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An ocular mass was noted in the right iridociliary region of a 13-year-old male beagle dog. The right eyeball was surgically removed. Histopathologically, the mass was located mainly in the ciliary body and appeared to be continuous with non-pigmented epithelial cells. The polygonal neoplastic cells proliferated in sheets with periodic acid-Schiff-positive basement membrane material. Neoplastic cells had small round nuclei with no atypia or mitosis, and abundant, pale eosinophilic, granular, and rarified cytoplasm displayed ballooning change. No invasive growth or metastasis was observed. Immunohistochemically, the cells were positive for vimentin, neuron-specific enolase, and S100, and negative for cytokeratin AE1/AE3, Iba-1, CD204, MelanA, Sox10, and PNL2. Based on these findings, the lesion was diagnosed as an iridociliary adenoma with ballooning change.

The control of digital dermatitis (DD) among cattle is crucial; however, effective and environmentally-sound control measures have yet to be identified. From the monitoring data of DD which were recorded during regular hoof trimmings in a farm in Hokkaido, Japan, we detected a decrease in the DD prevalence in a herd where an anaerobic bacterial fermentation enhancer (ABFE) was distributed. The possible effect of ABFE was analyzed using a retrospective repeated cross-sectional design. The prevalence of DD decreased over time in the ABFE-distributed group. Furthermore, a selected regression model indicated the time-dependent enhancement of the decreasing trend. While potential coincidental factors may influence, this study provides a basis for further research on the preventive effect of ABFE against DD.

We present the report of trismus due to hyperadrenocorticism-associated myotonia diagnosed by electromyography in a dog. An intact female Miniature Dachshund, 13 years and 9 months old, presented with stiff gait and trismus as well as polyuria and polydipsia. Abdominal ultrasonography showed enlarged adrenal glands. An adrenocorticotropic hormone stimulation test revealed an exaggerated response. Based on these findings, this case was diagnosed with hyperadrenocorticism. Electromyography revealed myotonic discharge in the temporalis muscle and limbs. Therefore, trismus was considered to be caused by hyperadrenocorticism-associated myotonia, and the case was treated with oral trilostane (1.3 mg/kg, once daily). During the 4-month follow-up period, despite the partial improvement in stiff gait, trismus did not recover. Long-term data on more cases are warranted to assess the prognosis and clinical characteristics of trismus due to hyperadrenocorticism-associated myotonia.

We investigated the prevalence of porcine circovirus 2 (PCV2) and 3 (PCV3) in wild boars captured or found dead in Nagano Prefecture in 2020. Based on PCR testing, 21 of 254 (8.3%) wild boars were PCV2-positive and 43 of 256 (16.8%) wild boars were PCV3-positive, 5 of 253 (2.0%) wild boars were both PCV2-positive and PCV3-positive. The frequency of detecting PCV3 in wild boars was significantly higher in adults than in juveniles (P=0.014). The PCV2-positive wild boars were found in all districts except for the North Alps and Hokushin, while PCV3-positive wild boars were found in all districts except for Suwa. This is the first report of PCV2 and PCV3 detected in wild boars in Japan.

Although feline idiopathic cystitis (FIC) distresses of many cats, its pathogenesis is unknown and the diagnosis is challenging. Polyunsaturated fatty acids (PUFAs) are metabolized into various lipid mediators. Lipid mediators such as prostaglandins (PGs) modulate inflammation and many of them are excreted into the urine. Thus, the investigation of the urinary lipid profile may reveal pathogenesis and help diagnosis of FIC. We collected urine samples from five FIC cats by spontaneous urination and analyzed 158 types of lipid mediators in urines using liquid chromatography-mass spectrometry. The urinary levels of PUFAs were higher in FIC compared to those of the healthy group. The excretions of a major inflammatory mediator, PGD 2 , were less in FIC. Other well-known inflammatory mediators such as PGE 2 , PGI 2 , and their metabolites did not show a difference. In contrast, the levels of PGF 2α and its 2 metabolites and PGF 3α were higher in FIC. These results may provide new insights into the future management of cat FIC.

Since gamma-glutamyl transpeptidase (GGT) is highly and locally expressed in human breast cancer, a GGT-enzymatically activatable fluorescent probe, gamma-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG), has been developed to detect the human breast cancer area with high performance. In this study, GGT expression and the efficacy of gGlu-HMRG on visualization were investigated in canine mammary gland tumors (MGT). Seventeen non-fixed fresh-frozen MGT specimens and each peritumoral control tissue were utilized. The GGT mRNA levels were highly observed in the tumor specimens compared with the control. GGT immunostaining was mostly observed on the cell membrane and cytosol of the alveolar and duct mammary epithelium of MGT tissues. These signals were strongly positive in several cases while they were mild to not observed in other cases. When gGlu-HMRG solution was dropped to the non-fixed tissue pieces of MGT or control tissues, the fluorescence intensities (FIs) were measured using Maestro in-vivo imaging device. FIs in MGT tissues were significantly higher than each control tissue 20 min after treatment. Based on Youden index method, the maximum sensitivity and specificity of FI was 82.4% and 82.4%. These findings suggest that GGT is highly expressed in several MGTs in dogs and gGlu-HMRG could visualize at least a part of MGT tissues in dogs. Nevertheless, it should be needed to assess the false-negative areas more carefully in canine than human cases.

The present study aimed to quantitatively evaluate muscle mass and gene expression in dogs with glucocorticoid-induced muscle atrophy. Five healthy beagles received oral prednisolone for 4 weeks (1 mg/kg/day), and muscle mass was then evaluated via computed tomography. Histological and gene expression analyses were performed using biopsy samples from the biceps femoris before and after prednisolone administration. The cross-sectional area of the third lumbar paraspinal and mid-femoral muscles significantly decreased after glucocorticoid administration (from 27.5 ± 1.9 to 22.6 ± 2.0 cm 2 and from 55.1 ± 4.7 to 50.7 ± 4.1 cm 2 , respectively; P<0.01). The fast- and slow-twitch muscle fibers were both atrophied (from 2,779 ± 369 to 1,581 ± 207 μm 2 and from 2,871 ± 211 to 1,971 ± 169 μm 2 , respectively; P<0.05). The expression of the growth factor receptor-bound protein 10 (GRB10) significantly increased after prednisolone administration (P<0.05). Because GRB10 suppresses insulin signaling and the subsequent mammalian target of rapamycin complex 1 activity, increased expression of GRB10 may have resulted in a decrease in protein anabolism. Taken together, 1 mg/kg/day oral prednisolone for 4 weeks induced significant muscle atrophy in dogs, and GRB10 might participate in the pathology of glucocorticoid-induced muscle atrophy in canines.

Bacterial cystitis is one of the feline lower urinary tract diseases (FLUTDs). Polyunsaturated fatty acids, such as arachidonic acid (ARA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), are oxidized into various lipid mediators that modulate inflammation. Since the profile of lipid metabolites excreted in urine is useful for assessing inflammatory body conditions, we analyzed 126 types of urinary lipid metabolites in cats with bacterial cystitis. Using LC-MS/MS, we found that the levels of 11 metabolites were higher in the urine of cystitis cats than in the urine of healthy cats. In detail, the urinary levels of ARA, EPA, and DHA and eight of their metabolites were increased in cystitis cats. Focusing on the lipid oxidation pathway, the urinary levels of four cyclooxygenase-, three lipoxygenase-, and one cytochrome P450-dependent oxidated metabolites were increased in bacterial cystitis. These urinary lipid profiles can provide some insight into the pathology and future diagnosis of bacterial cystitis.

Liver-type fatty acid-binding protein (L-FABP) is a biomarker for the early detection of renal diseases in humans. L-FABP is a cytotoxic oxidation product secreted from the proximal tubules under ischemic and oxidative stress conditions. First, L-FABP gene expression in the kidney and liver was evaluated. Next, the urinary L-FABP concentrations in dogs with or without renal diseases were measured using a novel enzyme-linked immunosorbent assay kit. Urinary L-FABP was normalized relative to urinary creatinine (uCre) concentrations (µg/g uCre). Finally, the relationships between urinary L-FABP and renal biomarkers used in canine medicine or serum alanine transaminase (ALT) as an indicator of liver damage were examined. Serum and urine samples from 94 client-owned dogs including 23 dogs with renal diseases and 71 dogs without renal diseases were used for analysis. Relative L-FABP gene expression was confirmed both in the liver and kidney. Dogs with renal diseases had a significantly higher urinary L-FABP than those without, and its predictive cutoff value was 26 µg/g uCre. Urinary L-FABP was significantly correlated with serum creatinine (r=0.4674, P<0.01), urea nitrogen (r=0.4907, P<0.01), urine specific gravity (r=-0.5100, P<0.01), and urine protein/creatinine ratio (r=0.7216, P<0.01), but not with serum ALT. Hence, dogs with a high urinary L-FABP value were more likely to have renal diseases.

Nasal papules and oral ulcers were observed in calves that were group-housed at a dairy farm. The calves were diagnosed with bovine papular stomatitis (BPS) due to parapoxvirus (PPV) infection based on virologic examinations using polymerase chain reaction to detect PPV. To prevent the spread of BPS, we isolated the affected calves, made procedural changes so that the affected herd was managed after the healthy herd, disinfected the bedding with slaked lime, disinfected the stalls and fences with invert soap, and changed the animals' feed to soft grass which does not damage the oral cavity. As a result, we succeeded in control the infection quickly.

The aim of this study was to compare changes in tissue blood flow and tissue oxygen tension in the masseter muscle and mandibular bone marrow induced by remifentanil under desflurane or sevoflurane anesthesia. Eleven male tracheotomized Japan White rabbits were anesthetized with desflurane or sevoflurane under mechanical ventilation. The order of the inhalation of desflurane or sevoflurane was randomized. Desflurane or sevoflurane was administered at 1.0 minimum alveolar concentration and remifentanil was infused at 0.4 µg/kg/min. Observed variables included heart rate (HR), blood pressure (BP), common carotid artery blood flow (CCBF), mandibular bone marrow tissue blood flow (BBF), masseter muscle tissue blood flow (MBF), mandibular bone marrow tissue oxygen tension (PbO 2 ), and masseter muscle tissue oxygen tension (PmO 2 ). Two way repeated measures ANOVA showed no interaction between volatile anesthetics and remifentanil infusion except for MBF. There were significant differences in HR, SBP, DBP, MAP and CCBF between desflurane and sevoflurane groups. There were also significant differences in HR, SBP, DBP, MAP, CCBF, BBF and PbO 2 before, during and after remifentanil infusion. Desflurane reduced tissue blood flow in the masseter muscle and mandibular bone marrow while better maintained HR and BP than sevoflurane. Under remifentanil infusion, although both anesthetics reduced tissue blood flow, tissue oxygen tension was maintained in masseter muscle and mandibular bone marrow.

A 1-year- and 11-month-old spayed female toy poodle had showed progressive ataxia and paresis in the hindlimbs since 11 months old. Magnetic resonance imaging revealed high signal intensity on T2-weighted and fluid-attenuated inversion recovery images at the thoracic and lumbar spinal cord. The dog's neurological condition slowly deteriorated and flaccid tetraparesis was exhibited. At 4 years and 11 months old, the dog died of respiratory failure. On postmortem examination, eosinophilic corkscrew bundles (Rosenthal fibers) were observed mainly in the thoracic and lumbar spinal cord. Histological features were comparable to previously reported cases with Alexander disease. This is a first case report to describe the clinical course and long-term prognosis of a dog with Alexander disease.

Polyunsaturated fatty acids including arachidonic acid (AA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), are converted to lipid mediators by oxidation. Unlike other mammals, cats cannot synthesize AA. Since their lipid metabolic features remain unknown, we qualitatively analyzed 118 types of urinary lipid metabolites in healthy neutered cats. Using LC-MS, we found 26 lipid metabolites in urines of all individuals. In detail, 20 AA-, 5 EPA- and 1 DHA-derived lipid mediators were detected. Focusing on oxidative pathway, 17 cyclooxygenase-metabolites and 5 metabolites produced by non-enzymatic pathway were detected. Of interest, few lipoxygenase- or cytochrome P450-metabolites were excreted. Thus, AA-derived cyclooxygenase-metabolites mainly composed the urinary lipid metabolites in cats.

In our previous studies, by simply inducing burn injuries on bullfrog hearts or partially exposing their surface to high-potassium (K + ) solution, we could reproduce a ST segment elevation in the electrocardiogram (ECG), which is a characteristic finding in human ischemic heart disease. In the present study, using our burn-induced subepicardial injury model, we could additionally reproduce "reciprocal" ST segment changes for the first time in frog hearts, mimicking those observed in human acute myocardial infarction. Immunohistochemistry demonstrated markedly decreased Na + /K + -ATPase protein expression in the ventricular surface after the burn injury. The loss of this pump expression in injured cardiomyocytes was thought to be responsible for the creation of "currents of injury" and the subsequent ST segment changes observed in acute myocardial infarction.

This study evaluated the monitoring methods in asymptomatic dogs with high serum cystatin C (Cys-C) concentrations. Ten dogs with high serum Cys-C were divided into two groups based on the owner's choice; one receiving clinical pathology-based monitoring at an animal hospital specialised in chronic kidney disease, and the other receiving symptom-based monitoring at home, partly because they showed no clinical symptoms. The dogs that received the clinical pathology-based monitoring led to an early treatment intervention, resulted in a longer survival period than dogs received the symptom-based monitoring (P<0.05). It became clear that early treatment intervention by clinical pathology-based monitoring extends the renal survival period even in asymptomatic dogs with increased serum Cys-C concentrations.

We monitored changes in serum leptin, adiponectin, and resistin concentrations in obese cats during weight loss. Six naturally developed obese cats were fed low-fat, high-fiber dry food during a 9-week experimental period. Serum leptin, adiponectin, and resistin concentrations were measured at week 0, 4, 8, and 9. Body weight became significantly lower week 4 onward than that at week 0 (P<0.05 or 0.01). At week 9, serum leptin concentrations were significantly lower than those at week 0 (P<0.05). Contrarily, serum adiponectin and resistin concentrations did not significantly differ within the 9 weeks. While serum leptin levels were strongly positively correlated with body weight (r=0.923, P<0.001), serum adiponectin levels were moderately negatively correlated with it (r=-0.529, P<0.01), with serum resistin having a no correlation with body weight. Serum leptin levels might be more closely related with pathogenesis of adiposity than serum adiponectin or resistin in cats.

The aim of the current study was to identify independent risk factors for thyroid axis alterations in dogs with non-thyroidal diseases. In this retrospective cross-sectional study, data and plasma samples from 207 dogs with non-thyroidal diseases was used. The involvement of various factors (disease severity, sex, age, breed, category and duration of disease, and medication) in the alteration of plasma thyroxine (T4) or thyroid-stimulation hormone (TSH) concentrations was analyzed using multivariate logistic regression. Among the 207 dogs analyzed, 99 (47.8%) had low plasma T4 concentrations, while 45 (21.7%) had high TSH concentrations. Intact male sex [odds ratio (OR), 3.25; 1.67-6.35; P<0.001], Labrador Retrievers (OR, 18.70; 2.32-151.00; P=0.006), moderate (OR, 2.39; 1.21-4.74; P=0.012) and severe diseases (OR, 6.84; 2.27-20.70; P<0.001) were associated with increased risk for low plasma T4 concentrations. Meanwhile, intact male (OR, 3.93; 1.51-10.30; P=0.005), spayed female (OR, 4.22; 1.59-11.20; P=0.004), older age (OR, 2.73; 1.28-5.84; P=0.009), and Miniature Dachshunds (OR, 5.39; 2.38-12.20; P<0.001) had increased risk for high plasma TSH concentrations. Disease severity had been determined as an independent risk factor for canine NTIS. In addition, sex, age and breed were also associated with thyroid axis alterations in dogs with non-thyroidal diseases.

An 8-year-old castrated male cat presented with acute ataxia and paresis in all four limbs. The cat also exhibited signs of autonomic nervous system impairment. Magnetic resonance imaging revealed swelling of the brachial plexuses bilaterally. Despite treatment, the cat died after 10 days of treatment. A postmortem examination revealed swollen radial nerves and cervical nerve roots in which infiltration of inflammatory cells was histologically confirmed. Additionally, lymphocytic infiltration was found around the blood vessels of the sciatic nerve bundle and the vagus nerve. Histological features were comparable to previously reported brachial plexus hypertrophic neuritis in a cat. Our case was unique in that the autonomic nerves were also involved in addition to the somatic nerves in all four limbs.

The safety and biological effects of a long-term dose of D-allulose were evaluated in healthy dogs. For 12 weeks, the dogs were administered D-allulose (0.2 g/kg) or placebo daily. Plasma total cholesterol concentrations in the D-allulose group were significantly lower than those in the control group at and after week 2 (P<0.05). D-Allulose administration did not cause clinical signs or changes in hematological and biochemical levels, except for lipids. D-Allulose administration also did not influence body weight. Plasma glucose and insulin concentrations in the glucose tolerance test, performed one day after the termination of D-allulose administration, were not different between groups, suggesting no cumulative effects of D-allulose on glucose metabolism in healthy dogs. In conclusion, long-term administration of D-allulose caused no harmful effects in dogs.

Canine degenerative myelopathy (DM) is an adult-onset, progressive neurodegenerative disease that occurs in multiple dog breeds. A DM-associated mutation of the canine superoxide dismutase 1 (SOD1) gene, designated as c.118G>A (p.E40K), has been implicated as one of pathogenetic determinants of the disease in many breeds, but it remains to be determined whether the c.118G>A mutation is responsible for development or progression of DM in Collies. Previously, a Rough Collie was diagnosed clinically and histopathologically as having DM in Japan, suggesting the possibility that the Collie breed may be predisposed to DM due to the high frequency of c.118G>A in Japan. In this study, accumulation and aggregate formation of SOD1 protein were retrospectively demonstrated in the spinal cord of the DM-affected dog by immunohistochemical analysis. Furthermore, a molecular epidemiological survey revealed a high carrier rate (27.6%) and mutant allele frequency (0.138) of c.118G>A in a population of Collies in Japan, suggesting that the Collie breed may be predisposed to DM associated with c.118G>A, and the prevention of DM in Collies in Japan should be addressed through epidemiological and genetic testing strategies.

Canine degenerative myelopathy (DM) is characterized by progressive degeneration of the spinal cord. Although atrophic changes in the intercostal muscles were previously reported in the late stage of DM in Pembroke Welsh Corgis (PWCs), changes in respiratory function have not yet been examined. In the present study, we performed an arterial blood gas analysis and measured respiratory movements over progressive disease stages to document changes in respiratory function in DM-affected PWCs. We found that respiratory dysfunction progressed during the later stages of DM and correlated with a change in respiratory movement to the abdominal breathing pattern. These results suggested that hypoventilation occurred due to dysfunctional changes in the intercostal muscles and resulted in hypoxemia in the later stages of DM.

Feline renal diseases are increasingly noted in veterinary practice. It is important to diagnose and identify the pathological basis of renal dysfunction accurately at an early stage, but there are only a few reports on this area in clinical veterinary medicine. We investigated the efficacy of measurement of urinary albumin (u-Alb) and urinary transferrin (u-Tf) for early diagnosis using 5-µl urine samples collected noninvasively by catheterization from normal (IRIS stage I) cats and cats with stage I chronic kidney disease (CKD). The u-Alb levels in normal and stage I CKD cats were 6.0 ± 4.5 and 11.2 ± 8.4 mg/dl, respectively, and the u-Tf levels were 0.09 ± 0.42 and 0.52 ± 0.79 mg/dl, respectively. Based on ROC curve analysis, the sensitivity and specificity of u-Alb and u-Tf were higher than those of the currently used biomarker, the plasma creatinine level. The sensitivity of u-Alb was higher than that of u-Tf, whereas the specificity of u-Tf was higher than that of u-Alb. The validity of the threshold albumin level (20 mg/dl) was confirmed by measurements using SDS-PAGE. Since leakage of u-Tf in urine precedes leakage of u-Alb, inclusion of u-Tf in biochemistry tests may be appropriate for IRIS staging as a diagnostic marker of early diagnosis of renal disorder in cats.

The aim of this study was to evaluate the prognostic value of concurrent measurement of serum phosphorylated neurofilament heavy subunit (pNF-H) concentration and intramedullary T2W hyperintensity in paraplegic to paraplegic dogs. Our hypothesis was that concurrent measurement of these would provide a more accurate prediction of functional outcome in dogs with thoracolumbar intervertebral disc herniation (IVDH). A prospective case-control clinical study was designed using 94 dogs with acute onset of thoracolumbar IVDH. The association of serum pNF-H concentration, T2W hyperintensity on sagittal MRI (T2H/L2), deep pain perception and surgical outcome were evaluated with logistic regression analysis after three months for all 94 surgically treated dogs. Sensitivity to predict non-ambulatory outcome was compared among pNF-H and T2H/L2 and combination of both. Logistic regression analysis indicated that serum pNF-H concentration and T2H/L2 were significantly correlated with surgical outcome (P<0.05); however, deep pain perception was not (P=0.41). The results of logistic regression analysis indicated that the odds ratios of unsuccessful long-term outcome were 2.6 for serum pNF-H concentration, 1.9 for T2H/L2 and 2.3 for deep pain sensation. The sensitivity and specificity to predict non-ambulatory outcome for using serum parameter pNF-H>2.6 ng/ml, using T2H/L2 value of>0.84 and using both serum pNF-H and T2H/L2, were 95% and 75.7%, 65% and 86.5%, and 90.0% and 97.5%, respectively. Therefore, combined measurements of serum pNF-H and T2H/L2 might be useful for predicting long-term outcome in dogs with thoracolumbar IVDH.

A 12-week-old female Wire-haired miniature dachshund presented with non-progressive ataxia and hypermetria. Due to the animal's clinical history and symptoms, cerebellar malformations were suspected. Computed tomography (CT) and magnetic resonance imaging (MRI) detected bilateral ventriculomegaly, dorsal displacement of the cerebellar tentorium, a defect in the cerebellar tentorium and a large fluid-filled cystic structure that occupied the regions where the cerebellar vermis and occipital lobes are normally located. The abovementioned cystic structure and the defect in the cerebellar tentorium were comparable to those seen in humans with Dandy-Walker syndrome. However, the presence of the cystic structure in the occipital lobe region was unique to the present case. During necropsy, the MRI findings were confirmed, but the etiology of the condition was not determined.

Guide dogs help visually impaired persons both physically and psychologically. More than half of all candidate dogs do not qualify, mainly for behavioral reasons. Improved training efficacy is desirable, and earlier prediction of qualification-related traits would be beneficial. In a previous study, we identified 'Distraction', assessed during the training period, as an important behavioral trait for judging the qualification of guide dogs at the Japan Guide Dog Association. As a second step, we aimed to develop an index that can predict during the puppy period. In this study, candidate guide dogs, 5-month-old Labrador retrievers, were assessed by puppy raisers using a newly developed questionnaire that consisted of 20 items. The same dogs were assessed later, at 15 months, by trainers to determine 'Distraction'. In principal components analysis, nine items, including excitability toward strangers, initiative while out for a walk, and exploration, composed the first principal component (PC1). When we compared PC1 points with 'Distraction' points, the two categories were positively correlated (n=110, r(s)=0.31, P=0.0009). Although the accuracy of the questionnaire should be increased, the results of the present study suggest that it may be possible to assess and predict 'Distraction', which is associated with disqualification for guide dogs, early in the puppy-raising period.

A 15-month-old male beagle dog used in a toxicity study had a primary renal mesenchymal tumor. Macroscopically, the tumor was a gray-white mass which was found in the right kidney, and extended from the capsule to a position slightly compressing the medulla. Microscopically, most of the tumor cells showed a myxoid pattern, in which the matrix was positive for alcian blue staining. In the other parts of the tumor, a fascicular and wavy pattern was observed, and the matrix was full of collagen fibrils. Immunohistochemically, tumor cells were positive for vimentin and fibronectin, and negative for cytokeratin, desmin, α-smooth muscle actin, Von Willebrand factor, cyclooxigenase-2 and myelin basic protein. As a result, we diagnosed this case to be a renal mesenchymal tumor. Based on the microscopic findings, interstitial characteristics and immunohistochemical features, the present case was classified as a congenital mesoblastic tumor.

Adult rat dorsal root ganglion (DRG) neurons cultured in the presence of 100 ng/ml NGF show spontaneous action potentials and fluctuations in their cytosolic Ca(2+) concentrations ([Ca(2+)](i)). In the present study, the Ca(2+) sources of the [Ca(2+)](i) fluctuations and the types of neurons whose excitability was affected by NGF were examined. In the subpopulation of NGF-treated neurons, obvious fluctuations of [Ca(2+)](i) were observed. The [Ca(2+)](i) fluctuations were inhibited by Ca(2+) removal or inhibitors of voltage-gated Ca(2+) channels. Regardless of the treatment with NGF, about half of the neurons responded to capsaicin and 10% of the neurons responded to icilin, and almost all icilin-responding neurons also responded to capsaicin. Fluctuations of [Ca(2+)](i) with large amplitudes were observed in 12 out of 131 NGF-treated neurons. Among these 12 neurons, 10 neurons responded to both capsaicin and icilin. The degree of the [Ca(2+)](i) fluctuations in the NGF-treated neurons responding to both capsaicin and icilin was significantly larger than in other neurons. These results suggest that neurons expressing both capsaicin- and icilin-sensitive TRP channels are susceptible to NGF and become hyperexcitable and that Ca(2+) influx through voltage-gated Ca(2+) channels is the major source contributing to the [Ca(2+)](i) fluctuations. Since such DRG neurons could play a physiological role as nociceptors, the NGF-induced spontaneous activity of DRG neurons may be the underlying mechanism of neuropathic pain.

Canine respiratory coronavirus (CRCoV), which is more closely related to the bovine coronavirus (BCoV), has recently been detected in dogs. In this study, we examined whether BCoV was capable of infecting and exhibiting pathogenicity in dogs. Three 1-month-old pups were oronasally given field isolates of BCoV, and were kept together with 2 control animals. As a result, increases in BCoV-neutralizing antibody titers were confirmed in all pups in the challenged and control groups. Moreover, the virus gene was also detected in oral and rectal swabs by RT-PCR. These results indicate that BCoV infects dogs, and easily infects other dogs that are kept together. However, no clinical symptoms such as respiratory symptoms and diarrhea were observed.