[en] The effects of low-doses of ionizing radiation in humans are of growing concern, especially in the context of current radiation techniques such as medical imaging. The biological response of healthy tissue to low dose of 1-10 cGy in vivo is largely unknown. In this project, we propose firstly to study the effects (long and short-term) of low-doses on cell proliferation, apoptosis, and capacity to obtain a cohesive and stratified epidermis after irradiation. Secondly, we will evaluate the carcinogenesis risk by measuring the modulation of the DNA repair/damage systems after low-dose exposure. For short-term radiosensitivity, cell viability was determined by MTT assay after 24, 48 and 72 h post irradiation, we also performed an in vivo colony-forming assay, which measures the radiation toxicity after 2 weeks. DNA repair system and damage was assessed by different techniques available in our laboratory (DNA repair chips, modified comet assay). Finally, organogenesis potential was determined by the capacity of normal exposed keratinocytes to form a pluri-stratified epithelium in 3D organo-typic cultures. We showed that low-dose of ionizing radiation increases 2 fold the oxidative DNA damage (p=0.01) without any activation of the base excision repair pathway, an important pathway to repair oxidative DNA damage. Moreover, we showed that low-dose affects the organogenesis potential of keratinocytes and impairs the proliferation-differentiation balance in the reconstructed skin. We postulate that when the dose or dose rate is very low the radiation damage sensors (ATM or ATR) are not activated, and the repair machinery is not induced. Hence damage could be accumulated in the genome of a cell until eventually it become malignant. (authors)

[en] Radioimmunofixation of human ferritin following isoelectric focusing of serum was developed to study the microheterogeneity of this protein in native serum without previous purification or concentration. This method requires only 2-10 μl of serum and can be used with levels of ferritin as low as 10 μg/l. In this way, the extensive microheterogeneity of this protein was revealed, since in some cases it produced as many as 35 bands with isoelectric points in a pH range of 4.95-5.9. Very different isoelectric focusing patterns (spectrotypes) of ferritin were observed during the investigation of pathological sera. The high sensitivity of this technique makes it useful for the investigation of serum ferritin in diseases involving modifications of the metabolism of this protein. (orig.)

[en] Epidermal keratinocytes are used as a cell source for autologous and allogenic cell transplant therapy for skin burns. The question addressed here is to determine whether the culture process may induce cellular, molecular, or genetic alterations that might increase the risk of cellular transformation. Keratinocytes from four different human donors were investigated for molecular and cellular parameters indicative of transformation status, including (i) karyotype, (ii) telomere length, (iii) proliferation rate, (iv) epithelial-mesenchymal transition, (v) anchorage-independent growth potential, and (vi) tumorigenicity in nude mice. Results show that, despite increased cell survival in one keratinocyte strain, none of the cultures displayed characteristics of cell transformations, implying that the culture protocol does not generate artefacts leading to the selection of transformed cells. We conclude that the current protocol does not result in an increased risk of tumorigenicity of transplanted cells.