1) STUDY OF MOLECULAR INTERACTIONS OF ENVIRONMENTAL MUTAGENS AND CARCINOGENS AT DNA LEVEL.
2) TO RELATE BIOLOGICAL CONSEQUENCES OF CHEMICAL MUTAGENS TO THE EXTENT AND SITE OF REACTION BETWEEN THE MUTAGENS AND DNA.
Methodology was developed to assess genotoxic damage in human individuals as a result of exposure to mutagenic or carcinogenic chemicals at the workplace or in the general environment. The methods used were based on the immunochemical and biochemical detection of deoxyribonucleic acid (DNA) adduct formation in peripheral blood leukocytes. In order to deal with biomonitoring studies in large populations, the immunochemical techniques used were adapted so that low levels of DNA damage could be analysed in a large number of samples.
1)FURTHER DEVELOPMENT AND APPLICATION OF METHODS TO ANALYSE THE INDUCTION OF DNA DAMAGE (ADDUCTS) IN SITU, IN THE NUCLEI OF FIXED CELLS, BY MEANS OF IMMUNOFLUORESCENCE MICROSCOPY WITH ADDUCT SPECIFIC ANTIBODIES. LASER-SCAN MICROSCOPY IS INTRODUCED TO DETECT AND QUANTIFY IMMUNOFLUORESCENCE SIGNALS IN NUCLEI OF CELLS TREATED WITH GENOTOXIC AGENTS.
2)THE FOLLOWING WORK IS CARRIED OUT:
I)THE OCCURRENCE OF HEMOGLOBIN MUTATIONS IN ERYTHROCYTES - THEIR PRESENCE IS DETECTED WITH MONOCLONAL ANTIBODIES RAISED AGAINST TWO WILDTYPE HEMOGLOBIN CHAINS.
II)THE INDUCTION OF SEX-LINKED RECESSIVE LETHAL MUTATIONS (SLRL) IN DROSOPHILA; LINKED WITH DNA ADDUCT INDUCTION AND POSSIBLE REPAIR IN GERM CELLS.
III)INDUCTION AND REPAIR OF DNA DAMAGE IN THE MOUSE GERM LINE; SIMILAR STUDY TO II) ABOVE, WITH RESPECT TO DOSIMETRY OF DNA DAMAGE IN MOUSE GERM CELLS.
IV)MOLECULAR DOSIMETRY OF ALKYL DAMAGE IN DNA BY MEANS OF ALKALINE ELUTION; THE EFFECTS OF FIVE ETHYLATING AGENTS ON CULTURED CELLS FROM VARIOUS MUTANT CELL LINES IS STUDIED WITH RESPECT TO THE LEVELS OF INITIAL DNA DAMAGE, AND SUBSEQUENT DNA REPAIR RESPONSE.
V)MOLECULAR DOSIMETRY OF DNA DAMAGE AT N7 OF GUANINE; BIOMONITORING METHODS WILL BE DEVELOPED BASED ON IMMUNODETECTION OF N7-ALKYL ADDUCTS.