An extraskeletal osteosarcoma in the auricle of a Wistar Hannover rat.

An extraskeletal osteosarcoma was detected in the auricle of a 110-week-old female Wistar Hannover rat. Grossly, the tumor, measuring 15 mm in size, was observed in the subcutis as a solid and hard mass. Histologically, the majority of the mass comprised mature, compact bone. It was surrounded by neoplastic cells showing a variety of histologies, such as sarcoma, not otherwise specified, and myxosarcoma away from the bone-forming region. However, these different histological regions were considered to be components of a single bone tumor, based on the common expression of osterix and a similar mixture of constituent cells in each region. The tumor was diagnosed as an extraskeletal osteosarcoma because of the presence of infiltrative growth and abnormal mitosis and its development in the auricle without attachment to the skeleton. The present case is a rare histological type of an extraskeletal osteosarcoma with independent and different histological elements in rats.

Subacute oral administration of folic acid elicits anti-inflammatory response in a mouse model of allergic dermatitis.

Folic acid (FA) deficiency is associated with several health problems, including megaloblastic anemia and fetal neural tube defects. Therefore, supplementation with FA is strongly recommended by governments worldwide. Recent published reports indicate that FA functions in immune system maintenance. The main objective of this study is to examine possible anti-inflammatory and antipruritic effects of FA using a mouse model of allergic dermatitis. The mouse model was developed by repetitive sensitization to the Th2-type hapten toluene-2,4-diisocyanate (TDI). During the development of allergic dermatitis, FA was orally administered to the mice at doses of 8, 160, 1000 or 10,000 µg/day for 5 weeks. The ear swelling response and scratching behavior were monitored after the TDI challenge. Serum, ear tissue and auricular lymph node samples were isolated for further analysis 24 h after the TDI challenge. The ear swelling response was reduced in a dose-dependent manner by FA administration, and a significant change was observed at a concentration of 10,000-µg/day group. Comparable results were obtained through histological evaluation and cytokine level measurement in the ear tissue samples. Oral administration of FA exhibited the inhibitory effect on T-cell infiltration and T-cell-related cytokine production in auricular lymph nodes. Scratching behavior was not altered by FA administration. The in vivo evidence was corroborated by in vitro results, which showed that FA treatment significantly interfered with T-cell proliferation in a dose-dependent manner. Our findings imply that subacute oral administration of FA elicits an anti-inflammatory response, mainly through inhibition of T-cell proliferation.

Spontaneous malignant myoid thymoma in an aged female Fischer 344 rat.

A whitish mass approximately 30 mm in diameter was noted in the anterior mediastinum of a 67-week-old female Fischer 344 rat. Histopathologically, two types of tumor cells were identified on the basis of morphologic features: epithelial tumor cells with a tubular or cord-like growth pattern and rhabdomyosarcomatous tumor cells characterized by the presence of cross-striations. Immunohistochemically, the epithelial tumor cells reacted positively for cytokeratin AE1/AE3, and some reacted positively for p63, which is expressed in normal thymic epithelial cells. The rhabdomyosarcomatous tumor cells stained positively for desmin, sarcomeric actin, and S-100 protein, which coincides with the stainability of normal thymic myoid cells. Since the tumor was also found to have malignant features such as high proliferative activity, cytologic atypia, and necrotic behavior, it was diagnosed as a malignant myoid thymoma. We believe that this is the first case report of such a tumor in a rodent.

A spontaneous myoepithelial carcinoma in the mammary gland of an aged female ICR (CD-1) mouse.

We report a female Crlj:CD1(ICR) mouse with a spontaneous mammary gland tumor composed of biphasic tumor cells, i.e., epithelioid and spindle-shaped myoepithelial cells. Macroscopically, a subcutaneous mass, approximately 3 cm in diameter was found in the lumbodorsal region. Histopathologically, the epithelioid cells proliferated in an alveolar or nest-like growth pattern, occasionally forming glandular-like structures. On the other hand, the spindle-shaped cells proliferated in a sarcomatous pattern. Normal mammary gland was observed in the vicinity of the tumor. Both types of tumor cells showed immunoreactivity for cytokeratin (wide spectrum screening), vimentin, S100, and p63. In addition, the epithelioid cells and spindle-shaped cells were immunopositive for glial fibrillary acidic protein and smooth muscle actin, respectively. Moderate atypia, high proliferative activity, massive necrosis, and partial infiltration to the surrounding tissues were also observed. We made a diagnosis of myoepithelial carcinoma, which is extremely rare in ICR mice.

2-Nitroanisole-induced oxidative DNA damage in Salmonella typhimurium and in rat urinary bladder cells.

2-Nitroanisole (2-NA) is used in the manufacturing of azo dyes and causes cancer, mainly in the urinary bladder. Previous in vivo genotoxic data seems to be insufficient to explain the mechanism through which 2-NA induces carcinogenesis, and several bladder carcinogens were reported to induce oxidative DNA damage. Thus, we examined the potential induction of oxidative DNA damage by 2-NA using bacterial strain YG3008, a mutMST-deficient derivative of strain TA100. Consequently, strain YG3008, when compared with strain TA100, was found to be more sensitive to 2-NA, indicating oxidative DNA damage in bacterial cells. For further investigation, we performed the comet assay using the urinary bladder and liver of rats, with and without human 8-oxoguanine DNA-glycosylase 1 (hOGG1), to confirm the potential of 2-NA for inducing oxidative DNA damage. Simultaneously, we conducted a micronucleus test using bone marrow from rats to assess the genotoxicity of 2-NA in vivo. 2-NA was administered orally to male Fischer 344 rats for 3 consecutive days. The rats were divided into 6 treatment groups: 3 groups treated with 2-NA at doses of 125, 250, and 500mg/kg; a group treated with the combination of 2-NA and glutathione-SH (GSH); a negative control group; and a positive control group. The comet assay without hOGG1 detected no DNA damage in the liver or urinary bladder, and the micronucleus test did not show clastogenic effects in bone marrow cells. However, the comet assay with hOGG1 was positive in the urinary bladder samples, indicating the induction of oxidative DNA damage in the urinary bladder for the group treated with 2-NA at 500mg/kg. Moreover, an antioxidant of GSH significantly reduced oxidative DNA damage caused by 2-NA. These results indicate that oxidative DNA damage is a possible mode of action for carcinogenesis in the urinary bladder of rats treated with 2-NA.

Spontaneous extraskeletal osteosarcoma with various histological growth patterns in the abdominal wall of an ICR mouse.

Extraskeletal osteosarcoma is extremely rare in mice. This case report demonstrates a spontaneous murine extraskeletal osteosarcoma that exhibited various histological growth patterns in an ICR mouse. At necropsy, the tumor mass was located in the abdominal wall and was 45 × 30 × 25 mm in size. Histopathologically, the tumor showed the following four growth patterns: a solid pattern of polygonal cells embedded in an osteoid eosinophilic matrix with calcification, an irregular sheet pattern of short spindle cells accompanying some eosinophilic multinucleated cells, a fascicular pattern of spindle cells and a cystic pattern lined by short spindle cells. Immunohistochemically, most of the tumor cells were positive for vimentin, proliferating cell nuclear antigen and osterix. The multinucleated cells mentioned above were desmin positive and were regarded as regenerative striated muscles but not tumor cells. Since no clear continuity with normal bone tissues was observed, the tumor was diagnosed as an "extraskeletal osteosarcoma."

Poorly differentiated salivary gland carcinoma with prominent squamous metaplasia in a pregnant Wistar Hannover rat.

A subcutaneous pale brown-colored mass was observed macroscopically in the ventral neck of a 16-week-old Wistar rat on day 18 of gestation. The mass was well demarcated from the adjacent tissues with partial invasion into connective tissues. Necrosis and hemorrhage were evident throughout the mass. The mass comprised a diffuse sheet and a nest-like structure of epithelial cells with prominent squamous metaplasia. The neoplastic cells tested immunopositive for keratin, vimentin, glial fibrillary acidic protein and p63. A portion of the neoplastic cells exhibited a similar immunoreaction of prominin-1 to the ductal and acinar cells in normal submandibular and parotid glands. Collectively, the tumor was diagnosed as a poorly differentiated carcinoma derived from epithelial/myoepithelial lineages in the submandibular and/or parotid glands.

Inhibition of lipopolysaccharide-induced liver injury in rats treated with a hepatic drug-metabolizing enzyme inducer p,p'-DDT.

Hepatocellular hypertrophy in association with drug-metabolizing enzyme induction is considered to be an adaptive change associated with drug metabolism. To improve our understanding of liver hypertrophy, we determined the effect of a single ip injection of either lipopolysaccharide (LPS) or vehicle in male F344 rats with hepatocellular hypertrophy induced by oral delivery of p,p'-DDT for 2 weeks. The rats were sacrificed 3h or 24h after LPS or vehicle injection. LPS induced a focal hepatocellular necrosis in rats fed the control diet. When rats pre-treated with p,p'-DDT were injected with LPS, necrotic foci surrounded by ballooned hepatocytes were observed in the liver. The change was consistent with reduced LPS-mediated increases in plasma hepatic biomarkers, neutrophil influx, and apoptosis, and also associated with hepatic mRNA levels of TNF-α, CYPs, and NOS2. By contrast, when combined with p,p'-DDT and LPS, faint hepatocellular fatty change was extended, together with a synergistic increase in total blood cholesterol. These results suggest that hepatocytes exposed to p,p'-DDT are protected from the cell-lethal toxic effects of an exogenous stimulus, resulting in cell ballooning rather than necrosis in association with reduced inflammation and apoptosis, but compromised by an adverse effect on lipid metabolism.

p,p'-DDT induces microcytic anemia in rats.

Emerging evidence suggests that chronic exposure to DDT and its derivatives is associated with a variety of human disorders such as anemia. The present study demonstrated that p,p'-DDT caused microcystic anemia in a dose-dependent manner (0, 5, 50, and 500 ppm) in the long-term study up to 2 years. To elucidate the mechanism(s) by which p,p'-DDT induces anemia, certain hematological parameters were assessed in rats fed specific doses of p,p'-DDT for 2 weeks, and the effect of lipopolysaccharide on anemia of inflammation was also examined in p,p'-DDT-treated rats. The parameters included the content of hemoglobin per reticulocyte, mean corpuscular volume of reticulocytes and mature erythrocytes, corpuscular hemoglobin concentration mean of mature erythrocytes, and saturation levels of transferrin and iron. During the 2-week treatment period, hypochromic microcytic reticulocytes and hypochromic normocytic mature erythrocytes were observed in p,p'-DDT-treated rats, with no evidence of alteration in plasma iron levels. p,p'-DDT enhanced microcytosis of reticulocytes, as well as mature erythrocytes, which occurred due to severe hypoferremia resulting from anemia of inflammation; however, plasma iron levels were attenuated probably through the inhibition of interleukin-6. Our data suggests that long-term treatment with p,p'-DDT induces microcytic anemia, possibly because of the impairment of iron utility in erythrocytes.

Malignant peritoneal mesothelioma with a sarcomatoid growth pattern and signet-ring-like structure in a female f344 rat.

We report a biphasic malignant mesothelioma in an aged female F344/DuCrlCrlj rat. Macroscopically, multiple pale brown nodules were observed in the abdominal cavity with retention of bloody ascites. Histopathologically, the tumor cells spread over the peritoneum and formed masses on the surface and underlying adipose tissues. The tumor cells dominantly proliferated in a solid, nodular or nest-like pattern with modest amount of fibrillar connective tissues, which contained hyaluronan. The tumor consisted of ovoid, polygonal or spindle-shaped cells that possessed eosinophilic cytoplasms including glycogen; some tumor cells showed a signet-ring-like structure. Multinucleated cells and mitosis were found frequently, and direct invasion to intra-abdominal organs and intravascular metastasis to the liver were observed. Immunohistochemically, keratin and mesothelin were strongly positive in most of tumor cells, while vimentin was mainly positive in spindle-shaped cells. Podoplanin was also positive, particularly in the cell membrane of tumor cells. Electron microscopically, tumor cells showed an intercellular desmosome-like structure, basement membrane and microvillus. We diagnosed the case as a malignant peritoneal mesothelioma with a sarcomatoid growth pattern and signet-ring-like structure.

Didecyldimethylammonium chloride induces pulmonary fibrosis in association with TGF-ß signaling in mice.

Didecyldimethylammonium chloride (DDAC) is a representative dialkyl-quaternary ammonium compound that is used as a disinfectant against several pathogens and is also used in commercial, industrial, and residential settings. We previously investigated toxicity on air way system following single instillation of DDAC to the lungs in mice, and found that DDAC causes pulmonary injury, which is associated with altered antioxidant antimicrobial responses; the inflammatory phase is accompanied or followed by fibrotic response. The present study was conducted to monitor transforming growth factor-ß (TGF-ß) signaling in pulmonary fibrosis induced by DDAC. Mice were intratracheally instilled with DDAC and sacrificed 1, 3, or 7 days after treatment to measure TGF-ß signaling. In order to further evaluate TGF-ß signaling, we treated isolated mouse lung fibroblasts with DDAC. Fibrotic foci were observed in the lungs on day 3, and were widely extended on day 7, with evidence of increased α-smooth muscle actin-positive mesenchymal cells and upregulation of Type I procollagen mRNA. Developing fibrotic foci were likely associated with increased expression of Tgf-ß1 mRNA, in addition to decreased expression of Bone morphogenetic protein-7 mRNA. In fibrotic lung samples, the expression of phosphorylated SMAD2/3 was considerably higher than that of phosphorylated SMAD1/5. In isolated lung fibroblasts, the mRNA levels of Tgf-ß1 were specifically increased by DDAC treatment, which prolonged phosphorylation of SMAD2/3. These effects were abolished by treatment with SD208 - a TGF-ßRI kinase inhibitor. The results suggest that DDAC induces pulmonary fibrosis in association with TGF-ß signaling.