9ZE TaBlE A8.—Outline of methods used in retrospective studies of smoking in relation to lung cancer (eunt.) Author, year, Sex of Number of persons and method of selection country, cases - - Collection of data reference Cases Controls Randig, M-F 448 lung cancer patients in a 512 patients with other diagnoses, matched Controls were interviewed at about the same 1954, number of West Berlin hospi- for age. time as the cases, each case-control pair Germany tals. by the same physician. (218). Wynder et al., F 105 patients with lung cancer in 1,304 patients at Memoria] Center with tu- Cases: Personal interview or questionnaire 1956, several New York City hospi- mors of sites other than respiratory or mailed to close relatives or friends. U.S.A. tals. upper alimentary. Controls: Personal interview. (311). Segi et al., M-F 207 patients with lung cancer in 5,636 patients free of cancer in 420 local Cases and controls by personal interview 1957, 33 hospitals in all parts of health centers, selected to approximate using long questionnaire on occupational Japan the country. the sex and age distributions of cases. and medical history and living habits. (250), Mills and M--F 578 residents of defined areas 3,310 population sample approximately pro- Cases: From death certificates, hospital rec- Porter, dying of respiratory cancer. portional to cases as regards areas of resi- ords, and close relatives or friends. 1957, dence, and 10 years or more in the area, Controls: Personal home visits or telephone U.S.A. calls, usually interviewing housewife. (187). Stocks, M-F 2,356 patients suffering from or 0,362 unselected patients of the same area Cases: Histories taken at the hospital from 1957, dying with lung cancer within admitted for conditions other than cancer. relatives by health visitors. England certain areas. Controls: Personal interview in hospital. (263). Schwartz and M 602 patients with bronchopul- 1,204 patients (3 groups) in same hospitals Personal interviews in the hospital; cases Denoix, monary cancer in hospitals. with other cancer, with nonmalignant ill- and controls at about the same time by 1957, ness, and accident cases, matched by age the same interviewer. France group. (247). LTE TABLE A3.—Outline of methods used in retrospective studies of smoking in relation to lung cancer (cont.) Author, year, Sex of country, cases reference Number of persons and method of selection Cases Controls Collection of data Haenszel and F Shimkin, 1958, U.S.A. (113). 158 lung cancer patients avail- able for interview in 29 hos- pitals. 339 patients in same hospital and service at same time, next older and next younger than each case. Personal interviews by resident, medical so- cial worker, or clinic secretary. Lombard and M Snegireff, 1959, U.S.A. (176). 500 men dying of lung cancer, microscopically confirmed. 4,238 controls in 7 groups including volun- teers, hospital and clinic patients, random population sample, and house-to-house sur- vey samples. Personal interviews by trained workers. Pernu, M-F 1960, Finland (277). 1,606 respiratory cancer patients in 4 hospitals and from cancer registry. 1,773 cancer-free persons recruited by Parish Sisters of 2 institutes in all parts of the country. Cases: From case histories or mailed ques- tionnaires. Controls: Questionnaires distributed by Par- ish Sisters. Haenszel M etal., 1962, U.S.A. (112). 2,191 sample of 10 percent of white male lung cancer deaths in the U.S.A. in 1958. 31,516 random sample from Current Popu- lation Survey. Cases: By mail from certifying physicians and family informants. Controls: Personal interview by census enu- merators. Lancaster, M 1962, Australia (158). 238 hospital patients with lung cancer. 476 in 2 groups, 1 with other cancer, 1 with some other disease, matched by sex and age. Personal interviews of both cases and con- trols in hospitals. Haenszel and F Taeuber, 1964, U.S.A. (115). 749 sample of 10 percent of white female lung cancer deaths in the U.S.A. in 1958 and 1959. 34,339 random sample from Current Popula- tion Survey used to estimate population base. Cases: Ry mail from certifying physicians and family informants. Controls: Personal interview by census enu- merators. 8ze TABLE A3.—Outline of methods used in retrospective studies of smoking in relation to lung cancer (cont.) Author, year, Sex of Number of persons and method of selection country, cases Collection of data reference Cases Controls Wicken, M-F 954 patients with primary lung 954 age and sex-matched controls from same Interviews with relatives. 1966, cancer. locale and deceased from nonrespiratory Northern diseases. Treland (308). Gelfand et al., M 1968, 32 patients with bronchogenic cancer. 32 age and sex-matched patients Hospitalization interviews. Rhodesia (98), Hitosugi, M-F 185 patients with lung cancer 491 persons sex-matched from similar air- Cases: Hospital interviews. 1968, pollution regions. Controls: Interviews by trained public health Japan nurses. (126), Bradshaw and M Schonland, 1969, South Africa (Natal) (41). 45 Zulu patients with lung can- eer. 341 Zulu patients without lung cancer. Interviewed by trained African social worker. Ormos et al., M-F 1969, Hungary (204). 118 patients with lung cancer. 3,089 control persons without data on health history. Cases: Data derived from case histories and interviews with relatives. Controls: Interviews with a random sample of train passengers. Wynder, et al., M-F 1970 U.S.A. (324). 240 patients with Kreyberg Type I lung cancer. 480 age and sex-matched patients Hospitalization interview. 6ZE TABLE A4.—Group characteristics in retrospective studies on lung cancer and tobacco use SM = Smokers. NS = Nonsmokers. —— _ _—_— _ reece Males Females Author, Cases Controls Cases Controls year, a Relative _—— “————— Relative Comments reference Percent Percent Percent Percent risk Percent Percent Percent Percent risk non- heavy non- heavy ratio non- heavy non- heavy ratio Number smokers smokers! Number smokers smokers! SM:NS?. Number smokers smokers! Number smokers smokers! SM:NS2 — ee Or Miiller, 86 3.5 65.1 86 16.3 36.0 95.4 (4) (4) (4) (4) (4) (4) a 1939 (196). — 7 ee oo eee Schairer and 93 3.2 31.2 270 15.9 9.3 55.7 (4) (4) (4) (4) (4) (4) ++. 16 female Schéniger, cases not 1943 (242). analyzed. So eee Se Potter and 43 7.0 30.2 2,804 26.0 23.0 34,1 (4) () (4) (4) (4) (4) tae Tully, 1945 (212). — a 192 AGB Wassink, 134 4.8 54.8 100 19.2 19.2 4.7 {*) (4) (4) {4) (4) (4) -+. Percentages 1948 (204). estimated from chart. ST a Be EEE Schrek et al., 82 14.6 18.3 522 23.9 9.2 1.8 (4) @) (4) (4) (4) (4) tee 1950 (246). ———_._ ~ — OT Mills and 444 7.2 ee 430 30.5 see 5.7 (4) (4) (4) a) (4) (*) see Porter, 1950 (186), Levin et al., 236 15.3 vee 481 21.7 wee 1.5 (*) (*) (4) (*) (*) (4) -+. Quantity 1950 (171). smoked not considered. — ‘780.~«ABSSSC*S dg tered Wynder and 605 1.3 51.2 780 14.6 19.1 13.0 40 57.5 25.0 552 79.6 1.2 2.9 Graham, 1950 (816). a Oce TABLE A4.—Group characteristics in retrospective studies on lung cancer and tobacco use (cont.) SM = Smokers. NS = Nonsmokers. Males Females Author, Cases Controls Cases Controls ; year, “ — Relative Relative Comments reference Percent Pereent Percent Percent risk Percent Percent Percent Percent risk non- heavy non- heavy ratio non- heavy non- heavy ratio Number smokers smokers! Number smokers smokers! SM:NS2 Number smokers smokers' Number smokers smokers! SM:NS? McConnell 93 54 38.5 186 6.5 23.2 1.2 7 BT. 14 78.6 wee 2.8 etal, 1952 (180). Doll and Hill, 1,357 0.5 25.1 1,357 45 13.4 9.4 108 37.0 11.1 108 54.6 0.9 2.1 Percentage 1952 (78). “heavy” smokers understated. Sadowsky 477 3k tee 615 13.2 3.9 (4) (4) (*) @) (*) (4) ... Gradient etal, with amount 1953 (232). smoked. Wynder and 63 41 67.6 133 20.6 29.3 36,1 (*) () (@) QO) (4) @) Cornfield, 1953 (814). Koulumies R12 0.6 58.9 300 18.0 25.0 36.0 (4) (4) (4) 1958 (152). Lickint 224 1.8 35.8 1,000 16.0 4.8 310.4 22 64.0 4.5 1,002 90.4 0.1 5.3 _ 1953 (170). Breslow et al., 493 3.7 TA. 518 10.8 42.7 3.2 1954 (42). Watson and 265 1.9 71.7 287 9.7 51.6 35.6 36 58.3 2.8 181 82.0 11 3.3 Conte, 1954 (305). Gsell, 135 0.7 68.1 135 16.0 14.0 326.8 (*) @) (4) (@) Q) @) 1954 (107). TABLE A4,—Group characteristics in retrospective studies on lun SM = Smokers. NS = Nonsmokers. g cancer and tobacco use (cont.) Males Females Author, Cases Controls Cases Controls year, Relative — Relative Comments reference Percent Percent Percent Percent risk Percent Percent Percent Percent risk Num- non- heavy non- heavy ratio Num- non- heavy non- heavy ratio ber smokers smokers! Number smokers smokers? SM: NS? ber smokers smokers! Number smokers smokers' SM:NS? Randig, 415 1.2 34.2 381 6.8 17.9 35.1 33 51.5 3.0 131 70.3 0 2.2 1954 (218). Wynder etal. (*) (4) (*) (4) (4) (4) 105 56.2 16.2 1,304 66.0 3.4 1.4 1956 (3112). Segi et al., 166 2,124 oe Quantities 1957 (250). smoked stated as averages only. Differences are statistically significant. Mills and 484 8.4 26.0 1,588 27.6 5.3 42 94 83.0 4.3 1,722 73.3 0.5 0.6 Percent “heavy” Porter, smokers under- 1957 stated. Only (187), 506¢ survey yesponse among female cases. Stocks, 2,101 1.9 28.2 5,960 8.7 22.3 49 255 57.6 17.2 3,402 68.6 10.7 1.6 1957 (262). Schwartz and 602 1.0 58.2 1,204 9.5 36.2 10.4 (4) (4) () (4) (@) (4) Denoix, 1957 (247). Haenszel and (*) @) (*) (‘) (4) (*) 158 51.9 14.6 339 69.6 8.2 2.5 Shimkin, 1958 (113). zee TABLE A4.—Group characteristics in retrospective studies on lung cancer and tobacco use (cont.) SM = Smokers. NS = Nonsmokers. Males Females Author, Cases Controls Cases Controls year, oa Relative Relative Comments reference Pereent Percent Percent Percent risk Percent Percent Percent Percent risk Num- non- heavy non- heavy ratio non- heavy non- heavy ratio ber smokers smokers' Number smokers smokers! SM:NS? Number smokers smokers! Number smokers smokers! SM:NS? Lombard and 500 1.6 sae 4,238 11.0 tee U9 (4) (4) (@) {4) @) (4) ... Authors’ Snegireff, calculations for 1959 (176). heavy smoking based on lifetime number of packs of cigarettes. Pernu, 1,477 6.6 34.5 713 37.2 20.8 8.4 129 $5.3 26.4 1,060 91.6 ; 0.7 1.9 Quantities 1960 (211). given only in grams per day. Haenszel 2,191 3.4 41.9 (*) 16.2 12.0 5.2 (4) (@) {*) (4) (4) (*4) .«» Population et al., sample of 1962 (1122). 31,516 used as base. Not a case- control study. Lancaster, 238 2.5 86.1 476 20.1 71.2 9.8 (4) @) () (*) «) ) 1962 (158). Haenszel and @) (4) (4) (*) (4) () see 749 60.9 11.5 (4) 67.3 2.5 1.3 Population Taeuber, sample of 1964 (115). 34,339 used as base. Not a case-control! study. a @ w TABLE A4.—Group characteristies in retrospective stuslies on lung cancer and tobacco use (cont.) SM = Smokers. NS = Nonsmokers. Males Females Author, Cases Controls Cases Controls year, Relative Relative Comments reference Percent Percent Percent Percent risk Percent Percent Percent Percent risk um- non- heavy non- heavy ratio non- heavy non- heavy ratio ber smokers smokers! Number smokers smokerst SM:NS? Number smokers smokers! Number smokers smokerst SM:NS? Wicken, 803 4.0 40.0 803 14.0 22.0 3.9 151 58.0 29.0 151 80.0 17.0 2.9 Heavy smokers— 1966 (308). greater than 23 a day. Gelfand et al., 32 6.3 ves 32 63.0 ee 525.3 () (4) (4) () () (4) 1968 (98). Hitosugi, 124 5.6 67.8 1,889 13.2 55.0 2.6 61 54.1 6.6 2,352 80.5 2.9 2.3 Air pollution 1968 (125). found to have no effect on lung cancer rates of non- smokers. Heavy smokers—great- er than 15 a day. Bradshaw and 45 0.0 wae 341 31.7 a tae (4) (4) @) (4) Q) (4) Schonland, 1969 (41). Ormos et al., 94 15 58.5 1,811 42.9 38.9 9.3 24 95.8 0.0 1,278 81.7 9.7 0.2 Heavy smokers— 1969 (204). greater than 15 a day. Wynderetal., 210 1.4 67.5 420 21.0 40.9 320.8 30 16.7 44.0 182 57.6 23.3 6.78 Heavy 1970 (3384). smokers~— greater than 20a day. 1 For this table, heavy smokers are defined as those smoking 20 or more 3 Based upon fewer than 5 case nonsmokers. cigarettes per day, unless otherwise stated. * Does not apply. = Computed according to method of Cornfield, J. (61). TABLE A7.—Grouping of pulmonary carcinomas Group I: A. Epidermoid carcinoma. B. Small cell anaplastic carcinoma (‘‘oat-cell”’ earcinoma). Group H: A. Adenocarcinoma. B. Bronchiolo-alveolar cell carcinoma. C. Carcinoid tumor. D. Mucous gland tumor. Extra (not included in I and II): A. Large cell undifferentiated carcinoma. B. Combined epidermoid und adenocarcinoma. Unsuitable for diagnosis. Source: Kreyberg, L. (1532). 334 see TABLE Al2.—Autopsy studies concerning the presence of radioactivity in the lungs of smokers NS = Nonsmokers. SM = Smokers. Author, year, Number country, of Results Comments reference cases Little et al., Po!” levels in various tisanes (pe/g tissue) Vertebral bodies, renal 1964, Peribronchia! Bronchial cortex, spleen, and UNS.A. (178). lymph nodeg Lung (average) epithelium urinary bladder showed NS wool. 5 0.011 0.001-2 negligible no differences. SM oo... eae 12 0.011 0.008 0.028-1.25 Hill, 1965, Mean Po? levels in various tissues (pe/kg tissue) The authors found no ULS.A. (128). Bronchial tree Alveolae Total lung Liver Kidney excessive concentrations NS wo. eee e eee eee 6 3.1 3.4 3.2 14.8 15.0 at bronchial bifurca- SM ow. eee. 4 7.3 9.9 2.6 20.0 20.5 tions. Little et al., Po’ levels in various epithelial tissue regions of lung (ne/g)t The authors noted con- 1965, Site: siderable interpersonal UVS.A. (174). NS 2... eee eee 8 Mainstem bronchus ..........000 0.00 .. ccc eeeece cece cess <0.2- 1.7 variation but did find a SM... 28 Lobar bronchus . 2.6.0... cece cee ee ss etee cece <0.2~ 1.0 trend relationship be- Basal segmental bronchus ...........0.000.. cece cece cues <0.2- 2.6 tween increased daily Upper segmental bifurcation ............... see ee eee eces LOS 7.8 consumption and in- Lower segmental bifurcation .......... 0.00.0... ccc e eee <0.5-13.9 creased Po”? levels in lung parenchyma. No such relationship was noted for age of indi- vidual at death or for total pack-years. tSmokers only, 9€e TABLE Al2.—Autopsy studies concerning the presence of radioactivity in the lungs of smokers (cont.) NS = Nonsmokers. SM = Smokers. Author, year, Number country, of Results Comments reference cases Ferri and Mean Po” levels in various tissues (pe/g wet tissue) Baratta, Lung Liver Kidney 1966, NS oe. eee 10 0.031 0.1038 0.080 U.S.A. (95). SM oo... cee ee aes 14 0.065 0.125 0.070 Rajewsky and Mean Po* levels in various tissues (pe/g) t Data not given. Smokers Stahlhofen, Lung parenchyma Bronchial tree Bronchial bifurcation were considered those 1966, NS o.. eee eee t 0.0025 0.0020 0.0012 using more than 1 pack Germany (217). SM . 12 0.0078 0.0077 0.0047 a day. The authors noted that their figures were con- siderably smaller than those of Little et al. (178, 174) and also disagreed with their data on bifurcation. Little and Mean Po*'? levels in various epithelial tissues (pc/g wet tissue) Radford, SM . 25 Bronchial wall and submucosa 1967, Bronchial] epithelium: U.S.A. (172). Pipe weve Z Trachea... 6. cee cee ce eee tenet e ete te eeetnneens Ex-cigarette ...... 1 Lobar bronchi ........0 00... cee ee ee eee Never ..........05 8 Segmental bifurcation ...........02.. 0000008 Lee TABLE A13,—Experiments concerning the e fects of the skin painting or subcutaneous injection of cigarette smoke condensate or its constituents upon animals Author, A. Method, year, Animal B. Frequency and/ country, and or duration, Results Comments reference strain C. Material Wynder CAF, mice A. Painting shaved skin. Pereent animals with: t Number in paren- et al., B. 3/week for 2 years. Treatment: Papillomast Cancert thesis represents 1953, C. Whole cigarette smoke “Tar” alone wees 59.0(81) 44.0(81) total in that U.S.A. condensate in acetone. “Tar’’ and croton oil 42.0(31) 9.7(31) experimental group. (317). Croton oil once/week. Acetone alone ............ 00000000. (30) (30) Skin-painting Acetone and croton oil (14) (14) experiments prior to 1953 are fully detailed in tab- ular form in this article. Passey 5 different A. Painting unshaven No malignant tumore noted in either group. etal., mouse skin. Papilloma noted on one animal (in whole ‘tar’ group) which later regressed. 1955, strains B. 2/week for 9 England (101). months. (209). C. Whole “tar” or neutral fraction. Orr et al., Mice of 2 A. Painting skin. Number animals with: 1955, strains. B. 1 or 2/week for Treatment: Papillomas England 18 months. Benzpyrene 1/week followed 4/30 at 18 months (separate group received only (205). C. 20 percent cigarette by ‘‘tar’’ 2/week. benzpyrene and showed no tumors). “tar” in acetone, “Tar” alone ............00. 0/50 at 18 months. 0.3 percent benzpyrene. see TaBLeE A18.—Experiments concerning the effects of the skin painting or subcutaneous injection of cigarette smoke condensate or its constituents upon animals (cont.) Author, Animal A. Method, year, and B. Frequency and/ country, strain or duration, Results Comments reference C. Material Wynder Mice of 4 A. Painting shaved skin. Strain Papillomas Carcinomas No tumors noted with etal., separate B. 3/week for 80 days. CBTIBL 2... ee cee cee eee eens 10/89 2/89 acetone alone. 1955, strains. C. Whole condensate Swiss .... 00... ccc ce eee cee cee tetera 22/86 12/86 Stresses U.S.A. in acetone. differences in (318). susceptibility of strain. Hamer and Outbred A. Painting unshaved Treatment: Papillomaa Woodhouse, albino skin. “Tar” 2/week ..... 0.0. ee ee 1/50 1956, strain B. Varied for 18 months. “Tar” and croton oil 1/week. ........ 2/30 U.S.A. mice. C. Whole “‘tar’’/acetone, B(a)P 3 times then “tar” 2/week ..... 4/30 (116). benzpyrene [B(a)P], Bla)P 3 times ........ 00. cee eee 0/30 croton oil. Sugiura, Rockland A. Painting unshaved Papillomas Carcinomas 1956, Swiss skin. 16/44 12/44 (only 44/60 U.S.A. albino B. 3/week for 2 years. lived from (266). mice (60). C. Whole “‘tar’’. 365-696 days). Graham Albino New A. Painting shaved skin. Treatment: Papillomas Carcinomas The authors review et al., Zealand B. 3/week for 6 years. Condensate ..........-.. 0.0002 ee eee 41/41 5/41 previous experiments 1957, rabbits. C. Whole condensate. Condensate and croton oil 1/week. 10/10 2/10 with rabbits in U.S.A. Croton oi] and acetone 1/week. 0/3 0/3 tabular form. (101). Acetone 1/week 2.0... ..... 000. e cee 0/7 0/7 Guerin and Mice A. Painting neck skin. Original number Survivors Papillomas Sarcomas ft Control group. Cuzin, (Pasteur 3B. 2/week for >1 year. TCO.112 6 eee eee eee 51 0/51 0/51 t+ Experimental group. 1957, strain.) C. Whole condensate. FE. 672 ool eee eee 220 10/220 5/220 U.S.A 6ee& TABLE A13.—Experiments concerning the effects of the skin painting or subcutaneous injection of cigarette smoke condensate or its constituents upon animals (cont.) Author, Animal A. Method, year, and B. Frequency and/ country, strain or duration, Results Comments reference Cc. Material Wynder Swiss mice A. Painting skin. Percent Percent etal., B. Varied for 12 Treatment: Number papillomas carcinomas 1957, months. B/hweek oo... cece eee eee eee 50 12.0 8.0 U.S.A. C. Whole condensate B/week so. cee cece eee 50 38.0 16.0 (828). in acetone. Q/week oo. cc cece ete eee 40 10.0 3.0 U/week 26... eee eee 40 6.0 Wynder and CAF, or A. Painting shaved skin. Percent Percent Swiss mice noted Wright, Swiss B. 8/week for lifespan. Treatment CAF: Number papillomas carcinomas to be more sus- 1957, mice. C. Whole ‘‘tar” or nicotine Whole ‘tar’ ........ 06... eee 30 53.0 27.0 ceptible. U.S.A. free ‘‘tar”’ derived Nicotine free “tar” .......... 40 73.0 25.0 Majority of carcino- ($28). from pipe and Cigarette “tar” ...........00- 30 30.0 30.0 gens noted to be cigarette tobacco. Pipe “tar? 0.0... cece eee 30 60.0 20.0 in neutral fraction Treatment Swiss: of condensate. Whole ‘tar’ .........-....00. 30 53.0 10.0 Nicotine free “tar” ........... 40 43.0 20.0 Cigarette “tar” .............. 30 63.0 33.0 Pipe “tar”? oo... cece eee eee 30 63.0 50.0 Gellhorn, Paris RII A. Painting shaved skin. Treatment: Papillomas Carcinomas 1958, mice B. Varied for 1-2 years. Benzpyrene (twice only) ............ 20/529 5/529 U.S.A. C. ‘Tar’ in acetone, Croton oi] (5/6 week) ..............4. 4/26 0/26 (99). pbenzpyrene, “Tar”? (5/6 week) .......... 0.20 cee 3/659 2/559 eroton oil. Acetone (5/6 week) ..............-. 0/30 0/30 “Tar” and croton oil (5/6 week) 10/175 0/175 Bock and Swiss A. Painting skin. Percent Moore, female B. 5/week for lifespan. Group: Number living at 6 months Skin tumors at 64 weeks 1959, mice C. Whole condensate Painted wo... ccc cece cece eee 49 13.0 U.S.A. irradiation. Painted and irradiated ......... 65 44.0 (28). Irradiated ........ 00 ce cee eee 36 Ove subcutaneous injection of cigarette smoke condensate or its constituents upon animals (cont.) TABLE A13.—-Hxperiments concerning the effects of the skin painting or Author, Animal A. Method, year, and B. Frequency and/ country, strain or duration, Results Comments reference C. Material Druckrey, Rats A. Subcutaneous Group: Sarcomas ¢ Control group. 1961, injection, tc. 1/75 t Experimental group. Germany B. 1/week for 60 weeks. TE lo eee ee ee 15/75 (78), €. Smoke condensate in tricaprylin and alcohol. Bock et al., ICR Swiss A. Painting shaved skin. Surviving Pereent Percent Skin 1962, mice B. 10/week for 1 year. Treatment: at £8 weeks Skin cancer neoplasia U.S.A, C. Cigarette “tar”. Standard cigarette 24/30 25.0 54.0 (31). Standard cigarette 21/30 5.0 57.0 Standard cigarette 18/30 33.0 44.0 Standard cigarette 13/30 23.0 62.0 Filter cigarette ..... 30/30 7.0 27.0 Filter cigarette 30/30 3.0 23.0 Acetone only 66/66 Control ...........202 0000200 65/65 Roe, Albino mice A. Painting shaved skin. Treatment: Survivors Percent skin tumors Author concluded 1962, B. 3/week for 84 weeks. “Tar” and 0.025 mg. B(a)P ..... 26 12.0 that cigarette U.S.A. C. Whole smoke ‘‘tar” “Tar” and 0.06 mg. B(a)P ...... 15 27.0 smoke contains (225). with added B(a)P “Tar” and 0.25 mg. B(a)P...... 15 13.0 cocarcinogens. in acetone. “Tar” and 1.25mg.Bla)P ...... 14 64.0 B(a)P 1.25 mg. ........... 0.00.0. 14 Druckrey and Rats A. Subcutaneous Treatment (BP mg./weck): Sarcomas Schildbach, injection. 30 .. 25/30 1963, BR. 1/week for 700 days. 10... 14/40 Germany C. Benzpyrene in 3... &/50 (82). tricaprylin. — (solv 2/75 TABLE A13.—Eaperiments concerning the effects of the skin painting or subcutaneous injection of cigarette smoke condensate or its constituents upon animals (cont.) Author, A. Method, year, Animal B. Frequency and/ country, and or duration, Results Comments reference strain C. Material Homburger CAF, mice A. Painting shaved skin. Com plei¢ Percent Percent etal. B. 2-3/week for 2 years. Condensate: autopsics Papillomas Carcinomas 1963, C. Various tobaeco Pipe tobacco ................, 77 35.0 15.0 U.S.A. condensates in Cigar tobacco ................ 84 27.5 15.0 (181). acetone. Cigarette tobacco ............. 82 27.0 15.0 Benzpyrene .................. 54 10.0 20.0 Acetone only ................. 62 Bock et al., Swiss ICR A. Painting clipped skin. Percent Percent 1965, mice B. 10/week for 11 weeks. Percent concentration of tar surviving Percent cancer and U.S.A, C. Various smoke (type cigarette) : tl weeks cancer papilloma (29), condensates in 9.2 (standard) .............., 96.0 30.0 67.0 acetone, 8.3 (standard) -. 93.0 27.0 67.0 7.9 (English standard) 90.0 24.0 58.0 8.7 (king) .........,.., tee 100.0 28.0 69.0 4.0 (filter) 22... 98.0 9.0 36.0 4.4 (filter) 22... 100.0 10.0 41.0 2.5 (filter) 22... el, 97.0 4.0 16.0 Acetone control .............. 94.0 Untreated control ..........., 100.0 Cumulative number of mice with + 7,12-dimethyl- Lve Van Duuren Swiss ICR/ A. Painting shaved skin. etal, Ha mice B. Initiating agent once— Initiator Promoter Papillomaa Carcinomas benz (a) anthracene. 1966, Promoter 3/week for DMBA ...Ether tobacco leaf extract 4/20 0/20 ULS.A. 12-14 months. (0 Ether tobacco leaf extract . 0/20 0/20 (296). C. DMBAT, tobacco DMBA .. Choloroform tubacco leaf extract ...... 1/20 0/20 extracts ciga- OL... Choloroform tobacco leaf extract 2.2... 0/20 0/20 rette “tar”. DMBA ...Cigarette “tar” 11/20 4/20 O ....... Cigarette “tar” 0/20 0/20 O ....... Acetone 0/20 0/20 ove TABLE A13.—Experiments concerning the effects of the skin painting or subcutancous injection of cigarette smoke condensate or its constituents upon animals (cont.) Author, A. Method year, Animal B. Frequency and/ country, and or duration, Results Comments reference strain C. Material Munozetal., Swiss ICR/ A. Painting shaved skin. Dark tobacco “tar” At risk Tumors Carcinomas The authors noted 1968, 4a mice B. Varied. 4.0 percent ...........0.0 00005 81 50 17 a shortened latent U.S.A. C. ‘'Tar” from dark 8.0 percent .........-......--- 71 46 16 period for dark and (Colombian) and Light tobacco tar: tebacce. Colombia light (U.S.A.) 4.0 percent ..............0-005 95 26 6 (197). tobaccos. 8.0 percent oe 938 54 20 Acetone . 0... ieee cece eens 91 0 0 Davies and Albino A. Painting shaved skin. Percent of carcinoma-bearing animals at 116 weeka The authors concluded Day, mice B. Varied regimen. Treatment: (actual number of animals in parentheses) that the lack of 1969, C. Cigarette and 300 mg. 150 mg. 75mg. 37.5 mg. difference in re- Great cigar condensate. Standard cigarette ........ 20.1(29) 13.2 (19) 0.7 (1) . sults from the first Britain Cigar oo... cee eee oe 27.1(39) 11.1(16) 2.1(3) and third groups (65). Cigar tobacco cigarette 18.9(10) . .. under treatment suggests that the increased tumori- genicity of cigar tobacco is due to physical processing factors. eve TABLE A14.—Experiments concerning the effect of cigarette smoke or its constituents on tissue and organ cultures Author, year, country, reference Bouchard and May, 1960, France (85). Awa etal., 1961, Japan (16). Thayer and Kensler, 1964, U.S.A. (275). Berwald and Sachs, 1965, Israel (20). Tissue or organ culture Mouse lung. Human fetal lung. KB mammalian tumor cells. SWR mice and golden hamster embryos. Results Material/delivery Increased number of mitotic abnormalities in the treated cultures; particularly in Tobacco smoke condensate perfusion for 24 hours the first 5-10 days after grafting. and subsequent grafting under renal capsule of mice. Paper smoke induced the most severe changes, consisting of cytoplasmic vacu- Direct exposure to smoke from: olization and nuclear pyknosis. Also noted were a decrease in the mitotic index a. Whole cigarettes. and an increase in abnormal divisions, more so with paper. smoke than with b. Tobacco alone. ec. Paper alone. the other two. Significant growth inhibition was shown in unfiltered smoke. Cytotoxic compo- Cigarette smoke condensate applica- nents were noted in both the gas and particulate phases. tion; filtered and unfiltered cigarettes. Benzo(s) pyrene caused increased cell transformation as manifested by: Direct application of benzo(a)pyrene a. Hereditary random growth pattern. b. Progressive growth as tumors after subcutaneous injection into adults. (B(a)P]. ec. Ability to grow continuously in culture. Application of Treated cultures revealed cellular metaplasia, basal cell hyperplasia, increased mitotic rate, and increased H*-thymidine incorporation proportional to the con- Crocker et al., Suckling rat trachea in organ culture. B(a) P in acetone. centration of material and duration of application. Inhibition of cell growth. 1965, ULS.A. (63). Diamond, 1965, U.S.A. (68). Various con- tinuous cell strains (mammalian). Application of B(a) Pin either dimethylsulfoxide (DMSO) or paraffin. yre TABLE Al4.—E'xperiments concerning the effect of cigarette smoke or its constituents on tissue and organ cultures (cont.) Author, year, country, reference Tissue or organ culture Material/delivery Results Borenfreund et al, 1966, U.S.A. (38). Guimard, 1966, France (110). Lasnitzki, 1068, England (160). Hamster Jung tissue. Chicken embryo muscular explants. Mice neonatal trachea. Application of B(a)P in either DMSO or dimethyl- formamide. Application of tobacco extract. Application of a hy- drocarbun-enriched fraction of whole smoke condensate. a. Increased appearance of new small chromosomes and telocentric chromosomes. b. Increased ability to grow in hamster cheek pouch and there become spindle- cell sarcomas. Increased mitotic activity and increased incidence of anomalous mitoses. . Increased basal cell hyperplasia and pleomorphism of newly formed cells. . Increased epithelial mitosis. on Lasnitzki, Human fetal lung Application of a hy- a. Cellular enlargement and promotion of growth of new bronchi. 1968, in organ culture. drocarbon-enriched b. Increased mitoses, bronchial epithelial hyperplasia, and squamous metaplasia. England (161). fraction of whole c. Inhibition of stromal growth. smuke condensate. Chan et al., Mouse lung Application of a. Cellular disorganization. 1969, bud embryonic B(a)P in DMSO. b. Cellular pyknosis; nuclear shape and size irregularities. U.S.A. (54). cultures. c. Increased epithelial mitotic rate and decreased mesenchymal mitotic rate in those cultures exposed to B(a)P versus those exposed to pyrene or DMSO. Leuchtenberger Mouse lung Exposure to fresh smoke: and and kidney a. Unfiltered. a. Decreased RNA production, pyknosis, and death of cells. Leuchtenberger, tissue and b. Activated b. Similar results, but changes were of minimal severity. 1969, organ cultures. charcoal filter. Switzerland (165), ec. Cigarette or cigar tobacco, c. Similar effects as group a., but less severe. SVE TABLE Al4.—Experiments concerning the effect of cigarette smoke or its constituents on tissue and Author, year, Tissue or country, organ culture Material/delivery Results reference Crocker, Various organ Application of Squamous metaplasia; frequent pleomorphic cells; 1970, cultures: B(a)P in serum. (inhibited by Vitamin A). U.S.A. (62). a. Whole suck- ling hamster tracheas. b. Whole bron- chia] tubes from late fetal dogs and monkeys. organ cultures (cont.) dedifferentiation of epithelium OPE TaBLe Al5.—Eaxperiments concerning the effect of the instillation or implantation of cigarette smoke or its constituents into the tracheobronchial tree of animals Author, A. Method year, Animal B. Frequency and/ country, and or duration Results reference strain ©. Material Blacklock, CR white A. Injection into 3,4-benzpyrene: Number with tumors/number exposed 1957, rats. lung parenchyma a. 3 mg. in olive oil 5/6 sarcoma. Great by thoracotomy. b. 3 mg. in olive oil with dead Tb bacilli 2/4 sarcoma, 4/8 squamous cell carcinoma. Britain B. Once. c. 5.75 mg. in cholesterol pellet 1/8 squamous cell carcinoma. 124). C. 3,4-benzpyrene Cigarette “‘tar’’: in olive oil, a. In olive oil 0/10. with dead Tb b. In olive oil with dead Tb bacilli 1/8 sarcoma, 1/8 squamous cell carcinoma. bacilli or in Controls: cholesterol, a. 0.15 cc. olive oil 0/4. cigarette “tar’’. b. 0.15 ce. olive oil with dead Tb bacilli 0/4 ec. Cholesterol pellets 0/4. Della Porta Syrian golden A. Direct tracheal Number of etal., hamsters. instilllation. hamsters with 1958, B. Weekly up to Survivors at 20 tracheobronchial U.S.A. 45 weeks. weeks/original carcinomas (67). C. 1 percent 7,12-dime- Material: Weeks number exposed at death thylbenz(a)anthra- a. DMBA 50 pe-/ week 45 10/20 2 cene (DMBA), b. “Tar” 200 pe-/week oe 32 11/21 — cigarette “‘tar’’ c. DMBA 50 pe./week wena 12 9/20 — concentrate. then “‘tar’’ 200. pe-/ week ..........4- 30 — — d. DMBA 100 ,2./week .... 17 7/20 4 e. DMBA 100 pe./week } and “tar" 500 b 20 9/20 3 pe./week ves fesse sseee ef} Rigdon, White Pekin A. Intratracheal No neoplastic changes noted in either the experimental or control groups. 1960, ducks. injection. U.S.A. Controls: 99 B. Daily for 721 days. (221). Experimental C. Tobaeco condensate group: 52 in liquid petrolatum. Lve TABLE Al5.— FE xperiments concerning the effect of the instillation or implantation of cigarette smoke or its constituents into the tracheobronchial tree of animals (cont.) Author, A. Method year, Animal B. Frequency and/ country, and or duration Results reference strain C. Material Blacklock, CB white rats. A. Inoculation at Number Percent with 1961, thoracotomy. of rate malignant tumors Great B. Once and sacrificed Controls oo... 0. eee eee ee cence ace ecuee 275 1.5 (1 carcinoma, 3 sarcomas). Britain at 1 week-2 years. Cigarette condensate . 72 11.1 (6 carcinomas, 2 sarcomas). (25). C. Cigarette tobacco Bucerin alone 0.2... eee ee cece 44 2.3 (1 sarcoma). smoke condensate in eucerin. Herroldand = Syrian golden A. Intratracheal Number of Number with Number of tracheo- Dunham, hamsters. inoculation. Material: hamsters tumors bronchial tumors 1962, B. 0.5 cc./week for B(a) in Tween60 ... 6 3 5 (3 papillomas, 2 carcinomas). U.S.A. 5/6 months. B(a)P in Tween60! 6 3 9 (4 papillomas, 5 carcinomas). (122). C. Benzo(a) pyrene Tween60 6 0 _— in Tween60 B(a)P in olive oil 6 0 — or olive oil, Olive oil ........... 6 0 —- Rockey et al., Dogs. A. Bronchial inoculation Squamous 1962, or stimulation, Pre- metaplasia U.S.A. B. 3-5 times/week Number Invasive Carcinoma- cancerous with atypical inflam- (224). for up to 5 Procedure: of dogs carcinoma in sity changes changea mation years. Controls ........ 27 — — 6 24 C. Cigarette smoke Manipulation of condensate. bronchus ..... 25 — — _ 9G 25 Smoke condensate 130 1 3 25 98 128 Tipton and Mongrel dogs. A. Bronchial Rapid induction of squamous metaplasia in condensate-exposed animals. No tabular Crocker, Control group and inoculation. data is presented. 1964, experimental B. Daily for 8 days. U.S.A. group—19. C. Cigarette smokc (277). condensate. TABLE A15.—Experiments concerning the effect of the instillation or implantation of a & cigarette smoke or its constituents into the tracheobronchial tree of animals (cont.) Author, A. Method year, Animal B. Frequency and/ country, and or duration Results reference strain C. Material Saffiotti et al, Syrian golden A. Intratracheal Percent tumor- 1966, hamsters. inoculation. Number of bearing of Total Total number U.S.A. B. Weekly for 15 weeks. Number autopsied: tumor-bearing survivors at number of respiratory (287). C. B(a)P (3 meg.) animals 15 weeks of tumors tract cancers attached to fine Male ............... 15 100.0 24 18 hematite dust. Female 11 100.0 17 16 Kuschner, Hamsters. A. Wire mesh pellet Number of Number of 1968, implantation survivors/original animals with ULS.A. into bronchus. Implant: number in group lung cancer (157). B. Lifetime. Wire mesh only 34/35 — C. Bla)P, MCA ............. 88/91 43 methyleholan- Bay P cece ec ce teen eee ee 89/91 57 threne (MCA). Saffiotti et al., Syrian golden A. Intratracheal Number of 1968, hamsters. inoculation. hamsters with U.S.A. B. Weekly for 15 Number autopsied respiratory (235), weeks. Tnoculate: tract tumora C. B(a)P attached Control Phebe este te a eres esse eeeuee 176 —_ to a fine B(a)Pin hematite 2.0.0.0... 0c cece eee eee ae 55 35 hematite dust. Hematite only 0.0... . ce eee 41 —_ Borisyuk, Wistar rats. A. Intratracheal Number final/ Duration of 1969, intubation. initial inoculation Russia B. Monthly up to Inoculate: (months) (84). 10 months. Controls 2... ee cect eee eens 11/20 12 C. Cigarette ‘tar’. Unfractionated ‘‘tar’’ 24/200 10 Denicotinized ‘‘tar’ ..... 9/45 8 (1/9 metaplasia) Neutral ‘‘tar’’ fraction 14/100 8 (2/14 carcinomas, This group also reccived one injection of urethane intraperitonealls 1/14 papillary adenoma). 6re TABLE Al6.--E'xperiments concerning the effect of the inhalation of cigarette smoke or its constituents upon the respiratory tract of animals (Figures in parentheses represent total number survivors in specific group) Author, year, Animal A. Type of exposure country, and B. Duration Results Comments reference strain C. Material Lorenz et al., Strain A mice: A. Chamber. E. No increase in tumor formation over that noted in controls. This strain of mice does 1943, TC. 97. B. Up to 693 hours. have a hereditary U.S.A. (177). TE. OT. C. Cigarette smoke. tendency to tumor formation. tC. Control. 0.01 level, Miihlbock, Hybrid (020 x A. Chamber. Percent with alveolar earcinomas No other type of 1955, DBA) mice: B. 2 hours per day for C. 31.0 lung tumors were Netherlands C. 32. up to 684 days. E. 79.0 found. (195). E, 29. C. Cigarette smoke. Leuchtenberger CF, albino mice: A. Chamber. 23 of the experimental mice showed: et al., C. and E. 275. B. To & cigarettes 15 basal cell hyperplasia. 1958, per day from 14 atypical basal cell hyperplasia. U.S.A. (166). 11-201 days. 7 dysplasia. C. Cigarette smoke. 2 squamous cel] metaplasia. Guerin, IC and Wistar A. Chamber. Percentage of rats with pulmonary tumors 1949, strain rats. B. 45 minutes C. 2.4 percent of 39 survivors. France (108). Cc. 40. per day from E. 5.1 percent of 68 survivors. E. 100. 2-6 months. C. Cigarette smoke. Ose TABLE Al6.--Experiments concerning the effect of the inhalation of cigarette smoke or its constituents upon the respiratory tract of animals (cont.) (Figures in parentheses represent total number survivors in specific group) Author, year, Animal A. Type of exposure country, and B. Duration Results Comments reference strain Cc. Material Leuchtenberger Female CF, mice: A. Chamber. Number with et. al., C. 243. B. 14-6 cigarettes severe bronchitis; 1960, E. 360. per day for 1 Exposure peribronchitis; U.S.A. (167). month to 2 years. Number Number of length atypical epithe- C. Cigarette smoke. of mice cigarettes (months) lial proliferation 151 we. 25-1,526 1-23 30 150... 0 0 2 36 ...... 100- 200 1-3 7 36 wo... 250- 500 4-8 7 34 ...... 600-1,600 9-23 8 51 100- 400 3-6 4 63 100- 400 3- 6 17 Leuchtenberger Female CF, mice: A. Chamber. Number Percent of mice Presence of tumors et al., Cc. 166. B. 4-8 cigarettes of mice Exposure with pulmonary showed an age- 1960, E. 281. per day for examined (days) adenomatous tumors relationship U.S.A. (168). 17-600 days. BL cece ee ee eee 0 56 independent of C. Cigarette smoke. Ch 17- 99 41 smoking exposure. 85 eee ee ee eee © 100-199 37 Bl wee eee ecco aes 200-600 66 Otto, 1963, Albino mice. A. Chamber. Number Exposure Number with Germany Cc. 60. B. Approximately of mice None. lung tumore (206). E. 189. 12 cigarettes per examined Varying 3 pulmonary adenomas. day for varying Cc. 60 up to 24 21 pulmonary adenomas. intervals. E. 189 months. 2 epithelial carcinomas. C. Cigarette smoke.