Delwin L. Bokelman

Spontaneous tumors in control F344 and charles river-CD rats and Charles River CD-1 and B6C3HF1 mice.

The incidence of spontaneous neoplasms in outbred, inbred and F1 hybrid strains was compared using the Charles River-CD rat and mouse, the F344 rat, and B6C3HF1 mouse. These strains are commonly used in carcinogenic studies. Each strain has a consistent pattern of tumor occurrence; testicular, pituitary and lymphoreticular neoplasms are common in F344 rats, mammary and pituitary neoplasms are common in Charles River-CD rats, liver neoplasms are uncommon in CD-1 mice, while hepatic tumors are frequent in male B6C3HF1 mice. There is considerable variation in tumor incidence in individual studies regardless of strain and there appeared to be greater variation in incidence between laboratories using the same strain than in the different laboratories using unlike strains. Therefore, the choice between these strains may be fortuitous or recommended by governmental agencies. Regardless of the strain selected, it is vital to develop sufficient historical tumor data on the strain used at the particular test laboratory.

Association between Adverse Maternal and Embryo-Fetal Effects in Norfloxacin-Treated and Food-Deprived Rabbits

Norfloxacin is a new antibiotic which caused embryo-fetal toxicity in association with maternotoxicity when given orally to rabbits at 100 mg/kg/day. The intestinal flora of rabbits is unusually sensitive to many antibiotics and it was suspected that the maternotoxicity and embryo-fetal toxicity caused by oral norfloxacin were secondary to an effect on the intestinal flora. To test this idea, a teratologic study was conducted in which rabbits were dosed on Days 6 to 18 of gestation with norfloxacin given orally at 100 mg/kg/day or subcutaneously at 20 mg/kg/day. The oral treatment caused decreased food consumption (to less than 15 g/day in some animals), body weight loss, an increased resorption rate, and decreased fetal weight. Among the females in the orally dosed group, there was a significant correlation (p less than or equal to 0.005) between the effects on maternal body weight and the resorption rate. The subcutaneous treatment caused little intestinal exposure (biliary excretion = only 2-4% of dose) and no maternotoxicity or embryo-fetal toxicity, even though blood levels of drug were at least as high as those in the oral group. Since the maternotoxicity and embryo-fetal toxicity were specific to the oral route and not correlated with the level of systemic exposure, the maternotoxicity may have been secondary to an effect on the intestinal flora and the embryo-fetal toxicity may have been secondary to the maternotoxicity. The decreased food consumption observed in the oral group may have contributed to the embryo-fetal toxicity since, in a separate study, it was found that lowering the amount of food provided to rabbits on Days 6 to 18 of gestation from 150 g/day to 50 or 15 g/day also caused adverse maternal and fetal effects including, at 15 g/day, fetal malformations.

Qualitative and quantitative evaluation of prostatic histomorphology in rats following chronic treatment with finasteride, a 5-alpha reductase inhibitor.

OBJECTIVE To determine any potential direct and/or indirect effects of elevated intraprostatic T levels on the prostates of rats chronically (1-2 years) exposed to high doses (160 mg/kg/day) of finasteride, a selective inhibitor of 5-alpha reductase. METHODS Sprague-Dawley male rats were administered daily finasteride by oral gavage. Prostates from all rats were weighed, fixed in 10% neutral buffered formalin, and processed for light microscopic examination. The volume fractions of the prostatic glandular and stromal compartments were quantitated by morphometric analysis. RESULTS Administration of finasteride at doses of 20, 40, and 80 mg/kg/day for one year resulted in a significant (P < or = 0.05) decrease in prostatic weight; prostatic atrophy was evident by light microscopy. Morphometric analysis of the prostate showed that chronic finasteride administration resulted in a significant (P < or = 0.001) decrease in the absolute volume of both glandular (-65.2%) and stromal (-57.1%) compartments of the prostate. Furthermore, the total number of epithelial and stromal cells per gland were significantly (P < or = 0.002) decreased in finasteride-treated rats compared with vehicle controls; the magnitude of mean decrease was 69.8 percent and 50.6 percent of controls in epithelial and stromal cells, respectively. In addition, prostates from all two hundred fifty rats in a two-year study were qualitatively evaluated by light microscopy. Administration of finasteride at doses ranging from 2.5 mg/kg/day to 160 mg/kg/day for two years did not result in an increase over the background incidence of prostatic focal hyperplasia or adenoma. No malignant tumors of the prostate were seen in any of the groups. CONCLUSIONS These studies have demonstrated that the expected pharmacologic effects of finasteride on the prostate are maintained following chronic treatment and that there was no evidence of a direct and/or an indirect effect of elevated intraprostatic T on prostatic morphology in rats.

Maternotoxicity and fetotoxicity of an angiotensin-converting enzyme inhibitor, enalapril, in rabbits

When enalapril, an angiotensin-converting enzyme (ACE) inhibitor, was orally administered to inseminated rabbits at dosages of 0.1 to 30 mg/kg/day for 13 days in a range-finding study, nephrotoxicity, as measured by elevated serum urea nitrogen concentrations, occurred at 1 mg/kg/day and higher dosages and significant (p less than or equal to 0.05) increases in fetal wastage were observed at dosages as low as 3 mg/kg/day. Saline supplementation during treatment prevented this rise in urea nitrogen. Fetal wastage was significantly (p less than or equal to 0.05) increased in the absence of maternotoxicity when saline-supplemented females were treated with enalapril at 30 mg/kg/day. A developmental toxicity study of enalapril in saline-supplemented rabbits produced no evidence of teratogenicity at 3, 10, and 30 mg/kg/day. The period of sensitivity of fetuses to the toxic effects of enalapril was found to be limited to middle-to-late gestation (Gestational Days 14-27). A single oral dose of enalapril (30 mg/kg) on Day 26 of gestation resulted in 100% fetal deaths. On the basis of the work done by Broughton Pipkin et al. [1982, J. Physiol. (London) 323, 415-422] and Broughton Pipkin and Wallace (1986, Brit. J. Pharmacol. 87, 533-542), which demonstrated that the sheep fetus becomes markedly hypotensive when the dam is treated with captopril or enalapril during late pregnancy, we believe that the observed fetotoxicity of enalapril in rabbits is also due to fetal hypotension.(ABSTRACT TRUNCATED AT 250 WORDS)

Potassium loss as a causative factor for skeletal malformations in rats produced by indacrinone: A new investigational loop diuretic

Indacrinone (MK-196), an investigational loop diuretic, was administered to female CRCD rats from Days 6 to 17 of gestation at doses of 40, 80, or 120 mg/kg/day. Dose-related increases in wavy ribs and malformations of the scapula and humerus were apparent in fetuses from all drug-treated groups. Studies on the individual enantiomers of MK-196 showed that the teratogenic potency was associated with the relative saluretic activity of the enantiomers. Furosemide, a widely used diuretic with pharmacologic activity similar to that of MK-196, was administered to rats on Days 6 to 17 at doses of 37.5, 75, 150, or 300 mg/kg b.i.d. Dose-related increases in wavy ribs occurred in all drug treatment groups. At 150 mg/kg b.i.d. there were malformations in the scapula and humerus identical to those described for MK-196. Providing a 1% solution of KCl during the dosing period reduced the incidence of wavy ribs in fetuses from MK-196-treated dams by 90% and eliminated the scapular and humeral malformations. Coadministration of amiloride was also found to antagonize the teratogenicity of MK-196. The results indicate that high doses of diuretics with similar pharmacologic activities can produce identical skeletal malformations in rats, and that this teratogenicity is related to hypokalemia.

The toxicity of a fluorinated-biphenyl HMG-CoA reductase inhibitor in beagle dogs.

L-645, 164, a potent inhibitor of hydroxymethylglutarylcoenzyme A (HMG-CoA) reductase, is a structurally unique, synthetic monofluorinated-biphenyl that was administered to beagle dogs at dosages of 2, 10, or 50 mg/kg/day for 14 weeks to evaluate its toxic potential. Previously tested HMG-CoA reductase inhibitors from this laboratory have either been semisynthetic or fermentation-derived products containing a hexahydronaphthalene ring structure (i.e., lovastatin and simvastatin). Administration of L-645, 164 produced a significant spectrum of lesions, some of which have been previously associated with compounds of this pharmacological class, while others were unique to this monofluorinated-biphenyl inhibitor. Subcapsular lenticular opacities were produced in six of eight of the dogs receiving 50 mg/kg/day of L-645, 164 within 8 weeks of dosing. One dog receiving this dosage level experienced increases in serum alanine aminotransferase activity to levels 10 times those in concurrent control dogs. Light and electron microscopy of a wedge biopsy obtained within 3 days of this transaminase elevation failed to reveal any significant changes and the elevation resolved spontaneously despite continued drug administration. Lesions of the optic nerve and acoustic-vestibular tract and trapezoid decussation were observed in several dogs receiving 50 mg/kg/day. In addition, similar changes were observed in the optic tract in several of the dogs receiving 50 mg/kg/day and in one dog receiving 2 mg/kg/day of L-645,164. These were unique to L-645,164 and have not been observed after the administration of other HMG-CoA reductase inhibitors in this laboratory. Optic tract changes were generally mild, consisting of small to medium vacuoles without apparent myelin loss. Lesions in the other areas ranged from very slight to prominent vacuolation. No clinical signs were observed. Peak plasma drug levels of L-645,164 at 50 mg/kg were greater than 5 micrograms/ml, about one order of magnitude greater than those attained after administration of pharmacologically equipotent doses of lovastatin and simvastatin. These findings support previous observations that HMG-CoA reductase inhibitors producing high plasma drug levels are associated with a significant degree of systemic toxicity. In addition, the drug-induced CNS lesions attributed to L-645,164 appear also to be related to its chemical structure since similar lesions have not been observed after the administration of other structurally unrelated HMG-CoA reductase inhibitors that produce high plasma drug concentrations and comparable degrees of serum cholesterol lowering.

Liver function: Postprandial urea nitrogen elevation and indocyanine green clearance in the dog

Abstract A method for evaluation of liver function, based on the postprandial elevation of urea nitrogen in the serum of beagles is described. Biopsy specimens were obtained and alkaline phosphatase, serum glutamic-oxalacetic transaminase, and indocyanine green clearance also were determined after the induction of liver injury by carbon tetrachloride or ethionine. The postprandial elevation of urea nitrogen was significantly reduced by both agents. In ethionine-treated animals the depression of postprandial urea nitrogen elevation was the most effective of the clinical diagnostic tests indicating the onset of hepatotoxicity. Indocyanine green clearance, however, was the most satisfactory test for determining residual hepatic damage during the recovery studies after carbon tetrachloride administration.

Exposure of Rats to Lead Nitrate in utero or Postpartum; Effects on Morphology and Behavior

Female rats were administered lead (Pb), as the nitrate salt, on day 17 of pregnancy (5 or 25 mg/kg i.v.) or throughout lactation (5 or 25 mg/kg/day p.o.). There were adverse effects on weights of females receiving Pb on day 17. At 25 mg/kg i.v. gestation was significantly prolonged. In both groups treated intravenously, average pup weight on day 1 postpartum was significantly reduced and there was a significantly higher mortality than in controls, and hydrocephalus occurred. Survival rate and weight gain of pups from dams that received Pb throughout lactation was not different from controls. Brain weights and histomorphology of all groups was normal. Behavior in male offspring, as measured by open-field activity, rotorod or passive avoidance tests, was unaffected by exposure to Pb.