Jeffrey Everitt

<jats:p>&lt;p&gt;HER2BOW transgene design&lt;/p&gt;</jats:p>

<jats:p>&lt;p&gt;Supplementary File 1.&lt;/p&gt;</jats:p>

<jats:p>&lt;p&gt;Long-term survival of an autochthonous p53/MCA mouse model in immunocompetent 129/SvJae wild-type mice induced by CRISPR/Cas9&lt;/p&gt;</jats:p>

Wy-14,643 was selected for inclusion in a series of studies on peroxisome proliferators because it is known to produce considerable peroxisome proliferation and hepatocarcinogenicity in rats. Male Sprague-Dawley rats were exposed to Wy-14,643 (greater than 98% pure) in feed for up to 3 months; male B6C3F1mice and male Syrian hamsters were exposed to Wy-14,643 in feed for 2 weeks or up to 3 months. Animals were evaluated for clinical pathology, plasma concentrations of Wy-14,643, reproductive system effects, cell proliferation and peroxisomal enzyme analyses, and histopathology. Single and multiple-dose toxicokinetic studies of Wy-14,643 were conducted in additional groups of male Sprague-Dawley and Wistar Furth rats, B6C3F1mice, and Syrian hamsters. Genetic toxicology studies were conducted in vivo in Tg.AC mouse peripheral blood erythrocytes. In the 2-week studies, groups of five mice were fed diets containing 0, 10, 50, 100, 500, or 1,000 ppm Wy-14,643 (equivalent to average daily doses of approximately 2 to 184 mg Wy-14,643/kg body weight). Groups of five hamsters were fed diets containing 0, 10, 100, 500, 1,000, or 5,000 ppm Wy-14,643 (equivalent to average daily doses of approximately 1 to 550 mg/kg). All animals survived to the end of the studies. The mean body weight gain of 500 ppm mice was significantly less than that of the controls; hamsters exposed to 100 ppm or greater lost weight during the study. Feed consumption by 500 ppm mice was greater than that by the controls. Liver weights of all exposed groups of mice and hamsters were generally significantly increased. In the 2-week studies, an increase in peroxisomal enzyme activity occurred in 10 ppm mice; increases in peroxisomal β-oxidation, carnitine acetyltransferase, catalase, and acyl CoA oxidase occurred in all exposed mice compared to controls. Significantly increased BrdU-labeled hepatocyte percentages occurred in 100 and 1,000 ppm mice and 500 and 5,000 ppm hamsters; peroxisomal β-oxidation of lipids was increased in all exposed groups of mice and hamsters. Gross lesions in the 2-week studies included liver foci in one 500 ppm mouse and one 1,000 ppm hamster and enlarged livers in one hamster in each of the 100 and 500 ppm groups and two 5,000 ppm hamsters. All 500 and 1,000 ppm mice had hepatocyte hypertrophy of the liver, and 1,000 ppm mice also had widespread individual cell necrosis. Minimal to mild multifocal vacuolation of the liver occurred in hamsters exposed to 500 ppm or greater. In the 3-month core studies, groups of 10 male rats, mice, or hamsters were fed diets containing 0, 5, 10, 50, 100, or 500 ppm Wy-14,643 (equivalent to average daily doses of approximately 0.3 to 34 mg/kg for rats, 0.9 to 135 mg/kg for mice, and 0.4 to 42 mg/kg for hamsters). Groups of 15 male rats, mice, or hamsters designated for special studies received the same concentrations of Wy-14,643 for up to 13 weeks. Groups of six male rats, 36 male mice, or 12 male hamsters designated for plasma concentration studies were fed diets containing 50, 100, or 500 ppm Wy-14,643 for up to 9 weeks. All core study animals survived to the end of the studies. Mean body weights were significantly decreased in all exposed groups except the 5 ppm groups and 10 ppm mice; hamsters in the 100 and 500 ppm groups lost weight during the study. Feed consumption by exposed rats and mice was generally similar to that by the controls; during week 14, hamsters exposed to 50 ppm or greater consumed slightly less feed than did the controls. The only clinical finding of toxicity was thinness of two 50 ppm and five 500 ppm hamsters. At all time points, the liver weights of exposed groups of core and special study rats, mice, and hamsters were generally significantly greater than those of the controls. Testis weights were significantly decreased in 500 ppm hamsters on day 34, in hamsters exposed to 5 ppm or greater at week 13 (special study), and in 100 and 500 ppm core study hamsters at the end of the study. In the sperm motility evaluation, the cauda epididymis weight of 500 ppm rats, epididymis weights of 100 and 500 ppm rats and mice, and the testis weight of 500 ppm mice were significantly less than those of the controls. For hamsters, cauda epididymis, epididymis, and testis weights; spermatid heads per testis; and spermatid counts were significantly decreased in all exposed groups evaluated for sperm motility. Epididymal spermatozoal motility and concentration in the 100 and 500 ppm groups and spermatid heads per gram testis in the 500 ppm group were also significantly decreased. Serum concentrations of estradiol were significantly decreased in all exposed groups of hamsters, and concentrations of testosterone and luteinizing hormone were decreased in groups exposed to 50 ppm or greater. At necropsy in the 3-month studies, liver foci were observed in three special study mice, including one 100 ppm mouse and one 500 ppm mouse on day 34 and one 100 ppm mouse at week 13. Liver discoloration and small testes were noted in 500 ppm hamsters on day 34, and hamsters exposed to 50 ppm or greater had enlarged livers and/or small testes at week 13 (special study) and at 3 months (core study). The incidences of cytoplasmic alteration in the liver were significantly increased in all exposed core groups of rats, mice, and hamsters; the severity of this lesion increased with increasing exposure concentration. The incidences of mitotic alteration of the liver in mice exposed to 50 ppm or greater and of liver pigmentation and oval cell hyperplasia in 500 ppm mice were significantly increased. Minimal regeneration of the corticomedullary junction of the renal tubule occurred in all exposed groups of rats. Significantly increased incidences of atrophy of the prostate gland, seminal vesicle, and testis occurred in 100 and 500 ppm hamsters. Degenerative myopathy of skeletal muscle was observed in the lumbar area and thigh of rats, mice, and hamsters and the lower leg of mice, primarily at 500 ppm. Following single-dose gavage exposure to Wy-14,643, plasma concentrations were generally higher in mice than in rats, which in turn were higher than those in hamsters. This pattern of plasma concentrations was usually attributed to high bioavailability in mice, medium bioavailability in rats, and low bioavailabilty in hamsters following an oral exposure to Wy-14,643. No increase in the frequency of micronucleated normochromatic erythrocytes was observed in the peripheral blood of male or female Tg.AC mice exposed to Wy-14,643 in feed or via dermal application for 6 months.

Nickel subsulfide is used in the manufacture of lidiium batteries and is a major component in the refining of certain nickel ores. Nickel subsulfide was nominated as part of a class study of nickel compounds, for which there was little information on the toxic and carcinogenic effects of inhalation exposure. Male and female F344/N rats and B6C3F, mice were exposed to nickel subsulfide (at least 97% pure; the mean value for the mass median aerodynamic diameter at each exposure concentration ranged from 2.0 to 2.2 μm) by inhalation 6 hours per day, 5 days per week, for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, and mouse peripheral blood samples were analyzed for frequency of micronucleated normochromatic erythrocytes. 16-DAY STUDY IN RATS Groups of five male and five female F344/N rats were exposed to atmospheres containing 0, 0.6, 1.2, 2.5, 5, or 10 mg nickel subsulfide/m3 (equivalent to 0, 0.44, 0.88, 1.83, 3.65, and 7.33 mg nickel/m3) 6 hours per day, 5 days per week for a total of 12 exposure days during a 16-day period. Additional groups of three male and three female rats were exposed to 0, 0.6, 2.5, or 10 mg/m3 for tissue burden studies. One male exposed to 10 mg nickel subsulfide/m3 in the core study died on day 14; all other rats survived until the end of the study. Final mean body weights and mean body weight gains of males exposed to 5 or 10 mg nickel subsulfide/m3 and females exposed to 2.5, 5, or 10 mg/m3 were significantly lower than those of the controls. Clinical findings of toxicity on day 5 of the study included labored respiration in 10 mg/m3 males and 5 and 10 mg/m3 females and dehydration in 5 and 10 mg/m3 females. Absolute and relative lung weights of 2.5, 5, and 10 mg/m3 males and all exposed groups of females were significantly greater than those of the controls, as was the absolute lung weight of 1.2 mg/m3 males. Inflammation of the lung and atrophy of the nasal olfactory epithelium occurred in all exposed groups. The concentrations of nickel in the lungs of exposed groups of rats increased with exposure concentration (males, 7 to 67 μg nickel/g lung; females, 9 to 77μg/g lung). 16-DAY STUDY IN MICE Groups of five male and five female B6C3F, mice were exposed to atmospheres containing 0, 0.6, 1.2, 2.5, 5, or 10 mg nickel subsulfide/m3 for 6 hours per day, 5 days per week for a total of 12 exposure days during a 16-day period. Additional groups of three male and three female mice were exposed to 0, 0.6, 2.5, or 10 mg/m3 for tissue burden studies. All male and female mice exposed to 10 mg nickel subsulfide/m3 in the core study died before the end of the study; the death of one female was accidental. One control male, one control female, and one 1.2 mg/m3 male also died before the end of the study. Final mean body weights and mean body weight gains of 5 mg/m3 males were significantly lower than those of the controls. Clinical findings at day 5 included labored respiration in 10 mg/m3 males and females. The absolute lung weight of 5 mg/m3 males, the absolute and relative lung weights of 10 mg/m3 males and 5 mg/m3 females, and the relative lung weight of 10 mg/m3 females were significantly greater than those of the controls. Inflammation of the lung occurred in 2.5, 5, and 10 mg/m3 male and female mice, fibrosis of the lung occurred 5 mg/m3 males and females, and lymphoid hyperplasia of the bronchial lymph nodes and atrophy of the nasal olfactory epithelium occurred in 1.2, 2.5, 5, and 10 mg/m3 males and females. Nickel concentrations in the lung of exposed male and female mice were greater than those of the controls (males, 10 to 20 μg nickel/g lung; females, 8 to 20 μg/g lung). 13-WEEK STUDY IN RATS Groups of 10 male and 10 female F344/N rats were exposed to atmospheres containing 0, 0.15, 0.3, 0.6, 1.2, or 2.5 mg nickel subsulfide/m3 (equivalent to 0, 0.11, 0.22, 0.44, 0.88, and 1.83 mg nickel/m3) 6 hours per day, 5 days per week for 13 weeks. Additional groups of 18 male and 18 female rats were exposed to 0, 0.15, 0.6, or 2.5 mg/m3 for tissue burden studies. All core study rats survived until the end of the study. Final mean body weights and mean body weight gains of 2.5 mg/m3 males were significantly lower than those of the controls; final mean body weights of all other exposure groups were similar to those of the controls. Chemical-related clinical findings included labored respiration in 2.5 mg/m3 males and females during weeks 2 through 7. In general, neutrophil and erythrocyte counts, hematocrit values, and hemoglobin concentrations were minimally increased in exposed rats. Absolute and relative lung weights of all exposed groups were significantly greater than those of the controls. Increases in the number of alveolar macrophages, interstitial infiltrates, or incidences of chronic inflammation of the lung occurred in all groups exposed to nickel subsulfide concentrations of 0.3 mg/m3 or greater; the severity of these lesions generally increased with increasing exposure concentration. Increases in the number of alveolar macrophages were observed in 0.15 mg/m3 males and females. Lymphoid hyperplasia of the bronchial and mediastinal lymph nodes was observed in rats exposed to 0.3 mg/m3 or greater. Most 0.6, 1.2, and 2.5 mg/m3 males and females had atrophy of the nasal olfactory epithelium, and the severity generally increased with increasing exposure concentration. Nickel concentrations in the lung increased with exposure concentration and were greater than those in the controls in rats exposed for 13 weeks (males, 5 to 18 μg nickel/g lung; females, 5 to 17 μg/g lung). 13-WEEK STUDY IN MICE Groups of 10 male and 10 female B6C3F1 mice were exposed to atmospheres containing 0, 0.15, 0.3, 0.6, 1.2, or 2.5 mg nickel subsulfide/m3 for 6 hours per day, 5 days per week for 13 weeks. Additional groups of six male and six female mice were exposed to 0, 0.15, 0.6, or 2.5 mg/m3 for tissue burden studies. Final mean body weights of all exposure groups were similar to those of the controls. No chemical-related clinical findings were observed. Lymphocyte counts in 1.2 and 2.5 mg/m3 males were minimally greater than that of the controls. Hemoglobin concentrations and erythrocyte counts in 0.3, 0.6, 1.2, and 2.5 mg/m3 females were minimally greater than those of the controls. Absolute and relative lung weights of 1.2 and 2.5 mg/m3 males and females were significantly greater than those of the controls. An increase in alveolar macrophages was present in mice from the 0.3 mg/m3 and higher exposure groups. Chronic inflammation and fibrosis were observed in the lung of 1.2 and 2.5 mg/m3 males and females. Interstitial infiltrates of lymphocytes were observed in mice exposed to 0.6, 1.2, or 2.5 mg/m3. Lymphoid hyperplasia of the bronchial lymph nodes was observed in groups exposed to 1.2 or 2.5 mg/m3. Atrophy of the nasal olfactory epithelium occurred in 0.6, 1.2, and 2.5 mg/m3 males and females, and incidences and severity generally increased with increasing exposure concentration. At 13 weeks, nickel concentrations in the lungs of exposed mice were greater than those of the controls (males, 3 to 17 μg nickel/g lung; females, 6 to 23 μg/g lung), and these concentrations increased with increasing exposure concentration. 2-YEAR STUDY IN RATS Survival, Body Weights, Clinical Findings, and Hematology Groups of 63 male and 63 female F344/N rats were exposed to 0, 0.15, or 1 mg nickel subsulfide/m3 (equivalent to 0, 0.11, or 0.73 mg nickel/m3) by inhalation for 6 hours per day, 5 days per week for 104 weeks. Survival of exposed males and female rats was similar to that of the controls. Mean body weights of males and females exposed to 0.15 mg/m3 were similar to those of the controls. Mean body weights of rats exposed to 1 mg/m3 were lower than those of the controls throughout the second year of the study. Chemical-related clinical findings included rapid and shallow breathing following exposure periods. Hematocrit values and hemoglobin concentrations in 1 mg/m3 males and females and the erythrocyte count in 1 mg/m3 males were mildly greater than those in the controls. Pathology Findings In general, the absolute and relative lung weights of exposed males and females were significantly greater than those of the controls at 7 and 15 months. There were exposure-related increases in the incidences of alveolar/bronchiolar adenoma in males, alveolar/bronchiolar carcinoma in males and females, and alveolar/bronchiolar adenoma or carcinoma (combined) in males and females at 2 years. Nonneoplastic lung lesions generally observed in exposed males and females included fibrosis; chronic active inflammation; focal alveolar epithelial hyperplasia, macrophage hyperplasia, and proteinosis; bronchial lymphoid hyperplasia; and interstitial inflammation. At 2 years, there were significant exposure-related increases in the incidences of benign pheochromocytoma, malignant pheochromocytoma, and benign or malignant pheochromocytoma (combined) in males and of benign pheochromocytoma in females. The incidence of adrenal medulla hyperplasia in 1 mg/m3 females was significantly greater than that of the controls. At 2 years, the incidences of chronic active inflammation of the nose in 1 mg/m3 females and of olfactory epithelial atrophy in 1 mg/m3 males and females were significantly greater than those of the controls. The incidences of lymphoid hyperplasia of the bronchial lymph node in exposed males at 7 and 15 months and in exposed males and females at 2 years were significantly greater than those of the controls. Incidences of macrophage hyperplasia in the bronchial lymph node of exposed males at 15 months and exposed males and females at 2 years were greater than those of the controls. Tissue Burden Analyses Nickel concentrations in the lung of exposed rats were greater than those of the controls at 7 months (males, 6 to 9 μg nickel/g lung; females, 6 to 9 μg/g lung) and 15 months (males, 4 to 3 μg nickel/g lung; females, 4 to 7 μg/g lung). 2-YEAR STUDY IN MICE Survival, Body Weights, Clinical Findings, and Hematology Groups of 80 male and 80 female B6C3F1 mice were exposed to 0, 0.6, or 1.2 mg nickel subsulfide/m3