We compared the immunoreactivity and numbers of tyrosine hydroxylase (TH) immunoreactive neurons and neuropil in the paraventricular nucleus (PVN) of the hypothalamus between the seizure sensitive (SS) and seizure resistant (SR) gerbils. The distributional pattern of TH immunoreactivity was similar in both groups: TH immunoreactivity was seen mainly in magnocellular neurons of the PVN. However, total TH immunoreactivity in the neurons and neuropil in the SS gerbils was significantly lower than that in the SR gerbils. In addition, the number of TH immunoreactive neurons in the SS gerbils was also much lower than those in the SR gerbils. These results indicate that SS gerbils have a low TH immunoreactivity in the hypothalamic PVN compared with that in SR gerbils.
Reverse transcription polymerase chain reaction (RT-PCR) of turkey astrovirus (TAstV) capsid and polymerase genes was applied to the bursa of Fabricius (BF), thymus (TH), spleen (SP) and cloacal swabs (CS) of young poults with "Poult enteritis complex" (PEC). The histological lesions included atrophy, lymphoid depletion, cellular infiltration and necrosis of the BF, TH and SP, respectively. The RT-PCR reactions were positive for the polymerase gene of TAstV-2 in all 100 CSs, 7 out of 10 of BFs and 10 out of 20 THs and SPs, respectively. Five out of 10 THs and SPs samples, considered to be negative by RT-PCR, were positive when specific primers designed for the TAstV-2 capsid gene were applied. This is the first description of turkey astrovirus infection presenting PEC in Latin America.
Salmonellaenterica isolates from poultry (n=125) and swine (n=132) in Thailand were investigated for antibiotic resistance, susceptibility to disinfectants (benzalkonium chloride (BKC), chlorhexidine digluconate (CHX), zinc chloride and copper sulfate) and cyclohexane tolerance. Forty-two percent were of multiple resistance to antibiotics. The minimum inhibitory concentrations (MICs) of all antibiotics against isolates from swine were higher than that against the isolates from poultry. There were generally few variations in MICs to all disinfectants, indicating that the isolates had either no or only a limited degree of developed resistance to the disinfectants tested. Only 5 isolates (1.9%) were tolerant to cyclohexane. The proton-dependent efflux systems did not play a major role in the reduced susceptibility to BKC and CHX, since susceptibility was not restored when an efflux inhibitor, carbonyl cyanide m-chlorophenylhydrazone (CCCP) was present. Successive exposure to subinhibitory concentrations of BKC and CHX generated mutants resistant to BKC and CHX. A spontaneous BKC-resistant derivative expressed cross-resistance to antibiotics, chloramphenicol and erythromycin. The mechanism responsible for cross-resistance between BKC and antibiotics was not driven by the proton motif force (PMF).
TRAIL is a member of the tumor necrosis factor family and engages apoptosis via recruitment and rapid activation of caspase-8. This study investigated the effect of carbonyl cyanide m-chlorophenylhydrazone (CCCP), a classic uncoupler of oxidative phosphorylation, on TRAIL-induced apoptosis in SNU-638 cells derived from human gastric cancer cells. It was found that treatment with CCCP followed by incubation with TRAIL markedly enhanced apoptosis by 2 fold compared with treatment with TRAIL alone. This effect was accompanied by reduction in mitochondrial transmembrane potential and generation of reactive oxygen species. This sensitization was inhibited by N-acetyl-l-cysteine, which restored the mitochondrial transmembrane potential and reduced reactive oxygen species generation. Treatment with N-acetyl-L-cysteine also inhibited expression of apoptotic proteins such as Bax and Smac and abrogated caspase-8 activation. Moreover, treatment with N-acetyl-L-cysteine prior to induction with TRAIL increased expression of the anti-apoptotic Bcl-2 protein. These data indicate that CCCP enhanced TRAIL-induced apoptosis by dissipation of mitochondrial transmembrane potential and reactive oxygen species, suggesting that treatment with CCCP combined with that with TRAIL can be an efficient method to induce death of tumor cells, particularly cells that are resistant to TRAIL-induced apoptosis.
A three-year-old dog with marked leukocytosis, lymphadenopathy, and diarrhea showed an increase in unidentified blasts in the peripheral blood, and they were proliferated in the bone marrow. The dog was diagnosed with myelomonocytic leukemia (M4) because the blast cells were demonstrated by cytochemical staining to be both myeloid and monocytic cells. Although the dog was treated with a multi-combination chemotherapy and induction therapy using vitamin K2, it died on day 47 after the first admission. This case is the first report of M4 in Japan.
The present study determined the cDNA and deduced amino acid sequences of ferret (Mustela putorius furo) inflammatory cytokines, interferon (IFN)-γ, interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor (TNF)-α. The homologies of the nucleotide sequences of IFN-γ, IL-1β, IL-6, IL-8 and TNF-α of the ferret to those from other mammalian species ranged from 64.3-92.9%, 73.0-83.9, 58.1-84.8%, 58.1-89.7% and 79.0-95.0%, respectively. As distinctive amino acid residues constituting various motifs and ligand-binding sites and cysteine residues were highly conserved in ferret inflammatory cytokine proteins, ferret cytokines may have fundamentally similar functions to those of other mammals. Phylogenetic analyses based on the deduced amino acid sequences revealed that all ferret inflammatory cytokines were more closely related to those of the Carnivora order, specifically dog and cat, than to other species.
The prokinetic effect of the 5-HT4 receptor agonist mosapride was evaluated in seven healthy thoroughbreds. Mosapride was orally administerd at doses of 0.5, 1.0 or 1.5 mg/kg. The breath 13CO2/12CO2 rate (Δ13CO2), an indirect indicator for the rate of gastric empting, was measured at appropriate points for 4 hr after drug administration. There was a significant increase compared with the control value at 15, 20 and 165 min for 0.5 mg/kg, 30 min at 1.0 mg/kg and 165 min for 1.5 mg/kg. The results suggest that mosapride may facilitate the gastric emptying in horses.
Interferon (IFN)-γ is essential but not sufficient to control leishmaniasis. It is known that IFN-γ is one of the major macrophage-activating cytokines, and the activated macrophages are a principal source of interleukin (IL)-12, which induces autocrine macrophage activation. In this study, the combined effect of IFN-γ and IL-12 on the susceptibility of macrophages to Leishmania major infection was evaluated. Macrophages pretreated with IFN-γ and/or IL-12 were infected with the parasites. Four hr post-infection (p.i.), the levels of infection and parasite load in the macrophages treated with the combination of IFN-γ and IL-12 (IFN-γ/IL-12) were significantly lower than those in the nontreated cells. However, the macrophages treated with either IFN-γ or IL-12 did not show resistance to L. major infection. In addition, 72 hr p.i., the IFN-γ/IL-12-treated and IFN-γ-treated macrophages showed significantly lower levels of infection and parasite load than the nontreated cells, and higher levels of resistance was observed in the IFN-γ/IL-12-treated macrophages than in the IFN-γ-treated macrophages. Although IFN-γ/IL-12 treatment of macrophages prior to the infection led to the induction of resistance, as described above, this resistance was not induced when these cytokines and the parasites were added simultaneously to the macrophage culture. These results suggest that IFN-γ/IL-12 treatment prior to the infection restricts the early phase of the infection.
In order to investigate the toxic effects of 6-mercaptopurine (6-MP) on placental development, we examined sequential morphology in the placentas from rats exposed to 6-MP. 6-MP was intraperitoneally administered at 60 mg/kg during gestation days (GDs) 11 to 12, and the placentas were sampled on GD 13, 15 or 21. In the 6-MP-treated group, maternal body weight suppression, increased death embryo/fetus ratio and some malformations were observed. The placenta weights were decreased on GDs 15 and 21. Macroscopically, placentas on GD 21 were small, brittle and thin with a white peripheral rim. Histopathologically, in the labyrinth zone, 6-MP treatment mainly evoked decreased mitosis on GDs 13 and 15, increased apoptotic cell on GDs 13, 15 and 21 and thinning on GDs 15 and 21. In the basal zone, 6-MP evoked decreased mitosis on GDs 13, and PAS-positive material in the spongiotrophoblasts was still detected on GD 15. Thickening of the basal zone was observed with cytolysis of glycogen cells, apoptosis and an increased number of composed cells on GD 21. In conclusion, 6-MP administration in pregnant rats induced growth arrest of the labyrinth zone and developmental delay in the basal zone, leading to small placentas. The fetotoxicity of 6-MP may be responsible for its direct anti-proliferative effects and resulting placental dysfunction.
Many small wild birds died in the 2005-2006 wintertime in Hokkaido. Thirteen birds were pathologically examined and it was attempted to detect West Nile and influenza viruses from their organs. Consecutive pathological changes were fresh hemorrhage and acute circulatory failure. Viral detections were negative. Selective occurrence in wintertime, literature review and the results of pathological and virological examinations suggested chemical deicer poisoning as the cause of wild bird death. Chicks treated orally with deicer showed acute death and their pathological changes were similar to those of the wild birds. Because the chicks showed significant elevation of plasma Na concentration, plasma electrolyte analysis of the affected wild birds might be crucial to confirm our tentative diagnosis.
A 3-year-old, female Spitz, was presented due to lack of response to therapies with a 6-month history of skin lesions characterized by diffuse erythema and scaling on the dorsal trunk. Physical examination revealed the dog was active and healthy. Skin culture isolated no fungus. Histological examination of skin biopsy specimens revealed interface dermatitis with hydropic degeneration of the basal layers, predominant plasmacytic perivascular accumulation in the dermis, and intensive plasma cell-rich interface mural folliculitis. Moderate CD3-positive lymphocytes infiltrated the superficial dermis. This report may provide unique information of canine discoid lupus erythematosus in an unusual breed with atypical cutaneous lesions.
The aim of the present study was to estimate whether rat sense exogenous electric field (EF) including one used in our previous studies. Employing a conditioned place aversion response paradigm based on an aversive behavior against light environment, alteration in both voluntary behavior of Wistar rat to a 50 Hz sinusoidal EF was examined. Following conditioning without EF, the times spent in white place in rats was significantly shortened (P<0.05). While, such changes were not shown in rats conditioned with EF. Thus, it was considered that the aversion response to light environment was interfered by exposure to EF. An interference in recognition of brightness via EF induced effect to visual system or in learning system via direct effect to central nerve system was considerable as a factor for EF-induced effect. In addition, it was remained that rat possibly sense exposure to EF as preferable. In order to confirm which factor functioned, further studies are needed.
Ubiquitin C-terminal hydrolase L1 is a component of the ubiquitin proteasome system, which evidences unique biological activities. In this study, we report the pattern of UCH-L1 expression, and show that it regulates bone mineralization in osteogenesis. UCH-L1 was expressed in osteoblasts, osteoclasts, and hematopoietic precursor cells of bone marrow in the metaphysis and diaphysis of the femora. To further assess the involvement of UCH-L1 in the regulation of bone mineralization, we evaluated the bone mineral density (BMD) rate of gad mice, using the Latheta computed tomography system. Male gad mice evidenced a significantly decreased BMD rate in the metaphysis and diaphysis of the femora. These findings of decreased BMD rate in the bones of gad mice may suggest that UCH-L1 function regulates bone mineralization during osteogenesis.
We attempted to accumulate the basic data for evaluation of activated lymphocyte therapy for small animal medicine. The peripheral blood mononuclear cells (PBMCs) collected from healthy dogs were activated using anti-CD3 antibody and human recombinant (hr) interleukin (IL)-2 and reactivated using hr interferon (IFN)-α and hr IL-2. The property of obtained cells was compared with PBMCs. The number of cells was shown to have increased approximately > 50 -fold by cultivation. The proportion of CD8+ cells was significantly increased, the cytotoxicity of the cultured cells was revealed to have been reinforced. Additionally, CD56 mRNA levels tended to have increased. The cells obtained by this method were confirmed to be activated lymphocytes. Furthermore, we investigated the effects of sequential administration of the obtained cells to healthy dogs. By sequential administration of the activated lymphocytes, the cell proliferative activity, proportion of CD4+ cells and CD8+ cells, and serum IFN-γ concentration were shown to have increased, and no severe adverse effects were observed. Consequently, activated lymphocytes could be induced using anti-CD3 antibody and IL-2 in healthy dogs, and sequential administration of activated lymphocytes reinforced the recipient's immunity.
A male six-year-old Pomeranian showed recurrent seizures and progressive left hemiparesis. MRI revealed a mass in the right paramedian frontal-temporal region with hyperintensity on T1-weighted (T1W) and mixed-intensity on T2-weighted (T2W) images. After gadolinium enhancement, the mass was enhanced homogenously and demonstrated the dural tail sign. Surgical resection of mass was performed and its histological diagnosis was meningioma. The meningioma of this case had a high cellular density with some lipid contents and intra- and extra-tumor hemorrhage, however, calcification was not found. These cellular characteristics may contribute to a higher signal intensity on T1W imaging.
We report a liposarcoma at the mesojejunoileum that was widely excised, and then the intrahepatic metastatic liposarcoma was found. The dog was treated by two single-agent chemotherapy separately. The patient was an 8-year-old, male Dachshund, and was referred to our hospital owing to the significant distention of the left abdomen. Neither radiography nor ultrasonography detected the actual association of the tumor with the abdominal viscera before surgery. A large-sized tumor mass that adhered to the mesojejunoileum was explored by laparatomy. Final diagnosis of a pleomorphic liposarcoma was made by the routine histology and further identified by Oil Red O stain. The two single-agent chemotherapy including doxorubicin followed by capecitabine, however, failed to cause remission of the intrahepatic metastatic lesions. The patient died 22 months after operation and the cause of death was supposed to be metastatic liposarcoma.
A nine-month-old domestic short haired cat was admitted with the history of acute vomiting, depression and shivering. Abdominal ultrasonography revealed minimum enlargement of the right uterine horn filled with anechoic fluid. On excretory urography, functionally and anatomically normal, enlarged left kidney was found, but right kidney was absent. It was preliminary diagnosed as hydrometra with right renal agenesis. Aiming at the correction of hydrometra, we performed ovariohysterectomy. During spaying, we found a missing segment of distal part of the right uterine horn and absence of ipsilateral kidney and ureter. Compressed uterine structure and segmental aplasia of right uterine horn were found in histopathological investigation. Taken together, it was diagnosed as a segmental aplasia of uterine horn with ipsilateral renal agenesis.
As well as H5 highly pathogenic avian influenza viruses (HPAIV), H7 HPAIV strains have caused serious damages in poultry industries worldwide. Cases of bird-to-human transmission of H7 HPAIV have also been reported . On the outbreak of avian influenza, rapid diagnosis is critical not only for the control of HPAI but also for human health. In the present study, a rapid diagnosis kit based on immunochromatography for the detection of H7 hemagglutinin (HA) antigen of influenza A virus was developed using 2 monoclonal antibodies that recognize different epitopes on the H7 HAs. The kit detected each of the tested 15 H7 influenza virus strains and did not react with influenza A viruses of the other subtypes than H7 or other avian viral and bacterial pathogens. The kit detected H7 HA antigen in the swabs and tissue homogenates of the chickens experimentally infected with HPAIV strain A/chicken/Netherlands/2586/03 (H7N7). The results indicate that the present kit is specific and sensitive enough for the diagnosis of HPAI caused by H7 viruses, thus, recommended for the field application as a pen-site test kit.
Infectious tracheobronchitis (ITB), also known as the kennel cough, is a respiratory syndrome of dogs and usually appears to be contagious among dogs housed in groups. Etiologic agent of ITB is multiple and sometimes complex. In the present study, 68 household dogs showing clinical signs of respiratory infection were examined, and 20 dogs (29.4%) were found to be positive for either of following agents. Bordetella bronchiseptica (B.b.) was most frequently detected from nasal and oropharynx sites of 7 dogs (10.3%). Among the viruses examined, canine parainfluenza virus (CPIV) was detected with the highest frequency (7.4%). Other pathogens included in the order of frequency group 1 canine coronavirus (4.4%), canine adenovirus type 2 (2.9%), group 2 canine respiratory coronavirus (1.5%), and canine distemper virus (1.5%). Only 2 cases showed mixed infections. Neither influenza A virus nor canine bocavirus (minute virus of canines) was found in any dogs examined. These results indicate that both B.b. and CPIV are likely to be the principal etiologic agents of canine ITB in Japan, and they may be considered as the target for prophylaxis by vaccination.
Bovine viral diarrhea virus 2 (BVDV-2) strains demonstrated in cattle, sheep, and adventitious contaminants of biological products have been evaluated by the palindromic nucleotide substitutions (PNS) method at the three variable loci (V1, V2 and V3) in the 5' untranslated region (UTR), to determine their taxonomical status. Variation in conserved genomic sequences was used as parameter for epidemiological evaluation of the species in relation with geographical distribution, animal host and virulence. Four genotypes, BVDV-2a, BVDV-2b, BVDV-2c, and BVDV-2d have been identified within the species. Taxonomical segregation corresponded to geographical distribution of genotype variants. Genotype 2a was present worldwide, and was the only circulating also in sheep, in addition to cattle. Genotypes 2b, 2c and 2d were restricted to South America. Contamination of biological products was related to genotypes 2a and 2d. Genetic variation could be related with chronological diffusion of the BVDV-2 species variants in different geographic areas. Chronologically, the species emerged in North America in 1978, spreading in UK and Japan, continental Europe, South America and New Zealand. Correlation between clinical features related with isolation of BVDV-2 strains and genetic variation indicated that subgenotype 1, variant 4 of genotype 2a was related with hemorrhagic syndrome. These observations suggest that evaluation of genomic secondary structure, by identifying markers for expression of virus biological activities and species evolutionary history, may be applied as useful tool for epidemiological evaluation of the BVDV-2 species, and possibly for other species of the genus Pestivirus.
Porcine circovirus type 2 (PCV2) has been recognized as the causal agent of postweaning multisystemic wasting syndrome and can be divided into two major genotypic groups. We developed a method of restriction fragment length polymorphism (RFLP) analysis of PCV2 open reading frame 2 for easy discrimination between the two major groups. Genotyping of PCV2 isolates from 10 Japanese commercial pig farms was performed, and the analysis revealed that both PCV2 groups and at least five RFLP types of PCV2 are prevalent in Japan. On two farms, the genotypes of the PCV2 isolates in the spring of 2007 were different from those in the autumn of 2006. One genotype may have become dominant within only six months on these farms.
On August 2007, we encountered equine influenza epidemic by Florida sub-lineage strain (H3N8) in Japan Racing Association's facilities where 4142 racehorses in total were stabled. The number of new febrile cases sharply increased, but the occurrence was rapidly calmed down within 2 weeks. The morbidity rate in these facilities was 12.8% and the subclinical infection rate of healthy racehorses examined by rapid antigen detection tests was 19.4% at the early stage of epidemic. The serological studies along with the low morbidity rate and the existence of numbers of asymptomatically infected racehorses, suggested that the cross-reactivity of the antibodies provided by vaccination against the epidemic strain contributed to reducing the morbidity rate and duration of epidemic.