez, Guillermo T, Cerda, Concha, Broberg, Karin, Lindh, Christian, Hossain, Mohammad Bakhtiar, Haghdoost, Siamak, Hu, Chiung-Wen, Chao, Mu-Rong, Wu, Kuen-Yuh, Orhan, Hilmi, Senduran, Nilufer, Smith, Raymond J, Santella, Regina M, Su, Yali, Cortez, Czarina, Yeh, Susan, Olinski, Ryszard, Loft, Steffen, Cooke, Marcus S, Barregard, Lars, Møller, Peter, Henriksen, Trine, Mistry, Vilas, Koppen, Gudrun, Rossner, Pavel, Sram, Radim J, Weimann, Allan, Poulsen, Henrik E, Nataf, Robert, Andreoli, Roberta, Manini, Paola, Marczylo, Tim, Lam, Patricia, Evans, Mark D, Kasai, Hiroshi, Kawai, Kazuaki, Li, Yun-Shan, Sakai, Kazuo, Singh, Rajinder, Teichert, Friederike, Farmer, Peter B, Rozalski, Rafal, Gackowski, Daniel, Siomek, Agnieszka, Saez, Guillermo T, Cerda, Concha, Broberg, Karin, Lindh, Christian, Hossain, Mohammad Bakhtiar, Haghdoost, Siamak, Hu, Chiung-Wen, Chao, Mu-Rong, Wu, Kuen-Yuh, Orhan, Hilmi, Senduran, Nilufer, Smith, Raymond J, Santella, Regina M, Su, Yali, Cortez, Czarina, Yeh, Susan, Olinski, Ryszard, Loft, Steffen, and Cooke, Marcus S

Abstract

Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability.

Published
2013

49. Measurement of DNA oxidation in human cells by chromatographic and enzymic methods

Author

Collins, Andrew, Gedik, Catherine, Vaughan, Nicholas, Wood, Sharon, White, Ann, Dubois, Jacques, Rees, Jean-François, Loft, Steffen, Møller, Peter, Poulsen, Henrik, Cadet, Jean, Douki, Thierry, Ravanat, Jean-Luc, Sauvaigo, Sylvie, Faure, Henry, Morel, Isabelle, Morin, Bénédicte, Epe, Bernd, Phoa, Nicole, Hartwig, Andrea, Schwerdtle, Tanja, Dolara, Piero, Giovannelli, Lisa, Lodovici, Maura, Olinski, Ryszard, Bialkowski, Karol, Foksinski, Marek, Gackowski, Daniel, Duracková, Zdena, Hlinciková, Lucia, Korytar, Peter, Sivonová, Monika, Dusinská, Mária, Mislanová, Csilla, Viña, José, Möller, Lennart, Hofer, Tim, Nygren, Jonas, Gremaud, Eric, Herbert, Karl, Lunec, Joseph, Collins, Andrew, Gedik, Catherine, Vaughan, Nicholas, Wood, Sharon, White, Ann, Dubois, Jacques, Rees, Jean-François, Loft, Steffen, Møller, Peter, Poulsen, Henrik, Cadet, Jean, Douki, Thierry, Ravanat, Jean-Luc, Sauvaigo, Sylvie, Faure, Henry, Morel, Isabelle, Morin, Bénédicte, Epe, Bernd, Phoa, Nicole, Hartwig, Andrea, Schwerdtle, Tanja, Dolara, Piero, Giovannelli, Lisa, Lodovici, Maura, Olinski, Ryszard, Bialkowski, Karol, Foksinski, Marek, Gackowski, Daniel, Duracková, Zdena, Hlinciková, Lucia, Korytar, Peter, Sivonová, Monika, Dusinská, Mária, Mislanová, Csilla, Viña, José, Möller, Lennart, Hofer, Tim, Nygren, Jonas, Gremaud, Eric, Herbert, Karl, and Lunec, Joseph

Abstract

The European Standards Committee on Oxidative DNA Damage (ESCODD) was set up to resolve problems in the measurement of DNA oxidation that have resulted in varying estimates of the extent of this damage in humans. HeLa cells, sent to members for analysis, were either untreated, or treated with light in the presence of a photosensitizer to induce different amounts of 8-oxo-7,8-dihydroguanine (8-oxoGua) in DNA. Laboratories employing HPLC with electrochemical detection were able to measure the induced damage with similar efficiency; dose response gradients for seven of the eight sets of results were almost identical. GC-MS and HPLC-MS/MS, employed in three laboratories, did not convincingly detect the dose response. An alternative approach to measuring base oxidation employs the enzyme formamidopyrimidine DNA N-glycosylase (FPG) to convert 8-oxoGua to strand breaks, which are then measured by alkaline unwinding, alkaline elution, or the comet assay. Ten laboratories used this approach; five were able to detect the dose response in cells treated with photosensitizer plus light (at lower doses than for chromatographic methods, because the enzymic methods are more sensitive and less prone to spurious oxidation). Median values for 8-oxoGua (or FPG-sensitive sites) in untreated cells were 4.01 per 106 guanines for chromatographic methods, and 0.53 per 106 guanines for techniques based on FPG. © 2003 Elsevier Inc., SCOPUS: ar.j, info:eu-repo/semantics/published

Published
2003

50. Measurement of DNA oxidation in human cells by chromatographic and enzymatic methods

Author

Collins, A., Gedik, Catherine, Vaughan, Nicholas, Wood, Sharon, White, Ann, Dubois, Jacques, Rees, Jean-François, Loft, Steffen, Moller, P., Poulsen, Henrik, Cadet, Jean, Douki, Thierry, Ravanat, Jean-Luc, Sauvaigo, Sylvie, Morel, Isabelle, Morin, Bénédicte, Epe, Bernd, Phoa, Nicole, Hartwig, Andrea, Schwerdtle, Tanja, Dolara, Piero, Giovannelli, Lisa, Lodovici, Maura, Olinski, Ryszard, Bialkowski, Karol, Foksinski, Marek, Gackowski, Daniel, Duracková, Zdena, Hlinciková, Lucia, Korytar, Peter, Sivonová, Monika, Dusinská, Mária, Mislanová, Csilla, Viña, José, Möller, Lennart, Hofer, Tim, Nygren, Jonas, Gremaud, Eric, Herbert, Karl, Lunec, Joseph, Wild, Chris, Hardie, Laura, Olliver, Joanna, Smith, Elizabeth, Collins, A., Gedik, Catherine, Vaughan, Nicholas, Wood, Sharon, White, Ann, Dubois, Jacques, Rees, Jean-François, Loft, Steffen, Moller, P., Poulsen, Henrik, Cadet, Jean, Douki, Thierry, Ravanat, Jean-Luc, Sauvaigo, Sylvie, Morel, Isabelle, Morin, Bénédicte, Epe, Bernd, Phoa, Nicole, Hartwig, Andrea, Schwerdtle, Tanja, Dolara, Piero, Giovannelli, Lisa, Lodovici, Maura, Olinski, Ryszard, Bialkowski, Karol, Foksinski, Marek, Gackowski, Daniel, Duracková, Zdena, Hlinciková, Lucia, Korytar, Peter, Sivonová, Monika, Dusinská, Mária, Mislanová, Csilla, Viña, José, Möller, Lennart, Hofer, Tim, Nygren, Jonas, Gremaud, Eric, Herbert, Karl, Lunec, Joseph, Wild, Chris, Hardie, Laura, Olliver, Joanna, and Smith, Elizabeth

Abstract

ESCODD, info:eu-repo/semantics/published

Published
2003

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