Since the first identification of bovine spongiform encephalopathy (BSE) in Japan in September 2001, a series of safety measures was introduced by the Ministry of Agriculture, Forestry and Fisheries, the Ministry of Health, Labour and Welfare and the Food Safety Commission of the Cabinet Office. These measures included blanket BSE testing and removal of specified risk materials at slaughterhouses, surveillance of risk animals and a ban on the use of meat-and-bone meals and traceability on all farms. The Japanese experience over the past five years has shed light on several issues in countries that have a low BSE incidence.

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The present study was designed to evaluate whether ethanol suppresses survival-signaling pathways in rat testes. Ethanol (1.5 g/kg or 3 g/kg i.p., 15% v/v in saline) was administrated to adult male rats for 10 days. Ethanol treatment significantly increased the number of TUNEL-positive cells in rat testes. Potential activation was measured by phosphorylation of Akt and Erk1/2 using Western blot analysis. Ethanol decreased the levels of activated survival kinases, pAkt and pErk1/2. The phosphorylation of Bad at Ser112 and Ser136 was decreased in ethanol-treated animals in comparison to saline-treated animals. Moreover, the interaction of pBad with 14-3-3 was decreased by ethanol exposure. In conclusion, our findings suggest that ethanol induces apoptotic cell death by suppressing the activation of survival kinases and the phosphorylation of their downstream targets in rat testes.

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Neural transplantation is one of the most promising treatments for neurodegenerative disorders. Survival rates of embryonic dopamine (DA) neurons following transplantation are low, between 2% and 20% in a number of animal models. To further establish survival changes of the transplanted gestational day 13.5 ventral mesencephalic (VM) cells into left intact adult rat striata so that design strategies of increasing survival of DA neurons, the tyrosine hydroxylase (TH) expression of VM-derived progenitor cells has been examined using immunohistochemistry and Western blot analysis. TH immunostaining revealed that the grafted VM cells developed to mature TH-positive neurons strongly at 3 weeks, peaked at 4 weeks, thereafter, gradually dropped following the degenerative expression of the grafted cells at both 5 and 6 weeks after transplantation. Western blot analysis also showed that the TH proteins were maximally expressed at 4 weeks post-grafting. Our finding suggested that the peak of surviving VM-derived TH positive cells occurred approximately 4 weeks after transplantation.

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This research assesses the relative contribution of splenic contraction and fluid shifts out of the vascular compartment to the increases in packed cell volume associated with Agility exercises. It also aims to evaluate the changes in the concentrations of electrolytes and markers of hydration state. Fifteen dogs of both sexes were subjected to an Agility exercise of an approximate duration of 100 s. Blood samples were obtained within the first 30 s after competition and at 5, 15, and 30 min of recuperation. Resting values were established previously. The following parameters were determined: packed cell volume (PCV), plasma lactate (LA), total plasma protein (TPP), albumin (ALB), urea (BUN), creatinine (CREA), chloride (Cl), calcium (Ca), phosphorus (P), sodium (Na) and potassium (K). Changes in plasma volume (PV), total RBC volume (V_RBC) and blood volume (BV) were calculated immediately after exercise and at 30 minutes of recovery. It was found that during Agility competition, BV, V_RBC and PV increased 12, 21 and 4% respectively, indicating that the spleen contraction was the main determinant on the increase of BV. In comparison with resting values, BV decreased after recuperation (-5%), due to the recapture of erythrocytes by the splenic reserve (V_RBC, -12%). Additionally, Agility exercise induced increases in plasma Cl and LA, with significant reductions of ALB, Ca and P and absence of modifications in Na, K, BUN and CREA concentrations.

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Lipid extracted from the ovary of skipjack tuna by the method that we developed is rich in phospholipid-type docosahexaenoic acid. The ovary lipid of skipjack tuna (OLS) was studied for its anti-stress activity in male Wistar rats, focusing on stress-related blood components: recovery from stress was examined after application of water immersion restraint stress. As a result, serum corticosterone (CORT) secretion was inhibited and decreased rapidly after stress application in rats given OLS compared with control rats. As CORT acts as a glucocorticoid, non-esterified fatty acid (NEFA) is expected to increase by stress application. However, the concentration tended to be lower in rats given OLS than in control rats. With respect to OLS concentration, OLS increased serum dehydroepiandrosterone, secretion concentration-dependently. In addition, as with the recovery study, it tended to inhibit the increase in NEFA. These results indicate that OLS may have an anti-stress activity against acute stress.

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We cloned a cDNA fragment encoding a feline homologue of L-selectin (CD62L). The extracellular region of the feline CD62L fragment contained a calcium-dependent (C-type) lectin domain, an epidermal growth factor-like domain, and two Sushi/CCP/SCR domains. The flow cytometric analysis confirmed that the feline CD62L molecule, which was expressed 293T cells, retained an epitope recognized by an anti-human CD62L monoclonal antibody (Leu-8).

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The antiviral effects of recombinant bovine interferon-τ (rboIFN-τ) on bovine leukemia virus (BLV) were examined in vitro and in vivo. In the in vitro experiments, BLV titers decreased in FLK-BLV cells and in peripheral blood mononuclear cells of BLV-infected cattle treated with rboIFN-τ at a concentration higher than 10² U/ml. In order to examine the in vivo effects of rboIFN-τ, 10 BLV-infected cattle were subcutaneously injected with rboIFN-τ. In the first experiment, 6 cows were administrated with 10⁵ U/kg body weight of rboIFN-τ 3 times per week for 4 weeks, while in the second experiment 4 cows were administrated with 10⁶ U/kg body weight of rboIFN-τ 3 times per week for 3 weeks. No adverse effects were observed after the administration of rboIFN-τ. In experiment No. 1, the mean BLV titers in cattle decreased in the post- rboIFN-τ administration period compared to the pre- rboIFN-τ administration period. In experiment No. 2, the mean BLV titers in cattle decreased in the rboIFN-τ administration period. These results suggest that rboIFN-τ decreases BLV titers in vitro and in vivo and that rboIFN-τ possibly reduces the degree of BLV titer in cattle without severe side effects.

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To evaluate the energy condition of cattle with growth retardation, propionate (PTT) and arginine tolerance tests (ATT) were carried out. The insulin/glucagon concentration ratio immediately before PTT or ATT in the cattle with growth retardation was lower than in the control. In the growth-retarded cattle, insulin-AUC_0-120min during PTT was lower than in the control, while glucagon- AUC_0-120min was the same as in the control. Insulin-AUC_0-120min during ATT in the cattle with growth retardation tended to be lower than in the control, whereas glucagon-AUC_0-120min was the same. Therefore, insulin-AUC_0-120min/glucagon-AUC_0-120min in the cattle with growth retardation was lower than in the control during both tolerance tests. The growth-retarded cattle showed lower insulin/glucagon ratio similar to that found in starved and lactating cattle, a suggesting lack of energy.

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A 5-month-old female Korean Sapsaree dog was presented with severe ascites, cyanosis, respiratory difficulty and exercise intolerance. Diagnostic imaging studies revealed a dextropositioned and over-riding aorta, pulmonary valvular stenosis, ventricular and atrial septal defects, and right ventricular hypertrophy. Based on these findings, the dog was diagnosed as a case of tetralogy of Fallot with atrial septal defect (pentalogy of Fallot). The dog was medically managed by use of diuretics and vasodilators and an occasional phlebotomy.

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Eight species of Japanese birds were found to be infected with Leucocytozoon species using microscopic analysis. We used PCR and sequence analysis of the mitochondrial cytochrome b gene (cyt b) to compare the genetic background among these detected protozoa species. In 20 individuals of 22 samples, a single amplified band was detected from 6 of 8 bird species; 9 Japanese rock ptarmigans (Lagopus mutus japonicus), 4 large-billed crows (Corvus macrorhynchos), 2 carrion crows (C. corone), 2 scops owls (Otus scops), 1 Japanese grosbeak (Eophona personata), and 2 brown-eared bulbuls (Hypsipetes amaurotis), respectively. Phylogenetic analysis based on the partial cyt b sequences revealed that all Leucocytozoon isolates in Japan closely grouped with other Leucocytozoon species previously reported in the literature. Among the Japanese isolates, the phylogenetic tree suggested that L. lovati from the Japanese rock ptarmigan may be basal to the parasites found in other bird species. Our study is the first to identify the molecular relationships among Leucocytozoon parasites in the avifauna of Japan.

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We identified two caspase-like genes from the midgut cDNA library of the hard tick, Haemaphysalis longicornis. Sequence analysis showed that these cDNAs encoded homologues of caspase-2 and caspase-8 that were categorized as apoptosis initiators. The H. longicornis caspase-2 (Hlcaspase-2) cDNA encodes 340 amino acid residues with a predicted molecular weight (Mw) of 38.5 kDa. Another cDNA identified as the H. longicornis caspase-8 (Hlcaspase-8) encodes 306 amino acid residues with an estimated Mw of 35.3 kDa. A catalytic active site was highly conserved in Hlcaspase-8 but not in Hlcaspase-2. RT-PCR analysis showed that both Hlcaspase-2 and Hlcaspase-8 were expressed in tick midgut and salivary glands. This is the first report of the molecular cloning of apoptosis-related genes in the tick.

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Nerve growth factor, retinoic acid, dibutyryl cAMP, ganglioside G_Q1b, and botulinum C3 exoenzyme were evaluated for their neural differentiating potential on human neuroblastoma GOTO cells. C3 exoenzyme is an ADP-ribosyltransferase inactivating Rho protein (a small GTP-binding protein). C3 exoenzyme caused the fastest differentiation of GOTO cells into neural cells and induced the strongest network of the cells. Fasudil, an inhibitor of Rho-kinase, induced outgrowth of the neurites in the GOTO cells. Calyculin A, an inhibitor of phosphatases including myosin phosphatase, counteracted C3 exoenzyme-induced neurite outgrowth of the cells. These findings suggest that differentiation of GOTO cells triggered by C3 exoenzyme is attained via inactivation of Rho protein, inhibition of Rho-kinase, and activation of myosin phosphatase. Because of the strong differentiating potential of C3 exoenzyme, the transduction pathway consisting of Rho protein, Rho-kinase, and myosin phosphatase seems to be main stream in the neural differentiation of GOTO cells. A single GOTO cell was observed continuously after treatment with C3 exoenzyme. The cell started to change shape from its original morphology only 15 min after treatment with C3 exoenzyme, and it was completely spherical within 60 min. Neurites appeared on the membrane of the cell 2 hr after the treatment and then gradual outgrowth began. These observations are fundamental information in elucidating the mechanism of neural differentiation, especially at an early stage.

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The anesthetic and cardiopulmonary effects of midazolam, ketamine and medetomidine for total intravenous anesthesia (MKM-TIVA) were evaluated in 14 horses. Horses were administered medetomidine 5 μg/kg intravenously as pre-anesthetic medication and anesthetized with an intravenous injection of ketamine 2.5 mg/kg and midazolam 0.04 mg/kg followed by the infusion of MKM-drug combination (midazolam 0.8 mg/ml-ketamine 40 mg/ml-medetomidine 0.1 mg/ml). Nine stallions (3 thoroughbred and 6 draft horses) were castrated during infusion of MKM-drug combination. The average duration of anesthesia was 38 ± 8 min and infusion rate of MKM-drug combination was 0.091 ± 0.021 ml/kg/hr. Time to standing after discontinuing MKM-TIVA was 33 ± 13 min. The quality of recovery from anesthesia was satisfactory in 3 horses and good in 6 horses. An additional 5 healthy thoroughbred horses were anesthetized with MKM- TIVA in order to assess cardiopulmonary effects. These 5 horses were anesthetized for 60 min and administered MKM-drug combination at 0.1 ml/kg/hr. Cardiac output and cardiac index decreased to 70-80%, stroke volume increased to 110% and systemic vascular resistance increased to 130% of baseline value. The partial pressure of arterial blood carbon dioxide was maintained at approximately 50 mmHg while the arterial partial pressure of oxygen pressure decreased to 50-60 mmHg. MKM-TIVA provides clinically acceptable general anesthesia with mild cardiopulmonary depression in horses. Inspired air should be supplemented with oxygen to prevent hypoxemia during MKM-TIVA.

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Minimally invasive cardiac output was determined using transthoracic bioimpedance (BICO), partial carbon dioxide rebreathing (NICO) and transesophageal Doppler echocardiography (TEECO) and compared to thermodilution (TDCO) in 6 beagle dogs. The dogs were 2 years old, weigh between 9.1-13.0 kg and were anesthetized with nitrous oxide-oxygen-sevoflurane. All dogs were administered a neuromuscular blocking drug and artificially ventilated during anesthesia. Thirty paired measurements of TDCO and each non-invasive method were collected during low, intermediate, and high values of cardiac output achieved by varying the depth of anesthesia and the administration of dobutamine. Cardiac output values ranged from 1.10-2.50 L/min for BICO compared to 0.81-4.88 L/min for TDCO; 0.70-2.60 L/min for NICO compared to 0.89-4.45 L/min for TDCO; and 0.59-4.37 L/min for TEECO compared to 0.57-4.15 L/min for TDCO. The limits of agreement and percentage error were -0.58 ± 1.56 L/min and ± 75.4% for BICO, -1.04 ± 1.08 L/min and ± 56.0% for NICO, and 0.03 ± 0.26 L/min and ± 12.3% for TEECO compared to TDCO. In conclusion, TEECO provided the best agreement to TDCO in sevoflurane anesthetized beagle dogs.

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An 11-year-old male Golden Retriever presented with progressive weight loss, tachycardia, hyperthermia, polyuria and polydipsia. A freely movable mass, 4.5 × 4 cm in size, was palpated at the cranioventral cervical region. Hormonal study revealed high levels of serum thyroid hormones, and a tentative diagnosis of hyperthyroidism due to a thyroid tumor was made. The tumor was removed surgically and diagnosed histopathologically as thyroid gland adenoma. Serum thyroid hormone levels decreased after surgery with improved clinical signs. At 12 months after surgery, the dog maintained a good physical condition with no evidence of recurrence.

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Three dogs had a diagnosis of maxillofacial rhabdomyosarcoma. These dogs were treated with surgery and/or radiotherapy, and had poor clinical responses. The tumor tissues in all three cases were observed around the upper premolar teeth with ulcerative lesions and CT examinations in each case revealed extensive bony involvement into the maxilla. Two cases were subjected to surgical excision of the tissues, followed by an external radiation therapy. The other case was only treated with palliative radiation. Outcomes of the treatment of all the cases were quite poor because of the invasive and refractory nature of the tumor cells, leading to the local recurrence and lung metastasis early in the clinical course. All dogs died within two months of the first admission.

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Increase in circulating vascular endothelial growth factor (VEGF) is suggested as a prognostic indicator in human patients with malignant tumors. The purpose of this study was to evaluate the clinical significance of circulating VEGF in dogs with mammary gland tumors (MGT). Both plasma and serum VEGF were significantly higher in dogs with MGT when compared with those in the healthy dogs. In dogs with MGT, the plasma and serum VEGF of the malignant group increased significantly compared with those of the benign group. Additionally, there was a significant difference between the plasma and serum VEGF in the groups with postoperative metastasis and no metastasis. Circulating VEGF is expected to be clinically available for the determination of prognosis in canine MGT.

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This report concerns a case of pancreatic carcinoma with widespread metastases to many organs including intracranial metastasis. An eleven-year-old, male, mixed-breed dog showed emaciation, ataxia, and multiple visible tumors within the neck. A MRI examination of the patient was conducted because of ataxia, and it was found that the intracranial invasive growth had resulted in compression of the brain stem. Necropsy was performed after the patient died. Based on gross and microscopic examination, the primary tumor cells were located in the left lobe of the pancreas and widespread metastasis was found into various organs, including the brain, lungs, liver, kidneys, tonsils, serosal surface of the esophagus, and submandibular, pulmonary hilar, mediastinal, and mesenteric lymph nodes. This case indicates that pancreatic adenocarcinoma should be included in the differential diagnosis list when cervical neck masses are detected.

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