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Abstract:
The assessment of cellular effects by the aqueous phase of human feces (fecal water, FW) is a useful biomarker approach to study cancer risks and protective activities of food. In order to refine and develop the biomarker, different protocols of preparing FW were compared. Fecal waters were prepared by 3 methods: (A) direct centrifugation; (B) extraction of feces in PBS before centrifugation; and (C) centrifugation of lyophilized and reconstituted feces. Genotoxicity was determined in colon cells using the Comet assay. Selected samples were investigated for additional parameters related to carcinogenesis. Two of 7 FWs obtained by methods A and B were similarly genotoxic. Method B, however, yielded higher volumes of FW, allowing sterile filtration for long-term culture experiments. Four of 7 samples were non-genotoxic when prepared according to all 3 methods. FW from lyophilized feces and from fresh samples were equally genotoxic. FWs modulated cytotoxicity, paracellular permeability, and invasion, independent of their genotoxicity. All 3 methods of FW preparation can be used to assess genotoxicity. The higher volumes of FW obtained by preparation method B greatly enhance the perspectives of measuring different types of biological parameters and using these to disclose activities related to cancer development.
Acknowledgments and Notes
This project was supported by the European Communities, specific RTD program “Quality of Life and Management of Living Resources,” Key Action 1 “Food, Nutrition and Health,” project number QLK1-1999-00346 -SYNCAN. We are grateful to Eva Gietl for technical assistance and to Claudia Lüdtke for excellent secretarial assistance.
Notes
†Each value represents the data of 7 volunteers from 3 independent experiments (mean ± SD).
∗Significantly different compared to NaCl control.
†Each value represents the data of one volunteer from 3 independent experiments (mean ± SD). TI = tail intensity.