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Inflammasomes play a pivotal role in gastrointestinal homeostasis and inflammation. However, it remains elusive whether the nucleotide-binding oligomerization domain-like receptor (NLR) family inflammasomes, such as NLR family pyrin domain-containing (NLRP) 3, NLRP6, and NLRP12, are involved in the pathogenesis of canine chronic enteropathy (CE), which includes antibiotic-responsive enteropathy (ARE), food-responsive enteropathy (FRE), immunosuppressant-responsive enteropathy (IRE), and non-responsive enteropathy (NRE). Thus, we measured mRNA expression of NLRP3, NLRP6, and NLRP12 in the intestinal mucosa of 35 dogs with CE (ARE, four dogs; FRE, 11 dogs; IRE and NRE, 20 dogs) and seven healthy dogs. As per real-time PCR analysis, significant increases in mRNA expression of NLRP3 and NLRP12 were noted in the colonic but not in the duodenal mucosa of dogs with FRE compared to healthy dogs. These findings suggested that the NLRP3 and NLRP12 inflammasomes might contribute to the development of colitis in dogs with FRE.

Background Lymphoma in the nasal cavity is the most common tumor of cats' upper respiratory tract. However, the effect of single-agent chlorambucil on nasal or nasopharyngeal lymphoma has not been evaluated in cats. Case description An 8-year-old, castrated male Scottish Fold weighing 3.5 kg presented with an 8-month history of nasal discharge, sneezing, and mild epistaxis. CT and rhinoscopy revealed nasal discharge and slight swelling of the nasopharyngeal mucosa, but no masses and local invasions were detected. Histopathological and immunohistochemical analyses of the nasopharyngeal mucosa demonstrated B-cell lymphoma in the cat. The treatment with chlorambucil led to long-term management of the cat without any side effects. No recurrences of clinical signs have been observed for 754 days. Conclusion The present case report suggests that chlorambucil can be a therapeutic option for feline localized nasopharyngeal B-cell lymphoma without masses and local invasions.

A 7-year 6-month-old, castrated male Shiba dog presented with a 1-month history of lethargy, anorexia, vomiting, and frequent watery diarrhea. Weight loss, hypoalbuminemia, anemia, and leukocytosis were detected at the first visit. The dog was diagnosed with non-responsive enteropathy (NRE) based on clinical and histopathological examinations. Since the dog did not respond to the immunosuppressive drugs, fecal microbiota transplantation (FMT) was performed during the treatment with chlorambucil. A single endoscopic FMT into the cecum and colon drastically recovered clinical signs and clinicopathological abnormalities and corrected dysbiosis in the dog. No recurrence or adverse events were observed. The present case report suggests that FMT, possibly together with chlorambucil, might be a treatment option for NRE in Shiba dogs that have poorer prognosis compared with other dog breeds.

Vincristine, one of the anti-cancer drugs used in veterinary practice, has adverse hematological and gastrointestinal effects in dogs. Juzen-taiho-to is a traditional Chinese medicine used for patients with anorexia in human medicine. However, the protective effects of Juzen-taiho-to against anti-cancer drug-induced toxicity in dogs have not been investigated. We therefore examined whether the administration of Juzen-taiho-to to dogs affects gastric motility, and vincristine-induced gastrointestinal and hematological toxicity. The study was composed of three trials. In the first trial, Juzen-taiho-to (450 mg/kg/day) was orally administered to five dogs. In the second and third trials, vincristine (0.75 mg/m2) was intravenously administered to each dog in the absence or presence of Juzen-taiho-to (450 mg/kg/day). During these trials, gastric motility and blood parameters were assessed. Juzen-taiho-to increased gastric motility and improved vincristine-induced gastrointestinal, but not hematological, adverse effects in dogs. This study suggested that Juzen-taiho-to may be applicable for gastrointestinal care in dogs receiving chemotherapy.

An entire, female, mixed-breed cat of unknown age was presented with a 6-week history of lethargy, anorexia and vomiting There was an increase in the number of white blood cells in the blood, including neutrophils and eosinophils;moderate anaemia;ascites;and possible mesenteric peritonitis Exploratory laparotomy revealed firm, multifocal small nodules in the mesentery As the nodules were surgically unresectable, they were biopsied Histologically, the nodules were composed of thin trabeculae of dense collagen fibres mixed with plump fibroblasts and numerous eosinophils, consistent with feline gastrointestinal eosinophilic sclerosing fibroplasia Bacteria were not detected on histological examination of the nodules and cytology of the ascites Remission of disease occurred following treatment with prednisolone and ciclosporin A for 22 days and antibiotics for 40 days After remission, ciclosporin A was administered for 236 days and then discontinued Eosinophilia also resolved after treatment with ciclosporin A The cat is still alive and in good condition on day 689 This report describes what may be an atypical case of feline gastrointestinal eosinophilic sclerosing fibroplasia, lacking involvement of the gastrointestinal tract, and was apparently cured by treatment that involved ciclosporin A Copyright © 2019 British Small Animal Veterinary Association

Background Mucosal imbalance of interleukin (IL)-1β and IL-1 receptor antagonist (Ra) has been reported in the duodenal mucosa of dogs with inflammatory bowel disease (IBD). However, the imbalance in the colonic mucosa and its role in duodenitis and colitis in IBD of dogs remain unclear. Objectives To measure the expression of IL-1β and IL-1Ra proteins in the colonic mucosa of dogs with IBD, and to determine the effect of IL-1β on expression of occludin (ocln) mRNA, a tight junction component, in the duodenal and colonic mucosa of dogs with IBD. Animals Twelve dogs with IBD and 6 healthy dogs. Methods IL-1β and IL-1 Ra proteins in the colonic mucosa were quantified by ELISA in 7 of the 12 dogs with IBD. Expression of ocln mRNA in the duodenal and colonic mucosa was examined in the 12 dogs by real-time PCR. Results The ratio of IL-1β to IL-1Ra in the colonic mucosa was significantly higher in dogs with IBD than in healthy dogs. The ex vivo experiment determined that IL-1β suppressed expression of ocln mRNA in the colonic mucosa, but not in the duodenal mucosa, of healthy dogs. Expression of ocln mRNA in the colonic mucosa, but not in the duodenal mucosa, was significantly lower in dogs with IBD than in healthy dogs. Conclusions and clinical importance A relative increase in IL-1β may attenuate ocln expression, leading to intestinal barrier dysfunction and promotion of intestinal inflammation in the colonic mucosa, but not in the duodenal mucosa, of dogs with IBD.

A genital ulcer can be associated with sexually transmitted infections, such as syphilis and herpes, and sexually transmitted organisms, such as Chlamydia trachomatis. We report a case of scrotal ulcer caused by Staphylococcus aureus clone USA300 in a 56-year-old man living with human immunodeficiency virus in Osaka, Japan. It is important to enhance the awareness and surveillance regarding the potential of spread of this organism in the community of men who have sex with men in this district.

It remains unclear whether epithelial cell-derived cytokines, including interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP), contribute to development of canine chronic enteropathy (CE), which includes antibiotic-responsive enteropathy (ARE), food-responsive enteropathy (FRE) and inflammatory bowel disease (IBD). In the present study, we examined mRNA expression of il-25, il-33 and tslp in the duodenal and colonic mucosae of dogs with ARE, FRE and IBD. Real-time PCR analysis revealed that mRNA expression of il-33 was significantly lower in the duodenum in dogs with FRE than in healthy dogs. The results suggest that epithelial cell-derived cytokines may not be an inducer of Th2-type immunity in the gut of dogs with CE, and decreased expression of IL-33 may be involved in induction of FRE. Further studies are required to clarify roles of epithelial cell-derived cytokines, especially IL-33, in the pathogenesis of canine CE.

Background Successful clinical outcomes of faecal microbiota transplantation (FMT) for recurrent Clostridium difficile infection have been reported in humans and a marmoset. However, it has been unclear whether oral FMT was effective for the treatment of C. difficile-associated diarrhoea in dogs. Case presentation An 8-month-old, intact male French bulldog was presented with a 4-month history of intermittent large bowel diarrhoea. Physical and clinical examinations did not identify any specific causes for diarrhoea. Real-time PCR analysis and immunochromatography detected C. difficile antigen and toxin A&B genes and proteins in a faecal sample. Based on these findings, diarrhoea in the dog was considered to be induced by C. difficile-associated colitis. The dog was treated with oral FMT, in which a faecal solution obtained from a healthy beagle was orally administered to the subject. Stool consistency and frequency and faecal blood and mucus became normal 2–3 days after oral FMT, and real-time PCR analysis and immunochromatography was negative for C. difficile antigen and toxin A&B genes and proteins. No adverse events were observed. Conclusion The present case report demonstrated that oral FMT was an effective treatment for C. difficile-associated diarrhoea in a dog. The findings in this report provide a rationale to evaluate clinical efficacy of oral FMT for other gastrointestinal diseases in dogs.

OBJECTIVE To characterize platelet-activating factor (PAF)–induced edema and erythema in the skin of dogs and compare those reactions with histamine-induced cutaneous reactions. ANIMALS 6 healthy Beagles. PROCEDURES Experiments were performed at ≥ 2-week intervals. Each dog received ID injections (5 μg/site) of PAF C16, PAF C18, lyso-PAF, and histamine. Edema (mean diameter) and erythema scores (none, mild, moderate, or severe) were assessed 30 minutes after the injections. Dogs received ID injections of PAF and histamine each with various concentrations of WEB 2086 (PAF receptor antagonist) or underwent ID testing with PAF and histamine before and 3 hours after oral administration of cetirizine hydrochloride or prednisolone (at 2 doses each). RESULTS ID injections of PAF C16 and PAF C18, but not lyso-PAF, induced comparable levels of edema and erythema. The PAF-induced edema and erythema peaked at 30 minutes and lasted for 6 hours after the injection; histamine-induced edema and erythema peaked at 30 minutes and lasted for 3 hours after the injection. Edema sizes and erythema scores were significantly smaller and lower, respectively, for PAF than for histamine. The WEB 2086 inhibited PAF-induced but not histamine-induced edema and erythema. Cetirizine slightly, but significantly, repressed PAF-induced edema and erythema as well as histamine-induced cutaneous reactions. Prednisolone suppressed both PAF-induced and histamine-induced edema and erythema. CONCLUSIONS AND CLINICAL RELEVANCE In canine skin, the duration of PAF-induced inflammation was longer than that of histamine-induced inflammation. The PAF- and histamine-induced cutaneous reactions were effectively suppressed by oral administration of prednisolone. The importance of PAF in dogs with anaphylaxis and allergic disorders warrants further investigation.

There have been limited reports on the prevalence of adverse food reactions among dogs suffering from chronic enteropathy (CE) in Japan. We examined the prevalence and histological features of food-responsive enteropathy (FRE) in a total of 32 dogs with history of CE. Fourteen of 18 cases (56.2%) diagnosed as FRE had lymphocytic-plasmacytic enteritis or eosinophilic enteritis by histopathological examination. Characteristic histopathological changes indicating FRE were not identified in 18 cases, though 4 cases did not show any abnormalities. Results collected from this study provided important information that can help to change the way dogs with CE are treated in the future.

Case summary A spayed female mixed breed cat of unknown age (presumably more than 9 years old), weighing 2.9 kg, was presented with a 4 day history of lethargy, anorexia and vomiting following oral administration of calcium fosfomycin (20 mg/kg q12h). A serum biochemical analysis revealed a dramatic increase in the levels of blood urea nitrogen (>140 mg/dl) and creatinine (15.3 mg/dl), and hypercalcaemia (13.1 mg/dl), hyperphosphataemia (13.3 mg/dl) and hyperkalaemia (6.1 mmol/l). The cat was hospitalised and treated with infusion therapy. However, the renal function and clinical signs did not improve with any treatment. The cat was euthanased upon the owner’s request. Histopathological analysis of the kidneys revealed acute tubular necrosis in the cortex. Relevance and novel information The present case report provides, for the first time, clinical and histopathological evidence for acute renal failure induced by oral administration of fosfomycin in an adult cat. It is highly advisable that fosfomycin should not be used in either young or adult cats.

A time-of-day-dependent variation in IgE-mediated passive systemic anaphylaxis was previously reported in ICR mice. In the present study, we investigated time-of-day-dependent variations in IgE-, histamine-, and platelet-activating factor (PAF)-mediated systemic anaphylaxis in C57BL/6, BALB/c, and NC/Nga mice at 9:00 h and 21:00 h, and evaluated the potential influence of glucocorticoids (GCs) on these variations. We found significant time-of-day-dependent variations in IgE-mediated systemic anaphylaxis in C57BL/6 mice, and in histamine- and PAF-mediated systemic anaphylaxis in BALB/c mice. Significant daily variations in IgE-, histamine-, and PAF-mediated systemic anaphylaxis were not observed in NC/Nga mice. Pretreatment with dexamethasone and adrenalectomy abolished the daily variations in IgE-mediated systemic anaphylaxis in C57BL/6 mice and in PAF-mediated systemic anaphylaxis in BALB/c mice, suggesting that GCs from adrenal glands are pivotal in regulating these variations. In contrast, pretreatment with dexamethasone and adrenalectomy did not abolish the daily variation in histamine-mediated systemic anaphylaxis in BALB/c mice, suggesting that GC-independent and adrenal gland-independent mechanisms are important for the variation. The present study demonstrated that time-of-day-dependent variations in systemic anaphylaxis differed among inbred mouse strains and with anaphylaxis-inducing substances. Thus, mouse strains, time of experiment, and anaphylaxis-inducing substances used must be considered to obtain appropriate experimental results.

A 9-year-old, spayed female Chihuahua was presented with a 1-week history of lethargy and anorexia. Abdominal ultrasonography and computed tomography found bilateral adrenal masses without metastasis. Serum cortisol levels that were sampled before and after an adrenocorticotropic hormone stimulation test were within reference ranges. Lethargy and anorexia completely resolved after short-term fluid therapy; the clinical signs did not occur for approximately 8 months until her sudden death. A postmortem examination revealed bilateral adrenocortical carcinomas and liver metastasis. Primary adrenocortical carcinomas developed in the dog met the definition of bilateral incidental adrenal gland masses (IAGMs). This is the first case report to demonstrate based on histological identification that adrenocortical carcinomas cause bilateral IAGMs in dogs.

Ultrapure soft water (UPSW) is water in which calcium and magnesium ions have been replaced with sodium ions using a cation-exchange resin. We recently demonstrated that washing with soap and UPSW reduced the clinical severity of dermatitis and improved the skin barrier function in NC/NgaTnd mice, a murine model for human atopic dermatitis. The purpose of this pilot study was to evaluate the efficacy of shampoo treatment with UPSW for dogs with pruritus. Eleven dogs with pruritus were randomly assigned to two groups depending on whether they received weekly shampoo treatment with UPSW or tap water for 4 weeks. After a washout period, the treatment protocol was switched such that each dog received both treatments. The pre-treatment and post-treatment values of the following were compared: pruritus scores assessed by the owners; dermatitis scores recorded by an investigator; and transepidermal water loss (TEWL). Shampoo treatment with UPSW significantly decreased pruritus and dermatitis scores in the dogs, whereas shampoo treatment with tap water did not. In addition, shampoo treatment with UPSW, but not with tap water, significantly reduced TEWL in the dogs. Adverse events due to the treatment were not observed in the dogs. Furthermore, we found that topical application of UPSW for barrier-disrupted skin caused by tape stripping in healthy dogs decreased TEWL more rapidly than topical application of tap water. Our findings suggest that shampoo treatment with UPSW promotes skin barrier recovery and thus could be considered as a possible therapeutic option in the management of pruritus and dermatitis in dogs.

There has been a need for improvement of the elimination diet used for diagnosis of adverse food reaction (AFR) in dogs. Recently, a novel elimination diet composed of a mixture of amino acids and potatoes was developed. We evaluated the efficacy of the elimination diet for diagnosis of AFR in dogs. Twenty dogs that were suspected to have allergic dermatitis were enrolled in a 2-month food elimination trial using the diet. Before and after the trial, the clinical symptoms were evaluated based on the change in canine atopic dermatitis extent and severity index (CADESI), pruritus score and medication score. Of the 20 dogs, 15 completed the food elimination trial. The remaining 5 dogs were removed from the trial because of diet unpalatability, skin disease progression or diarrhea. On the basis of evaluation of the clinical scores, we observed that the clinical symptoms improved in 11 of the 15 dogs that completed the food elimination trial. Provocative challenge was performed in 10 of the 11 dogs that showed improvement in their clinical symptoms. Of the 10 dogs, 7 were diagnosed as having AFR against food ingredients such as pork, beef, chicken and wheat because their skin symptoms reappeared after intake of these ingredients. The results of the food elimination trial and the provocative challenge indicated the usefulness of the novel elimination diet for diagnosis of AFR.

Prevalence of allergies has increased during the last two decades. Alteration of the gut microbiota composition is thought to play a crucial role in development of atopic diseases. Oral administration of probiotics has been reported to treat and/or prevent symptoms of atopic diseases in infants, but the results are still controversial. We investigated the potential efficacy of dietary interventions by a probiotic strain on prevention and treatment of atopic dermatitis (AD) in a human-like AD model, NC/NgaTnd mice by perinatal administration. Pregnant NC/NgaTnd mice were orally treated with the probiotic strain Lactobacillus rhamnosus CGMCC 1.3724 (LPR), which was followed by treatment of pups until 12 weeks of age. LPR-treated mice exhibited significant lower clinical symptoms of dermatitis, reduced scratching frequency, lower levels of plasma total Immunoglobulin E and higher levels of interferon-gamma in skin biopsies, compared with untreated mice. The protective effect was also observed when mice started to be treated at weaning time (5 weeks of age) even with limited supplementation period of 2 weeks. However, treatment of mice with the probiotic starting 1 week after the onset of the disease (8 weeks of age) had limited effects. The usefulness of LPR for primary prevention of AD was supported.

We previously demonstrated the presence of IgE directed to fetal calf serum (FCS) in the sera from dogs that developed allergic reactions after vaccination. In this study, by an immunoblot analysis, we investigated the IgE-reactive components of FCS using sera from 16 dogs that exhibited allergic reactions after vaccination. The immunoblot analysis revealed that several FCS proteins of approximately 25-, 50-, 66-, 75-, 120-, and 175-kDa strongly reacted with IgE in the sera from dogs that showed post-vaccination allergic reactions. The 66-kDa band was detected in the sera from 14 of the 16 dog serum samples analyzed in the immunoblot analysis for FCS, and it was speculated to be albumin based on its molecular weight; however, serum IgE reactivity to bovine serum albumin could be detected in only four of the 14 dog samples. These findings demonstrated that a variety of FCS components including albumin could function as allergens in dogs that developed allergic reactions after vaccination.

The objectives of this study were to determine daily variation in intradermal reactivity to histamine in dogs and to evaluate a potential influence of glucocorticoids on reactivity. Wheal sizes formed after intradermal injections of histamine were measured every 6 h during a single 24 h period in six healthy dogs. To determine whether glucocorticoids were implicated in daily variation, intradermal reactivity to histamine was evaluated at 9:00 h and at 21:00 h during a single day in dogs that received oral prednisolone (a synthetic glucocorticoid) or oral trilostane (an inhibitor of endogenous glucocorticoid synthesis). Finally, the time required for the histamine reaction to diminish after an intravenous injection of hydrocortisone was also assessed. A significant time-of-day-dependent variation in intradermal reactivity to histamine was detected in dogs, with a larger wheal size observed at 9:00 h than at 21:00 h. Administration of prednisolone or trilostane disrupted this variation. Intradermal reactivity to histamine was significantly reduced 6 h after an intravenous injection of hydrocortisone. These results suggest that glucocorticoid secretion from the adrenal glands could be involved in the regulation of daily variation in histamine-mediated reactions in dogs.

Scratching and skin barrier dysfunctions are pivotal features and therapeutic targets of atopic dermatitis (AD); however, time-of-day-dependent variations of these characteristics remain unclear. NC/Tnd mice have been shown to exhibit severe scratching behavior and skin barrier disruption together with the development of spontaneous atopic dermatitis when they are raised under air-uncontrolled environment. In the present study, time-of-day-dependent variations of scratching behavior and transepidermal water loss (TEWL) were evaluated in NC/Tnd mice that developed moderate to severe AD. Analysis of the mice for 24 hr revealed that scratching frequency and duration were increased from in the afternoon to the nocturnal period when locomotor activity was low, and scratching behavior was decreased in the morning. The highest scratching frequency and duration were 3.8- and 4.1-fold increases in the lowest scratching frequency and duration, respectively. In addition, TEWL on the dorsal skin lesion was decreased in the diurnal period, while that was increased in the nocturnal period. The highest TEWL was a 1.3-fold increase in the lowest TEWL. Significant daily variations were detected in scratching frequency and duration and TEWL. These results indicate that NC/Tnd mice are an appropriate mouse model to investigate time-of-day-dependent variations of scratching behavior and skin barrier dysfunctions associated with AD.

Background Effects of probiotics on the prevention of atopic diseases have been proposed recently. Although we have already reported the suppressive effects of the probiotic, ImmuBalance™, on a mouse model for peanuts allergy, its influence on atopic diseases remains unclear. Objective Potential efficacy of ImmuBalance™, which is the fermented soy product, on treatment of atopic dermatitis (AD) was investigated using a mouse model for human AD, NC/Tnd mice. Methods For in vivo study, ImmuBalance containing chow or a control diet were fed to NC/Tnd mice with moderate dermatitis for 2 weeks. Topical application of FK506 ointment was used as a positive control. Clinical skin severity scores, scratching behaviors, trans-epidermal water loss (TEWL), and histological features were analyzed. For in vitro study, suppressive effect of ImmuBalance™ on nerve growth factor (NGF)-activated neurite outgrowth of PC12 cells was examined. Results Clinical skin severity scores of the mice fed with ImmuBalance containing chow were gradually reduced as well as the mice treated with FK506. Feeding with ImmuBalance completely inhibited the increase in scratching behavior of NC/Tnd mice. The value of TEWL of NC/Tnd mice fed with ImmuBalance was significantly decreased. In addition, histological examination revealed that application of ImmuBalance decreased the number of PGP9.5-positive neuronal fibers in the lesional skin. When ImmuBalance extract was added to the culture, NGF-activated neurite outgrowth of PC12 cells was diminished through the inhibition of the phosphatidylinositol 3-kinase phosphorylation. Conclusion ImmuBalance could exhibit favorable alterations on AD symptoms, particularly through down regulation of the itch sensation.

Adverse reactions to vaccines were examined in 311 canine cases reported to the Ministry of Agriculture, Forestry and Fisheries in Japan during the period of 6 years from April of 1994 to March of 2000, and classified according to their clinical symptoms. There were 27 cases of adverse reactions to rabies virus vaccines. Gastrointestinal symptoms were the most frequently observed (26%), followed by respiratory and/or cardiovascular symptoms (22%) and dermatologic symptoms (11%). There were 284 cases of adverse reactions to non-rabies monovalent vaccines and mixed vaccines. Dermatologic symptoms were the most frequently observed (53%), followed by gastrointestinal symptoms (16%) and respiratory and/or cardiovascular symptoms (14%). Of the total 311 cases, 11 (3.5%) died of adverse reactions to vaccines.

Aims Intermittent claudication (IC) is one of the serious symptoms of peripheral arterial disease (PAD) and is characterized by pain in the legs or buttocks that worsens with exercise and subsides with rest. The concept of ‘therapeutic angiogenesis’ for PAD has been widely proposed; however, the methodology, including cell transplantation, is still unclear. In this study, we examined the clinical efficacy of silencing the int6 gene, which encodes a protein that stabilizes hypoxia-inducible factor (HIF)-2α, on angiogenesis in PAD. Methods and results An animal model for IC was established in Sprague–Dawley rats by external iliac artery ligation and evaluated by quantitative analysis of gait disturbance. Next, we explored the therapeutic effects of int6 siRNA injected into the adductor magnus muscle on IC. Recovery of hindlimb function occurred in the early stages after int6 siRNA injection. The number of blood vessels showed an obvious increase in the int6 siRNA-treated muscles. Angiography revealed the recovery of peripheral circulation at the affected sites. Early up-regulation of HIF-2α and other angiogenic factors, including basic fibroblast growth factor and hepatocyte growth factor, was also apparent in the int6 siRNA-treated sites. We also confirmed the up-regulation of HIF-2α and its translocation to the nucleus in the int6 siRNA-injected muscle. Conclusion A single injection of int6 siRNA promoted angiogenesis via up-regulation of HIF-2α-related angiogenic factors in the muscles of the affected hindlimb and reduced gait disturbance. The int6 gene may be a novel therapeutic target for the treatment of IC in patients with PAD.

NC/Tnd mice, a spontaneous model for human atopic dermatitis, are also useful animal models for various corneal disorders accompanying allergic diseases. The purposes of the current study were to investigate the development of retinal degeneration in NC/Tnd mice.Histological examination was performed to determine time-dependent alterations of the retina in NC/Tnd from 8 to 28 days of age. Apoptotic cells were determined by TUNEL assay. Retinal function was examined by electroretinography. Fundoscopy was performed in NC/Tnd mice at 8 weeks of age. Melanin contents in whole-eye extracts were measured by spectrophotometry. Since the retinal degeneration 1 (rd1) mutation in the rod photoreceptor cyclic guanosine monophosphate phosphodiesterase 6 β-subunit (Pde6b(rd1)) has been identified in laboratory mice, the possible existence of the rd1 mutation was analyzed with PCR genotyping and gene sequencing. C57BL/6, WB, and C3H/HeN mice were used as controls.Histological examination revealed rapid postnatal retinal degeneration in NC/Tnd mice. The number of apoptotic cells in the outer nuclear layer (ONL) increased with aging, and finally the ONL disappeared. Histological abnormality was not obvious in the inner nuclear layer or the ganglion cell layer. Electroretinography shows no response in adult NC/Tnd mice. Fundoscopic observation revealed hypopigmentation in the retina, and melanin contents in the eye were significantly reduced when compared with other inbred strains. Insertion in the rd1 allele was confirmed and a nonsense mutation of Pde6b(rd1) gene was determined in NC/Tnd mice.NC/Tnd mice also preserve the Pde6b(rd1) gene mutation resulting in the rapid postnatal retinal degeneration similar to that in C3H/HeN mice. Unlike C3H/HeN mice, since melanin contents of the retina in NC/Tnd mice was decreased, unknown defects may be present in the process of melanin composition in retinal pigment epithelial cells during fetal development of NC/Tnd mice.

Allergic reactions after vaccination are considered as an important practical problem in dogs; however, their immunological mechanism has not been well understood. The present study was designed to investigate the relationship between IgE reactivity to the vaccines and immediate-type allergic reactions after vaccination in dogs. Sera from 10 dogs that developed immediate-type allergic reactions such as circulatory collapse, cyanosis, dyspnea, facial edema, and vomiting within 1h after vaccination with non-rabies monovalent or combined vaccines and sera from 50 dogs that did not develop allergic reactions after vaccination were collected. Serum IgE reactivity to the injected vaccines was measured by fluorometric ELISA using a mouse monoclonal anti-dog IgE antibody. Then, IgE reactivity to fetal calf serum (FCS) and stabilizer proteins (gelatin, casein, and peptone) included in the vaccines was measured in sera that had high levels of IgE to the vaccines. Levels of serum specific IgE to the vaccines in dogs with immediate-type allergic reactions (59-4173 fluorescence units [FU], mean +/- S.D.: 992.5 +/- 1181.9 FU) were significantly higher than those in control dogs (38-192 FU, 92.4 +/- 43.3 FU) (P < 0.001). Of the eight dogs that developed immediate-type allergic reactions and had high levels of serum specific IgE to the vaccines, seven had specific IgE directed to FCS. The IgE reactivity to the vaccines in sera from these dogs was almost completely inhibited by FCS. The other one dog had serum IgE directed to gelatin and casein included in the vaccine as stabilizers. The results obtained in this study suggest that immediate-type allergic reactions after vaccination in dogs were induced by type I hypersensitivity mediated by IgE directed to vaccine components. In addition, FCS, gelatin, and casein included in vaccines could be the causative allergens that induced immediate-type allergic reactions after vaccination in dogs.

Background Hardness in water is consisted with mineral salts, including calcium (Ca++) and magnesium (Mg++). The mineral salts react with soap to form an insoluble precipitate known as metallic soap (soap scum). Since metallic soap remains tightly on the skin and is hard to rinse off, it may become one of irritants that exacerbate dermatitis. In this study, we used cation-exchange resin to prepare ultra-pure soft water (UPSW) excluding both Ca++ and Mg++, and investigated effect of UPSW rinsing on skin conditions in subjects with atopic dermatitis. Furthermore, antigenic activity of metallic soap was investigated by in vivo experiments. Methods Stratum corneum was collected from arms of healthy volunteers who rinsed soap in tap water or UPSW, and the quantity of remained soap scum was determined with a gas chromatography. After 4 weeks of bathing in UPSW, the water content of the stratum corneum and transepidermal water loss (TEWL) of volunteers with mild atopic dermatitis were measured. With atopic NC/Tnd mice, a model for atopic dermatitis, we attempted to confirm results obtained from atopic volunteers. Plasma total IgE was measured by an ELISA after immunization of mice with metallic soap. Results On skins rinsed in UPSW, soap was disappeared immediately and remained metallic soap was significantly reduced when compared to that on skins rinsed in tap water. In skins of atopic volunteers who used UPSW for bath, the water content in stratum corneum was increased and TEWL was decreased. Skin dryness and itch scores were reduced in most volunteers. After washed with soap and rinsed in UPSW for 2 weeks, severe dermatitis of NC/Tnd mice were reduced as well as TEWL. On the other hand, dermatitis in NC/Tnd mice rinsed in tap water became worse. Plasma total IgE was increased in mice that were immunized with metallic soap. Conclusions UPSW protected skins from residue of metallic soap. Metallic soap has antigenic activity and increased plasma IgE levels. Clinical symptoms and the skin barrier function were improved by the use of soap with UPSW. UPSW is beneficial for the skin of patients with atopic dermatitis.

Background Although elevated specific IgG and IgE are observed in sera of patients with atopic dermatitis (AD), their involvement in the pathogenesis of AD remains to be determined. In this study, we investigated the contribution of the immunoglobulin in AD by using Fc receptor common γ-chain (FcRγ)-deficient NC/Tnd mice. Methods NC/Tnd mice spontaneously develop the AD skin lesion when they are raised in air-unregulated conventional circumstances, but not when maintained under air-regulated specific pathogen-free conditions. We established FcRγ - NC/Tnd mice; those mice lacked FcRγ-mediated immune responses initiated by specific IgG and IgE. The clinical skin severity score and scratching behavior were evaluated in FcRγ-deficient mice and wild-type (WT) littermates. To examine histological features and distribution of mast cells, tissue sections of the lesional skin were stained with hematoxylin-eosin and toluidine blue, respectively. With regard to inflammatory cytokine production, the mRNA expression was detected in the dorsal skin and the axillary lymph node by real-time RT-PCR. Results Although the absence of FcRγ did not affect production of the immunoglobulin, the clinical skin severity scores were lower in FcRγ-/- NC/Tnd mice by half than in conventional WT mice. On the other hand, there were no differences in both scratching behavior elicited by dermatitis and Th1/Th2 cytokine production between 2 groups of mice. In the skin lesion of FcRγ null mice, mild epidermal hyperplasia and immune cell infiltration were observed. Particularly, mast cell numbers and their degranulation were significantly decreased in the skin of FcRγ-deficient mice. Conclusions FcRγ was not critical to the onset of AD, because FcRγ-deficient mice exhibited moderate dermatitis and scratching behavior comparable to WT. On the other hand, although scratching behavior induced the mechanical destruction of skin barriers and mast cell activation, the absence of FcRγ markedly attenuated the skin severity, immune cell recruitment including lymphocytes, and mast cell degranulation. These results indicated that the FcRγ-mediated immune response by specific IgG and IgE regulate the exacerbation of clinical symptoms in AD.

A dog with Japanese cedar (Cryptomeria japonica, CJ) pollinosis had oral allergy syndrome (OAS) after ingesting fresh tomato. The dog showed specific IgE to both CJ and tomato allergens. As a negative control, twenty dogs without atopic dermatitis that had no exposure to tomato and no specific IgE to CJ allergen were used. They had no specific IgE to tomato allergen. Furthermore, IgE cross-reactivity was observed between CJ and tomato allergens in the dog. We found that OAS induced by tomato exists in the dog and there is a relationship between CJ and tomato allergens.

ALLERGIC reactions, including anaphylaxis, have been observed in dogs after vaccination (Brooks 1991, Greene 1998). Anaphylaxis is the most severe and life-threatening adverse event after vaccination. It is a systemic reaction of immediate type I hypersensitivity caused by allergen-specific immunoglobulin E, which causes the release of various mediators from mast cells and basophils (Greene 1998). In a previous retrospective study of 311 dogs in Japan with adverse reactions after vaccination, 46 of the dogs showed anaphylaxis, and seven of the 46 cases of anaphylaxis resulted in death (Ohmori and others 2002). Although allergic reactions, including anaphylaxis, after vaccination in dogs are a problem in small animal practice, little is known about them. The aim of this study was to obtain epidemiological information regarding allergic reactions after vaccination in dogs. Information on 85 dogs that had showed suspected allergic reactions within 24 hours after non-rabies vaccine injections was collected from 40 private small animal veterinary practitioners in Japan between January 2001 and July 2002; information on one to eight dogs (mean 2·1 dogs) was obtained from each practice. For the purposes of the study, the suspected allergic reactions were defined as clinical signs associated with type I hypersensitivity, such as hypotensive shock, dyspnoea, facial oedema, pruritus, weakness and diarrhoea, occurring after vaccination, as described previously by Greene (1998). The information on the 85 dogs comprised their history, the types of vaccine used, the history of previous vaccination, the clinical signs, and the time of onset of clinical signs after vaccination; the dogs were classified into three groups based on their clinical signs. Group 1 consisted of dogs with predominantly cardiovascular and/or respiratory signs, group 2 comprised those with predominantly dermatological signs, and group 3 consisted of dogs showing cardiovascular/respiratory and dermatological signs simultaneously. Of the 85 dogs, 83 were purebred and two were crossbred; there were 31 miniature dachshunds, seven shih tzus, six Maltese, six pugs, four labrador retrievers, four papillons, four Yorkshire terriers, three beagles, three miniature schnauzers, three Shetland sheepdogs, three toy poodles, two chihuahuas, two golden retrievers, two Welsh corgis and one akita, one Bernese mountain dog and one pomeranian. The dogs’ ages ranged from two months to 16 years; 43 were aged up to one year, 39 were over one year old, and the age of three of them was not clear. There were 42 males and 43 females. The following four types of vaccines are commercially available in Japan: a monovalent live parvovirus vaccine (type 1); monovalent inactivated parvovirus or Leptospira vaccines (type 2); combined live vaccines composed of canine parvovirus, distemper virus, adenovirus type 2 and/or parainfluenza virus (type 3); and combined live and inactivated vaccines composed of live canine parvovirus, distemper virus, adenovirus type 2, parainfluenza virus and/or coronavirus vaccines, and inactivated coronavirus and/or Leptospira vaccines (type 4). Of the 85 dogs studied, clinical signs of suspected allergic reactions were observed in two recipients of type 1 vaccine, 28 recipients of type 3 vaccines and 53 recipients of type 4 vaccines; no dogs had been injected with type 2 vaccines. Information on the types of vaccine used in two dogs was not obtained. Of the 85 dogs, 24 were classified into group 1: they showed predominant cardiovascular/respiratory signs consistent with anaphylaxis, such as circulatory collapse, cyanosis, bradycardia, hypotension, hypothermia, dyspnoea and/or tachypnoea. Of these 24 dogs, five showed concurrent gastrointestinal signs such as vomiting and/or diarrhoea. In addition, one dog, a seven-year-old male miniature dachshund, died of anaphylactic shock after vaccination. Fifty-nine of the 85 dogs showed dermatological signs such as facial oedema, erythema, pruritus and/or urticaria, and were classified into group 2; 55 of these dogs showed facial oedema, which was the most common dermatological sign. Of the 59 dogs in group 2, 11 dogs also showed concurrent gastrointestinal signs. Two of the 85 dogs showed cardiovascular/respiratory and dermatological signs at the same time (group 3), without concurrent gastrointestinal signs. The cardiovascular and/or respiratory signs in the 24 dogs in group 1 were observed within 60 minutes of vaccination, whereas the dermatological signs in the 59 dogs in group 2 appeared within 60 minutes in 22 dogs, but appeared from one to 24 hours after vaccination in the other 37 dogs (Table 1). Concurrent cardiovascular/respiratory and dermatological signs in the two dogs of group 3 were observed within 60 minutes after vaccination (Table 1). The results suggest that the cardiovascular/respiratory signs could have been caused by an immediate-type reaction, and the dermatological signs could have consisted of both immediateand non-immediate-type reactions, similar to the allergic reactions observed in human beings after vaccination (Kumagai and others 1997, Ohsaki and others 1999). Of the 85 dogs studied, 24 had received one vaccine injection, and 40 had received two or more vaccine injections, before the last vaccination which had provoked the suspected allergic reactions. Unexpectedly, 16 dogs had not received any vaccine injections before the vaccination which induced the reaction. In the other five dogs, the number of previous vaccinations was unknown. The 16 dogs with suspected allergic reactions after the first vaccination may have already been sensitised to some allergens included in the vaccines before the vaccination. The nature of such allergens in dogs is unclear, but may relate to the use of gelatin or bovine serum in the vaccines, as has been suggested in human beings (Bonin and others 1973, Erdos and others 1975, Kelso and others 1993, Sakaguchi and others 1995). It is also possible that anaphylactoid reactions induced the clinical signs in these 16 dogs, without sensitisation to allergens in the vaccines. In the present study, it was not possible to calculate the incidence of suspected allergic reactions because the total number of dogs that had received vaccination at the 40 veterinary practices was not obtained. Further studies, including the gathering of such information, will be necessary to invesShort Communications

Background Scratching behavior is a pivotal clinical sign and a therapeutic target of atopic dermatitis; however, due to the lack of an appropriate animal model, circadian regulation of this behavior remains unclear. Methods NC/Tnd mice, a mouse model for human atopic dermatitis, have been shown to exhibit severe scratching behavior together with the development of spontaneous atopic dermatitis when they are raised under air-uncontrolled environment. In this study, scratching behavior of the mice was measured by a SCLABA-Real system, a newly developed equipment that provides us with the real-time, long-term, quick and accurate quantification of murine scratching behavior by analyzing images with the high-speed camera and the invisible near-infrared light. Results Analysis by a SCLABA-Real system demonstrated that the frequency and duration of scratching behavior significantly correlated with the exacerbation of dermatitis in the mice, indicating that this system was able to measure scratching behavior without putting stress upon the mice. Twenty-four hours analysis revealed that the frequency and duration of scratching behavior increased in from the afternoon to the midnight and decreased in the morning. In addition to scratching behavior, transepidermal water loss (TEWL) also changed during a day in the mice. TEWL increased in the midnight and decreased in the morning. Conclusions These results indicate that scratching behavior and skin barrier function in atopic dermatitis exhibited circadian rhythm. In addition, NC/Tnd mice are considered to be an appropriate mouse model to investigate circadian rhythm of scratching behavior associated with atopic dermatitis.

Rationale Since allergic diseases such as atopic dermatitis (AD) and food allergy (FA) in dogs show many clinical features similar to those in humans, dogs appear to be useful as an animal model to study allergic diseases in humans. In order to examine the immunological characteristics in dogs with allergic diseases, expression of CC chemokine receptor 4 (CCR4) in peripheral CD4+ cells was investigated in dogs with allergic diseases. Methods Seventeen dogs with AD and 12 dogs with FA had compatible clinical findings including recurrent and chronic pruritus with superficial pyoderma and positive results in allergy tests. Ten healthy dogs, used as controls, did not have any clinical signs associated with allergic diseases. Peripheral blood mononuclear cells (PBMCs) were obtained from these dogs. Populations of CCR4+ cells in CD4+ cells were evaluated by flow cytometry and were compared among these dog groups. Results The proportions (mean ± SEM) of CCR4+ cells in CD4+ cells were 40.4 ± 3.3% in AD group, 41.6 ± 4.0% in FA group and 23.6 ± 4.3% in normal dog group, respectively. The proportions of CCR4+ cells in dogs with allergic diseases (AD and FA) were significantly higher than that in normal dogs ( P Conclusions The present results indicated that CCR4+ cells were predominant in peripheral blood CD4+ cells in dogs with allergic diseases, suggesting that CCR4 plays important roles in immunopathogenesis of allergic diseases in dogs as well as in humans.