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Armen K.Nersesyan 3000–5000 (9) and even, in the opinion of Belien et al. (10), 10 000 exfoliated cells should be studied, due to a low baseline Laboratory of Carcinogenesis, Cancer Research Centre, 76 Fanardjian MN frequency. The second reason may be the use of Giemsa Street, Yerevan 75052, Armenia staining instead of DNA-specific stains (acridine orange, Email: genetik@ysu.am Hoechst 33258 plus pyronin, propidium iodine or DAPI) because the latter can eliminate most of the artefacts associated Dear Sir with use of DNA non-specific stains (9). I read with great interest the research findings of Dr Rajeswari In conclusion, I suggest that, along with real MN, Rajeswari and co-workers (1). I draw attention to the extremely high et al. (1) registered artefacts due to the few cells scored and background micronucleus (MN) level in exfoliated buccal the use of Giemsa staining. mucosa cells of untreated cancer patients and their first degree female relatives (3.41 and 1.65%, respectively) compared with References controls (0.27%). This means that in buccal cells of cancer 1.Rajeswari,N., Ahuja,Y.R., Malani,U., Chandrashekar,S., Balakrishna,N., patients and their relatives MN frequencies were 12.6and Rao,K.V. and Khar,A. (2000) Risk assessment in the first degree female 6.1-fold higher than in control subjects. relatives of breast cancer patients using the alkaline Comet assay. Analysis of some recent publications has shown that after Carcinogenesis, 21, 557–561. 2.Gomez-Arroya,S., Diaz-Sanchez,Y., Meneses-Perez,M.A., Villalobosexposure to environmental mutagens, such as pesticides (2), Pictrini,R. and De Leon-Rodriguez,J. (2000) Cytogenetic biomonitoring organic solvents and lead-containing pigments (3), radioactive in Mexican floriculture workers exposed to pesticides. Mutat. Res., 466, iodine (4) and antineoplastic drugs (5), MN level increased 117–124. only 2.7–3.7 times (2,3) or did not increase significantly (4,5). 3.Pinto,D., Ceballos,J.M., Garcia,G., Guzman,P., Del Razo,L.M., Vera,E., Gomez,H., Garcia,A. and Gonsebatt,E. (2000) Increased cytogenetic It is extremely unlikely that the difference in MN content damage in outdoor painters. Mutat. Res., 467, 105–111. in the same healthy population may differ by 6.1 times (female 4.Ramirez,M.J., Surralles,J., Galofre,P., Creus,A. and Marcos,R. (1999) FISH relatives and controls) while after exposure to mutagens analysis of 1cen-1q12 breakage, chromosome 1 numerical abnormalities the differences between exposed subjects and controls were and centromeric content of micronuclei in buccal cells from thyroid cancer 2.7–3.7. and hyperthyroidism patients treated with radioactive iodine. Mutagenesis, 14, 121–127. My opinion is supported by some recent papers concerning 5.Burgaz,S., Karahalil,B., Bayrak,P., Taskin,L., Yavuzaslan,F., Bokesoy,I., cytogenetic disturbances in lymphocytes of breast cancer Anzion,R.B.M., Bos,R.P. and Platin,N. (1999) Urinary cyclophosphamide patients and their female relatives (6,7). The differences in the excretion and micronuclei frequences in peripheral lymphocytes and in levels of chromosomal aberrations in lymphocytes of patients exfoliated buccal epithelial cells of nurses handling antineoplastics. Mutat. Res., 439, 97–104. and their relatives were 1.4 (6) and 1.7 (7) and between 6.Roy,S.K., Trivedi,A.H., Bakshi,S.R., Patel,R.K., Shukla,P.H., Patel,S.J., relatives and controls was 1.5 (6) (data concerning control Bhatavdekar,J.M., Patel,D.D. and Shah,P.M. (2000) Spontaneous subjects was not presented in Trivedi et al.; ref. 7). Moreover, chromosomal instability in breast cancer families. Cancer Genet. a paper by Jyothish et al. (8) showed no differences (neither Cytogenet., 118, 52–56. constitutional nor structural) in lymphocytes of breast cancer 7.Trivedi,A.H., Roy,S.K., Bhachech,S.H., Patel,R.K., Dalal,A.A., Bhatavdekar,J.M. and Patel,D.D. (1998) Cytogenetic evaluation of 20 patients and their relatives compared with control subjects. sporadic breast cancer patients and their first degree relatives. Breast However, it is well established that use of MN in exfoliated Cancer Res. Treat., 48, 187–190. buccal mucosa cells to detect chromosomal damage induced 8. Jyothish,B., Ankathil,R., Chandini,R., Vinodkumar,B., Nayar,G.S., by various mutagens is less efficient than that of both MN and Roy,D.D., Madhavan,J. and Nair,M.K. (1998) DNA repair proficiency: a potential marker for identification of high risk members in breast cancer chromosomal aberrations in lymphocytes (3,5). Thus, if the families. Cancer Lett., 124, 9–14. difference in chromosomal aberration levels in lymphocytes 9.Albertini,R.J., Anderson,D., Douglas,G.R., Hagmar,L., Hemminki,K., of relatives and control subjects is 2, the difference in MN Merlo,F., Natarajan,A.T., Norppa,H., Shuker,D.E.G., Tice,R., Waters,M.D. level in buccal mucosa cells should be at least the same, but and Aitio,A. (2000) IPCS guidelines for the monitoring of genotoxic no more. effects of carcinogens in humans. Mutat. Res., 463, 111–172. 10.Belien,J.A., Copper,M.P., Braakhuis,B.J., Snow,G.B. and Baak,J.P. (1995) I suggest that the extremely high levels of cells with MN Standardization of counting micronuclei: definition of a protocol to (1) were due to two causes. Rajeswari et al. (1) analyzed only measure genotoxic damage in human exfoliated cells. Carcinogenesis, 16, 1000 cells from each subject, although for this purpose at least 2395–2400.

carcinogenesis

Re: Rajeswari,N., Ahuja,Y.R., Malani,U., Chandrashekar,S., Balakrishna,N., Rao,K.V. and Khar,A. (2000) Risk assessment in the first degree female relatives of breast cancer patients using the alkaline Comet assay. Carcinogenesis, 21, 557–561