This study was carried out to investigate the changes of orexin-A (OXA) and neuropeptide Y (NPY) expression in the hypothalamus of the obese and lean Zucker Diabetic Fatty (ZDF) rats which have a missense mutation in the leptin receptor gene. The mean body weights (MBW) between the obese and lean ZDF rats were significantly different at 28 and 70 postnatal days. However, at 14 postnatal day, there was no significant difference in the MBW between the obese and lean ZDF rats in both male and female. The OXA immunoreactivities were not significantly different between the obese and lean ZDF rats in both sexes at 14, 28, and 70 postnatal days, respectively. The NPY immunoreactivity was higher in the obese than in the lean ZDF rats in both male and female at 28 and 70 postnatal days, whereas there was no significant difference between the obese and lean ZDF rats at 14 postnatal day. These results indicate that both OXA and NPY might halt their roles for food intake in the obese phenotype of the male and female ZDF rats in the preweaning period of 14 postnatal day, whereas NPY might play a main role in the obesity of these rats in the weaning period of 28 and 70 postnatal days.
Expression of neurofilament 200 (NF200)-like immunoreactivity was examined in the main olfactory system and the vomeronasal system of the Japanese newt, Cynops pyrrhogaster, using anti-porcine NF200 monoclonal antibody (clone N52) to investigate the differences in phenotypical characteristics between these systems. The entire nasal cavity was a flattened single chamber consisting of the main nasal chamber (MNC) and the lateral nasal sinus (LNS) communicating with each other. The olfactory epithelium (OE) was present in the MNC, and the vomeronasal epithelium (VNE) was in the LNS. The OE possessed only a small number of NF200-like immunoreactive receptor neurons. The olfactory nerve and the olfactory nerve layer of the main olfactory bulb also contained a small number of NF200-like immunoreactive axons. In contrast, the VNE possessed many NF200-like immunoreactive receptor neurons. The vomeronasal nerve and the vomeronasal nerve layer of the accessory olfactory bulb contained many NF200-like immunoreactive axons. These findings in the Japanese newt indicate that NF200-like immunoreactive receptor neurons constitute a major subpopulation in the VNE and a minor subpopulation in the OE. In addition, NF200-like immunoreactivity seems to be a useful marker to distinguish the vomeronasal system from the other nervous systems including the main olfactory system in the Japanese newt. The localization of a few NF200-like immunoreactive receptor neurons in the OE might indicate that pheromone-sensitive receptor neurons are intermingled in the OE of the Japanese newt.
Salivary secretory immunoglobulin A (sIgA) concentrations of prospective guide dogs for the blind were determined to clarify whether salivary sIgA is useful in evaluating the potential suitability of guide dogs for the blind. Saliva was collected from 73 prospective guide dogs in the kennel on day 1 (the day of separation from volunteer puppy-raisers), 2, 3, 7 and 14 during the estimation period (at about 1 year old). We selected particularly suitable dogs (superior dogs) and unsuitable dogs (inferior dogs) on the basis of the trainers' estimation. All dogs were divided into two groups, those were acceptable dogs would become the guide dogs and rejected dogs could not become guide dogs. The sIgA concentrations in superior dogs gradually increased from day 1 to 14 and those in inferior dogs remained at low levels. Moreover, the sIgA concentrations on day 14 in the acceptable dogs were significantly higher than those in rejected dogs. The cut-off point of sIgA concentrations on day 14 using an ROC curve was 90 EU/ml, and the specificity of the estimation at this point (70.4%) was higher than that of trainers' estimation (50%). Moreover, parallel testing using both trainers' estimation and sIgA estimation, showed that specificity was further improved (79.5%). The present study showed that sIgA concentration was extremely useful in estimating the adaptive ability for guide dogs for the blind.
We have identified various lactoferrin (Lf) molecules in mastitic mammary gland secretions (MGSs), and these Lf molecules were examined for their physiological function in MG. These Lf molecules were isolated by Con A affinity chromatography, and then analyzed by various electrophoresis methods and N-terminal amino acid sequencing. The low Con A affinity Lf was found to have low molecular peptides as compared with the 86 kDa of the high Con A affinity Lf, which is usually detected in healthy MGSs. The N-terminal amino acid sequence of each of the small molecular Lfs were confirmed as fragments of 86 kDa Lf. This low Con A affinity Lf stimulated spleen adherent cells to produce more O2- than 86 kDa Lf. Furthermore, the low Con A affinity Lf showed low antibacterial activity against E. coli and S. aureus, and had decreased iron-binding capacity in comparison with 86 kDa Lf. Moreover, the 86 kDa Lf could stimulate bovine T cells or macrophages to produce IFN-γ, IL-4, IL-6, and IL-1α. However low Con A affinity Lf induced the production of TNFα, but not physiological T cell or macrophage cytokines. It was also found that when the healthy MGs of dry cows were injected with the low Con A affinity Lf, there was an increase in polymorphonuclear cells together with TNFα, MCP-1, and IL-8 production. These results suggested that low Con A affinity Lf in mastitic MGSs differed from 86 kDa Lf in physiological characteristics, and, that it induced an inflammatory reaction in MGs.
Rapid plasma reagin (RPR) titers were periodically examined during and after treatment in three rabbits clinically diagnosed with rabbit syphilis. RPR titers remained positive after clinical recovery and then gradually declined. Of the two rabbits inoculated experimentally, one showed clinical signs of the disease, while the other did not. RPR titers were also periodically evaluated before and after inoculation in these two rabbits. The trends in RPR titers reflected the course of infection, both in the spontaneous and in the experimental cases. An inapparent case and cases without clinical signs after clinical recovery showed low titers for long period of time. Useful information for interpretation of RPR titers measured clinically was obtained by this survey.
Giardia intestinalis antigen in fecal samples was examined in 361 dogs of 14 breeding kennels located at various areas in Japan, using a commercial enzyme-linked immunosorbent assay (ELISA) kit. G. intestinalis antigen was detected in 37.4% of the fecal specimens. All of the 14 breeding kennels were positive for G. intestinalis antigen with the range from 6.7 to 59.3%. The prevalence in puppies (54.5%) was significantly (p < 0.01) higher than that in adults (30.9%). There was no difference in prevalence between males and females, and between the puppies from the mother dogs positive and negative for Giardia antigen. In conclusion, G. intestinalis widely invaded the breeding kennels in Japan.
Between the laminar epidermis and the laminar dermis of laminar region (LR) in equine foot, it can be observed the basement membrane zone (BMZ), which is composed of a basement membrane and its accompaniments like the hemidesmosome and anchoring fibril. Alteration in the BMZ in equine laminitis is possibly related with not only development but also recovery outcome and recurrence of this disease. However, there is little known about the structure of the BMZ during the recovery phase of this disease. To assess the condition of the BMZ of LR affected by chronic laminitis, the tissue was examined in three cases at two weeks, four weeks and three months after the onset of laminitis, using pathological, immunohistochemical and electron microscopic techniques. Histologically in all laminitis cases, there was a regenerated laminar epidermis with proliferating keratinocytes between the Stratum medium and the dermis, but it included the undeveloped secondary epidermal laminae (ud-SELs) structure in one side of the primary epidermal laminae, especially in the part of the deep area of LR. Immunohistochemical results were positive for the anti-type IV collagen, anti-type VII collagen and anti-laminin 5 antibodies in the most BMZs. However, partial BMZs adjacent to the ud-SELs were negative for the anti-type VII collagen and anti-laminin 5 antibodies. Ultrastructurally, in the BMZ of the ud-SEL, the lamina densa and the lamina lucida were present. In contrast, the anchoring fibrils and the hemidesmosomes were either absent, or present at lower than normal levels. In conclusion, the present study indicated that the part of regenerated LR in chronic laminitis was not able to fully restore to construct the BMZ for a long time, especially in the unilateral side of laminar epidermis. It might be related with recurrence of this disease.
Seven of 92 lactating Holstein cows on a dairy farm developed urticaria with alopecia and decreased milk production, and three of the seven died over the course of 7 to 18 days. Pathologic examination of the three cows, including the two dead and one euthanized cow, revealed that the skin, liver, spleen, kidneys, heart, salivary glands, pancreas, adrenal glands, mammary glands, lymph nodes, and trigeminal ganglia had lymphocytic to lymphogranulomatous inflammation. Inflammation predominated by lymphocytic infiltration was prominent in the heart, pancreas, mammary glands, adrenal gland, and trigeminal ganglia. Severe granulomatous inflammation with multinucleated giant cells was present in the spleen and kidneys. These lesions and their distributions were most similar to those seen in suspected cases of citrus pulp toxicosis and hairy vetch toxicosis. The outbreak of this disease resolved with the elimination of Citrus pulp from the feed. Immunohistochemical detection of lymphocytes and macrophage markers confirmed dramatic hyperplasia of CD3-positive T lymphocytes in these lesions. This strongly suggested that a type 4 hypersensitivity reaction played a role in the development of the lesions.
Reproductive failure associated with porcine circovirus type 2 (PCV2) was observed in a farm, and a weak-born 8-day-old piglet was examined pathologically. Focal to locally extensive lesions, including infiltration of macrophages and lymphocytes, fibrosis and degeneration of the myocardium were observed in the heart. PCV2 antigens and nucleic acids were detected in degenerated cardiomyocytes and macrophages infiltrating the heart by immunohistochemical staining and in situ hybridization. Depletion of lymphocytes with occasional infiltration of multinucleated giant cells was seen in the lymphoid organs and PCV2 antigens were demonstrated in histiocytic cells. Crystalline arrays of viral particles were observed in infiltrated macrophages in the heart and, rarely, in cardiomyocytes by electron microscopy. Although myocarditis is a common finding in aborted or stillborn piglets in reproductive failure due to PCV2, it was also observed in the 8-day-old piglet with PCV2 association.
There are several benefits to a high-fat diet for animals exposed to cold, including improved tolerance to severe cold conditions and increased survival rates in cold environments. It is therefore of interest to examine whether animals exposed to cold will selectively consume lipids. We examined the intake of safflower oil (SO) by rats exposed to cold (4 ± 2°C) under a feeding condition in which the rats were given free access to SO. Rats exposed to cold consumed more SO than those housed at 25 ± 2°C. This finding suggests that rats prefer SO in a cold environment. There was no significant difference in the ratio of calories of SO ingested to that of matter (standard laboratory chow plus SO) ingested between rats exposed to cold and those at 25 ± 2°C. The high SO intake also affected cold tolerance and metabolite kinetics in the rats. Factors that affected the SO intake of rats exposed to cold are also discussed.
The effects of various selective phosphodiesterase (PDE) inhibitors on muscle contractility and cyclic nucleotide contents in the guinea pig gall bladder were investigated. Various selective PDE inhibitors, vinpocetine (type 1), erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA, type 2), milrinone (type 3), Ro20-1724 (type 4), and zaprinast (type 5), inhibited CCh-induced contractions in a concentration-dependent manner. The rank order of potency for the gall bladder was Ro20-1724 > vinpocetine > EHNA > milrinone > zaprinast, which was different from that of the trachea, taenia coli, and aorta. In the presence of CCh (0.3 μM), vinpocetine, milrinone, and Ro20-1724 each increased cAMP content, but not cGMP. By contrast, zaprinast increased cGMP content, but not cAMP, and EHNA increased both cAMP and cGMP contents. These results suggest that vinpocetine-, milrinone-, and Ro20-1724-induced relaxation was correlated with cAMP, zaprinast-induced relaxation was correlated with cGMP, and that EHNA-induced relaxation was correlated with cAMP and cGMP in the guinea pig gall bladder. In conclusion, the effect of PDE inhibitors in the guinea pig gall bladder was different from those in smooth muscles, such as the trachea, taenia coli, and aorta.
Hemilaminectomy was performed to treat cervical disc disease in 18 small dogs. Cervical spinal cord compression was characterized by ventral and/or lateral compression on myelograms. The duration of follow-up examinations ranged from 2 to 72 months. The optimal response time after surgery ranged from 2 days to 3 months. The outcome was determined to be excellent if clinical signs resolved and the dog had completely improved. The outcome was determined to be good if the dog improved, but was not clinically normal or if the degree of the owner's satisfaction was insufficient. Fourteen dogs achieved complete neurologic recovery without complications. One dog was initially neurologically worse after surgery, but ultimately improved to normal neurologic status. These outcomes were judged to be excellent. In the remaining 3 dogs, 2 dogs had relapse of neck pain and one dog remained mildly ataxic. These outcomes were judged to be good. These results suggest that hemilaminectomy is an effective option for surgical treatment of spinal cord compression secondary to cervical disc disease in small dogs.
We conducted somatosensory evoked potential (SEP) monitoring on 3 dogs with cervical spinal cord dysfunction caused by various diseases throughout operative procedures to examine whether the intraoperative SEP monitoring was effective for prediction of spinal cord conductive function. The SEP was recorded on the scalp via stimulation of the ulnar nerve. Stable SEP was recorded in all animals examined. Its amplitude was decreased by surgical manipulations of the regio vertebralis, but the amplitude gradually recovered once the manipulations were halted. The latency showed small variation throughout the operations. This evidence suggests that intraoperative SEP monitoring may provide continuous and instantaneous information regarding the functional integrity of the central nervous system.
Two Holstein Friesian calves were referred to the Animal Medical Center of Chonbuk National University with severe lameness on the hind limb(s), flexed stifle joint and an inability to walk since birth. Based on the clinical history, clinical findings and radiography of the stifle joint, the cases were diagnosed as grade III lateral patellar luxation (LPL). Trochleoplasty along with medial retinacular reinforcement and lateral release were performed in both calves. The calves recovered 3 to 4 weeks after surgery. Trochleoplasty along with medial retinacular reinforcement and lateral release can be an effective technique for the treatment of LPL in calves.
Intraoperative ultrasonography was performed during spinal surgery in two dogs. An extradural lesion was demonstrated by myelography in case 1, and an extradural tumor was found during surgery. Case 2 was diagnosed with a spinal subarachnoid cyst by myelography and magnetic resonance imaging. Ultrasonographically, pathologic conditions of the spinal canal or cord were adequately imaged during surgery in both cases. Intraoperative ultrasonography was a superior method that non-invasively revealed intradural and extradural conditions and thus constitutes a valuable diagnostic mean to ensure safe and precise spinal surgery.
Factor XI deficiency is an autosomal recessive coagulopathy in Holstein cattle. Affected cows have a tendency to show repeat breeding. Forty repeat breeding Holstein Friesian cows were selected and tested for the Factor XI mutation. Genomic DNA was isolated from the blood of the cows (n=40). Exon 12 of the Factor XI gene of the cows was amplified by PCR. One repeat breeding cow was heterozygous to the Factor XI mutation as indicated by the presence of two DNA fragments of 320 bp and 244 bp. The insertion of the 76 bp in the heterozygous cow was confirmed by DNA sequencing. The heterozygous cow was in her fourth lactation. She gave birth to male twins at the last calving. She was inseminated artificially four times after the last calving. Factor XI deficiency in cattle has been reported in different countries. However, no case was reported in Japan. This might be the first to report Factor XI mutation in Holstein cattle in Japan.
Japanese horse chestnut seed extract (HCSE) dose-dependently inhibited the autooxidation of linoleic acid (IC50: 0.2 mg/ml), and the inhibition was almost complete at a concentration of 1 mg/ml. The HCSE scavenged DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals and superoxide anions with EC50s of 0.65 and 0.21 mg/ml, respectively. However, it had no effect on hydrogen peroxide. The HCSE inhibited the genotoxicities of furylfuramide, N-methyl-N-nitrosourea, methyl methanesulfonate, mitomycin C, 2-aminoanthracene and aflatoxin B1 at a concentration of 1 mg/ml or more. Total polyphenol content of the HCSE was 21 mg/g (13 mg/g-seeds). These results indicate that the Japanese horse chestnut seed is an antioxidative and antimutagenic botanical resource.
A molecular epidemiological analysis was performed in 19 rabies viruses (RVs) isolated from haematophagous, frugivorous and insectivorous bats, in Sao Paulo, Brazil. The authors carried out RT-PCR for amplification of the RV nucleoprotein (N) gene, and determined 1,335 nucleotide sequences of N gene by direct sequencing method. Phylogenetic analysis, which was based on the N gene of Brazilian RV isolates identified presently and previously, revealed that RVs isolated from bats were genetically divided into four lineages had a tendency to depend on the host bat species. The first lineage consisted mainly of haematophagous bat (Desmodus rotundus) isolates, including frugivorous bat (Artibeus spp.) isolates. Other three lineages consisted of insectivorous bat isolates; mainly Eptesicus spp., Molossus spp. and Nyctinomops spp. isolates, respectively. These results indicate a possibility of that there are bat species-specific RV variants in Brazil.