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European Standards Committee of Urinary (DNA) Lesion Analysis

There is growing evidence that, rather than simply being a non-invasive marker of whole body oxidative stress, measurement of urinary lesions, such as 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), may, to some extent, reflect the repair of oxidised DNA and deoxynucleotides, although their precise provenance is far from clear. A discrepancy in basal urinary 8-oxodG levels has been noted when comparing chromatographic techniques (e.g. GC/MS following prior HPLC pre-purification, or LC-MS/MS, or LC-EC) with ELISA, although all techniques have been shown to discriminate between diseased and healthy subjects, and possess good within-technique agreement. ELISA has received widespread use, and is clearly amenable to the greatest number of laboratories, however, this discrepancy continues to raise questions regarding its utility. Understanding the basis of this discrepancy will aid our understanding of the significance of urinary lesions.

Furthermore, performing inter-laboratory validation of assays for urinary 8-oxodG measurement would provide robust methods for widespread dissemination and application. Hitherto, this has been performed in a limited fashion, and the discrepancies remain unaddressed. Finally, there is growing clinical interest in the measurement of urinary 8-oxodG, as a means to determine the role of oxidative stress in disease, and evaluate intervention strategies. As with other clinical parameters, a reference range must first be determined.

Objectives of ESCULA:

To compare a multiple (n > 4), different methods for the analysis of urinary 8-oxodG.
To compare methods for urinary creatinine concentration
Evaluate correcting urinary lesion measurements for (i) creatinine, (ii) collection over a 24 hr period.
To identify consensus between labs and/or techniques and investigate the basis for any discrepancy.
To achieve a better understanding of the sources of DNA lesions in urine
Develop a biobank of urine samples from healthy individuals, in order to establish a reference range for urinary 8- oxodG determined by the above methods.
Examination of how levels of other urinary lesions compare to urinary 8-oxodG levels.
Examination of the significance of urinary 8-oxodG levels in urine, in healthy individuals, and those with disease, as a prelude to application in large, multi-centre trials.
Public engagement in science. Develop a user-friendly link from the ESCULA website informing the general public, in lay terms, and main EU languages: what are reactive oxygen species (ROS)?; ROS and disease; combating free radicals; ROS research

Project leads

Dr. Marcus S. Cooke (UK), Prof. Ryszard Olinski (PL), Prof. Steffen Loft (DK)

Steering Committee

Prof. Henrik Poulsen (DK)
Prof. Jean Cadet (FR)
Prof. Andrew R. Collins (NO)
Prof. Peter Farmer (UK)

Participants include:

Prof. Radim J. Sram (CZ)
Dr. Jagadeesan Nair (DE)
Prof. Francesco S. Violante (IT)
Dr. Guillermo T. Saez (ES)
Prof. Dan Segerback (SE)
Prof. Mat Harms-Ringdahl (SE)
Prof. Hiroshi Kasai (JP)
Drs. Mu-Rong Chao & Chiung-Wen Hu (TW)
Prof. Regina Santella (USA)
Prof. Lawrence Marnett (USA)
Dr. Andrew Jenner & Prof. Barry Halliwell (SG)
Dr. Hilmi Orhan(TR)
Dr. John Meerman (NL)
Dr. Kuen-Yuh Wu (TW)
Prof. Lars Barregård (SE)
Dr. Blánaid White (IE)
Dr. Christiaan Leeuwenburgh (USA)
Dr. Peter Svoboda (SE)
Prof. Hauh-Jyun Candy Chen (TW)

Industrial participants

Kronos Science Laboratory
ESA Laboratories Inc.
Japan Institute for the Control of Aging (JaICA)
OXOTOX Co.

Improving our understanding of the sources of urinary DNA lesions

Participants:

University of Leicester, Leicester, UK: Dr. M.D. Evans (project lead), Prof. P.B. Farmer, Dr. M.S. Cooke
Nicolaus Copernicus University, Collegium Medicum in Bydgoszcz, Poland: Professor R. Olinski
Institute of Cancer Research, Sutton, U.K.: Professor D. Phillips
DKFZ, Heidelberg, Germany: formerly Dr. J. Nair
Medical Institute of Bioregulation, Fukuoka, Japan: Prof. Y. Nakabeppu
Leiden University Medical College, Leiden, Netherlands: Dr. L. Mullenders
Lawrence Livermore Laboratory, California, USA: Dr. P.T. Henderson

Project description:

Greater biological understanding of measurements of urinary DNA lesions has been hampered by potential influences of diet or cell death, thus elimination of these confounding factors, ideally conducted on a lesion-by-lesion basis, is essential if such urinary measurements are to have any useful biological meaning. In the first instance, examination of the relative roles of diet, cell death and DNA repair is being established for 8-oxodG.

Objectives:

To determine the contribution of diet to urinary 8-oxodG.
To examine the impact of cell death on urinary 8-oxodG.
To determine the contributions of MTH1 and NER activity to the production of urinary 8-oxodG.

ESCULA Publications

Cooke, MS., Barregard, L., Mistry, V., Potdar, N., Rozalski, R., Gackowski, D., Siomek, A., Foksinski, M., Svoboda, P., Kasai, H., Konje, JC., Sallsten, G., Evans, MD. and Olinski, R. (2009) Inter-laboratory comparison of methodologies for the measurement of urinary 8-oxo-7,8-dihydro-2’-deoxyguanosine. Biomarkers. 14, 103-110.
Cooke, MS., Loft, S., Olinski, R. and members of the European Standards Committee on Urinary (DNA) Lesion Analysis. (2008) Measurement and meaning of oxidatively-modified DNA lesions in urine. Cancer Epid. Biomarkers & Prevent. 17, (1) 3-14.

If you are interested in joining ESCULA, please contact

This work is partly supported by ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility) , a network of excellence operating within the European Union 6th Framework Program, Priority 5: "Food Quality and Safety" (Contract No 513943).