Histopathological and immunohistochemical features of atypical epithelial tumours of the gland of the third eyelid in seven dogs.

This report documents the histopathological and immunohistochemical features of atypical epithelial tumours of the gland of the third eyelid (GTE) in seven dogs. Cases 1 and 2 were diagnosed as myoepithelioma, comprising of compressive proliferations of interlacing bundles of neoplastic spindle cells expressing cytokeratin 14, p63, calponin and α-smooth muscle actin. Cases 3, 4 and 5 were diagnosed as complex carcinomas comprising of atypical glandular cells expressing cytokeratin 8/18, together with spindle-shaped or round neoplastic cells expressing cytokeratin 14, p63, calponin and α-smooth muscle actin. Cases 6 and 7 were diagnosed as basal cell adenocarcinomas (BCACs) comprising of a mixed proliferation of glandular and basal-type cells expressing cytokeratin 14 and p63. Therefore, in addition to glandular components, these tumours may include neoplastic cells with a myoepithelial or basal cell phenotype. Hence, there is diversity in the features of epithelial neoplasia of the GTE in dogs, similar to tumours in human salivary and lacrimal glands.

Teratoma of the umbilical cord in a giraffe (Giraffa camelopardalis reticulata).

A 12-year-old pregnant female giraffe (Giraffa camelopardalis reticulata) died approximately 2 months prior to her anticipated parturition date. At necropsy, a mass measuring approximately 20 x 36 x 20 cm was observed, attached to the umbilical cord, the latter being otherwise normal in appearance. Histologically, the mass contained 3 germinal tissue components with areas of squamous epithelium, respiratory epithelium, primitive neural tissues, glial tissue, peripheral nerve, adipose tissue, cartilage, and smooth muscle. Based on these findings, the tumor was diagnosed as a teratoma originating from the umbilical cord. This is possibly the second reported case of umbilical cord teratoma in animals.

Expression of vascular endothelial growth factor, basic fibroblast growth factor, and their receptors (flt-1, flk-1, and flg-1) in canine vascular tumors.

Expression of vascular endothelial growth factor (VEGF), its receptors (flt-1 and flk-1), and basic fibroblast growth factor (bFGF) in canine hemangiosarcoma (HSA) and hemangiomas was investigated by immunohistochemical analysis. In addition, expression of the mRNAs of VEGF, flt-1, flk-1, and flg-1 (a receptor for bFGF), was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) with cRNA probes. VEGF, bFGF, flt-1, and flk-1 were immunohistochemically detected in the neoplastic cells in HSAs; the staining intensity was stronger in HSAs than in hemangiomas. On the other hand, the neoplastic cells in hemangiomas exhibited very weak or no expression of VEGF, although they showed moderate expression of flt-1 and flk-1. The mRNAs of VEGF, flt-1, flk-1, and flg-1 were detected in the neoplastic cells in HSAs by ISH and RT-PCR. However, VEGF mRNA was not detectable in the neoplastic cells in hemangiomas by ISH, although it was detected in the inflammatory cells in the tumors by RT-PCR. Moreover, the HSAs that showed intense staining for flk-1 had a high proliferative activity, which was reflected as a high Ki-67 positive index. These results suggest that the expression of the growth factors and their receptors, especially flk-1, might be associated with the malignant proliferation of HSAs.

Morphological and immunohistochemical features of Cryptosporidium andersoni in cattle.

Light and electron microscopic features and immunohistochemical features of Cryptosporidium andersoni (C. andersoni) and host reaction in the mucosa were studied. Although the affected cattle demonstrated no apparent clinical signs, a severe infection of C. andersoni was observed in the abomasum. C. andersoni were round in shape, measured 6-8 microm in size and were mainly observed to be freely located in the gastric pits, being attached in occasional cases to the surface of the abomasum epithelium. Frequent inflammatory cells had infiltrated the lamina propria of the affected mucosa, and frequent mitotic figures were observed in epithelial cells at the dilated isthmus. To access the cell kinetics, the number of epithelial cells infected with C. andersoni were counted and compared with noninfected cattle. The number of gastric pit cells in infected cattle was significantly higher than that in the controls. The number of proliferative cells determined by the Ki-67 antigen in C. andersoni infected cattle was also significantly higher than that in the controls. Transmission electron microscopy and scanning electron microscopy revealed that the morphology of the C. andersoni organism was common to those of other Cryptosporidium spp. Immunohistochemically, several commercial antibodies against Cryptosporidium spp. showed positive reactions at the wall of these oocysts or parasitophorous vacuoles. This report is possibly the first to discuss the prominent hyperplasia of the abomasum mucosa, as well as morphologic features of C. andersoni in cattle.

Assessment of proliferative potentials of canine osteosarcomas and chondrosarcomas by MIB-1 immunohistochemistry and bromodeoxyuridine incorporation.

The proliferative potential of 17 canine osteosarcomas (OSs) (13 osteoblastic, two anaplastic, one fibroblastic and one chondroblastic), 18 chondrosarcomas (CSs) (13 mesenchymal and five ordinary), three osteomas, and one chondroma was evaluated immunohistochemically by labelling Ki-67 antigen with MIB-1 antibody, and incorporated bromodeoxyuridine (BrdU) with anti-BrdU antibody. The location of BrdU-positive cells in OSs and CSs was similar to that of MIB-1 positive cells, and the mean value of the BrdU labelling index (BrdU LI) and the MIB-1 positive index (MIB-1 PI) in each case were significantly correlated (rs = 0.942, P < 0.05 with Spearman rank correlation coefficient; r = 0.779 P < 0.05 with linear regression analysis). The mean MIB-1 PI of OSs was 29.5%, which was approximately 2.5 times that of CSs, and the highest MIB-1 PI was 34.8% +/- 1.8 S.E.M. in areas without osteoid. In CS cases, the survival rate after 24 months was significantly higher than in OS cases. The high MIB-1 PI therefore supports the view that OSs are clinically more aggressive than CSs in dogs. On the other hand, the highest MIB-1 PI values of mesenchymal CS components occurred in transitional areas, which were composed of poorly differentiated cells embedded in a myxomatous matrix between the chondroidal and mesenchymal regions. The MIB-1 PI was 21.3% +/- 3.0 S.E.M. P < 0.001 in transitional areas. Proliferative markers may be useful in diagnosis and prognosis.

Expression of myogenic regulating factors, Myogenin and MyoD, in two canine botryoid rhabdomyosarcomas.

Myogenin and MyoD regulate the development of skeletal muscle, and their expressions are specific to the stages of myogenesis. Therefore, these myogenic regulatory proteins could be considered as sensitive and specific markers for rhabdomyosarcoma. In this report we investigated the immunohistochemical reactivities of myogenin and MyoD in two canine bladder botryoid rhabdomyosarcomas that were different in the degree of differentiation. MyoD was stained in the Ki-67 antigen-positive undifferentiated mesenchymal cells, which had proliferative activity similar to myoblasts differentiated from mesoblasts. In contrast, multinucleated neoplastic cells were positive for myogenin and alpha-sarcomeric actin but not for Ki-67 antigen, similar to the myotubes differentiated from myoblastic cells. The expressions of myogenin and MyoD were closely correlated to the histologic features of myogenic neoplastic cells.

Involvement of apoptosis and cyclin D1 gene repression in growth inhibition of T-47D human breast cancer cells by methylglyoxal bis(cyclopentylamidinohydrazone).

Polyamines are considered to be important intracellular molecules for the proliferation of the cancer cells. In this study, effects of methylglyoxal bis(cyclopentylamidinohydrazone) (MGBCP), a potent inhibitor of the polyamine biosynthetic pathway, on the growth and cell cycle of T-47D human breast cancer cells were investigated. MGBCP dose-dependently inhibited the growth of T-47D cells, in which the contents of spermine, spermidine and putrescine decreased concomitantly. The gene expression of cyclin D1 was also repressed by the MGBCP treatment. The MGBCP-treated cells clearly exhibited morphological changes indicating the blebbing and chromatin condensation which are characteristic of apoptosis. Flow cytometric analysis showed hypo-diploid subpopulations due to apoptotic cells, and characteristic oligonucleosomal-sized DNA fragments were clearly observed for MGBCP-treated cells as the concentration of the drug was increased. These findings suggest that the inhibition of polyamine synthesis results in the repressions of cyclin D1 expression and cell cycle progression, eventually inducing apoptosis in these human breast cancer cells.