124 same location suggest environmental (animate or inanimate) reservoir for the legionnaires’ disease organism.
years apart in association with the
an
We thank Mr Robert Pollard, Bureau of Epidemiology, C.D.C. for assistance; Dr Charles Shepard, Bureau of Laboratories, C.D.C., for the I.F.A. studies; Dr Frank White, Laboratory Center for Disease Control, Health and Welfare Canada, Ottawa, for the clinical data and pathology specimens from a Canadian citizen; and Dr John Blackmon, Bureau of Laboratories, C.D.C., for reviewing the pathology specimens. Merton Smith of Grand Junction, Colorado, first alerted us to the illness among members of the Independent Order of Odd Fellows. Requests for reprints should be addressed to W.T., Enteric Diseases Branch, Bacterial Diseases Division, Bureau of Epidemiology, Center for Disease Control, Atlanta, Georgia 30333, U.S.A.
view of the known mutagenicity and several hair-dye constituents.
carcinogenicity of
Introduction
computer
REFERENCES 1.
Fraser, D. W., Tsai, T. F., Orenstein, W., Parkin, W. E., Beecham, J. H., Sharrar, R. G., Harris, J., Maliison, G. F., Martin, S. M., McDade, J. E., Shepard, C. C., Brachman, P. S. New Engl. J. Med. 1977, 297, 1189. 2. McDade, J. E., Shepard, C. C., Fraser, D. W., Tsai, T. F., Redus, M. A.,
Dowdle, W. R. ibid. p. 1197. Pike, M. C., Morrow, R. H. Br. J. prev. soc. Med. 1970, 24, 42. 4. Thacker, S., Bennett, J. V., Tsai, T. F., Fraser, D. W., McDade, J. E., Shepard, C. C., Williams, K. H., Stuart, W. H., Dull, H. B., Eickhoff, T. C. J. infect. Dis. (in the press).
3.
CHROMOSOMAL DAMAGE AND HAIR DYES S. D. LAWLER J. KIRKLAND Department of Cytogenetics and Immunogenetics, Institute of D.
Cancer Research, Fulham Road, London SW3
S. VENITT Division of Chemical Carcinogenesis, Institute
are mutagenic and cause chromosomal damage and allied effects when tested in short-term laboratory tests designed to detect potential carcinogens and mutagens. There is also evidence that some of these compounds and some commercial hair-dye formulations are carcinogenic when tested in rodents.’ These compounds readily penetrate the skin and have been detected in the urine of people who have dyed their hair under normal conditions of use.1-3 Several epidemiological studies suggest an increased risk of cancer among hairdressers and beauticians.’1 The disquieting possibility must be entertained, therefore, that very large numbers of people are exposed, either voluntarily or occupationally, to mixtures of potentially carcinogenic chemicals. Cytogenetic examination of peripheral-blood lymphocytes is, at present, the only practicable method of detecting genotoxic effects in groups of nominally healthy exposed individuals. With this technique elevated rates of chromosomal damage have been observed in people exposed to such diverse chemicals as vinyl chloride,4 metronidazole,s cadmium,6 and sulphonylurea drugs.7 We now report a similar study of hair tinters-a professional group whose work consists largely of colouring hair with a variety of permanent, semi-permanent, and temporary hair-dye preparations.
anisoles)
of Cancer
Research, Pollards Wood Research Station, Nightingales Lane, Chalfont St. Giles, Bucks An investigation of the potentially genotoxic effects of hair dyes in man produced the following findings: (i) no significant differences in chromosomal damage were found in cultured peripheral-blood lymphocytes from 60 professional hair colorists compared with 36 control subjects closely matched for age and sex; (ii) when age-matched women were re-grouped according to whether their own hair was dyed or not, there was a statistically significant excess of chromosomal damage (mainly chromatid breaks) in women with dyed hair; (iii) men (mean age 22·9 years) with dyed hair had significantly less chromosomal damage than men (mean age 31·5 years) whose hair was not dyed. Possible explanations are that: (i) most tinters wear gloves when applying hair dyes, and even without gloves percutaneous absorption of hair-dye constituents may be effectively impeded by the horny surface of the hands and by the lack of sebaceous glands in the palms; (ii) hair-dye constituents are readily absorbed through the scalp, which contains numerous sebaceous glands; and (iii) the lower frequency of chromosome aberrations in young men with dyed hair compared with that in slightly older men without dyed hair is probably due to an age effect, the frequency of hairdye use by the younger men being too low to reverse this effect. This preliminary indication that hair dyes may cause genotoxic effects in man warrants further study in
Summary
hair-dye constituents diamino-substituted benzenes, toluenes, and
THERE is evidence that many
(mainly
Volunteers 60 hair tinters and 36 controls were studied. The tinters were volunteers from many salons, situated mainly in central London and Surrey. Controls were drawn both from within the hairdressing profession (stylists, manicurists, receptionists, caterers) and from other occupations. All volunteers completed a detailed questionnaire covering smoking habit, alcohol consumption, medical history, use of medicinal drugs (including contraceptive pills), drugs of abuse, infections, vaccinations, X-ray exposure, and whether or not their own hair was dyed, either by themselves or professionally. Letters were sent to each volunteer’s general practitioner informing him or her of the study and asking for any information which might be thought relevant (e.g., prescription of drugs, or recent illness): less than 3% of the general practitioners replied. In addition, detailed information of occupational exposure to hair dyes was gathered (period and frequency of tinting; method of application and use of gloves for permanent and semipermanent tints, rinses, bleaches, and permanent waves; preparation of tints; washing off after tinting). Any volunteers who had ever experienced occupational exposure to hair dyes (currently or previously) were included in the "tinter" group.
Methods Blood samples taken from each subject at the time of interview were heparinised and coded. A sample was sent to the Department of Hasmatology, Royal Marsden Hospital, London, for routine hamiatological investigation (haemoglobin, full white-cell count, and differential count), and the rest was retained for peripheral-lymphocyte culture: 0.5ml of whole blooa from each subject was cultured for 48 h at 37°C in
125
gassed (5% CO2 in air) air-tight universal bottles containing 8 ml of ’TC199’ medium (Burroughs Wellcome), 2 ml fetal-calf serum (Gibco-Biocult), from a single batch reserved for the whole study, and 0.1 ml phytohaemagglutinin (Burroughs Wellcome). Chromosome spreads were prepared and stained with Giemsa by conventional methods from cultures arrested in metaphase by colchicine (0.5 µg/ml) for 1 h before harvest. Two investigators each examined 50 consecutive well-spread metaphases, and the results were pooled. Only cells with 45 or more chromosomes were analysed. Abnormalities were classified as follows: chromatid and chromosome gaps, chromatid and chromosome breaks, acentric fragments, dicentrics, other structural rearrangements, hyperdiploidy, polyploidy, and endoreduplication. A lesion was classified as a break rather than a gap if it satisfied either or both of the following conditions : (a) the length of the lesion exceeded the width of the chromatid, or (b) there was obvious displacement. All studies were carried out "blind" and the cytogeneticists did not discuss their findings with the interviewers until the
study was complete. The cytogenetic data were analysed statistically with x.2 (with Yates’ correction) for 2x2contingency tables of total abnormalities and comparison of Poisson distributions for data calculated as means. Student’s in comparisons of age distributions.
t test was
used
Results Tinters versus Controls Table i shows the cytogenetic findings for all subjects included in the study, grouped by sex and occupation, and disregarding a recent history of X-ray exposure and/or virus infection. There was no significant difference in the frequency and type of chromosome aberrations between controls and tinters except for an increase in the number of gaps recorded in female tinters (p
years apart in association with the
an
We thank Mr Robert Pollard, Bureau of Epidemiology, C.D.C. for assistance; Dr Charles Shepard, Bureau of Laboratories, C.D.C., for the I.F.A. studies; Dr Frank White, Laboratory Center for Disease Control, Health and Welfare Canada, Ottawa, for the clinical data and pathology specimens from a Canadian citizen; and Dr John Blackmon, Bureau of Laboratories, C.D.C., for reviewing the pathology specimens. Merton Smith of Grand Junction, Colorado, first alerted us to the illness among members of the Independent Order of Odd Fellows. Requests for reprints should be addressed to W.T., Enteric Diseases Branch, Bacterial Diseases Division, Bureau of Epidemiology, Center for Disease Control, Atlanta, Georgia 30333, U.S.A.
view of the known mutagenicity and several hair-dye constituents.
carcinogenicity of
Introduction
computer
REFERENCES 1.
Fraser, D. W., Tsai, T. F., Orenstein, W., Parkin, W. E., Beecham, J. H., Sharrar, R. G., Harris, J., Maliison, G. F., Martin, S. M., McDade, J. E., Shepard, C. C., Brachman, P. S. New Engl. J. Med. 1977, 297, 1189. 2. McDade, J. E., Shepard, C. C., Fraser, D. W., Tsai, T. F., Redus, M. A.,
Dowdle, W. R. ibid. p. 1197. Pike, M. C., Morrow, R. H. Br. J. prev. soc. Med. 1970, 24, 42. 4. Thacker, S., Bennett, J. V., Tsai, T. F., Fraser, D. W., McDade, J. E., Shepard, C. C., Williams, K. H., Stuart, W. H., Dull, H. B., Eickhoff, T. C. J. infect. Dis. (in the press).
3.
CHROMOSOMAL DAMAGE AND HAIR DYES S. D. LAWLER J. KIRKLAND Department of Cytogenetics and Immunogenetics, Institute of D.
Cancer Research, Fulham Road, London SW3
S. VENITT Division of Chemical Carcinogenesis, Institute
are mutagenic and cause chromosomal damage and allied effects when tested in short-term laboratory tests designed to detect potential carcinogens and mutagens. There is also evidence that some of these compounds and some commercial hair-dye formulations are carcinogenic when tested in rodents.’ These compounds readily penetrate the skin and have been detected in the urine of people who have dyed their hair under normal conditions of use.1-3 Several epidemiological studies suggest an increased risk of cancer among hairdressers and beauticians.’1 The disquieting possibility must be entertained, therefore, that very large numbers of people are exposed, either voluntarily or occupationally, to mixtures of potentially carcinogenic chemicals. Cytogenetic examination of peripheral-blood lymphocytes is, at present, the only practicable method of detecting genotoxic effects in groups of nominally healthy exposed individuals. With this technique elevated rates of chromosomal damage have been observed in people exposed to such diverse chemicals as vinyl chloride,4 metronidazole,s cadmium,6 and sulphonylurea drugs.7 We now report a similar study of hair tinters-a professional group whose work consists largely of colouring hair with a variety of permanent, semi-permanent, and temporary hair-dye preparations.
anisoles)
of Cancer
Research, Pollards Wood Research Station, Nightingales Lane, Chalfont St. Giles, Bucks An investigation of the potentially genotoxic effects of hair dyes in man produced the following findings: (i) no significant differences in chromosomal damage were found in cultured peripheral-blood lymphocytes from 60 professional hair colorists compared with 36 control subjects closely matched for age and sex; (ii) when age-matched women were re-grouped according to whether their own hair was dyed or not, there was a statistically significant excess of chromosomal damage (mainly chromatid breaks) in women with dyed hair; (iii) men (mean age 22·9 years) with dyed hair had significantly less chromosomal damage than men (mean age 31·5 years) whose hair was not dyed. Possible explanations are that: (i) most tinters wear gloves when applying hair dyes, and even without gloves percutaneous absorption of hair-dye constituents may be effectively impeded by the horny surface of the hands and by the lack of sebaceous glands in the palms; (ii) hair-dye constituents are readily absorbed through the scalp, which contains numerous sebaceous glands; and (iii) the lower frequency of chromosome aberrations in young men with dyed hair compared with that in slightly older men without dyed hair is probably due to an age effect, the frequency of hairdye use by the younger men being too low to reverse this effect. This preliminary indication that hair dyes may cause genotoxic effects in man warrants further study in
Summary
hair-dye constituents diamino-substituted benzenes, toluenes, and
THERE is evidence that many
(mainly
Volunteers 60 hair tinters and 36 controls were studied. The tinters were volunteers from many salons, situated mainly in central London and Surrey. Controls were drawn both from within the hairdressing profession (stylists, manicurists, receptionists, caterers) and from other occupations. All volunteers completed a detailed questionnaire covering smoking habit, alcohol consumption, medical history, use of medicinal drugs (including contraceptive pills), drugs of abuse, infections, vaccinations, X-ray exposure, and whether or not their own hair was dyed, either by themselves or professionally. Letters were sent to each volunteer’s general practitioner informing him or her of the study and asking for any information which might be thought relevant (e.g., prescription of drugs, or recent illness): less than 3% of the general practitioners replied. In addition, detailed information of occupational exposure to hair dyes was gathered (period and frequency of tinting; method of application and use of gloves for permanent and semipermanent tints, rinses, bleaches, and permanent waves; preparation of tints; washing off after tinting). Any volunteers who had ever experienced occupational exposure to hair dyes (currently or previously) were included in the "tinter" group.
Methods Blood samples taken from each subject at the time of interview were heparinised and coded. A sample was sent to the Department of Hasmatology, Royal Marsden Hospital, London, for routine hamiatological investigation (haemoglobin, full white-cell count, and differential count), and the rest was retained for peripheral-lymphocyte culture: 0.5ml of whole blooa from each subject was cultured for 48 h at 37°C in
125
gassed (5% CO2 in air) air-tight universal bottles containing 8 ml of ’TC199’ medium (Burroughs Wellcome), 2 ml fetal-calf serum (Gibco-Biocult), from a single batch reserved for the whole study, and 0.1 ml phytohaemagglutinin (Burroughs Wellcome). Chromosome spreads were prepared and stained with Giemsa by conventional methods from cultures arrested in metaphase by colchicine (0.5 µg/ml) for 1 h before harvest. Two investigators each examined 50 consecutive well-spread metaphases, and the results were pooled. Only cells with 45 or more chromosomes were analysed. Abnormalities were classified as follows: chromatid and chromosome gaps, chromatid and chromosome breaks, acentric fragments, dicentrics, other structural rearrangements, hyperdiploidy, polyploidy, and endoreduplication. A lesion was classified as a break rather than a gap if it satisfied either or both of the following conditions : (a) the length of the lesion exceeded the width of the chromatid, or (b) there was obvious displacement. All studies were carried out "blind" and the cytogeneticists did not discuss their findings with the interviewers until the
study was complete. The cytogenetic data were analysed statistically with x.2 (with Yates’ correction) for 2x2contingency tables of total abnormalities and comparison of Poisson distributions for data calculated as means. Student’s in comparisons of age distributions.
t test was
used
Results Tinters versus Controls Table i shows the cytogenetic findings for all subjects included in the study, grouped by sex and occupation, and disregarding a recent history of X-ray exposure and/or virus infection. There was no significant difference in the frequency and type of chromosome aberrations between controls and tinters except for an increase in the number of gaps recorded in female tinters (p
