ESPEN 2012 - Late breaking abstract submission

Topic: Late Breaking Abstract

Abs n°:ESPEN12-1914

Abs Title: OXIDATIVE DNA DAMAGE IN PATIENTS WITH SEPSIS

P. Fikrova^1,*M. Hronek¹R. Štětina²E. Havel³Z. Zadák³

¹FACULTY OF PHARMACY, CHARLES UNIVERSITY, ²UNIVERSITY DEFENCE, ³UNIVERSITY HOSPITAL, HRADEC KRALOVE, Czech Republic

Rationale: Oxidative stress is connected with a large number of pathological states and plays an important role in patients in critical condition with sepsis. The aim of our study was to use the comet assay method to compare DNA damage in these patients with that of a control group of healthy individuals.

Methods: Comet assay is a method used to measure DNA damage at the level of individual cells. The principle of the method is that the cells are immobilized in agarose on a microscope slide, lysed, and then subjected to electrophoresis. We performed tests for DNA damage, specifically a test for SSB (single strand breaks), and tests for oxidative DNA damage which was measured by enzymatic modification of the comet assay using Endonuclease III (Endo III; detects oxidized pyrimidines) and DNA-Formamidopyrimidine glycosylase (FPG; detects oxidized purines). For statistical analysis GraphPad Prism was used.

Results: We compared DNA damage to peripheral blood lymphocytes in 5 patients with sepsis with that of 5 healthy controls. We found a significant higher level of DNA damage in the patient group in all types of measurement. P value in SSB and FPG measurement was p<0.0001 and in Endo III measurement P value was <0.0003.

^{mean ± SE}

	SSB	Endo III	FPG
mean % tail DNA - patients	35,192 ± 2,52	50,026 ± 3,48	51,03 ± 3,78
mean % tail DNA - controls	9,882 ± 2,53	8,062 ± 3,87	9,946 ± 3,92

Conclusion: In this study, we demonstrated the possibility of using the comet assay method for measuring DNA damage in clinical conditions. In further studies, we would like to focus on the repair of DNA and its variability among patients, which can influence the level of DNA damage.

This work was supported by Foundation for the development in the field of artificial feeding, metabolism and gerontology, project 00179906 (Ministry of Health) and Faculty of Pharmacy SVV/2012/265 003.

Disclosure of Interest: None Declared

Keywords: None