2008

Isoproterenol-induced Cardiotoxicity in Sprague-Dawley Rats: Correlation of Reversible and

Irreversible Myocardial Injury with Release of Cardiac Troponin T (cTnT) and Roles of iNOS in
Myocardial Injury A Type I response, potential reversible myocardial alterations associated with slight
increases in serum cTnT. The second type of response. irreversible myocyte alterations, together with
significant increases in serum cTnT. By 48hrsThere was interstial fibrosis cTnT was in normal levels.
Increases in iNOS immunoreactivity correlated with the
lesion severity. These findings suggest that low doses of Iso exert complex effects on the myocardium
and that the generation of NO through increased expression of iNOS could be an important factor in the
pathogenesis of myocyte injury.

Fibro-osseous (FOL) and Degenerative Joint Lesions in Female Outbred NIH Black Swiss Mice
NIH Black Swiss mice (Cr;NIH BL[SJ) revealed a high inc ~dence of fibro-osseouslesions (POL; 89%)
and degenerative joint lesions (90%). FOL in female Black Swiss was often accompanied by reproduc
tive-tract lesions, including ovarian atrophy and uterine cervical dysplasia with hydrometra

Summary of Chemically Induced Pulmonary Lesions in the National Toxicology Program (NTP)
Toxicology and Carcinogenesis Studies The lung is the second most common target site of neoplasia of
chemicals tested by the National Toxicology Program (NTP). The most commonly induced lung tumors
were alveolar/bronchiolar (A/B) adenoma and/or carcinoma for both species (rats and mice). The liver
was the most common primary site of origin of metastatic lesions to the lungs of mice; however, skin was
most often the primary site of origin of metastatic lesions to the lungs of rats. site of origin of metastatic
lesions to the lungs of mice; however, skin was most often the primary site of origin of metastatic lesions
to the lungs of rats. In summary, AJB adenoma and carcinoma were the most frequently diagnosed
chemically induced tumors in the lungs of both rats and mice.

Spontaneous and Age-Related Testicular Findings in Beagle Dogs Hypospermatogenesis,

characterized by randomly scattered missing spermatids and/or spermatocytes within seminiferous
tubules, was observed in 75% of dogsAtrophy/hypoplasia of seminiferous
tubules, Retained spermatids, multinucleate giant cells, intracytoplasmic vacuoles (presumably in Sertoli
cells), and swollen spermatocytes were common findings of minimal severity. If evaluation
ofspermatogenesis is critical, the incidental findings can be minimized by using males over twelve
months of age.

Characterization of Age- and Gender-related Changes in the Spleen and Thymus from Control
Cynomolgus Macaques Used in Toxicity Studies The thymus had a trend toward increased interlobular
fat infiltration with increasing age in both males and females. Thymic delineation of the cortex and
medulla was significantly decreased in the seven-to-fifteen-years age group for males only. The cortex-to-
medulla ratio was significantly lower only in males in the seven-to-fifteen-years age group. B

Histopathology of the Urinary Bladders of Cynomolgus Monkeys Treated with PPAR Agonists The
urinary bladder in the monkey has two very distinctive features that are not seen in rodents, dogs, or
humans. The first of these is the presence of eosinophilic granules. Second, the monkey bladder has a
wide range in the number of mononuclear inflammatory cells normally present in the submucosa and
urothelium.

Systemic Inflammatory Response Syndrome in Nonhuman Primates Culminating in Multiple

Organ Failure, Acute Lung Injury, and Disseminated Intravascular Coagulation
The systemic inflammatory response syndrome (SIRS) is a clinicopathological manifestation of
overexuberant acute-phase inflammation caused by infectious or noninfectious etiologies. The diagnosis
of SIRS should be considered when evaluating nonhuman primate (NHP) cases ofMOP manifesting with
histological evidence of vascular leakage.
2009

Carcinogenic Effects of MGP-7 and BjajP on the Hamster Cheek Pouch

The short-term study showed that B[a]P induced p53 protein accumulation, indicative of genotoxic
damage, as well as increased cell proliferation, hyperplasia, and inflammation, which is usually associated
with promotional activity. In contrast, the MGP-7 presented only marginal p53 accumulation and
induction of BrdU incorporation. In the long-term experiments, all the animals treated with B[a]P had a
100% incidence of squamous cell carcinoma (SCC). MGP-7 showed only weak hyperplasia and a low
proliferation rate and accumulation of p53 protein through long-term treatment.

Spontaneous Cardiomyopathy in Cynomolgus Monkeys (Macaca Fascicularis) Microscopically, the

apical to mid-ventricular myocardium to subendocardium had foci of cardiomyocyte disarray with
cytoplasmic pallor to stippling and karyomegaly, vacuolization of the perimyseal connective tissue, a
meshwork of fibrous tissue that concentrated around medium-sized blood vessels and dissected between
or less often replaced affected cardiomyocytes; and a minimal, predominantly macrophage infiltrate. The
features were reminiscent of chronic eatecholamine-induced experimental cardiomyopathy in monkeys.

Histopathological Features of Capillaria hepatica Infection in Laboratory Rabbits

Capillaria hepotica is a nematode parasite of wild rodents and other mammals. Large granulomas
(macrogranulomas) were evident in portal areas and involved the bile ducts. Macrogranulomas contained
collections of characteristic C. hepatica eggs, macrophages, eosinophils, and lymphocytes. Small
granulomas (microgranulomas), characterized by epithelioid macrophages surrounded by lymphocytes
and eosinophils, were also identified. C. hepotica eggs were also observed in the lumina of the bile ducts
and gall bladder. No adult C. hepatica worms were identified.
Life cycle:
After ingestion by the definitive (i.e., final) host, the embryonated egg containing the infective first-stage
larva (L1) hatches in the small intestine or The released L1 larva penetrates the intestinal mucosa and
passes to the liver via the portal or mesenteric veins. The L1 larvae are found in the liver at day 2/3 post-
infection. All the moults of the larval stages occur in the liver: L1 to L2 (160 to220 urn long) at day 3-7
post-infection; L2 to L3 (270 to670 urn long) at four or five days post-infection; and L3 to L4 (1100 to
3600 um long) at seven to sixteen days after infection. The moulting of the L4 larva to produce the male
or femaleL5 larva occurs at eighteen to twenty days post-infectionThe fertilized adult female worm
moves through a syncytium of host liver cells depositing clusters of uncleaved eggs; these eggs
subsequently become enclosed by the host in localized granulomas (macrogranulomas)

Strain-related Differences in Urine Composition of Male

Rats of Potential Relevance to Urolithiasis HSD rats had significantly higher urine calcium than SD or
WI rals, primarily as calcium phosphate-containing precipitate. When compared to SD rats, HSD rats had
lower urine volume, higher urine protein, and a comparable (week 4) to lower (week13) burden of
MgNH4PO4 aggregates. Relative to WI rats, HSD rats had higher urine protein and magnesium and
lower serum and urine citrate. Overall, the susceptibility to urolithiasis in male rats was HSD> SD > WI;

Phytol-induced Hepatotoxicity in Mice Phytanic acid is a branched-chain, saturated fatty acid present in
high concentrations in dairy products and ruminant fat. Some other dietary fats contain lower levels of
phytol, which is readily converted to phytanic acid after absorption. Phytanic acid is a peroxisome
proliferator binding the
nuclear transcription factor peroxisome proliferator-activated receptor a (PPA.Rre) to induce expression
of genes encoding enzymes of fatty acid oxidation in peroxisomes and mitochondria. Administration of
dietary phytol (0.5% or 1%) to normal mice for twelve to eighteen days caused consistent PPARrc-
mediated responses, such as lower body weights, higher liver weights, peroxisome proliferation,
increased catalase expression, and hepatocellular hypertrophy and hyperplasia. Female mice fed 0.5%
phytol and male and female mice fed 1% phytol exhibited midzonal hepatocellularnecrosis, periportal
hepatocellular fatty vacuolation, and corresponding increases in liver levels of the phytol metabolites
phytanic acid and pristanic acid. These results suggest
that phytol may cause selective midzonal hepatocellular necrosis in mice, an uncommon pattern of
hepatotoxie injury, and that the greater suscept ibility of female mice may reflect a lower capacity to
oxidize phytanic acid because of their intrinsically lower hepatie expression of sterol carrier protein-x
(SCP-x) males have 5X.

Cardiac Valvular Pathology: Comparative Pathology and Animal Models of Acquired Cardiac Valvular
Disease.
Mouse: Spontaneous valve disease has been described in the Swiss CD-i mouse. The alteration
was usually on the free edge of the leaf. The pulmonic valve was commonly affected. In the rat, the
incidence was greatest in the mitral valve. In the Dog, The condition is referred to as myxomatous
atrioventricular valvular degeneration, chronic valvular fibrosis, or endocardiosismost common in the
mitral valve, inflammation is rare, Hemorrhage and mineralization may be present, No valvular disease in
populations of laboratory nonhuman primates has not been reported. rheumatic heart disease in rats is
associated with streptococcal M protein. Carcinoid heart disease occurs in most patients with carcinoid
syndrome, a metastatic neoplasm of serotonin-producing gastrointestinal endocrine cells, and is
characterized by fibrous thickening of cardiac valves. [LDLr(-/-)ApoB(l00/l00)] develop aortic valve
calcification. Watanabe rabbits and diet-induced hypercholesterolemic and atherosclerotic
models in wild-type rabbits. Valvular disease, primarily aortic valvular stenosis, has been described.

Evaluation of Subchronic Toxicity of Pet Food Contaminated with Melamine and Cyanuric Acid in
Rats: food contained significant levels of cyanuric acid and melamine in a ratio of 1:6.8. Rats fed the diet
of 50%–100% for three months exhibited elevated serum BUN and creatinine, as well as dose-dependent
melamine/cyanuric acid crystal-induced nephrotoxicity. In conclusion, the combination ratio of cyanuric
acid to melamine and the acidic urine content were two factors that, upon repeated exposure, determined
the severity of the nephrotoxicity. Numerous MC crystals of various sizes were mixed with the necrotic
cell debris in both proximal and distal renal tubules. Slight to severe inflammatory cell infiltration was
accompanied by renal tubular dilation and epithelial cell regeneration with interstitial fibrosis.

Dietary Fat–Influenced Development of Colon Neoplasia in ApcMin Mice Exposed to

Benzo(a)pyrene: Consumption of well-done red meat and saturated fats rich in polycyclic aromatic
hydrocarbons may be one of the causative factors for sporadic colon cancer. ApcMin is a mutant mouse
with multiple intestinal neoplasias. It has a mutated adenomatous polyposis coli (Apc), a tumor-
suppressor gene, similar to that in patients with familial adenomatous polyposis. ApcMin mouse mimics the
rapid development of adenomatous polyps that affect humans. Among the several environmental
toxicants, benzo(a)pyrene (B[a]P), a polycyclic aromatic hydrocarbon (PAH) compound, has generated
the most interest, as this chemical is formed in red meat cooked at high temperatures. Treatment consisted
of 50 and 100 µg B(a)P/kg body weight dissolved in peanut or coconut oil (representatives of unsaturated
and saturated fats, respectively. An increased prevalence of adenomas in colons of mice that ingested
B(a)P through saturated dietary fat compared to unsaturated fat and controls

The Metrial Gland in the Rat and Its Similarities to Granular Cell Tumors (Tol Pat l 2009; 37; 474)
Metrial glands are normal structures located in the mesometrial triangle of the pregnant rat uterus from
gestational day (GD) 8 through termination of pregnancy. Metrial glands are composed of a dynamic
mixed cell population of granulated metrial gland (GMG) cells, endometrial stromal cells,
trophoblasts, blood vessels, and fibroblasts. GMG cells are approximately 40 um in diameter and have
abundant distinct eosinophilic glycoprotein granules that are periodic acid-Schiff (PAS) stain positive and
diastase resistant. GMGs are frequently binucleated, exhibit some mitotic activity, and can have a high
rate of apoptosis. Positive immunohistochemical staining for perforin.GD 8-GMG cells are distributed as
isolated, individualized cells located in the mesometrial triangle. GD 11-GMG cells become clearly
organized into angiocentric aggregates. GD 15-GMG cells increase in number; there is vascular
remodeling and increased vascularity within the gland; the metrial gland reaches maximum size.
Thereafter, GMG cells undergo apoptosis, retreat to a peripheral location in the gland, and allow for
interstitial and endovascular trophoblasts to invade into the central part of the metrial gland.

Spontaneous Hibernomas in Sprague-Dawley Rats (Toxicol Pathol 2009; 37; 547) Hibernomas are
rare neoplasms originating in brown adipose tissue of humans and other animal species, including
laboratory animals. Between April 2000 and April 2007, however, sixty-two hibernomas (an overall
prevalence of 3.52%) were observed in a total of 1760 Sprague-Dawley rats assigned to three
carcinogenesis bioassays at two separate research laboratories. Tumors (29 benign and 33 malignant)
were randomly distributed among test article–treated and control groups and were considered to be
spontaneous. Most tumors originated in the thoracic cavity, and they were usually described as soft,
mottled to tan masses with nodular to lobulated profiles. Immunohistochemical procedures for uncoupling
protein 1 (UCP1) confirmed brown adipose tissue as the site of origin rather than white fat. The marked
increase in hibernomas in our studies suggests that greater numbers of spontaneous hibernomas may be
sporadically encountered in future carcinogenesis studies with Sprague-Dawley rats.

Chronic cor pulmonale secondary to pulmonary atherosclerosis in an African Grey parrot: The
heart was globoid appearance with marked dilatation of the right ventricle. The ratio of the thickness of
the right ventricular free wall to thickness of the left ventricular free wall was approximately
1:1.Ascending aorta and main pulmonary artery had diffuse and irregular thickening with lumens
narrowing. The liver was diffusely pale gray in color but of normal consistency. HIsto: Arterial
atheroscelosis, myocardial fibrosis, hepatic sinusoidal congestion.

Nephrotic Syndrome Induced by Dibasic Sodium Phosphate Injections for Twenty-eight Days in
Rats: Sprague-Dawley rats received once daily tail-vein injections of 360 mM dibasic sodium phosphate
solution at 8 mL/kg for fourteen or twenty-eight days. Clinical examination revealed persistent proteinuria
and severe proteinuria from eight days. Animals developed lipemia, hypercholesterolemia, anemia, higher
serum fibrinogen levels, and lower serum albumin/globulin ratios on day 29. Histopathologically,
mineralization in whole glomeruli, glomerular capillary dilatation, partial adhesion of glomerular tufts to
Bowman’s capsule, and mesangiolysis.; tubular regeneration and dilatation, protein casts, mineralization
in the basement membrane, focal interstitial inflammation, and fibrosis in the cortex. These clinical and
morphological changes were similar to features of human nephrotic syndrome.

Eosinophilic Inclusions in Rat Clara Cells and the Effect of an Inhaled Corticosteroid: Large
eosinophilic cytoplasmic inclusions (ECIs) are occasionally seen in untreated rat Clara cells. Following
inhalation exposure to a corticosteroid, the number of ECIs was increased. . ECIs were strongly positive
for surfactant protein D and weakly positive for Clara cell specific protein. The ECI increased with age
and dose of corticosteroid.

N-Methyl-N-Nitrosourea (MNU): A Positive Control Chemical for p53+/– Mouse Carcinogenicity

Studies: A single injection of MNU produces a high incidence of thymic malignant lymphoma with
frequent involvement of spleen, lymph nodes, liver, and other parenchymal organs.
Temporal Profile of Clinical Signs and Histopathologic Changes in an F-344 Rat Model of Kainic
Acid–induced Mesial Temporal Lobe Epilepsy: Kainic acid (KA)-induced F-344 rat is used as model
of mesial temporal lobe epilepsy (Most common type of temporal lobe epilepsy in humans). Neuronal
degeneration occurred on Day 3 in the hippocampal CA1, CA3 (lesions in CA1 occurred before CA3,
similar to human condition), dentate hilar regions; amyg-daloid and thalamic nuclei. KA is anlog of L-
glutamate and an agonist of ionotropic, non-NMDA glutamate AMPA and KA receptors.

Spontaneous Aortitis in the Balb/c Mouse. Balb/c mice have been widely used in genetic, tumor,
andimmunological studies and as an animal model for induction of vasculitis 20 underwent ligation of the
femoral artery; incidences of aortic root inflammation were compared to those observed in controls..
Although a cumulative incidence of 12.5% was found, no difference was seen in the overall incidence
rates between the control and the surgically treated groups. The presence of background vascular lesions
in this mouse strain is of special interest following its use in models for inducing a vascular lesion.
Femoral artery ligation did cause an increase in blood levels of the inflammatory cytokine IL-6, but this
finding was not correlated with an increase in the incidence of vasculitis.