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Dose-dependent genotoxic effect of pan masala and areca nut in the Salmonella typhimurium assay
Fd Chem. Toxic. Vol. 31, No. 6, pp. 439442, 1993
0278-6915/93 $6.00 + 0.00 Copyright © 1993 Pergamon Press Ltd
Printed in Great Britain. All rights reserved
D O S E - D E P E N D E N T G E N O T O X I C E F F E C T OF P A N M A S A L A A N D A R E C A N U T IN THE SALMONELLA TYPHIMURIUM ASSAY K. POLASA*, S. BABU and I. S. SHENOLIKAR National Institute of Nutrition, Jamai-Osmania, Hyderabad 500 007 (A.P.), India
(Accepted 4 February 1993) Abstract--Aqueous extracts of different brands of pan masala and scented supari were tested for mutagenicity by the Salmonella typhimurium assay using tester strains TA98 and TA100. These extracts were found to be mutagenic to both tester strains. The mutagenic effects of pan masala and scented supari extracts were similar to that produced by areca nut extract. The addition of 500 ppm saccharin to the supari extracts did not alter the mutagenic response.
pan masala. 'Supari' is a form of processed areca nut with or without flavourings and spices. Some brands of 'supari' that are commercially available were also assessed for mutagenicity.
INTRODUCTION
The habit of chewing betel quid is very common in India today. A moderate estimate of the number of betel quid chewers in India is around 20 million. In fact, it outnumbers smokers. Epidemiological studies have associated the chewing of betel quid with tobacco with an increased risk of carcinoma of the oral cavity (IARC, 1985). Several reports also indicate other adverse effects of tobacco chewing, including increase in pulse rate and blood pressure, increased frequency of bronchitis, effects on reproduction and prenatal toxicity in women, and the occurrence of preneoplastic changes in the buccal cavity (leukoplakia) (IARC, 1985; Jussawalla and Deshpande, 1971; Shirname et al., 1983). Besides tobacco, another major ingredient of betel quid is the areca nut, which contains alkaloids such as arecoline and arecaidine. These substances have been shown to be mutagenic to bacteria (Shirname et al., 1983) and mammalian cells in culture (Shirname et al., 1984). Catechu, another ingredient of betel quid, has been shown to induce sister chromatid exchanges (SCE) and dominant lethality in male mice (Giri et al., 1987; Mukherjee and Giri, 1991). Several ingredients are used in the preparation of betel quid and its preparation is time consuming. Therefore, using traditional Indian technology, the industry has developed a product ready for consumption packed in small convenient sachets. This product is 'pan masala'. It has almost all of the ingredients that are present in betel quid except the betel leaf. There is one report in the literature demonstrating that in vitro the aqueous extract of pan masala produces SCE and chromosomal aberrations in Chinese hamster ovary cells (Adhvaryu et al., 1989). We report here the mutagenicity of some varieties of *To whom correspondence should be addressed.
MATERIALS AND METHODS
Eight varieties of pan masala and six varieties of supari were purchased from the local market. 100 g of the sample was powdered and mixed with 200 ml glass-distilled water on a mechanical shaker for 1 hr. The solution was filtered through Whatman No. 1 filter paper under vacuum and lyophilized. About 2.2-5.3g dry matter was obtained from a 100-g sample. The extract was stored at - 2 0 ° C until analysis. For the mutagenicity studies, extracts of pan masala were tested at 100, 200 and 300 #g/plate, and supari extract was tested at 200 pg/plate. An extract of areca nut without added flavourings or spices was also tested at 100, 200 and 300 pg/plate. Mutagenicity testing. All the chemicals for the assay were obtained from Sigma Chemical Company (St Louis, MO, USA). Salmonella typhimurium TA98 and TA100 were kindly provided by Dr B. N. Ames (Berkeley, CA, USA). S-9 was prepared from the livers of Aroclorinduced rats according to the procedure of Maron and Ames (1983). To the test substance, 0.1 ml of the bacterial culture, 0.5 ml S-9 mixture and 2.5 ml top agar were added and plated on Vogel Bonner minimum medium plates. For testing the samples without metabolic activation the same procedure was followed except that the S-9 mixture was omitted from the reaction mixture. These plates were incubated at 37°C for 48 hr in the dark and the numbers of revertant colonies were counted. Benzo[a]pyrene (5/~g/plate) and sodium azide (1/~g/plate) were used as the positive control for TA98 and TA100, respectively.
439
K. POLASA et al.
440
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(a) TA98 -$9
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TA100 -$9
500
400
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o
200
100
1
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6
V a r i e t i e s o f Pan M a s a l a
7
8
9
V a r i e t i e s of Pan Masala
Fig. 1. Dose-dependent mutagenicity of pan masala extracts in Salmonella typhimurium: (a) TA98 without S-9, (b) TA98 with S-9, (c) TAI00 without S-9 and (d) TA100 with S-9 all tested at 100 (~/), 200 ( O ) and 300 (11)/~g extract/plate. Values depicted are means of triplicate plate assays. Variety 9 represents areca nut extract. All of the values are significantly higher than the negative control values (horizontal line) given in Table I (Student's t-test; P < 0.01) except variety no. 4 tested at 100 # g extract/plate.
8
9
Mutagenicity of pan masala and areca nut extracts
441
Table I. Reversion frequency induced by supari extracts (200 #g/plate) in S. typhimurium TA 98 Sample of supari (areca nut) Sample 1 Sample 1 + 500 ppm saccharin Sample 2 Sample 3 Sample 4 Sample 4 + 500 ppm saccharin Sample 5 Sample 6 Negative control Positive controls: B[aIP (5 #g/plate) Sodium azide (1 #g/plate)
- S-9 43 _+ 15 49 £ 14 46 ± 11 46 ± 9* 48 ± 3*** 35 ± 4 39 ± 4* 38 ± 9 29 £ 3
TA 100 + S-9 66 ± 20 54 ± 13 61 ± 7** 63 ± 19" 64 ± 11 40 ± 3 54 ± 8* 45 ± 4* 35 ± 4
- S-9 206 ± 34* 198 ± 40* 244 ± 26*** 212 ± 28** 132 ± 17 210 ± 19"* 184 ± 11"* 198 ± 28* 132 ± 12
+ S-9 396 ± 30*** 492 ± 28*** 432 ± 52*** 409 ± 31"** 392 ± 42*** 400± 40*** 452 ± 45*** 392 ± 57*** 158 ± 12
2762 ± 220 626 ± 32 -426 ± 16 --BIa]P = benzo[a]pyrene Values are means + SD and those marked with asterisks differ significantly(Student's t-test) from the corresponding value for the negative control (*P < 0.05; **P < 0.01; ***P < 0.001). ---
RESULTS The m u t a g e n i c activities of the p a n masala a n d supari as q u a n t i t a t e d by reversion frequencies in S. t y p h i m u r i u m TA98 and TA100 are s h o w n in Fig. 1 a n d Table 1. The p a n masala extracts were tested at three c o n c e n t r a t i o n s (100, 200 a n d 300/~g/plate) whereas the areca nut (supari) extract was tested at a c o n c e n t r a t i o n of 200 #g/plate. Virtually all of the samples of p a n masala were mutagenic, with an e n h a n c e d effect seen with activation. A clear dosedependent mutagenic response was noted, particularly in T A I 0 0 with metabolic activation (Fig. ld). These results are similar to the mutagenic response shown by all the varieties of scented supari extracts (Table 1). The mutagenic effects of pan masala a n d scented supari extracts was similar to that produced by areca n u t extract (sample 9 in Fig. 1). Some of the supari extracts c o n t a i n e d saccharin but its presence did not seem to influence the mutagenic potential of the supari. DISCUSSION The m a j o r c o m p o n e n t of pan masala is the areca nut, which has at least six reduced pyridine alkaloids (Ashby et al., 1979). The m a j o r ones are arecoline, arecaidine, arecolidine, guvacoline a n d guvacine. Arecaidine a n d its methylester arecoline have been suggested to have mutagenic/carcinogenic properties since they are capable of reacting with cysteine in vivo and in vitro to produce cysteine/3-alkylation adducts (Boyland a n d Nery, 1969). The results of our mutagenicity studies of pan masala are significant because mutagenicity is highly correlated to carcinogen±city. A l t h o u g h there are no reports o f carcinogen±city induced by pan masala in the literature, one of the c o m p o n e n t s of pan masala, namely areca nut, has produced precancerous lesions 'leukoplakia' in Syrian hamsters (Suri et al., 1971). Likewise, the polyphenolic fraction of betel quid induced tumours in mice (Shivapurkar, 1980). A paste o f p a n masala applied to the oral cavity of albino rats resulted in increased keratosis, parakeratosis, inflam-
m a t o r y cell infiltration a n d vascularity indicating a mild leukoplakia (Khrime et al., 1991). C o m p o n e n t s of p a n masala, namely betel nuts, catechu and lime, have also been reported to produce this condition (Sirsat a n d K h a n o l k a r , 1962). The contents of betel quid have been clearly d e m o n s t r a t e d to produce a toxic effect o n cultured h u m a n buccal epithelial cells as well as o n the buccal m u c o s a of b a b o o n s a n d hamsters (Sundqvist et aL, 1989). There is also a positive correlation between oral cancer and precancerous lesions in ' p a n chewers' (Dayal et al., 1978). Besides the genotoxic effects, hepatotoxic effects Of p a n masala have been reported by S a r m a et al. (1992). A l t h o u g h we have studied the mutagenic potential of aqueous extracts of different samples of pan masala on the basis o f weight of extract (100, 200 and 300/~ g extract), these results have to be interpreted in terms of the content of arecoline and arecaidine in these samples. Moreover, plain supari a n d areca nut do not contain substances such as tobacco, catachu, flavourings and spices that are present in p a n masala. These additives m a y also lead to habitual chewing by virtue of e n h a n c i n g the taste of the product. This may also result in an increase in the prevalence of betel quid chewing. In view of the evidence available in the literature on the genotoxic potential of chewing substances such as betel quid (Nair et al., 1987; Stich et al., 1981), areca nut (Panigrahi a n d Rao, 1982), a n d tobacco, which is further supported by the dose-response effect reported here, caution needs to be exercised on the indiscriminate use of these products. Acknowledgements--The authors are grateful to Dr Vinodin± Reddy, Director, for her keen interest in the investigation. They also acknowledge the technical assistance of Mrs B. R. Annapurna and Mrs Amulya Rao. REFERENCES
Adhvaryu S. G., Dave B. J. and Trivedi A. H. (1989) An in vitro assessment of the genotoxic potential of pan masalas. Indian Journal o f Medical Research 90, 131-134. Ashby J., Styles J. A. and Boyland E. (1979) Betelnuts, arecaidine and oral cancer. Lancet i, 112.
442
K. POLASAet al.
Boyland E. and Neri R. (1969) Mercapturic acid formation during metabolism of arecoline and arecaidine in the rat. Biochemical Journal 113, 123-130. Dayal P. K., Mani N. J. and Bhargava K. (1978) Prevalence of oral cancer and precancerous lesions in "Pan" supari chewers. Indian Journal of Public Health 22, 234-245. Giri A. K., Banerjee T. S., Talukdar G. and Sharma A. (1987) Induction of sister chromatid exchange and dominant lethal mutation by Katha (catechu) in male mice. Cancer Letters 36, 189 196. IARC Working Group (1985) IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. Vol. 37. Tobacco Habits Other than Smoking; Betel-Quid and Areca-Nut Chewing; and Some Related Nitrosamines. International Agency for Research on Cancer, Lyon. Jussawalla D. J. and Deshpande V. A. (1971) Evaluation of cancer risk in tobacco chewers and smokers: an epidemiological assessment. Cancer 28, 244-252. Khrime R. D., Mehra Y. N., Manu S. B. S., Mehta S. K. and Chakraborti R. N. (1991) Effects of instant preparation of betelnut (pan masala) on the oral mucosa of albino rats. Indian Journal of Medical Research 94, 119-124. Maron D. M. and Ames B. N. (1983) Revised methods for the Salmonella mutagenicity test. Mutation Research 113, 173-215. Mukherjee A. and Giri A. K. (1991) Sister chromatid exchanges induced by pan masala (a betel quid ingredient) in male mice in vivo. Food and Chemical Toxicology 29, 401-403. Nair U. J., Floyd R. A., Nair J., Bussachini V., Friesen M. and Bartsch H. (1987) Formation of reactive oxygen species and hydroxydeoxyguanosine in DNA in vitro with betel quid ingredients. Chemico-Biological Interactions 63, 157 169.
Panigrahi G. B. and Rao A. R. (1982) Chromosome breaking ability of arecoline a major betelnut alkaloid in mouse bone marrow cells in vivo. Mutation Research 103, 197-204. Sarma A. B., Chakrabarti J., Chakrabarti A. C., Banerjee T. S., Roy D., Mukherjee D. and Mukherjee A. (1992) Evaluation of pan masala for toxic effects on liver and other organs. Food and Chemical Toxicology 30, 161 163. Shirname L. P., Menon M. M., Nair J. and Bhide S. V. (1983) Correlation of mutagenicity and tumorigenicity of betel quid and its ingredients using mammalian test systems. Nutrition and Cancer 5, 87 91. Shirname L. P., Menon M. M. and Bhide S. V. (1984) Mutagenicity of betel quid and its ingredients using mammalian test system. Carcinogenesis 5, 501-503. Shivapurkar N. M., Ranadive S. N., Gothoskar S. V., Bhide S. V. and Ranadive K. J. (1980) Tumorigenic effect of aqueous and polyphenolic fractions of betelnut in Swiss strain of mice. Indian Journal of Experimental Biology' 18, 1159-1161. Sirsat S. M. and Khanolkar V. R. (1962) Submucous fibrosis of the palate and pillars of fauces. Indian Journal of Medical Sciences 16, 189. Sundqvist K., Lin Y., Nair J., Bartsch H., Arvidson K. and Graftstr6m R. C. (1989) Cytotoxic and genotoxic effects of arecanut related compounds in cultured human buccal epithelial cells. Cancer Research 49, 5294 5298. Suri K., Goldman H. M. and Wells H. (1971) Carcinogenic effect of dimethyl sulfoxide extract of betelnut. Nature, London 230, 383-384. Stich H. F., Stich W. and Lam P. P. S. (1981) Potentiation of genotoxicity by concurrent application of compounds found in betel qui~' arecoline, eugenol, quercetin, chlorogenic acid and Mn ~+. Mutation Research 90, 355 363.
0278-6915/93 $6.00 + 0.00 Copyright © 1993 Pergamon Press Ltd
Printed in Great Britain. All rights reserved
D O S E - D E P E N D E N T G E N O T O X I C E F F E C T OF P A N M A S A L A A N D A R E C A N U T IN THE SALMONELLA TYPHIMURIUM ASSAY K. POLASA*, S. BABU and I. S. SHENOLIKAR National Institute of Nutrition, Jamai-Osmania, Hyderabad 500 007 (A.P.), India
(Accepted 4 February 1993) Abstract--Aqueous extracts of different brands of pan masala and scented supari were tested for mutagenicity by the Salmonella typhimurium assay using tester strains TA98 and TA100. These extracts were found to be mutagenic to both tester strains. The mutagenic effects of pan masala and scented supari extracts were similar to that produced by areca nut extract. The addition of 500 ppm saccharin to the supari extracts did not alter the mutagenic response.
pan masala. 'Supari' is a form of processed areca nut with or without flavourings and spices. Some brands of 'supari' that are commercially available were also assessed for mutagenicity.
INTRODUCTION
The habit of chewing betel quid is very common in India today. A moderate estimate of the number of betel quid chewers in India is around 20 million. In fact, it outnumbers smokers. Epidemiological studies have associated the chewing of betel quid with tobacco with an increased risk of carcinoma of the oral cavity (IARC, 1985). Several reports also indicate other adverse effects of tobacco chewing, including increase in pulse rate and blood pressure, increased frequency of bronchitis, effects on reproduction and prenatal toxicity in women, and the occurrence of preneoplastic changes in the buccal cavity (leukoplakia) (IARC, 1985; Jussawalla and Deshpande, 1971; Shirname et al., 1983). Besides tobacco, another major ingredient of betel quid is the areca nut, which contains alkaloids such as arecoline and arecaidine. These substances have been shown to be mutagenic to bacteria (Shirname et al., 1983) and mammalian cells in culture (Shirname et al., 1984). Catechu, another ingredient of betel quid, has been shown to induce sister chromatid exchanges (SCE) and dominant lethality in male mice (Giri et al., 1987; Mukherjee and Giri, 1991). Several ingredients are used in the preparation of betel quid and its preparation is time consuming. Therefore, using traditional Indian technology, the industry has developed a product ready for consumption packed in small convenient sachets. This product is 'pan masala'. It has almost all of the ingredients that are present in betel quid except the betel leaf. There is one report in the literature demonstrating that in vitro the aqueous extract of pan masala produces SCE and chromosomal aberrations in Chinese hamster ovary cells (Adhvaryu et al., 1989). We report here the mutagenicity of some varieties of *To whom correspondence should be addressed.
MATERIALS AND METHODS
Eight varieties of pan masala and six varieties of supari were purchased from the local market. 100 g of the sample was powdered and mixed with 200 ml glass-distilled water on a mechanical shaker for 1 hr. The solution was filtered through Whatman No. 1 filter paper under vacuum and lyophilized. About 2.2-5.3g dry matter was obtained from a 100-g sample. The extract was stored at - 2 0 ° C until analysis. For the mutagenicity studies, extracts of pan masala were tested at 100, 200 and 300 #g/plate, and supari extract was tested at 200 pg/plate. An extract of areca nut without added flavourings or spices was also tested at 100, 200 and 300 pg/plate. Mutagenicity testing. All the chemicals for the assay were obtained from Sigma Chemical Company (St Louis, MO, USA). Salmonella typhimurium TA98 and TA100 were kindly provided by Dr B. N. Ames (Berkeley, CA, USA). S-9 was prepared from the livers of Aroclorinduced rats according to the procedure of Maron and Ames (1983). To the test substance, 0.1 ml of the bacterial culture, 0.5 ml S-9 mixture and 2.5 ml top agar were added and plated on Vogel Bonner minimum medium plates. For testing the samples without metabolic activation the same procedure was followed except that the S-9 mixture was omitted from the reaction mixture. These plates were incubated at 37°C for 48 hr in the dark and the numbers of revertant colonies were counted. Benzo[a]pyrene (5/~g/plate) and sodium azide (1/~g/plate) were used as the positive control for TA98 and TA100, respectively.
439
K. POLASA et al.
440
80
(a) TA98 -$9
70
60
=
50
O O
N
4o
o
30
--
w/
20 /
I
tO
1
600
-
2
3
4
5
6
(c)
7
8
9
l
2
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TA100 -$9
500
400
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o
200
100
1
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5
6
V a r i e t i e s o f Pan M a s a l a
7
8
9
V a r i e t i e s of Pan Masala
Fig. 1. Dose-dependent mutagenicity of pan masala extracts in Salmonella typhimurium: (a) TA98 without S-9, (b) TA98 with S-9, (c) TAI00 without S-9 and (d) TA100 with S-9 all tested at 100 (~/), 200 ( O ) and 300 (11)/~g extract/plate. Values depicted are means of triplicate plate assays. Variety 9 represents areca nut extract. All of the values are significantly higher than the negative control values (horizontal line) given in Table I (Student's t-test; P < 0.01) except variety no. 4 tested at 100 # g extract/plate.
8
9
Mutagenicity of pan masala and areca nut extracts
441
Table I. Reversion frequency induced by supari extracts (200 #g/plate) in S. typhimurium TA 98 Sample of supari (areca nut) Sample 1 Sample 1 + 500 ppm saccharin Sample 2 Sample 3 Sample 4 Sample 4 + 500 ppm saccharin Sample 5 Sample 6 Negative control Positive controls: B[aIP (5 #g/plate) Sodium azide (1 #g/plate)
- S-9 43 _+ 15 49 £ 14 46 ± 11 46 ± 9* 48 ± 3*** 35 ± 4 39 ± 4* 38 ± 9 29 £ 3
TA 100 + S-9 66 ± 20 54 ± 13 61 ± 7** 63 ± 19" 64 ± 11 40 ± 3 54 ± 8* 45 ± 4* 35 ± 4
- S-9 206 ± 34* 198 ± 40* 244 ± 26*** 212 ± 28** 132 ± 17 210 ± 19"* 184 ± 11"* 198 ± 28* 132 ± 12
+ S-9 396 ± 30*** 492 ± 28*** 432 ± 52*** 409 ± 31"** 392 ± 42*** 400± 40*** 452 ± 45*** 392 ± 57*** 158 ± 12
2762 ± 220 626 ± 32 -426 ± 16 --BIa]P = benzo[a]pyrene Values are means + SD and those marked with asterisks differ significantly(Student's t-test) from the corresponding value for the negative control (*P < 0.05; **P < 0.01; ***P < 0.001). ---
RESULTS The m u t a g e n i c activities of the p a n masala a n d supari as q u a n t i t a t e d by reversion frequencies in S. t y p h i m u r i u m TA98 and TA100 are s h o w n in Fig. 1 a n d Table 1. The p a n masala extracts were tested at three c o n c e n t r a t i o n s (100, 200 a n d 300/~g/plate) whereas the areca nut (supari) extract was tested at a c o n c e n t r a t i o n of 200 #g/plate. Virtually all of the samples of p a n masala were mutagenic, with an e n h a n c e d effect seen with activation. A clear dosedependent mutagenic response was noted, particularly in T A I 0 0 with metabolic activation (Fig. ld). These results are similar to the mutagenic response shown by all the varieties of scented supari extracts (Table 1). The mutagenic effects of pan masala a n d scented supari extracts was similar to that produced by areca n u t extract (sample 9 in Fig. 1). Some of the supari extracts c o n t a i n e d saccharin but its presence did not seem to influence the mutagenic potential of the supari. DISCUSSION The m a j o r c o m p o n e n t of pan masala is the areca nut, which has at least six reduced pyridine alkaloids (Ashby et al., 1979). The m a j o r ones are arecoline, arecaidine, arecolidine, guvacoline a n d guvacine. Arecaidine a n d its methylester arecoline have been suggested to have mutagenic/carcinogenic properties since they are capable of reacting with cysteine in vivo and in vitro to produce cysteine/3-alkylation adducts (Boyland a n d Nery, 1969). The results of our mutagenicity studies of pan masala are significant because mutagenicity is highly correlated to carcinogen±city. A l t h o u g h there are no reports o f carcinogen±city induced by pan masala in the literature, one of the c o m p o n e n t s of pan masala, namely areca nut, has produced precancerous lesions 'leukoplakia' in Syrian hamsters (Suri et al., 1971). Likewise, the polyphenolic fraction of betel quid induced tumours in mice (Shivapurkar, 1980). A paste o f p a n masala applied to the oral cavity of albino rats resulted in increased keratosis, parakeratosis, inflam-
m a t o r y cell infiltration a n d vascularity indicating a mild leukoplakia (Khrime et al., 1991). C o m p o n e n t s of p a n masala, namely betel nuts, catechu and lime, have also been reported to produce this condition (Sirsat a n d K h a n o l k a r , 1962). The contents of betel quid have been clearly d e m o n s t r a t e d to produce a toxic effect o n cultured h u m a n buccal epithelial cells as well as o n the buccal m u c o s a of b a b o o n s a n d hamsters (Sundqvist et aL, 1989). There is also a positive correlation between oral cancer and precancerous lesions in ' p a n chewers' (Dayal et al., 1978). Besides the genotoxic effects, hepatotoxic effects Of p a n masala have been reported by S a r m a et al. (1992). A l t h o u g h we have studied the mutagenic potential of aqueous extracts of different samples of pan masala on the basis o f weight of extract (100, 200 and 300/~ g extract), these results have to be interpreted in terms of the content of arecoline and arecaidine in these samples. Moreover, plain supari a n d areca nut do not contain substances such as tobacco, catachu, flavourings and spices that are present in p a n masala. These additives m a y also lead to habitual chewing by virtue of e n h a n c i n g the taste of the product. This may also result in an increase in the prevalence of betel quid chewing. In view of the evidence available in the literature on the genotoxic potential of chewing substances such as betel quid (Nair et al., 1987; Stich et al., 1981), areca nut (Panigrahi a n d Rao, 1982), a n d tobacco, which is further supported by the dose-response effect reported here, caution needs to be exercised on the indiscriminate use of these products. Acknowledgements--The authors are grateful to Dr Vinodin± Reddy, Director, for her keen interest in the investigation. They also acknowledge the technical assistance of Mrs B. R. Annapurna and Mrs Amulya Rao. REFERENCES
Adhvaryu S. G., Dave B. J. and Trivedi A. H. (1989) An in vitro assessment of the genotoxic potential of pan masalas. Indian Journal o f Medical Research 90, 131-134. Ashby J., Styles J. A. and Boyland E. (1979) Betelnuts, arecaidine and oral cancer. Lancet i, 112.
442
K. POLASAet al.
Boyland E. and Neri R. (1969) Mercapturic acid formation during metabolism of arecoline and arecaidine in the rat. Biochemical Journal 113, 123-130. Dayal P. K., Mani N. J. and Bhargava K. (1978) Prevalence of oral cancer and precancerous lesions in "Pan" supari chewers. Indian Journal of Public Health 22, 234-245. Giri A. K., Banerjee T. S., Talukdar G. and Sharma A. (1987) Induction of sister chromatid exchange and dominant lethal mutation by Katha (catechu) in male mice. Cancer Letters 36, 189 196. IARC Working Group (1985) IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. Vol. 37. Tobacco Habits Other than Smoking; Betel-Quid and Areca-Nut Chewing; and Some Related Nitrosamines. International Agency for Research on Cancer, Lyon. Jussawalla D. J. and Deshpande V. A. (1971) Evaluation of cancer risk in tobacco chewers and smokers: an epidemiological assessment. Cancer 28, 244-252. Khrime R. D., Mehra Y. N., Manu S. B. S., Mehta S. K. and Chakraborti R. N. (1991) Effects of instant preparation of betelnut (pan masala) on the oral mucosa of albino rats. Indian Journal of Medical Research 94, 119-124. Maron D. M. and Ames B. N. (1983) Revised methods for the Salmonella mutagenicity test. Mutation Research 113, 173-215. Mukherjee A. and Giri A. K. (1991) Sister chromatid exchanges induced by pan masala (a betel quid ingredient) in male mice in vivo. Food and Chemical Toxicology 29, 401-403. Nair U. J., Floyd R. A., Nair J., Bussachini V., Friesen M. and Bartsch H. (1987) Formation of reactive oxygen species and hydroxydeoxyguanosine in DNA in vitro with betel quid ingredients. Chemico-Biological Interactions 63, 157 169.
Panigrahi G. B. and Rao A. R. (1982) Chromosome breaking ability of arecoline a major betelnut alkaloid in mouse bone marrow cells in vivo. Mutation Research 103, 197-204. Sarma A. B., Chakrabarti J., Chakrabarti A. C., Banerjee T. S., Roy D., Mukherjee D. and Mukherjee A. (1992) Evaluation of pan masala for toxic effects on liver and other organs. Food and Chemical Toxicology 30, 161 163. Shirname L. P., Menon M. M., Nair J. and Bhide S. V. (1983) Correlation of mutagenicity and tumorigenicity of betel quid and its ingredients using mammalian test systems. Nutrition and Cancer 5, 87 91. Shirname L. P., Menon M. M. and Bhide S. V. (1984) Mutagenicity of betel quid and its ingredients using mammalian test system. Carcinogenesis 5, 501-503. Shivapurkar N. M., Ranadive S. N., Gothoskar S. V., Bhide S. V. and Ranadive K. J. (1980) Tumorigenic effect of aqueous and polyphenolic fractions of betelnut in Swiss strain of mice. Indian Journal of Experimental Biology' 18, 1159-1161. Sirsat S. M. and Khanolkar V. R. (1962) Submucous fibrosis of the palate and pillars of fauces. Indian Journal of Medical Sciences 16, 189. Sundqvist K., Lin Y., Nair J., Bartsch H., Arvidson K. and Graftstr6m R. C. (1989) Cytotoxic and genotoxic effects of arecanut related compounds in cultured human buccal epithelial cells. Cancer Research 49, 5294 5298. Suri K., Goldman H. M. and Wells H. (1971) Carcinogenic effect of dimethyl sulfoxide extract of betelnut. Nature, London 230, 383-384. Stich H. F., Stich W. and Lam P. P. S. (1981) Potentiation of genotoxicity by concurrent application of compounds found in betel qui~' arecoline, eugenol, quercetin, chlorogenic acid and Mn ~+. Mutation Research 90, 355 363.