Conjugated linoleic acid and black currant seed oil in the treatment of canine atopic dermatitis: a preliminary report

Although conjugated linoleic acid (CLA) shows inhibitory effects on histamine release, eicosanoid production and pruritus in laboratory rodents, its use in canine atopic dermatitis (AD) has not been reported. The aims of this study were to assess the efficacy of CLA, black currant seed oil (BSO) or a combination of both, compared to placebo, in dogs with AD and to evaluate any changes in fatty acid metabolism with these treatments. Twenty-four dogs with AD were randomly allocated to four groups, and were treated orally each day for two months with either 1 mL/10 kg CLA (80% purity), 1 mL/10 kg pure BSO, 1 mL/10 kg CLA+1 mL/10 kg BSO, or 1 mL/10 kg sugar syrup (placebo). Serum was obtained on days 0, 30 and 60 for analysis of CLA metabolites, linoleic acid (LA), gamma-linolenic acid (GLA), dihomo-gamma-linolenic acid (DGLA) and arachidonic acid (AA). At the same time point, the owners were asked to assess pruritus and the veterinarians evaluated any skin lesions present. Although the best clinical results occurred with BSO treatment alone, improvement of clinical signs and pruritus was not significant with any of the treatments. Serum levels of GLA and DGLA significantly increased in BSO-treated dogs, but not in the CLA+BSO group. CLA at the dosage used in this study was not efficacious in treating canine AD, whereas BSO may help some dogs with AD, although further studies are necessary before this can be recommended as a treatment.     

Endothelial cells as active participants in veterinary infections and inflammatory disorders

Endothelial cells were once viewed as relatively inert cells lining the vasculature. They are now recognized as active and responsive regulators of coagulation, platelet adhesion, fluid homeostasis, wound healing, leukocyte extravasation and vascular tone. Endothelial cells play a key role in the host response to infectious agents by regulating leukocyte trafficking, producing inflammatory cytokines and presenting antigen in association with major histocompatibility class II (MHC II) molecules. A number of infectious agents have a tropism for endothelial cells. Infection of endothelial cells can promote thrombosis, vascular leakage, and increased adherence and emigration of leukocytes. Furthermore, activation of a systemic inflammatory response, in the absence of direct endothelial cell infection, can also lead to endothelial cell dysfunction. The purpose of this review is to highlight the interactions between endothelial cells and infectious or inflammatory agents that contribute to coagulation disturbances, vasculitis and edema. A select group of viral and bacterial pathogens will be used as examples to demonstrate how endothelial cell dysfunction contributes to the pathogenesis of infectious and inflammatory disorders.     

Pituitary apoplexy-like disease in 4 dogs

BACKGROUND: Pituitary apoplexy in humans is a clinical syndrome resulting from sudden infarction, hemorrhage, or both in a normal or an adenomatous pituitary gland. OBJECTIVE: Describe a clinical syndrome in dogs similar to pituitary apoplexy in humans. ANIMALS: Four dogs exhibiting a sudden onset of neurologic signs. METHODS: A retrospective study was used, including clinical examination, computed tomography (CT), postmortem examination, and histopathology of the brain. Pituitary tissue from 3 of the dogs was subjected to immunocytochemistry. RESULTS: Four dogs (2 Mongrels, 1 Bordeaux Dog, and 1 Cocker Spaniel; median age, 11 years; median body weight, 20.5 kg) presented with acute neurologic signs including depression (n = 3), behavioral changes (n = 1), vision loss (n = 1), seizures (n = 1), and collapse (n = 1). CT disclosed suprasellar infarction, hemorrhage, or both associated with a pituitary macroadenoma in 3 dogs and a frank hemorrhage in a nonadenomatous pituitary gland in 1 dog. CT findings were correlated with postmortem findings, and pituitary apoplexy was confirmed by histopathology and immunocytochemistry of the pituitary tissue. CONCLUSIONS AND CLINICAL IMPORTANCE: This study provides histopathologic evidence of pituitary apoplexy in dogs. The results are relevant for future diagnosis and treatment of pituitary disease in dogs.     

A neurologic syndrome in Golden Retrievers presenting as a sensory ataxic neuropathy

BACKGROUND: A sensory ataxic neuropathy has been observed in Swedish Golden Retrievers recently. ANIMALS: Twenty-one affected Golden Retrievers. METHODS: Clinical and neurologic status, electrophysiologic, and pathologic status as well as pedigree analyses were evaluated. RESULTS: Clinical signs had an insidious onset between 2 and 8 months of age and a slowly progressive course. Affected dogs were ataxic and dysmetric. They had abnormal postural reactions and decreased spinal reflexes but no apparent muscle atrophy. Clinical pathology, radiography, and electrophysiology of motor systems were all within reference values. Sensory nerve conduction results of affected dogs were significantly different from those of a group of control dogs. Necropsy revealed a chronic progressive central and peripheral sensorimotor axonopathy; the proprioceptive pathways were most severely affected. CONCLUSIONS AND CLINICAL IMPORTANCE: This disease in these Golden Retrievers is distinct from other canine breed-related neurodegenerative diseases or hereditary neurodegenerative diseases described in humans. Pedigree analyses indicated a hereditary background, but the mode of inheritance could not be established.     

Role of the hyaluronan receptor CD44 during porcine oocyte maturation

Previous our studies have shown that CD44, the principal receptor for hyaluronan, is present on cumulus cells during oocyte maturation. Although hyaluronan-CD44 interaction has been implicated in cumulus expansion and/or oocyte maturation, the full significance of CD44 remains unknown. The objective of the present study was to further investigate the role of CD44 in cumulus expansion and oocyte maturation in pigs. We demonstrate here in that CD44 has a key role in oocyte maturation but not in cumulus expansion. Previous studies have reported the physiological significance of cumulus expansion in oocyte maturation. However, our results suggest that cumulus expansion is a necessary condition for oocyte maturation, but that it is not sufficient on its own. Furthermore, western blot analysis demonstrated that the CD44 of the in vitro-matured cumulus-oocyte complexes (COCs) had a larger molecular weight and more terminal sialic acid, which has been proven to inhibit the hyaluronan-binding ability of the receptor, than the CD44 of the in vivo-matured COCs, indicating that the hyaluronan-CD44 interactions during in vitro maturation might be insufficient compared with those in vivo. The insufficient interactions of hyaluronan-CD44 during in vitro maturation may cause the inferior capacity of fertilization and development of oocytes matured in vitro.     

Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer

The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.     

Androgen induces production of male effect pheromone in female goats

Previously we showed that the primer pheromone responsible for the "male effect" was produced in specific skin regions of castrated male goats by androgen treatments. In the present study, we examined whether androgen can also induce production of the male effect pheromone in female goats. Capsules containing dihydrotestosterone (DHT) or testosterone (T) were subcutaneously implanted into six ovariectomized (OVX) goats for 28 days. Small skin samples were collected from the head and rump regions, and the pheromone activity of their ether extracts was examined using a bioassay that monitors the electrophysiological manifestation of the hypothalamic gonadotropin-releasing hormone pulse generator as multiple-unit activity. Behaviors of OVX goats towards ovary-intact estrous goats were also examined before and at the end of DHT or T treatment. Before androgen treatment, neither the head nor rump skin samples in OVX goats showed pheromone activity. DHT treatment induced pheromone activity in the head skin sample of six OVX goats and in the rump skin sample of two OVX goats. Similar results were obtained by T treatment. In addition, OVX goats treated with T showed masculine-type sexual behaviors such as courtship and mounting behaviors towards the estrous goats. These results demonstrate that androgen is capable of inducing primer pheromone activity in the female and suggest that the synthesis pathway of the male effect pheromone exists in both sexes in the goat.     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.     

Anti-oxidative/oxidative status of rat liver after ovariectomy

The lack of protective action of oestrogens which appears during menopausal period may be the reason of serious metabolic disturbances including oxidative stress. The hypothesis was stated that ovariectomy may induce the variations of antioxidant/oxidant status which can be detected in rat liver. A total of 102 healthy Wistar female rats were included in the experiment and divided into control (CON; n = 6), sham-operated (SHO; n = 48) and ovariectomized (OVX; n = 48) groups. Animals from SHO (n = 6) and OVX (n = 6) groups were killed every week during 8 weeks of experiment to detect dynamic changes in examined parameters. Anti-oxidative enzyme activities [glutathione peroxidase (GSH-Px); superoxide dismutase (SOD)] as well as total antioxidant capacity (TAC) and the intensity of lipid peroxidation, measured by the concentration of N,N,diethyl-p-phenylene diamine (DEPPD) radical, were determined in liver homogenates by the use of spectrophotometric methods. Wave-like patterns of examined parameters within 8 weeks of experiment were detected. GSH-Px activity tended to be higher in OVX animals and was significantly lower at 8th week when compared with 1st week of experiment. SOD activity was higher in SHO animals and showed significant differences between 3rd, 4th, 7th and 1st week. TAC values were significantly higher in OVX when compared with SHO groups in 2nd, 4th, 5th week and significantly lower in 3rd, 6th and 7th week of the experiment. The concentration of DEPPD radicals tended to increase in OVX group. In conclusion, ovariectomy which leads to oestrogen insufficiency is reflected as well in variation of anti-oxidative/oxidative parameters in rat liver homogenates.