Histopathological evaluation of proliferative liver lesions in rats fed trans-anethole in chronic studies

In a two-year study of rats fed trans-anethole--a flavouring that is Generally Recognized As Safe by the US Food and Drug Administration--a slight increase in proliferative lesions of the liver was observed. These results were reviewed by a Pathology Working Group (PWG) and their findings are detailed here. The increased incidence of non-neoplastic lesions in male and female rats fed 0.25, 0.5 or 1.0% trans-anethole was considered by the PWG to be treatment related. Male and female rats fed 0.25% trans-anethole in their diet (25 to 29 times the postulated maximum human intake) had no significant treatment-related microscopic lesions in the liver, although there was a slight increase in parenchymal cell hyperplasia. Rats fed 0.5% trans-anethole had hepatic changes consistent with those caused by chronic exposure of rodents to compounds with enzyme-inducing activity. Male and female rats fed 1.0% trans-anethole revealed increased incidence and severity of hepatic changes as seen in the mid-dose rats, and a low incidence of hepatocellular neoplasms. These neoplasms were not increased in male rats in any dietary group or in female rats of the low- (0.25%) and mid-dose (0.5%) groups. The slightly increased incidence of hepatocellular neoplasms in the high-dose females is not considered to be of significance to human safety and it is concluded that trans-anethole does not constitute a carcinogenic risk for man. The slightly increased tumour incidence was seen only in the highest dose group of one sex; it has not been observed in studies of mice fed trans-anethole--the metabolism of trans-anethole in the mouse is similar to that in man. The extremely high exposure was associated with other liver changes which may have contributed to the findings.     

Suppressive effect of irsogladine maleate on N-methyl-N-nitro-N-nitrosoguanidine (MNNG)-initiated and glyoxal-promoted gastric carcinogenesis in rats

The modifying effect of irsogladine maleate (IRG) on N-methyl-N-nitro-N-nitrosoguanidine (MNNG)-initiated and glyoxal-promoted gastric carcinogenesis was examined in male Wistar rats. Six-week-old rats were divided into ten groups. Groups 1 through 6 were given MNNG (100 mg/l in drinking water) for 25 weeks from the start of the experiment, whereas groups 7 through 10 received distilled water in the initiation phase as the vehicle treatment. Groups 1 and 8 were kept on the basal diet and distilled water throughout the experiment (55 weeks). Groups 2-8 were given 0.5% glyoxal in the drinking water for 30 weeks from 26th week of the experiment. Group 3 was fed the diet mixed with 100 ppm IRG for 25 weeks from the start of experiment. Groups 4 and 8 were fed the diet mixed with 100 ppm IRG for 30 weeks from 26th week of experiment. Groups 5 and 9 or 6 were given 100 or 25 ppm IRG containing diet, respectively throughout the experiment. Group 10 was given the basal diet and distilled water as the vehicle treated control. Tumors of upper digestive tracts (stomach and duodenum) were developed in groups: 1 (12/17 rats, 71%), 2 (11/12 rats, 92%), 3 (9/16 rats, 56%), 4 (5/12 rats, 42%), 5 (6/15 rats, 40%) and 6 (7/12 rats, 58%). High dose of IRG in initiation and/or promotion phase significantly reduced the incidence of tumors of the upper digestive tracts. The average numbers of the digestive tracts neoplasms in groups 3,5 and 6 given glyoxal and IRG were less than those in group 2 which received only glyoxal. These results suggest that IRG could be a preventive agent against the occurrence of neoplasms of the upper digestive tract.     

Promotion of thyroid carcinogenesis by para-aminobenzoic acid in rats initiated with N-bis(2-hydroxypropyl)nitrosamine.

Sulfonamide analogues of para-aminobenzoic acid (PABA), a precursor of folate synthesis, have beneficial effects as antifolate, but thyroid peroxidase inhibition has been reported as a side effect that results in promotion of rat thyroid carcinogenesis. In the present study, effects of PABA itself on F344 rat thyroid carcinogenesis after initiation with N-bis(2-hydroxypropyl)nitrosamine (DHPN) were evaluated. In experiment 1, rats in groups 1-4 received a single subcutaneous injection of DHPN at 2800 mg/kg, and groups 5 and 6 received vehicle saline alone. From 1 week after DHPN initiation, rats in groups 2, 3, 4, and 6 were fed basal diet containing 0.25%, 0.5%, 1.0%, and 1.0% PABA, respectively, for 40 weeks. Rats in groups 1 and 5 received basal diet alone throughout the experiment. The final incidence of thyroid follicular cell adenomas and adenocarcinomas was significantly (p < 0.05 or 0.01) increased in groups 3 and 4 as compared to group 1. No thyroid tumors were found in groups 5 and 6. In experiment 2, animals in group 1 were fed basal diet alone, while groups 2 and 3 were given 0.5% and 1.0% PABA in the diet, respectively, for 2 weeks. Thyroid weights in group 3, and serum thyroid stimulating hormone level and proliferative activity of follicular cells in groups 2 and 3 were significantly (p < 0.05 or 0.01) elevated. In addition, the serum thyroxine level in group 3 was significantly (p < 0.05) depressed. These results clearly indicate that PABA exerts promotion/progression effects on rat thyroid carcinogenesis as a result of hypothyroidism followed by negative-feedback via the thyroid-pituitary axis.     

Effect of urinary pH on the progression of urinary bladder tumours

Systemic alkalosis has been postulated to enhance tumorigenesis, whereas systemic acidosis has been implicated to exert a favourable influence on tumour control and regression. In the present study the urinary pH was influenced by feeding acid-forming or base-forming diets, and the effect of alkaline or acid urine on the early and late progression phase of urinary bladder carcinogenicity was investigated in male Wistar rats. Bladder lesions were initiated by N-butyl-N-(4-hydroxybutyl) nitrosamine (0.05% BBN in the drinking water during 4 weeks) and promoted by sodium bicarbonate (3.4% NaHCO₃ in the diet during 15 or 25 weeks). After short- (15 week) and more long-term (25 week) promotion with NaHCO₃, groups of 20 rats were fed a diet containing the acidifying salt ammoniumchloride (2.1% NH₄Cl) or the control diet. All surviving rats were killed after a total study duration of 52 weeks. Additional control groups were, after initiation, fed diets containing NaHCO₃ and killed after 15 wk or 25 wk of promotion, or at the end of the study. In rats fed diets with added salts, water intake and the amount of urine produced were increased and the urinary density was decreased compared to rats fed control diet. During NaHCO₃ feeding, urinary pH and sodium concentration were increased. During NH₄Cl feeding, urinary pH was decreased and urinary chloride and calcium concentrations were increased. Initiation by BBN followed by treatment with NaHCO₃ caused a high incidence of papillary/nodular hyperplasia, papillomas and carcinomas of the bladder epithelium. These lesions progressed with time or longer duration of NaHCO₃ promotion. A tumour protective effect of urinary acidification by NH₄Cl was not found. In fact, both acidification and prolonged alkalinization tended to aggravate the malignancy of bladder carcinomas.     

Effects of sodium salts of phenobarbital and barbital on development of bladder tumors in male F344/NCr rats pretreated with either N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide or N-nitrosobutyl-4-hydroxybutylamine

Promoting effects of sodium salts of phenobarbital (NaPB) and barbital (NaBB) on the development of bladder tumors were investigated in F344 male rats initiated with N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) or N-nitrosobutyl-4-hydroxybutylamine (BBN). To initiate with FANFT, rats were fed 0.2% FANFT mixed in diet for either 2 or 6 weeks and 2 weeks later were offered diet containing 1000 ppm of NaPB or NaBB. Rats were killed either at 52 or 68 weeks of age. To initiate with BBN, rats were given 0.05% BBN in drinking water for 4 weeks and beginning 1 day later were fed NaBB mixed in diet at 1000 ppm for up to 52 weeks. NaBB promoted bladder carcinogenesis initiated by either FANFT or BBN; the incidence and average number of simple or preneoplastic nodular (PN) hyperplasias, papillomas, and carcinomas per 10 cm of urothelium were significantly increased in the groups receiving NaBB following exposure to FANFT for 6 weeks (p less than 0.05) or BBN for 4 weeks (p less than 0.01). No such effect was seen in rats fed FANFT for only 2 weeks. NaPB also significantly increased (p less than 0.05) the frequency of preneoplastic PN hyperplasias but not the average number of papillomas and carcinomas per 10 cm of urothelium in rats fed FANFT for 6 weeks. NaBB was an effective promoter of bladder carcinogenesis under these experimental conditions, as expected from its known promoting effect on transitional epithelium of the renal pelvis, but NaPB in contrast did not affect the incidence or multiplicity of bladder papillomas or carcinomas under these conditions. NaPB could be considered a promoter for bladder urothelium only by the less rigorous criterion that it increased the frequency of preneoplastic PN hyperplasia.     

Dose-threshold for thyroid tumor-promoting effects of orally administered kojic acid in rats after initiation with N-bis(2-hydroxypropyl) nitrosamine.

In order to evaluate the threshold dose of thyroid tumor-promoting effects of KA, male F344 rats were initiated with N-bis(2-hydroxypropyl) nitrosamine (DHPN; 2000 mg/kg body wt., single s.c. injection) and, starting 1 week later, received pulverized basal diet containing 0%, 0.002%, 0.008%, 0.03%, 0.125%, 0.5% or 2%KA for 20 weeks. Five rats each in the 0%, 0.125%, 0.5% and 2%KA groups were sacrificed at week 12, and 10 rats each in all groups at week 20. As an additional experiment, three groups without DHPN initiation received basal diet, a diet containing 0.5% or 2%KA for 20 weeks. The serum T4 levels were significantly decreased in the DHPN-initiated groups given 0.125%KA or more at week 12. No significant decreases in serum T3 levels were observed in the groups treated with DHPN + KA and a significant increase was evident in the 2%KA-alone group at week 20. Some rats in the DHPN + 2%KA group at weeks 12 and 20 and the 2%KA-alone group at week 20 showed pronounced elevation of serum TSH. Thyroid weights were significantly increased in the DHPN-initiated groups receiving 0.5% and 2%KA at weeks 12 and 20 and in the 2%KA-alone group at week 20. Histopathologically, the incidences of focal thyroid follicular cell hyperplasias in the DHPN-initiated groups treated with 0.125%, 0.5% and 2%KA at week 20 were 5/10, 10/10 and 8/8 rats, respectively. At week 20, adenomas were observed in 7/10 rats in the DHPN + 0.5%KA group and 8/8 rats in the DHPN + 2%KA group, and carcinomas were observed in 6/8 rats in the DHPN + 2%KA group. In the groups without DHPN initiation, only focal follicular cell hyperplasia was observed in 1/9 rats in the 2%KA-alone group. These results suggest that the no-observed-adverse effect for the thyroid tumor-promoting effect of KA is 0.03% (15.5 mg/kg/day) under the present experimental conditions, and that KA possesses weak tumorigenic activity in rats due to continuous serum TSH stimulation by a non-genotoxic mechanism.     

Ethanol-extracted propolis enhances BBN-initiated urinary bladder carcinogenesis via non-mutagenic mechanisms in rats

Ethanol-extracted propolis (EEP) is used for medical, dietetic and cosmetic purposes. In this study, the effects of EEP on urinary bladder carcinogenesis, its underlying mechanism and in vivo genotoxicity were investigated. In experiment 1, rats were treated with N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) for 2 or 4 weeks followed by dietary administration of 0.125, 0.25, 0.5 or 1% EEP for 4 or 32 weeks, respectively. At week 6, the mRNA levels of top2a, cyclin D1 and survivin were significantly elevated in the 0.5 and 1% EEP groups. At week 36, the incidence and multiplicity of urothelial carcinomas and total tumors were markedly elevated in all EEP groups. In experiment 2, rats were fed basal diet or the 1% EEP diet for 13 weeks without carcinogen initiation. Increases in urinary precipitate, cell proliferation and incidence of simple hyperplasia were observed in the 1% EEP group. In experiment 3, dietary administration of 2.5% EEP to gpt delta rats for 13 weeks did not induce any obvious mutagenicity in the urinary bladder urothelium. Taken together, EEP enhanced BBN-initiated rat urinary bladder carcinogenesis in a non-genotoxic manner through increasing formation of urinary precipitate, enhancing cell proliferation and inhibiting apoptosis during the early stages of carcinogenesis.     

The effects of reduced dietary intake upon the body and organ weights, and some clinical chemistry and haematological variates of the young Wistar rat

Two groups, one of each sex, of 5-week-old Wistar rats were fed ab lib for 5 weeks with a commercial diet (LAD 2). Four other groups of each sex, were given the same diet daily; each male received 21, 19, 17 or 15 g and each female 17, 16, 15 or 13 g. In a second experiment, in addition to the groups fed ad lib, each day for 5 weeks males were given 30 ('daily ad lib' group), 23, 22 or 21 g and females 25 ('daily ad lib' group), 19, 18 or 17 g. Rats, particularly males, fed ad lib showed differences, e.g. in body weight, with rats fed 'daily ad lib' even though food consumption was similar. Rats fed reduced amounts of diet had smaller livers, increased haemoglobin concentration, erythrocyte count, haematocrit, and plasma chloride. These rats excreted an increased volume of more alkaline urine of lower specific gravity with more precipitated triple phosphate than rats fed ad lib. In the comparisons of the modestly reduced food intake groups with the ad lib groups the significant decreases found in plasma protein, urea and alkaline phosphatase activity probably arose mainly from differences in post-prandial intervals. In groups where the reduction in food intake was greater, the decreases observed were not wholly dependent upon the post-prandial interval.     

Studies on the pharmacological bases of fetal toxicity of drugs. III. Fetal toxicity of potassium nitrate in 2 generations of rats

The fetal toxicity of potassium nitrate (KNO3) used widely as a food additive was studied in Wistar rats. The pregnant rats were fed a diet containing 2.5, 0.5 or 0.1% of KNO3 from day 7 to 14 of pregnancy. Neither maternal nor fetal toxicity including external malformations were observed at term in any group. After spontaneous delivery, the offspring were reared until 13 weeks after birth. No harmful effects were detected in any group. The female offspring (F1) of all groups were mated with the male (F1) of the same group. Good reproductive performances were shown in all groups. The pregnant rats were fed the same diet, which their mothers (F0) had been fed, from day 7 to 14 of pregnancy. Various types of malformations such as exencephaly, cleft lip and palate, polydactyly and micro- or anophthalmia were observed in 6 of 133 fetuses and 7 of 63 newborns from dams treated with 2.5% KNO3, but no external malformations were observed in other groups. The male offspring (F2) of the treated groups showed slow growth until 13 weeks after birth. These results suggest that a dose of 2.5% KNO3 is toxic to the F2 generation, but not to the F1 generation.     

Comparison of the Hepatotoxicity of Coumarin in the Rat, Mouse, and Syrian Hamster: A Dose and Time Response Study

The effects of coumarin treatment have been compared in maleSprague-Dawley CD rats, male CD-1 mice, and male Syrian hamsters.Rats were fed 0–0.75% coumarin for 1 and 4 weeks and 0–0.5%coumarin for 13 weeks, whereas mice and Syrian hamsters werefed 0–0.5 and 0–1.0% coumarin, respectively, forperiods of 1, 4, and 13 weeks. In the rat, coumarin produceddose-related hepatotoxic effects which included vacuolar degeneration,apoptosis, and bile duct proliferation. These effects were particularlymarked at dose levels of 0.3 and 0.5%, where liver tumors havebeen observed in a chronic study. Coumarin administration torats also increased serum bilirubin content and both serum andhepatic -glutamyltransferase activity. While levels of hepatictotal glutathione were increased by coumarin administration,microsomal cytochrome P450 content and ethylmorphine N-demethylaseactivity were reduced. Such effects were either less markedor absent in the mouse and Syrian hamster. Replicative DNA synthesiswas studied by implanting osmotic pumps containing 5-bromo-2'-deoxyuridineduring Study Weeks 0–1, 3–4, and 12–13. Inthe rat, coumarin administration for 4 and 13 weeks at doselevels of 0.3 and 0.5% produced a sustained stimulation of hepatocytereplicative DNA synthesis. No such effects were observed inthe mouse and Syrian hamster. These results demonstrate markedspecies differences in coumarin-induced hepatotoxicity. Whiletumor formation in the rat appears due to chronic hepatotoxicityassociated with a sustained regenerative hyperplasia, such effectswere not observed in the CD-1 mouse and Syrian hamster. In assessingthe hazard of coumarin to humans, account needs to be takenof both levels of exposure and species differences in response.     

Influence of dietary protein on the response of rats receiving toxic levels of warfarin

Toxic doses of racemic [¹⁴C]warfarin (3-(α-acetonylbenzyl)-4-hydroxycoumarin) were administered (0.8 mg/kg, po) for 5 days to rats fed one of three different purified diets, 25% casein, 8% casein, and casein-free. Four rats from each dietary group were bled at 6, 30, 54, 78, and 102 hr after the initial dose. Rats consuming a casein-free diet maintained higher total plasma warfarin levels than rats fed a 25 or 8% casein diet. Additionally, percentage free warfarin levels of 0% casein rats were greater than those of 25 and 8% groups through the first 78 hr. An increase in percentage unbound drug was therefore most likely associated with a decrease in total plasma clearance. The protein-free rats had a more rapid increase in prothrombin time and lower plasma albumin levels than rats fed either a 25 or 8% casein diet. A significant correlation was found between the percentage free fraction of warfarin and the prothrombin response in each of the three dietary groups. A negative correlation was found between percentage free plasma warfarin and plasma albumin concentration in each of the three dietary groups. A correlation was found between total plasma warfarin and prothrombin time for the rats on the 0% protein diet. It was concluded that the quantity of dietary protein was an important factor in determining the response of rats to toxic levels of warfarin.     

Cadmium-induced testes oxidative damage in rats can be influenced by dietary zinc intake.

We tested the hypothesis that zinc deficient animals would be characterized by an increased sensitivity to cadmium-induced oxidative damage to the testes. Weanling male rats were given free access to either a control (25 microg Zn/g) or a zinc deficient (0.5 microg Zn/g) diet; or restricted access to the 25 microg Zn/g diet at a level of intake similar to that of rats fed the 0.5 microg Zn/g diet. After 14 days on the diets, animals were injected s.c. with either saline or CdCl2 (2 mg Cd/kg body weight) solutions, and killed 24 h later. In the zinc-deficient group, testes weight and testes/body weight were higher in the cadmium-injected rats than in the saline-injected rats. The extent of hemorrhages, as indicated by high hemoglobin and testes iron concentrations was higher in the cadmium-treated zinc deficient group than in the cadmium-injected controls. In the zinc-deficient group, cadmium injection was associated with higher levels of lipid peroxidation (33% higher TBARS content) and protein oxidation (17% lower glutamine synthetase activity). Cadmium injection did not influence these parameters in the zinc-adequate groups. Extracellular superoxide dismutase activity was lower in the zinc-deficient group than in the zinc-sufficient groups; there was a trend (P < 0.06) for a lower activity in the cadmium- versus the saline-injected rats. These results support the concept that zinc deficiency increases the susceptibility of testes to cadmium-mediated free radical damage.     

Modulation by dietary factors of BHA-induced alterations in cell kinetics of gastro-intestinal tract tissues in rats.

To determine the effects of dietary ethanol or fibre on 2(3)-tert-butyl-4-hydroxyanisole (BHA)-induced alterations in cell kinetics in gastro-intestinal tract tissues, groups of six male Wistar rats were fed diets containing 0% (control) or 1.5% BHA for 2 wk. One group fed 1.5% BHA and one pair-fed control group received 10% ethanol in the drinking-water; two similarly fed groups received drinking-water only. Another group fed 1.5% BHA and a pair-fed control group received a diet supplemented with 20% cellulose; two similar groups received no fibre supplementation. Cell kinetics in the forestomach, glandular stomach and oesophageal tissue were determined, after 14 days, by bivariate 5-bromo-deoxyuridine/DNA analysis using immunocytochemistry and flow cytometry. In the fibre experiment, colorectal tissue was also examined. In both experiments the labelling indices in all the gastro-intestinal tract tissues were significantly altered in the BHA-fed groups compared with the corresponding control groups. In the ethanol experiment no statistically significant difference in the labelling indices was observed in the forestomach or glandular stomach between the two control groups or between the two BHA-fed groups. However, intake of ethanol-supplemented drinking-water induced increases in oesophageal labelling indices in rats fed a BHA-free diet. Thus 14 days of simultaneous ethanol administration has no effect on BHA-induced alterations in cell kinetics in the oesophagus, glandular stomach or forestomach of rats. In the forestomach and colorectal tissue, a high-cellulose diet resulted in a significant decrease in the BHA-induced elevation of labelling indices. Thus dietary cellulose provides a partial protection against the proliferation-enhancing effects of BHA in the rat gastro-intestinal tract.     

Long-term toxicity of ortho-toluenesulfonamide and sodium saccharin in the rat

In a two-generation lifetime feeding study, 50 male and 50 female weanling Sprague-Dawley rats were included in each of the following dietary treatment groups: control; 2.5 mg ortho-toluenesulfonamide (o-TS, more than 99.9% pure)/kg/day; 25 mg o-TS/kg/day; 250 mg o-TS/kg/day; 250 mg o-TS/kg/day with 1% NH₄Cl in the drinking water: or 5% sodium saccharin, which contained no detectable amount of o-TS. After 3 months on test, the F₀ rats were bred and 50 pups of each sex, from every group, were weaned onto the parental diets which both generations received for their lifetime. Rats from both generations fed diets providing 250 mg o-TS/kg with 1.0% NH₄Cl in the drinking water or containing 5% sodium saccharin had decreased growth rates, but only the former two had lowered feed consumption. There were no treatment-related effects upon reproduction, longevity, or hematological parameters. The animals were free of the bladder parasite Trichosomoides crassicauda. A few animals developed grossly visible bladder and kidney stones but the incidence of these was not treatment related. The incidence of bladder tumours in male rats fed the 5% saccharin diet was significantly increased in both generations compared to their respective control groups. An evaluation of individual feed consumption data for animals fed saccharin-containing diets did not reveal any statistical difference between the amount of feed consumed by animals which had bladder tumors and those that did not. The incidence of bladder tumors in the o-TS-treated groups and in the female rats fed the 5% saccharin diet was not significantly different from that in control animals.     

Chronic toxicity tests of pyrilamine maleate and methapyrilene hydrochloride in F344 rats

Methapyrilene hydrochloride was administered at levels of 125 or 250 ppm in the diet to groups of male and female F344 rats. The closely analogous antihistaminic drug pyrilamine, as the maleate, was given at 2000 ppm in the diet or at 2 g/litre drinking-water to groups of male and female F344 rats. Almost all of the rats given the higher dose of methapyrilene had either carcinomas or neoplastic nodules of the liver, whereas at 125 ppm 40% of the rats had neoplastic nodules in the liver. Among the 20 male and 20 female rats treated with pyrilamine maleate mixed into the diet, two males and two females had hepatocellular carcinomas and, in addition, five males and eight females had neoplastic nodules in the liver. The incidence of liver neoplasms in the rats given pyrilamine in the drinking-water did not differ from that in the untreated controls, of which five males and three females had neoplastic nodules in the liver.     

Effects of aminothiols in 2-acetylaminofluorene-treated rats. I. Damage and repair of liver DNA, hyperplastic foci, and Zymbal gland tumors

One hundred and seventy-nine male Wistar rats were divided into 6 groups and fed with a standard diet supplemented with 0.05% 2-acetylaminofluorene (2AAF) and/or 0.1% glutathione (GSH) or N-acetyl-L-cysteine (NAC). Each treatment cycle lasted for 3 weeks, followed by 1 week of standard meal. After 4 cycles, survival was 100% in the 3 control groups, and 86.0, 100 and 91.7%, in the groups receiving 2AAF, 2AAF plus GSH, and 2AAF plus NAC, respectively. After an additional 4-8 weeks, all the 5 surviving rats fed with 2AAF exhibited deforming ear tumors, which on histological examination were classified as sebaceous squamocellular carcinomas of Zymbal glands. No such tumors were detectable in control groups, nor in the 16 surviving rats fed with 2AAF plus GSH or NAC. In the liver, 2AAF produced significant DNA damage at the 3rd week of each cycle, which was partially repaired during the week of standard meal feeding. Moreover, 2AAF determined the appearance of gamma-glutamyl transpeptidase-positive foci, which tended to increase with time both in number and in size. GSH and NAC exerted similar protective effects on these phenomena, but only at early stages of the experimental model used.     

Enhanced lung toxicity of intratracheally instilled cadmium chloride in selenium-deficient rats

In order to further characterize biochemical correlates of CdCl₂-induced lung injury, CdCl₂ lung toxicity was studied in Se-deficient rats. A solution of 0.5 μmol/kg of CdCl₂ was intratracheally instilled into chronic respiratory disease-free 70-day-old rats fed a basal Se-deficient diet or a basal diet supplemented with 0.5 or 2.0 ppm of Se for 40 days. Within 3 days Se-deficient rats showed signs of marked respiratory distress and 25% had died, whereas rats from Se-supplemented groups all survived this time period. As evidenced by gross examination and lung water contents, Se-deficient rats exhibited a greater degree of inflammatory pulmonary edema than did rats from Se-supplemented groups. The results of the various enzymatic and biochemical determinations may be interpreted as being consistent with greater damage occurring in the Se-deficient rats.     

Beneficial effects of NTP-2000 diet on growth, survival, and kidney and heart diseases of Fischer 344 rats in chronic studies.

Diet is one of the most important environmental factors influencing growth, survival, and appearance of age-associated diseases in rodents. NIH-07 open formula rodent diet was the selected diet for the National Toxicology Program studies from 1980 to 1994. After a number of experimental diets were evaluated, a new one designated as NTP-2000 was selected for rodents in NTP studies beginning in 1994. This report summarizes the results of dosed feed and inhalation studies for differences in growth, survival, and severity of kidney and heart lesions in Fischer 344 rats fed NTP-2000 or NIH-07 diets. In the dosed feed studies, male rats group housed and fed the NTP-2000 diet grew slightly slower, attained maximum body weight later, and lost less body weight by the end of the 2-year studies compared to the groups fed NIH-07. Female rats group housed and fed the NTP-2000 diet in dosed feed studies had significantly slower growth, with lower maximum body weight compared to the groups fed the NIH-07 diet. In the inhalation studies, male rats individually housed and fed the NTP-2000 diet had slightly higher maximum body weight and significantly higher final body weight, with lower loss of weight when compared to similarly housed groups fed the NIH-07 diet. In inhalation studies, female rats fed the NTP-2000 diet and individually housed had significantly slower growth. The NTP-2000 diet significantly increased the survival of male and female rats, with a dramatic increase in survival of males in inhalation studies. This diet also caused significant decreases in severity of nephropathy and cardiomyopathy, and the decrease was marked in males. These observations indicate that diets balanced for nutrients, such as the NTP-2000, could markedly improve the health and increase survival of the rats used in chronic studies.     

Effect of peroxisome proliferators on glutathione-dependent sulphobromophthalein excretion.

1. The effect of pretreatment of rats with the peroxisome proliferator 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) on sulphobromophthalein excretion from the isolated perfused rat liver has been investigated and compared with the effect of clofibrate which is also a peroxisome proliferator. 2. Rats fed 2,4,5-T at a dose of 0.25% in the diet showed a decrease in food intake, compared with controls and clofibrate-fed rats. 3. Treatment with either 2,4,5-T or clofibrate was associated with significant inhibition of the biliary excretion of unchanged, conjugated, and total sulphobromophthalein from perfused rat liver, compared with diet-matched controls. 4. There was a decrease in bile flow in the clofibrate-treated group, but not in the 2,4,5-T-treated group. 5. The results of the present study confirm previous studies that have shown an association between peroxisome proliferation treatment and inhibition of glutathione S-transferase-mediated sulphobromophthalein excretion.     

The inhibitory effect of thioproline on carcinogenesis induced by N-benzylmethylamine and nitrite

Thioproline, which is readily nitrosated to form nitrosothioproline, is expected to act as a nitrite scavenger. The effect of thioproline as an inhibitor of the carcinogenesis induced by N-nitroso-N-benzylmethylamine precursors was examined. Two groups of male F-344 rats were given diet containing 0.25% N-benzylmethylamine (group I) or 0.25% N-benzylmethylamine plus thioproline (0.25% until wk 17 and then 0.5%; group II). Both groups were given drinking-water containing sodium nitrite (0.1% until wk 17 and then 0.2%). The experiment was continued for 717 days. Squamous cell carcinoma of the forestomach developed in six out of seven rats in group I and in significantly fewer, two out of nine rats, in group II. The degree of invasion by the tumours was also less in group II rats, given thioproline, than in group I. Thus thioproline suppressed carcinogenesis induced by N-benzylmethylamine and nitrite, possibly by inhibiting the in vivo nitrosation of N-benzylmethylamine.