A new 90-day drinking water study of sodium cyanide in rats to further evaluate National Toxicology Program findings and inform risk assessment.

BACKGROUND: The National Toxicology Program (NTP, 1993) reported male reproductive effects in a sodium cyanide (NaCN) drinking water study. The critical effect, decreased cauda epididymis weights, was used by U.S. Environmental Protection Agency for their hazard characterization and risk assessment of hydrogen cyanide and cyanide salts. To further investigate potential male reproductive effects, we conducted a new 90-day drinking water study. METHODS: Our study expanded evaluations of testes and thyroid. Male F344 rats received NaCN in drinking water at 0, 0 (water restricted; paired to top dose), 3, 10, 30, 100, and 300 ppm for 13 weeks, followed by 10-weeks recovery. RESULTS: Plasma thiocyanate increased dose-dependently but returned to baseline during recovery. NaCN caused neither effects on survival, body weight, food consumption, hematology, serum chemistry, urinalysis, thyroid hormones, testes or epididymides weights, sperm motility/viability, sperm morphology, or sperm production; nor clinical, ophthalmic, or histopathologic findings. Increased organ weights in thyroid/parathyroid and liver occurred at 300-ppm but were recoverable. No changes occurred in male reproductive organs. CONCLUSIONS: Absent adverse effects, the NOAEL was 300 ppm (21.66 mg/kg/day; highest dose tested). Based on organ weight increases at 300 ppm, the NOEL was 100 ppm (7.46 mg/kg/day).

A 90-day oral (dietary) toxicity and mass balance study of corn starch fiber in Sprague Dawley rats.

The potential toxicity of corn starch fiber was assessed and compared to polydextrose, a commonly used bulking agent with a long history of safe use in the food supply. Groups of male and female Crl:CD(SD) rats were fed 0 (control), 1,000, 3,000, or 10,000 mg/kg-bw/day corn starch fiber in the diet for 90 days. The polydextrose reference article was offered on a comparable regimen at 10,000 mg/kg-bw/day. Following a single gavage dose of [14C]-corn starch fiber on study day 13 or 90, the mass balance of the test article was assessed by analysis of excreta samples collected from 0 to 168 h post-dose. There were no toxicologically or biologically relevant findings in any of the test article-treated groups. The few minor differences observed between the corn starch fiber and polydextrose exposed groups were considered to be due to normal biological variation. Following [14C]-corn starch fiber dosing, nearly complete excretion of the administered dose occurred over 168 h post-dosing, with the majority excreted in the feces. The dietary no-observed-adverse-effect level of corn starch fiber after 90 days was 10,000 mg/kg-bw/day. Similar toxicity profiles for corn starch fiber and polydextrose were observed due to the structural and compositional similarities of these materials.

A 90-day dietary study of a (2R,4R)-monatin salt in Beagle dogs.

(2R,4R)-Monatin salt (Na/K) [sodium/potassium (2R,4R)-2-amino-4-carboxy-4-hydroxy-5-(3-indolyl) pentanoate, hereafter "R,R-monatin"] was administered in the diets of groups of Beagle dogs (4/sex/group) at concentrations of 0 (basal diet), 5000, 20,000, or 35,000 ppm for 13 weeks. There were no effects on survival, clinical observations, body weight and body weight gain, feed consumption and feed efficiency, functional observational battery, ophthalmic examination, and electrocardiographic evaluation. No adverse effects on hematology, serum chemistry, and urinalysis parameters were reported. A statistically significant decrease in testicular weights associated with germ cell hypocellularity and reduced luminal sperm in the epididymides was reported in all treated male groups. Based on these findings, the dietary no-observed-adverse-effect level (NOAEL) of R,R-monatin for 90 days was considered 35,000 ppm for female dogs (approximately 1101 mg/kg bw/day) and <5000 ppm for male dogs (approximately <151 mg/kg bw/day).

Airway trefoil factor expression during naphthalene injury and repair.

While the role of trefoil factors (TFF) in the maintenance of epithelial integrity in the gastrointestinal tract is well known, their involvement in wound healing in the conducting airway is less well understood. We defined the pattern of expression of TFF1, TFF2, and TFF3 in the airways of mice during repair of both severe (300 mg/kg) and moderate (200 mg/kg) naphthalene-induced Clara cell injury. Quantitative real-time PCR for tff messenger RNA expression and immunohistochemistry for protein expression were applied to airway samples obtained by microdissection of airway trees or to fixed lung tissue from mice at 6 and 24 h and 4 and 7 days after exposure to either naphthalene or an oil (vehicle) control. All three TFF were expressed in normal whole lung and airways. TFF2 was the most abundant and was enriched in airways. Injury of the airway epithelium by 300 mg/kg naphthalene caused a significant induction of tff1 gene expression at 24 h, 4 days, and 7 days. In contrast, tff2 was decreased in the high-dose group at 24 h and 4 days but returned to baseline levels by 7 days. tff3 gene expression was not significantly changed at any time point. Protein localization via immunohistochemistry did not directly correlate with the gene expression measurements. TFF1 and TFF2 expression was most intense in the degenerating Clara cells in the injury target zone at 6 and 24 h. Following the acute injury phase, TFF1 and TFF2 were localized to the luminal apices of repairing epithelial cells and to the adjacent mesenchyme in focal regions that correlated with bifurcations and the bronchoalveolar duct junction. The temporal pattern of increases in TFF1, TFF2, and TFF3 indicate a role in cell death as well as proliferation, migration, and differentiation phases of airway epithelial repair.

Abdominal fluid from a dog.

A 2-year-old intact female mixed-breed dog with a 1-month history of lethargy and anorexia was evaluated for abdominal distension and an abdominal mass. The dog's last heat cycle, her third, was 1 month prior to presentation, and no reproductive cycle abnormalities were noted at any time. Hematologic and serum biochemical abnormalities were consistent with hemorrhage and inflammation. Ultrasonographic examination confirmed a large midabdominal mass and a moderate amount of abdominal fluid. Cytologically, the fluid showed evidence of pyogranulomatous inflammation, hemorrhage, and mesothelial reactivity, as well as ciliated columnar cells and free cilia that were interpreted as likely of oviductal origin. The mass was removed surgically, and the histopathologic interpretation was oviductal hamartoma with marked stroma formation and acute hemorrhage. To the authors' knowledge, this is the first reported case of oviductal hamartoma in any species and the first reported case detailing the finding of ciliated columnar epithelial cells in the abdominal fluid of a dog. Ciliated columnar epithelial cells in abdominal fluid should be considered indicative of a likely underlying oviductal lesion.