Comparison of tyrosinase-related protein-2, S-100, and Melan A immunoreactivity in canine amelanotic melanomas

Tyrosinase-related protein-2 (TRP-2) is a highly conserved melanogenic enzyme expressed in both pigmented and unpigmented melanomas of the mouse. To determine whether TRP-2 would be a good diagnostic marker for amelanotic melanomas of the dog, we performed immunohistochemistry for TRP-2, S-100, and Melan A on 21 canine tumors identified as amelanotic melanomas based on routine histopathologic examination. Thirteen of the tumors were TRP-2 positive, 10 were Melan A positive, and 19 were S-100 positive. TRP-2 was expressed in the cytoplasm of tumor cells in both primary and metastatic melanomas. S-100 staining was positive in all of three schwannomas and two of three gastrointestinal stromal tumors (one fibrosarcoma and one leiomyosarcoma) tested. Neither Melan A nor TRP-2 antibodies reacted with these tumors. Our findings indicate that staining for TRP-2 is a sensitive and specific method for confirming the diagnosis of amelanotic melanoma in dogs.     

Pax-7 immunoreactivity in the post-natal chicken central nervous system

In this immunocytochemical study on the constitutive expression of Pax-7 protein in the postnatal chicken brain, Pax-7 showed region and cell type specific expression. In the optic tectum, only cells in grey matter showed positive immunoreactivities (IRs), whereas those in the white matters did not show any IRs. In thalamic nuclei and several pontine nuclei, we also localized Pax-7 positive IRs. On the contrary, in the cerebellum, Pax-7 was mainly localized within the Bergmann glia, whereas Purkinje cells did not show any IRs. In double immunolabelling studies, most of the Pax-7 IRs did not originate from neuroglial cells such as oligodendrocytes, microglia or astrocytes, but from neurons, with the exception of Bergmann glia in the cerebellum. The presence of Pax-7 IRs in the adult chicken brain could suggest that Pax-7 might play a role in maintaining normal physiological function in some postnatal chicken brain cells.     

Monoclonal antibody-based competitive ELISA for simultaneous detection of rinderpest virus and peste des petits ruminants virus antibodies

An experimental competitive enzyme-linked immunosorbent assay (morbillivirus cELISA) using a recombinant N antigen (rRPV N) expressed in a baculovirus and a ruminant morbillivirus (RPV and PPRV)-specific monoclonal antibody (P-13A9) was developed for simultaneous detection of rinderpest virus (RPV) and peste des petits ruminants virus (PPRV) antibodies and its diagnostic performance was evaluated. A set of known reference antisera against RPV and PPRV belonging to different lineages, experimental sera from cattle vaccinated for a RPV of Asian lineage, and field sera from cattle and sheep/goat populations known to be positive (West Africa) and negative (Korea) for RPV and PPRV were used for the evaluation. Morbillivirus cELISA results on the panel of experimental RPV and PPRV antisera showed high correlation (r=0.97) between the whole virus and the rRPV N antigens, suggesting that the rRPV N contains a ruminant morbillivirus-specific antigenic determinant recognized by the P-13A9 and it may be suitable as an ELISA antigen in place of the whole virus. Morbillivirus cELISA detected anti-RPV and anti-PPRV antibodies in all reference RPV and PPRV antisera containing VN titers >/=1:8, suggesting that the assay can simultaneously detect antibodies against RPV and PPRV. Anti-RPV antibody was detected by morbillivirus cELISA in vaccinated cattle as early as the VNT and continued to be detectable by both the cELISA and the VNT until termination of the study. When applied to field samples from Africa, morbillivirus cELISA showed good agreement with a RP cELISA kit (kappa value of 0.86) in bovine sera and with a peste des petits ruminant cELISA kit (kappa value of 0.81) in caprine/ovine sera. Usefulness of morbillivirus cELISA using the rRPV N protein was discussed.     

Evaluation of canine gastric motility with ultrasonography

For the evaluation of canine gastric motility with ultrasonography, contraction number of pyloric antrum and gastric emptying time (GET) by area and volume method developed by Bolondi et al.'s method were studied in 14 dogs. All experimental dogs were administered with saline and soup solution (10 ml/kg, B.W.). The mean values of contraction number of pyloric antrum in saline and soup group were 4.19 +/- 1.30/min and 4.82 +/- 0.65/min before feeding, and overall mean values were 4.66 +/- 1.37/min and 5.13 +/- 1.71/min, respectively. The mean values of the GET by area and volume method were 36.73 +/- 11.27, 40.00 +/- 8.87 min in saline group and 61.35 +/- 17.58, 59.11 +/- 14.46 min in soup group. In the GET in saline and soup groups, there was no significant difference between the area and volume method (p>0.05). Therefore, Bolondi et al.'s method by ultrasound can be used to evaluate the antropyloric motility and gastric emptying time with area and volume methods. The area method is easier to determine the GET than the volume method, but the latter is more accurate.     

A case of chronic wasting disease in an elk imported to Korea from Canada

A seven-year-old male elk (Cervus elaphus nelsoni) was euthanized and necropsied after having a 3-week history of body weight loss, emaciation, excessive salivation, teeth grinding, fever, anorexia, and respiratory distress. The elk was imported into Korea from Canada on March 9, 1997. Gross pathologic findings were restricted to a diffuse fibrinous pneumonia. Microscopic lesions included mild neuronal vacuolation and spongiform change in the neuropil of selected brain stem nuclei and generalized astrocytosis. Immunohistochemistry for protease-resistant prion protein (PrP(res)) was positive in all brain sections but more pronounced in the section of the obex of the medulla. And the PrP(res) was also detected by western immunoblotting in the brain and spinal cord. All the remaining elk and deer that had been in contact with this elk were destroyed and negative for chronic wasting disease (CWD). To our knowledge, this is the first case of CWD occurring outside of the U.S.A. and Canada.     

Parameter estimates for direct and maternal genetic effects on yearling, eighteen-month, and slaughter weights of Korean native cattle

Data collected by the National Livestock Research Institute of the Rural Development Administration of Korea were used to estimate genetic parameters for yearling (YWT, n = 5,848), 18-mo (W18, n = 4,585), and slaughter (SWT, n = 2,279) weights for Korean Native cattle. Nine animal models were used to obtain REML estimates of genetic parameters: DP-2 included genetic, uncorrelated dam, and residual random effects; DQ-2 included genetic, sire x region x year-season interaction, and residual random effects; DPQ-2 was based on DQ-2 but included both interaction and dam effects; DMP-2 was based on DP-2 but with dam effect partitioned to include maternal genetic and permanent environmental effects; and DMPQ-2 was based on DMP-2 but also included sire interaction effects. Those five models included two fixed factors: region x year-season and age of dam x sex effects. Models DP-3, DQ-3, DPQ-3, and DMPQ-3 were based on DP-2, DQ-2, DPQ-2, and DMPQ-2 but included as a third fixed factor whether or not identification of the sire was known. Estimates of heritability with DMPQ-3 for YWT, with DPQ-3 for W18 and SWT when analyzed with single-trait analyses were .14, .11, and .17, respectively, and were nearly the same with bivariate analyses. Estimate of maternal heritability for YWT from single-trait analysis was .04, with estimates for other traits near zero. For bivariate analyses, the estimate for YWT was .01. With single trait analysis, estimate of the direct-maternal genetic correlation for YWT was negative (-.81). Estimates of direct genetic correlations between YWT and W18, YWT and SWT, and W18 and SWT were .99, 1.00, and .97, respectively. Estimates of environmental correlations varied from .60 to .81; the largest was between W18 and SWT. Including a fixed factor for whether sire identification was missing or not missing reduced the estimate of heritability for slaughter weight. The results suggest that the sire x region x year-season interaction is important for yearling weight and may be needed in a model for slaughter weight. Maternal effects may be of slight importance for yearling weight but of no importance for W18 and SWT. Models for national cattle evaluations for Korean Native cattle for YWT should be considered that include maternal genetic and permanent environmental as well as sire x region x year-season interaction effects, but those effects seem not to be needed for models for W18 and SWT. Not much reranking of sires occurred when ranked was based on the different models for W18 and SWT.     

Sequence analyses of feline B7 costimulatory molecules

Using RT-PCR amplifications with mRNA from mitogen-stimulated feline peripheral blood mononuclear cells, cDNA of feline B7-1 (CD80) and B7-2 (CD86) were cloned. The cDNA were sequenced and putative translated protein sequences compared with known counterpart sequences. Hydrophilicity patterns of the feline CD80 and CD86 which were only 26.8% identical at the amino acid sequence were very distinct from each other, but similar to the putative human CD80 and CD86 proteins, respectively. The feline CD80 gene encoded a protein of 292 amino acids and the CD86 gene encoded a protein of 329 amino acids. Amino-terminal signal sequences, extracellular Ig V- and Ig C-like domains, transmembrane domains, and carboxyl cytoplasmic domains were identified in both molecules. Although the most conserved domain among the CD80 sequences was the Ig C-like domain, the most conserved domain among the CD86 sequences was the Ig V-like domain. Among the known sequences, the bovine CD80 and the porcine CD86 sequences available for comparisons were identified as most closely related to the feline CD80 (63.3%) and CD86 (67.5%), respectively. The mouse molecules were the least identical (43.6 and 43.6%, respectively) with the feline CD80 and CD86 proteins. The human CD80 and CD86 molecules were 56.3 and 57.0% identical with the feline molecules.