Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of CD19+ B lymphocyte rather than CD4+ or CD8+ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

The present study was performed to evaluate the relationship between the neurotoxicity of acrylamide and the differential gene expression pattern in mice. Both locomotor test and rota-rod test showed that the group treated with higher than 30 mg/kg/day of acrylamide caused impaired motor activity in mice. Based on cDNA microarray analysis of mouse brain, myelin basic protein gene, kinesin family member 5B gene, and fibroblast growth factor (FGF) 1 and its receptor genes were down-regulated by acrylamide. The genes are known to be essential for neurofilament synthesis, axonal transport, and neuro-protection, respectively. Interestingly, both FGF 1 and its receptor genes were down-regulated. Genes involved in nucleic acid binding such as AU RNA binding protein/enoyl-coA hydratase, translation initiation factor (TIF) 2 alpha kinase 4, activating transcription factor 2, and U2AF 1 related sequence 1 genes were down-regulated. More interesting finding was that genes of both catalytic and regulatory subunit of protein phosphatases which are important for signal transduction pathways were down-regulated. Here, we propose that acrylamide induces neurotoxicity by regulation of genes associated with neurofilament synthesis, axonal transport, neuro-protection, and signal transduction pathways.

In this study, a medicinal herbal plant, Meliae fructus, was examined and screened for anti-Helicobacter (H.) pylori activity. Seventy percent ethanol was used for herbal extraction. For anti-H. pylori activity screening, inhibitory zone tests as an in vitro assay and in vivo study using a Mongolian gerbil (Meriones unguiculatus) model were performed. Also, the safety of herbal compounds was evaluated by animal study. As a result of inhibitory zone test, Meliae fructus extract demonstrated strong anti-H. pylori activities. Also, as results of in vivo animal studies, Meliae fructus demonstrated strong therapeutic effects against H. pylori infection according to the criteria of histological examination and rapid urease test. As results of the safety study, after 28 days treatment of the Meliae fructus extract, the animals were not detected any grossly and histological changes. These results demonstrate that it can be successfully cured against H. pylori infection and protected from H. pylori-induced pathology with Meliae fructus. It could be a promising native herbal treatment for patients with gastric complaints including gastric ulcer caused by H. pylori.

Objective: To identify gait characteristics during trotting on a treadmill in nonlame Labrador Retrievers presumed predisposed or not predisposed to cranial cruciate ligament disease (CCLD). Animals: Clinically normal Labrador Retrievers presumed predisposed (n = 10) or not predisposed (7) to CCLD. Procedures: The right hind limb of each dog was classified by use of a predictive score equation that combined tibial plateau angle and femoral anteversion angle as presumed predisposed (high score [> −1.5]) or not predisposed (low score [≤ −1.5]) to CCLD. Tarsal joint, stifle joint, and hip joint kinematics, net moments, and powers were computed. Results: The stifle joint was held at a greater degree of flexion in limbs presumed predisposed to CCLD (130.9° vs 139.3°). More power was generated by muscles acting on the stifle joint in the early stance phase of limbs presumed to be predisposed to CCLD (2.93 vs 1.64 W/kg). The tarsal joint did not reach the same degree of extension in limbs presumed predisposed to CCLD, compared with that in limbs presumed not predisposed to CCLD (179.0° vs 161.0°). Velocity, stance time, vertical and craniocaudal forces, angular velocities, and net joint muscle moments did not differ between groups. Conclusions and Clinical Relevance: Gait mechanics of dogs with high (> −1.5) and low (≤ −1.5) tibial plateau angle and femoral anteversion angle scores were characterized on a treadmill, which may help in the identification of dogs predisposed to CCLD.

Objective: To determine the effects of ocular administration of ophthalmic 2% dorzolamide hydrochloride solution on aqueous humor flow rate (AHFR) and intraocular pressure (IOP) in clinically normal cats. Animals: 20 clinically normal domestic shorthair cats. Procedures: Following an acclimation period, IOP was measured in each eye of all cats 5 times daily for 3 days to determine baseline values. Fifteen cats received 1 drop of 2% dorzolamide solution and 5 cats received 1 drop of control solution in each eye every 8 hours for 5 days (treatment phase). The IOP of each eye was measured 5 times during each day of the treatment phase. Prior to and after the treatment phase, AHFR in both eyes of each cat was measured via fluorophotometry. Results: Prior to treatment, AHFR or IOP did not differ between the treatment and control groups. In dorzolamide-treated cats, mean AHFR after the treatment phase (3.47 ± 1.5 μL/min) was significantly lower than the value prior to treatment (5.90 ± 2.2 μL/min) and mean IOP during the treatment phase (11.1 ± 1.0 mm Hg) was significantly lower than the baseline mean IOP (14.9 ± 1.0 mm Hg). In the control group, IOP values did not differ before or during the treatment phase and AHFRs did not differ before and after the treatment phase. Conclusions and Clinical Relevance: Ocular administration of 2% dorzolamide solution significantly decreased AHFR and IOP in clinically normal cats. Application of 2% dorzolamide solution may be an effective treatment in cats with glaucoma.

Objective: To investigate the influence of dietary supplementation with l-carnitine on metabolic rate, fatty acid oxidation, weight loss, and lean body mass (LBM) in overweight cats undergoing rapid weight reduction. Animals: 32 healthy adult neutered colony-housed cats. Procedures: Cats fattened through unrestricted ingestion of an energy-dense diet for 6 months were randomly assigned to 4 groups and fed a weight reduction diet supplemented with 0 (control), 50, 100, or 150 μg of carnitine/g of diet (unrestricted for 1 month, then restricted). Measurements included resting energy expenditure, respiratory quotient, daily energy expenditure, LBM, and fatty acid oxidation. Following weight loss, cats were allowed unrestricted feeding of the energy-dense diet to investigate weight gain after test diet cessation. Results: Median weekly weight loss in all groups was ≥ 1.3%, with no difference among groups in overall or cumulative percentage weight loss. During restricted feeding, the resting energy expenditure-to-LBM ratio was significantly higher in cats that received l-carnitine than in those that received the control diet. Respiratory quotient was significantly lower in each cat that received l-carnitine on day 42, compared with the value before the diet began, and in all cats that received l-carnitine, compared with the control group throughout restricted feeding. A significant increase in palmitate flux rate in cats fed the diet with 150 μg of carnitine/g relative to the flux rate in the control group on day 42 corresponded to significantly increased stoichiometric fat oxidation in the l-carnitine diet group (> 62% vs 14% for the control group). Weight gain (as high as 28%) was evident within 35 days after unrestricted feeding was reintroduced. Conclusions and Clinical Relevance: Dietary l-carnitine supplementation appeared to have a metabolic effect in overweight cats undergoing rapid weight loss that facilitated fatty acid oxidation.

Objective: To determine the prevalence and types of tooth resorption in dogs with oral tumors and to compare findings with those for control dogs. Animals: 101 dogs with oral tumors and 128 control dogs that did not have oral tumors and for which dental radiographs were available. Procedures: Exclusion criteria for dogs included systemic disease, long-term administration of anti-inflammatory drugs, traumatic occlusion, severe semigeneralized or generalized periodontitis, and endodontic disease. For each dog with an oral tumor, histologic sections of biopsy specimens of tumors were examined. Dental radiographic images of dogs were examined, and the presence and type of tooth resorption were determined for each tooth. Statistical analyses were performed to compare data regarding prevalence of tooth resorption. Results: Teeth at tumor sites in dogs with nonodontogenic tumors were significantly more frequently affected with external inflammatory resorption, compared with teeth at tumor sites in dogs with odontogenic tumors. Teeth at sites distant from tumors in dogs with oral tumors were 3.2 times as likely to have external surface resorption (OR, 3.2; 95% confidence interval, 1.3 to 7.9) and 83.4 times as likely to have external inflammatory resorption (OR, 83.4; 95% confidence interval, 9.7 to 719.6) as teeth in control dogs. Conclusions and Clinical Relevance: Resorption of teeth at tumor sites and at sites distant from tumors was common in dogs with oral tumors. Results of the present study will contribute to an understanding of the complex effects of oral tumors on local and distant hard tissues.

Objective: To investigate contrast-enhanced ultrasonography as a minimally invasive method for the subjective and quantitative assessment of pancreatic and duodenal perfusion in healthy adult dogs, with reference to perfusion in adjacent liver tissue. Animals: 8 clinically normal adult dogs. Procedures: Contrast-enhanced ultrasonograms of the right pancreatic limb, proximal portion of the descending duodenum, and adjacent liver were acquired after IV administration of a microbubble contrast medium. Following subjective evaluation, quantitative time-intensity curves were generated from regions of interest in the pancreas, duodenum, and liver. Five contrast medium characteristics representing perfusion parameters were determined for each organ and used for statistical analysis: interval to arrival, inflow rate, peak intensity (PI), time of peak intensity (TPI), and outflow rate. Results: Significant associations between pancreatic and duodenal values were found for interval to contrast medium arrival, PI, TPI, and outflow rate. Pancreatic and duodenal inflow rates were not correlated. Inflow and outflow rates were significantly faster and TPI significantly shorter for the pancreas and duodenum, compared with values for the liver. There was no significant difference among all 3 organs for interval to arrival and PI of contrast medium. Subjective evaluation findings corresponded to quantitative analysis results. Conclusions and Clinical Relevance: Results suggested that contrast-enhanced ultrasonography may be a useful, minimally invasive method for evaluating pancreatic and duodenal perfusion in dogs. The data from healthy dogs reported here could aid in the assessment of pancreatic and duodenal conditions and their response to medical treatment.

Objective-To assess the effects of ischemia and reperfusion on indicators of oxidative stress, activation of eosinophils, and apoptosis in the large colonic mucosa of horses. Animals-40 horses. Procedures-In 1 or two 20-cm-long segments of the pelvic flexure, ischemia was induced for 1 or 2 hours followed by no reperfusion or 30 minutes and 18 hours of reperfusion in anesthetized horses. Mucosal specimens were collected before (controls; n = 20 horses) and after each period of ischemia, and full-thickness tissue samples were collected after each period of reperfusion. Sections of colonic tissues were stained for histomorphometric analysis or assessment of eosinophil accumulation. Nitrotyrosine was identified immunohistochemically, and severity of apoptosis was determined via the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method. Results-Numbers of mucosal eosinophils were similar before induction of ischemia, after ischemia, and after ischemia-reperfusion. Eosinophil nitrotyrosine production increased significantly during ischemia and continued through 30 minutes of reperfusion; production was decreased at 18 hours of reperfusion but remained greater than that of the controls. In other leukocytes, nitrotyrosine generation peaked at 1 hour of ischemia and again at 18 hours of reperfusion. Compared with control findings, epithelial apoptosis increased gradually at 1 through 2 hours of ischemia with no further progression after reperfusion. Conclusions and Clinical Relevance-Results suggested that resident eosinophils in the large colon of horses react to mucosal injury from ischemia and reperfusion and may undergo oxidative stress under those conditions. Epithelial apoptosis could contribute to tissue damage.

Objective: To describe the effect of systemically administered oxytetracycline on the viscoelastic properties of rat tail tendon fascicles (TTfs) to provide a mechanistic rationale for pharmacological treatment of flexural limb deformities in foals. Sample: TTfs from ten 1-month-old and ten 6-month-old male Sprague-Dawley rats. Procedures: 5 rats in each age group were administered oxytetracycline (50 mg/kg, IP, q 24 h) for 4 days. The remaining 5 rats in each age group served as untreated controls. Five days after initiation of oxytetracycline treatment, TTfs were collected and their viscoelastic properties were evaluated via a stress-relaxation protocol. Maximum modulus and equilibrium modulus were compared via a 2-way ANOVA. Collagen fibril size, density, and orientation in TTfs were compared between treated and control rats. Results: Viscoelastic properties were significantly decreased in TTfs from 1-month-old oxytetracycline-treated rats, compared with those in TTfs from 1-month-old control rats. Oxytetracycline had no effect on the viscoelastic properties of TTfs from 6-month-old rats. Collagen fibril size, density, and orientation in TTfs from 1-month-old rats did not differ between oxytetracycline-treated and control rats. Conclusions and Clinical Relevance: Results confirmed that systemically administered oxytetracycline decreased the viscoelastic properties of TTfs from 1-month-old rats but not those of TTfs from 6-month-old rats. The decrease in viscoelastic properties associated with oxytetracycline treatment does not appear to be caused by altered collagen fibril diameter or organization. The age-dependent effect of oxytetracycline on the viscoelastic properties of tendons may be related to its effect on the maturation of the extracellular matrix of developing tendons.