Search

The toxicological relevance of the micronucleus (MN) test is well defined: it is a multi-target genotoxic endpoint, assessing not only clastogenic and aneugenic events but also some epigenetic effects, which is simple to score, accurate, applicable in different cell types. In addition, it is predictive for cancer, amenable for automation and allows good extrapolation for potential limits of exposure or thresholds and it is easily measured in experimental both in vitro and in vivo systems. Implementation of in vitro micronucleus (IVMN) assays in the battery of tests for hazard and risk assessment of potential mutagens/carcinogens is therefore fully justified. Moreover, the final draft of an OECD guideline became recently available for this test. In this review, we discuss the prerequisites for an acceptable MN assay, including the cell as unit of observation, importance of cell membranes, the requirement of a mitotic or meiotic division and the assessment of cell division in the presence of the test substance. Furthermore, the importance of adequate design of protocols is highlighted and new developments, in particular the in vitro 3D human skin models, are discussed. Finally, we address future research perspectives including the possibility of a combined primary 3D human skin and primary human whole blood culture system, and the need for adaptation of the IVMN assays to assess the genotoxic potential of new materials, in particular nanomaterials.

BACKGROUND: The use of cancer-related biomarkers in newborns has been very limited. OBJECTIVE: We investigated the formation of micronuclei (MN) in full-term and preterm newborns and their mothers from the Rhea cohort (Crete), applying for the first time in cord blood a validated semiautomated analysis system, in both mono-and binucleated T lymphocytes. METHODS: We assessed MN frequencies in peripheral blood samples from the mothers and in umbilical cord blood samples. We calculated MN in mononucleated (MNMONO) and binucleated (MNBN) T lymphocytes and the cytokinesis block proliferation index (CBPI) in 251 newborns (224 full term) and 223 mothers, including 182 mother-child pairs. Demographic and lifestyle characteristics were collected. RESULTS: We observed significantly higher MNBN and CBPI levels in mothers than in newborns. In newborns, MNMONO and MNBN were correlated (r = 0.35, p

A family pilot study was conducted in the Czech Republic to test the hypothesis that exposure to air pollution with particulate matter (PM) in children results in detectable effects indicated by a number of biomarkers of exposure and early effects. The frequency of micronuclei (MN) in peripheral blood lymphocytes (PBLs) was analysed to assess the cytogenetic effects in children and mothers living in two different areas. From each area two groups of children from a total of 24 families (mean age: 6.0+/-0.6 and 9.0+/-1.2 years) in a total of 47 children and 19 mothers (mean age: 33.6+/-3.9 years) participated. Chromosome aberrations determined with fluorescence in situ hybridization (FISH) painting for chromosomes #1 and #4 were analysed in 39 children and 20 parents. Teplice, a mining district, in Northern Bohemia was selected for the analyses of the effects in a population exposed to high levels of air pollution, especially during winter, and compared with a population from the rural area of Prachatice in Southern Bohemia. Significant higher frequencies of MN were found in the younger children living in the Teplice area as compared with those living in the Prachatice area (7.0+/-2.3 per thousand versus 4.9+/-2.0 per thousand, p=0.04). Higher levels of MN were also measured in the older children and the mothers from the Teplice area (9.2+/-3.7 per thousand versus 6.6+/-4.4 per thousand) and (12.6+/-3.4 per thousand versus 10.1+/-4.0 per thousand). The increased MN frequency may be associated with elevated carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) concentration of the PM(2.5) measured in the ambient Teplice air, but other factors like genotoxic compounds from the diet or protective effect of micronutrients, which was not addressed in this pilot study, may also differ between the two areas. MN frequencies were found to increase with age in children. Lower MN frequency was found in boys as compared to girls. The result of the FISH analyses showed a low number of individuals with detectable levels of aberrations and no significant increases in genomic frequency of stable chromosome exchanges (F(G)/100) were found in children or parents from the Teplice area in comparison with those from the Prachatice area. The family pilot study indicates that MN is a valuable and sensitive biomarker for early biological effect in children and adults living in two different areas characterised with significant exposure differences in c-PAHs concentrations during winter.

The use of biomarkers of early genetic effects, predictive for cancer, such as micronuclei (MN) in lymphocytes, may help to investigate the association between diet and cancer. We hypothesised that the presence of mutagens in the diet may increase MN formation. A 'pooled' standardised analysis was performed by applying the same experimental protocol for the cytokinesis block micronucleus assay in 625 young healthy women after delivery from five European study populations (Greece, Denmark, UK, Spain and Norway). We assessed MN frequencies in mono- and binucleated T-lymphocytes (MNMONO and MNBN) and the cytokinesis blocked proliferation index using a semi-automated image analysis system. Food frequency questionnaires (FFQs) were used to estimate intake of fatty acids and a broad range of immunotoxic and genotoxic/carcinogenic compounds through the diet. Pooled difference based on delivery type revealed higher MNMONO frequencies in caesarean than in vaginal delivery (P = 0.002). Statistical analysis showed a decrease in MNMONO frequencies with increasing calculated omega-6 PUFA concentrations and a decrease in MNBN frequencies with increasing calculated omega-3 PUFA concentrations. The expected toxic compounds estimated by FFQs were not associated with MN formation in mothers after delivery. In pregnant women, an omega-3 and -6 rich diet estimated by FFQ is associated with lower MN formation during pregnancy and delivery.

Recognition of the potential vulnerability of children and newborns and protection of their health is essential, especially regarding to genotoxic compounds. Benzo(a)pyrene B(a)P a commonly found carcinogen, and its metabolite BPDE, are known to cross the placenta. To investigate how well newborns are able to cope with BPDE-induced DNA damage, a recent developed nucleotide excision repair cell phenotype assay was applied in a pilot study of 25 newborn daughters and their mothers, using the Alkaline Comet Assay and taking demographic data into account. Newborns seemed to be less able to repair BPDE-induced DNA damage since lower repair capacity levels were calculated compared to their mothers although statistical significance was not reached. Assessment of repair capacity in combination with genotypes will provide important information to support preventive strategies in neonatal care and to define science based exposure limits for pregnant women and children.