................................................................................................................................................................ 5 RIASSUNTO ............................................................................................................................................................... 8 GENERAL INTRODUCTION ................................................................................................................................ 11 1. CRANIO-FACIAL DEFECTS ................................................................................................................... 12 2. CRANIOFACIAL DEVELOPMENT ........................................................................................................ 13 3. AZOLE FUNGICIDES TERATOLOGY .................................................................................................. 20 4. ETHANOL TERATOLOGY ...................................................................................................................... 25 5. STATE OF THE ART ON THE RISK ASSESSMENT FROM EXPOSURE TO MIXTURES OF CHEMICALS .............................................................................................................................................. 27 6. ALTERNATIVE MODELS IN DEVELOPMENTAL TOXICICTY EVALUATION ......................... 35 7. AIM OF THE WORK ................................................................................................................................. 44 8. LEGEND OF ABBREVIATIONS .............................................................................................................. 47 PART 1: PERTURBATION OF THE RETINOIC ACID PATHWAY ............................................................... 48 Introduction .......................................................................................................................................................... 49 Materials and methods ......................................................................................................................................... 51 Results .................................................................................................................................................................... 53 Discussion .............................................................................................................................................................. 57 Tables and Figures ................................................................................................................................................ 61 PART 2: DEVELOPMENT OF AN INTEGRATED SYSTEM BIOLOGY MODEL FOR PREDICTING MIXTURE OF CHEMICALS ACTING ON THE SAME PATHWAY ............................................. 76 Introduction .......................................................................................................................................................... 77 Materials and methods ......................................................................................................................................... 81 Results .................................................................................................................................................................... 86 Discussion .............................................................................................................................................................. 88 Tables and Figures ................................................................................................................................................ 94 PART 3: THE MIXTURE EFFECT OF CO-EXPOSURE TO ETHANOL AND FLUCONAZOLE: A STUDY IN AN ALTERNATIVE DEVELOPMENTAL MODEL, THE ASCIDIAN CIONA INTESTINALIS (AET) .......................................................................................................................... 102 Introduction ........................................................................................................................................................ 103 Materials and methods ....................................................................................................................................... 104 Discussion ............................................................................................................................................................ 109 Tables and Figures .............................................................................................................................................. 111 GENERAL DISCUSSION...................................................................................................................................... 117 ACKNOWLEDGEMENT ...................................................................................................................................... 127 REFERENCES ........................................................................................................................................................ 128 APPENDIX: Papers under review......................................................................................................................... 138