Enciso-Gadea, J. M. (José Manuel)

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    Comet assay modifications for its application in food safety
    (2019-01-16) Enciso-Gadea, J. M. (José Manuel); Lopez-de-Cerain, A. (Adela); Vettorazzi, A. (Ariane); Azqueta, A. (Amaya)
    Information on genotoxicity is of a key importance for the toxicological characterisation of different compounds. In this vein, and due to its various advantages, the comet assay is currently included in the genotoxicity testing strategy used in the food safety area. However, improvement points of particular interest have been identified. Thereby, the main objective of the present work was to evaluate some critical points of the comet assay, such as the time of lysis, in vitro, and the methodology used in the freezing/thawing procedures of tissue samples, their stability and the application of the Fpg-modified assay, in vivo. In addition, the in vivo comet assay was applied to frozen kidney samples obtained in a previous repeated-dose toxicity study of the food contaminant ochratoxin A. Finally, the knowledge derived from these objectives resulted in the development of standard operating procedures for both the in vitro and in vivo comet assays, which could be applied in good laboratory practice studies.
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    Sex differences in ochratoxin a toxicity in F344 rats after 7 and 21 days of daily oral administration
    (Elsevier, 2018) Pastor, L. (Laura); García-Jalón, J.A. (José Antonio); Enciso-Gadea, J. M. (José Manuel); Lopez-de-Cerain, A. (Adela); Vettorazzi, A. (Ariane); Gonzalez-Peñas, E. (Elena); Monreal, J.I. (José Ignacio)
    Ochratoxin A (OTA) is a potent renal carcinogen in male rats but not in females. The mechanisms underlying these differences are unknown. The sex-dependent response of F344 rats after a repeated OTA oral administration for 7 (0.50 mg/kg bw) or 21 days (0.21 and 0.50 mg/kg bw) was evaluated. General toxicity, sex and thyroid hormones and histopathology were studied. OTA was quantified (HPLC-FLD) in plasma, kidney and liver and the expression of kidney transporters (RT-qPCR) was studied. After 7 days, kidney histopathology showed more pronounced signs of toxicity in males than in females. After 21 days, a higher toxicity was observed but sex differences disappeared. OTA concentration in plasma and tissues was similar in both sexes. Downregulation was the general OTA-induced effect. Oats' downregulation was slow in males and Oat3 did not change in females. Oatp1 was strongly downregulated in males after 21 days. An opposite effect was observed in Bcrp after 21 days: downregulation in males and upregulation in females. Females showed a dose- and time-dependent decrease of progesterone. Despite the sex differences, the final balance in OTA toxicokinetics at renal cell level does not seem to support a higher accumulation of OTA in male kidneys.