Antiserum against culture filtrate is cross-protective for various serovars of Erysipelothrix rhusiopathiae

The protective effect of porcine antiserum prepared against culture filtrate (CF) of an attenuated strain of Erysipelothrix rhusiopathiae (serovar 2) in mice to challenge with 20 virulent strains of 18 serovars and one type N was investigated. Passively immunized mice survived after challenge with serovars 1a, 1b, 2, 5, 6, 8 (strain Goda), 11, 12, 15, 16, 21 or type N, but 10-30% mortality occurred in immunized mice challenged with each strain of serovars 4, 7, 8 (strain 911), 9, 18 or 19 and 70% mortality to serovar 10 (strain 2179). All immunized mice died after challenge with serovar 20 (strain 2553). Non-treated control mice died after challenge with all serovars and the type tested.     

Induction of T cell anergy by the treatment with IL-10-treated dendritic cells

In order to apply for reducing graft versus host disease in allogeneic stem cell transplantation, the study concerning the induction of specific T cell anergy was designed. Normal allogeneic lymphocytes, which were co-cultured with IL-10-treated immature dendritic cells in the first mixed leukocyte culture (MLC), were cultured with mature dendritic cells of the same origin as IL-10-treated immature dendritic cells in the second MLC. By co-culturing with IL-10-treated immature dendritic cells, the response of normal lymphocytes to mature dendritic cells cultured from the same individual as that of IL-10-treated dendritic cells was markedly reduced, compared with the lymphocytes cultured with non-treated dendritic cells or IL-10-treated dendritic cells from a third party individual. The present study demonstrated that antigen specific T cell anergy was generated by priming allogeneic lymphocytes with IL-10-treated immature dendritic cells. These data suggested the applicability of IL-10-treated recipient dendritic cells for the induction of recipient cell-specific donor T cell anergy in donor graft.     

Phenotypic analysis of hepatic T lymphocytes in a dog with chronic hepatitis

We evaluated hepatic T lymphocyte phenotypes in a dog with chronic hepatitis. Before treatment, numerous CD3+ lymphocytes were demonstrated in the liver, and the ratio of CD4+/CD8+ was remarkably high (2.96; reference range, 0.33+/-0.12). After treatment, CD3+ lymphocyte infiltration in the liver was reduced, and the ratio of CD4+/CD8+ decreased to 0.31. Therefore, hepatic T lymphocytes, especially CD4+ lymphocytes, might play a central role in the pathogenesis of this dog with chronic hepatitis.     

Characterization of spheres derived from canine mammary gland adenocarcinoma cell lines

There is increasing evidence for the presence of cancer stem cells in several solid tumors, and these cancer stem cells have a potential role in tumor initiation, aggression, and recurrence. The stem cell-like properties of spheres derived from canine mammary tumors remain largely elusive. We attempted to induce sphere formation using four cell lines of canine mammary adenocarcinoma, and characterized the spheres derived from a CHMp line in vitro and in vivo. The CHMp-derived spheres showed predominantly CD44⁺CD24⁻ population, higher expression of stem cell-related genes, such as CD133, Notch3 and MDR, and higher resistance to doxorubicin compared with the CHMp-derived adherent cells. Xenograft transplantations in nude mice demonstrated that only 1 × 10⁴ sphere cells were sufficient for tumor formation. Use of the sphere assay on these sphere-derived tumors showed that sphere-forming cells were present in the tumors, and were maintained in serial transplantation. We propose that spheres derived from canine mammary adenocarcinoma cell lines possess a potential characteristic of cancer stem cells. Spheres derived from canine mammary tumors could be a powerful tool with which to investigate novel therapeutic drugs and to elucidate the molecular and cellular mechanisms that underlie tumorigenesis.     

Apoptosis in the porcine uterine endometrium during the estrous cycle, early pregnancy and post partum

The mammalian uterus changes dramatically during the estrous cycle, pregnancy, and involution post partum. Dynamic changes in the uterine endometrium are a type of homeostasis and proceed with proliferation and exclusion of cells. Homeostasis of the uterus is closely related to apoptosis involving various hormones and cytokines. The objective of the present study was to determine the morphological features and occurrence of apoptosis in the porcine endometrium during the estrous cycle, early pregnancy, and post partum. Cyclic changes in the morphology of the surface epithelium were observed during the estrous cycle. The heights of surface epithelia were significantly high on day 4 of the estrous cycle and the early pregnancy. The heights of the surface epithelium remained low from days 1 to 31 post partum. We then used terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling (TUNEL) of the 3'-terminal of fragmented DNA, which is effective for detection of apoptosis in various tissues. We found that apoptosis in the porcine endometrium contributed to homeostasis of the endometrium during the estrous cycle through control of cell proliferation and exclusion. Conversely, apoptosis on days 4 and 8 of gestation before the implantation window depended on the plasma estrogen and progesterone levels; however, suppressive homeostasis of apoptosis occurred at the time of implantation on days 15, 18 and 21 of gestation. Our study is the first to demonstrate apoptotic cell death in the porcine endometrium directly by TUNEL method. The results strongly suggest that uterine homeostasis is mainly controlled by apoptosis during the estrous cycle and early pregnancy.     

PMSG profiles in superovulated and anti-PMSG antiserum treated mice and heifers with enzymeimmunoassay

A sandwich enzymeimmunoassay (EIA) for pregnant mare serum gonadotropin (PMSG) using a microtiter plate was developed. Sensitivity of the assay to PMSG was 15.6 mIU/ml (0.2 ng/well). The PMSG levels in serum were measured with the EIA in superovulated and anti-PMSG rabbit antiserum treated mice and heifers. In mice, the PMSG blood level was measurable in the serum 4-6 days after intraperitoneal injection of 5-30 IU of PMSG. The administration of anti-PMSG antiserum at the same dose level as PMSG caused a rapid decrease in the PMSG blood level, declining to undetectable levels within 17 hours. In heifers, the PMSG level was measurable at 10-11 days after the injection of 2500 or 3000 IU of PMSG. When antiserum was injected 48 hours after the PMSG injection, the clearance rate of PMSG was affected by the route of the administration. The administration of 3000 units of anti-PMSG antiserum intravenously caused a rapid decline and the disappearance of circulating PMSG within 17 hours. When 3000 units of anti-PMSG antiserum was injected intra-muscularly, the PMSG blood level also decreased and became unmeasurable 24 hours after administration; however, it was still detectable for up to 17 hours. These results indicate that the administration of anti-PMSG antiserum at the proper timing and dosage could lead to successful superovulation through the improvement of hormonal conditions.     

A cat with acute myeloblastic leukemia without maturation (M1) treated with combination chemotherapy

A 2-year-old domestic shorthair cat was presented to us with decreased activity and anorexia. Hematologic findings revealed a mild non-regenerative anemia, thrombocytopenia, and leukocytosis with an increase in blast cells. Bone marrow aspirates also revealed a marked increase of blasts. The blastic cells were shown to be positive for peroxidase. Acute myeloblastic leukemia without maturation (M1) was diagnosed according to the FAB classification. Chemotherapy was initiated with cyclophosphamide, vincristine, prednisolone, and cytosine arabinoside. The cat responded partially. In total, the cats were given 7 blood transfusions. The cat died 14 weeks after first being presented to us.     

Molecular typing and antimicrobial resistance of Salmonella enterica subspecies enterica serovar Choleraesuis isolates from diseased pigs in Japan

Salmonella enterica serovar Choleraesuis (S. Choleraesuis) isolates derived from diseased pigs in Japan during 2001 and 2005 were analyzed for biotype, based on H(2)S production and dulcitol fermentation, pulsed-field gel electrophoresis (PFGE) profile, and antimicrobial resistance profile. S. Choleraesuis biotype Choleraesuis (biotype Choleraesuis) was classified into one genotype, while varietas Kunzendorf (var. Kunzendorf) was classified into two genotypes. The isolates of var. Kunzendorf belonging to one genotype were isolated in a limited area of Japan. Variation in the antimicrobial resistance pattern was observed in isolates of both biotypes Choleraesuis and var. Kunzendorf. We have also shown that the PFGE profile was associated with the biotype and isolation region of each isolate.     

Intranasal hemangiosarcoma in a dog

Magnetic resonance (MR) was conducted for an 8-year-old, intact male Spitz with sneezing, serous discharge and epistaxis from the left nasal cavity. MR imaging showed a nasal cavity-occupied mass of iso-intensity on T1WI , high-intensity on T2WI and markedly enhanced on contrast-enhanced T1WI at parts of rostal to medial ocular angle in the left cavity. After Surgery and intraoperative radiation, the mass was diagnosed intranasal hemangiosarcoma by histopathology. Although the dog showed the finding, which suggested recurrence after the treatment ending, about 30 months later, it maintained good conditions without evidence of metastasis.