Robert M. Kovatch

Gross and microscopic lesions in the female SENCAR mouse skin and lung in tumor initiation and promotion studies.

The skin and lung tissues from SENCAR mice used as part of the Environmental Protection Agencys (EPAs) Carcinogenesis Testing Matrix were examined. This study included SENCAR mice used in three different short-term bioassay protocols in which the skin papilloma assay was used to identify initiators, promoters, and complete carcinogens. Also included were the pathology findings from SENCAR mice used in the combined bioassay in which the skin assay and the lung adenoma assay were conducted simultaneously. The gross and microscopic features of treatment-associated and spontaneous lesions of the skin and lung of the SENCAR mouse used in these studies are defined and the lesions most commonly observed are described. Generally, gross observations and microscopic findings in both the skin and lung tissues were poorly correlated. Although there are several definite criteria on which gross interpretations of the various skin and lung lesions can be made, with the exception of pedunculated squamous cell papillomas and the classic squamous cell carcinomas, the various lesion types had a wide variety of clinical presentations that severely compromised the accuracy of gross diagnosis. Further, in the case of benign skin neoplasms, malignant transformation of these tumors most often occurred at the base of the lesion and was initially hidden from gross observation. As a result, approximately 50% of the neoplasms interpreted clinically as benign tumors (papillomas and keratoacanthomas) were actually malignant neoplasms. Moreover, many lesions determined grossly to be nontumorous were in fact found to be neoplastic when examined microscopically. The SENCAR mouse was found to be more responsive in the lung adenoma assay than other strains examined with exception of the Strain A. Although accurate interpretation of the lung lesions in the SENCAR was compromised by nonneoplastic treatment-associated and/or spontaneous lesions, the feasibility of using the SENCAR skin and lung as target tissues in two-stage combined carcinogenesis studies merits further consideration. ImagesFIGURE 1.FIGURE 2.FIGURE 3.FIGURE 4.FIGURE 5.FIGURE 6.FIGURE 7.FIGURE 8.FIGURE 9.FIGURE 10.FIGURE 11.FIGURE 12.FIGURE 13.FIGURE 14.FIGURE 15.FIGURE 16.FIGURE 17.

Tumorigenicity of the tobacco-specific carcinogen 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone in infant mice

The tobacco-specific nitrosamine, 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK), is a potent carcinogen in adult rodents and variably effective transplacentally, depending on species. In pursuit of the thesis that human infants may be especially vulnerable targets for tumor initiation by tobacco smoke constituents, we tested the efficacy of NNK as a tumor initiator in infant mice. Cr:NIH(S) (NIH Swiss outbred) mice were given 50 mg/kg NNK i.p. on postnatal days 1, 4, 7, 10 and 14, with saline to controls. At an average age of 13-15 months, 57% of the NNK-exposed male offspring had hepatocellular tumors, with a multiplicity of 1.15 +/- 1.4, including 4 with carcinoma. Liver tumors including 2 carcinomas were found in 8 (14%) of the NNK-exposed female offspring. There were no hepatocellular neoplasms in any control. A significant increase in primary lung tumors also occurred in the NNK-treated males, with an incidence of 30/55 (57%) and a multiplicity of 0.7 +/- 0.2, vs. 7/33 (21%), multiplicity 0.3 +/- 0.6, in controls (P less than 0.025). An apparent increase in the incidence of lung tumors in NNK-treated females, 21/57 (37%) vs. 7/32 (22%) in controls, approached significance (P less than 0.1). Thus NNK was a moderately potent neonatal carcinogen for liver and lung in infant Swiss mice and more efficacious in this regard than when received transplacentally by mice of the same strain.

Carcinogenesis in F-344 rats induced by nitrosohydroxyalkyl-chloroethylureas.

SummaryThe two isomeric N-nitroso derivatives of 1-chloroethyl-3-(2-hydroxyethyl)-urea and of 1-chloroethyl-3-(2-hydroxypropyl)-urea were prepared and isolated. They were given by gavage in ethyl acetate/corn oil to groups of 20 male and female F-344 rats. The two nitroso-1-chloroethyl compounds were nephrotoxic and most animals died within 20 weeks; no neoplasms were seen in any of these animals. Nitroso-1-hydroxyethyl-3-chloroethylurea was given at 2 concentrations, 21 and 10.5 mg/ml; in both groups almost all animals died with neoplasms related to the treatment. These included hepatocellular and cholangiocellular neoplasms of the liver; many of the former metastasized. Many rats also had tubular cell neoplasms in the kidney. Nitroso-1-(2-hydroxypropyl)-3-chloroethylurea was a less potent carcinogen at equimolar doses, inducing fewer liver neoplasms than the nitrosohydroxyethyl analog and only few kidney neoplasms. Both of these carcinogens were less effective in female rats than in males, although the females, which were smaller, received a higher dose per unit body weight. The spectrum of neoplasms induced by the nitrosohydroxyalkyl-chloroethylureas was quite different from that induced by equimolar doses of each corresponding nitrosohydroxyalkylurea, neither of which induced neoplasms of the liver, although they were potent inducers of neoplasms in other organs.

Carcinogenicity studies of some analogs of the carcinogen methapyrilene in F344 rats

SummaryFour antihistaminic drugs similar in structure to the rat liver carcinogen methapyrilene were administered to comparable groups of male and female F344 rats in their drinking water for most of their lifetime (80–108 weeks). The concentrations were 0.1% or 0.05% and the total doses received by the animals were comparable with that of methapyrilene which induced 100% incidence of liver neoplasms. No increase in incidence of liver neoplasms was observed after treatment with any of the four compounds, thenyldiamine, chlorothen, methafurylene, or methaphenilene, although each differed structurally from methapyrilene only in one atom or one position of substitution. There were a few animals with neoplasms not usually found in untreated F344 rats, but none of these was found in statistically significant numbers. These results suggest that none of the four analogs of methapyrilene was carcinogenic under the conditions of this study, and that the property of inducing liver neoplasms in rats was confined to the intact methapyrilene molecule.

Carcinogenesis and nucleic acid alkylation by some oxygenated nitrosamines in rats and hamsters.

A comparison has been made of the carcinogenic effects of nitroso-2,6-dimethylmorpholine and several hydroxylated acyclic nitrosodialkylamines derived from it or related to it in rats and Syrian hamsters. In rats nitrosodimethylmorpholine was the most potent, inducing mainly esophageal tumors. Nitrosodiethanolamine was the weakest of the five nitrosamines in both rats and hamsters. Tumors of the pancreas ducts were induced by four of the five compounds, but only in hamsters, and esophageal tumors appeared only in rats. Most of the nitrosamines induced tumors of liver and lung in both rats and hamsters. A study of alkylation of nucleic acids of the liver following treatment of rats and hamsters with the radiolabeled nitrosamines showed that nitrosodiethanolamine alkylated liver nucleic acids in rats to only a very small extent. The other four nitrosamines all gave rise to 7-methylation and O6-methylation of guanine residues in DNA of hamster liver and all but nitrosodimethylmorpholine in rat liver DNA, which corresponded quite well with the induction of liver tumors in the two species. Quantitatively, however, there was not a good correlation between liver DNA alkylation and the potency of the nitrosamine in inducing tumors.

Carcinogenesis by nitrosobis-(2-oxopropyl)amine labeled with deuterium and by nitroso-2-hydroxypropyl-2-oxopropylamine in rats and hamsters

The effects of the labeling with deuterium of the alpha-methylene groups of the carcinogen nitrosobis-(2-oxopropyl)amine (NBOP) on its carcinogenic effectiveness in rats and hamsters have been studied. The greater strength of the C-D bond compared with the C-H bond often leads to slower metabolism and lesser carcinogenic activity. When NBOP and NBOP-d4 were given to male and female rats in drinking water at equimolar doses, the mortality rate from tumors was lower in the rats given the deuterium-labeled compound, although the results were statistically significant (P = 0.012) only in males. The incidences of tumors of several groups was similar for NBOP and NBOP-d4, but there was a marked difference between males and females, females having a high incidence of liver tumors, and males very few. When NBOP and NBOP-d4 were given by gavage to rats or Syrian hamsters at identical doses there was no difference in rate of mortality from tumors, or in the pattern of tumors induced by either compound. In rats, both compounds were given at two dose rates, and in neither was a difference seen. To complement the studies with NBOP, a normal reduction product formed metabolically in vivo, nitroso-(2-hydroxypropyl) (2-oxopropyl)amine (NHPOP) was administered to rats in drinking water at the same dose rate. In male rats, the mortality rate was lower with NHPOP than with NBOP, while with female rats the opposite was the case (P less than 0.01 in both cases) and there was little difference in the pattern of tumors induced in either sex. NHPOP appears to have a quite distinct carcinogenic effect from NBOP, suggesting that the metabolic conversion of one to the other does not play a large role. The weak deuterium-isotope effect of NBOP-d4 given to rats in drinking water, but not detected in rats or hamsters treated by gavage, suggests that alpha-oxidation of NBOP is not likely to be a rate-limiting step in carcinogenesis by NBOP.

Carcinogenesis by oxygenated nitrosomethylpropylamines in Syrian hamsters.

SummaryThree oxygenated propylnitrosomethylamines were administered to female Syrian hamsters at doses similar to those which had induced high incidences of esophageal neoplasms in rats. Nitrosomethyl-2-oxopropylamine (NMOP) given at the rate of 2 mg/animal/week, whether as one application of 2 mg or two applications of 1 mg, led to early death of the animals, mostly with liver neoplasms; administration of 1 mg/animal/week led to longer survival, but most animals died with both liver neoplasms and neoplasms of the nasal mucosa. Only one hamster treated with NMOP had a neoplasm of the pancreatic duct. Of the 14 hamsters treated with the higher dose of nitrosomethyl-2-hydroxypropylamine (NMHP) and surviving beyond 6 weeks, most had liver neoplasms and nine had neoplasms of the pancreatic ducts. At the lower dose of NMHP, most hamsters developed neoplasms of the nasal mucosa, as did those receiving the same dose of NMOP, and seven animals had hemangioendothelial tumors of the liver, but only one animal had a carcinoma of the pancreatic duct. Nitrosomethyldihydroxypropylamine (NMDHP) was a much weaker carcinogen than the other two compounds and induced mainly neoplasms of the nasal mucosa, with little shortening of life.