Antitumor Activity of Mycobacterial Cell Wall‐DNA Complex (MCC) Against Canine Urinary Bladder Transitional Cell Carcinoma Cells

Introduction:  Mycobacterial cell wall‐DNA complex (MCC) is a bifunctional anticancer agent that induces cancer cell apoptosis and stimulates cytokine synthesis by immune cells. Intravesical MCC is currently being evaluated in humans with high‐grade urinary bladder cancer. Evaluation of MCC in dogs with transitional cell carcinoma (TCC) will allow mechanistic studies in a natural animal model of TCC, and a potentially beneficial therapy for dogs with this cancer. In this study, we have determined the anticancer activity of MCC against canine TCC cells in vitro .
Methods:  Canine TCC cells (K9TCC cell line) were incubated with MCC (0.05–100 μg/ml, 0.5–72 hours). Cellular proliferation was measured by MTT reduction. Cell cycle was analyzed by flow cytometry with propidium iodide. Apoptosis was identified by flow cytometry using anti‐active‐caspase‐3/PE and anti‐cleaved‐PARP/FITC antibodies. Apoptosis‐inducing activity of 100 μg/ml MCC in combination with piroxicam (0.1–1.0 uM) was evaluated.
Results:  MCC inhibited K9TCC cell proliferation in a concentration‐dependent manner (maximal activity – 45% at 100 μg/ml MCC) in association with the presence of activated caspase‐3 and cleaved PARP. Inhibition of proliferation and apoptosis‐inducing activities of MCC were independent of cell cycle phase. A thirty‐minute exposure of MCC was sufficient for optimal activity. Piroxicam (0.5 uM) enhanced apoptosis‐inducing activity of MCC.
Conclusions:  MCC induces apoptosis in K9TCC cells. This activity is potentiated by piroxicam. Following positive results in vitro , in vivo studies have been initiated. One dog, treated to date, has had a minor reduction in tumor volume following the first course of treatment with no treatment‐related toxicity.     

Effect of diet composition and slaughter weight on animal performance, carcass and meat quality, and fatty acid composition in veal calves

The effect of true milk use in the diet of Spanish Brown Swiss male calves on animal performance, carcass and meat quality, and fatty acids composition was studied. In experiment 1, the effect of milk intake [ad libitum continuous (ADLIB) feed vs. restricted 0.7 during 75 days followed by ad libitum feed (RESTR)] and slaughter endpoint (225 kg vs. 5 month) were studied. In experiment 2, ad libitum concentrate feeding [grain-fed (GF)] was compared with milk supplementation until slaughter [milk-fed (MF)] in calves slaughtered at 345 kg. As regards to milk intake, carcass weight and degree of fatness were higher in the ADLIB group (P<0.05). The RESTR group revealed a higher percentage of saturated fatty acids (P<0.05). In experiment 2, the MF group exhibited a higher fat percentage (P<0.05), lower press and cooking losses (P<0.05), and higher scores for tenderness and juiciness (P<0.05) than the GF group. The percentage of saturated fatty acids was higher in the MF group (P<0.05). The results suggest that true milk use in veal production could be an advantageous alternative in terms on production costs, animal performance, and carcass and meat quality.     

First Established Pregnancies in Mediterranean Italian Buffaloes (Bubalus bubalis) Following Deposition of Sexed Spermatozoa near the Utero Tubal Junction

At the time of AI following Ovsynch protocol, a total of 51 buffaloes were randomly divided in a first group (n = 30) subjected to conventional AI into the uterine body with 20 million non-sex sorted frozen-thawed spermatozoa, while a second group (n = 21) was inseminated near the utero-tubal junction (UTJ) ipsilateral to the ovary carrying the preovulatory follicle with 2.5 million live (4 million total) sex-sorted frozen-thawed spermatozoa. The semen used for flowcytometric sorting was collected and processed on a farm in Italy, and then shipped to a laboratory in Germany. Eleven buffaloes were inseminated with X-chromosome bearing spermatozoa and 10 with Y-chromosome bearing spermatozoa. Conception rates after conventional and UTJ inseminations were 43.3% (n = 13) and 42.8% (n = 9) respectively (p = 0.97). Eight of the nine foetuses obtained after insemination with sexed spermatozoa corresponded to the sex as predicted by the cell sorting procedure (five male and four female foetuses by ultrasound vs six male and three female foetuses by cell sorting). In conclusion, for the first time buffalo semen has been successfully subjected to procedures for flowcytometric sperm sorting and freezing. Low doses of sexed spermatozoa have been deposited near the UTJ giving conception rates similar to those of conventional AI with full dose.     

THE EFFECTS OF PATIENT POSITIONING AND SLICE SELECTION ON CANINE ACETABULAR ANGLE ASSESSMENT WITH COMPUTED TOMOGRAPHY

Acetabular angles (AAs) and dorsal acetabular rim angles acquired by computed tomographic (CT) imaging have been used to assess patient response to juvenile pubic symphysiodesis surgery. The purpose of this study was to evaluate the effects of patient positioning and slice selection on these angles, and an attempt was made to devise a repeatable method of measuring these angles that would eliminate positioning effects. We found significant variation in AAs with small differences in pelvic tilt and slice selection. Dorsal acetabular rim angles were not affected. As a result of positioning differences from one CT study to the next, every attempt should be made to standardize pelvic tilt, or eliminate its effect on AAs by standardizing gantry angle in relation to an anatomic landmark that will not change over time. The floor of the sacral vertebral canal may be a reasonable landmark for this purpose and deserves further study.     

Mixed models applied to the study of variation of grower-finisher mortality and culling rates of a large swine production system

Large scale production systems for swine are frequently organized in a hierarchical structure. Consequently, important production parameters, such as mortality and culling, can be analyzed at different levels. The major aims of this study were to assess variance components (VC) of mortality and culling rates attributed to sites and to barns within a site, and subsequently to investigate the impact of average entry weight, days on feed (length of the production turn), and season on the magnitude of the VC. Then, data from a large farm with 3 sites were collected during 5 y. In total, 1720040 pigs distributed in 1502 all-in/all-out grower-finisher groups were included. Linear mixed models were fitted for mortality and culling rates. The barn was modeled as the residual component (barn-to-barn variations) with production turn and site nested within production turn as random intercept variance components. Barn-to-barn pig group variation was the largest VC for mortality (63.08%), when no predictors were included in the models. Predictors, such as pigs placed on quarters 2 and 3, low average entry weight, and shorter production turn length, were associated together with higher mortality. The explained proportion of variance due to these predictors was about 12.05% and the VC for barn, site, and production turn were 67.6%, 17.6%, and 14.8%, respectively. Barn-to-barn variation was also the largest VC for culling rate (46.2%), but the same predictor mentioned above explained only about 1.4% of the variation. The VC for barn, site, and production turn were 46.8%, 21.3%, and 31.8%, respectively. Since the variability among barns far exceeded the variability among sites, the barn should be used as experimental unit in studies with grower-finisher mortality, culling rate, or both, as outcome variables.     

Occurrence and Distribution of Lactate Dehydrogenase in Knee Joint Carilage in Healthy and Osteoarthrotic Dogs

According to clinical studies, degenerative diseases of canine joints lead to higher lactate dehydrogenase (LDH) levels in synovial fluid. The goal of the present study was to examine the intraarticular distribution of LDH in healthy and osteoarthrotic knee joints in order to identify possible sources of LDH in synovial fluid. As synovial LDH concentrations neither correlate with the number of leukocytes nor with synovitis, our investigation focused on the articular cartilage. Samples from healthy and osteoarthrotic knee joints were fixed and processed for transmission electron microscopy (TEM), immunohistochemistry (IHC), and immunocytochemistry (ICC). In addition, fresh cartilage samples were investigated cytochemically by the tetrazolium‐formazan reaction. Analyses of blood and synovial fluid samples were used to confirm the absence of inflammatory disease. Morphology of articular cartilage was assessed macroscopically and by means of TEM. IHC revealed highest levels of LDH in chondrones and a diffuse labelling of the matrix with a distinctive decrease in signal from superficial to deeper cartilage layers. Ultrastructural localization by ICC showed LDH to be present in the cytoplasm of all chondrocytes and confirmed the density gradient in the matrix. Labelling was absent from nuclei and from pericellular rims. Cytochemistry confirmed the distribution pattern and, thus, expanded our findings beyond immunological evidence by providing proof of enzymatic activity of LDH in articular cartilage. The present results indicate that LDH is transferred from chondrocytes to the cartilaginous matrix. We suggest, therefore, that LDH found in synovial fluid originates from the articular cartilage and that osteoarthrotic processes promote LDH release from the cartilaginous matrix.     

Distribution and Activity of Fatty Acid Amide Hydralase (FAAH) in the Gastrointestinal Neurons of Mouse

The endocannabinoid anandamide may regulate intestinal motility through activation of CB1 receptors. Anandamide is then inactivated by fatty acid amide hydrolase (FAAH), a membrane bound enzyme. Under pathological conditions, inactivation of such enzymatic activity may lead to inhibition of the intestinal motility. Here, preliminary reports on the distribution of Fatty Acid Amide Hydrolase (FAAH) immunoreactivity in the mouse gastrointestinal neurons, and the pharmacological effects of N‐arachidonoylserotonin (AA‐5HT), a selective inhibitor of FAAH, are reported. FAAH was revealed by an indirect immunofluorescence. Laminar preparations containing the myenteric or the submucous plexus adhered, were peeled off after the whole gut wall had been stretched out and fixed in 4% paraformaldehyde. They were subsequently incubated with a polyclonal anti‐serum directed against a region near the N‐terminus of the human FAAH and revealed by a FITC‐conjugated goat anti‐rabbit secondary anti‐serum. FAAH‐immunoreactive neurons were observed within the myenteric ganglia throughout the GIT. The positive nerve cells varied in size and density of immunoreactivity. Stomach and large intestine showed the highest neuronal density. AA‐5HT significantly reduced both gastric emptying and gastrointestinal tract transit. Such inhibitory effect was reduced by the C1 receptor antagonist SR141716A. Both morphological and pharmacological results suggest that FAAH may play a critical role in controlling gut anandamide levels.     

Serodiagnosis of Burkholderia mallei Infections in Horses: State‐of‐the‐art and Perspectives

Burkholderia mallei causes glanders or farcy in solipeds, a disease that must be reported to the OIE (Office International des Epizooties, Paris, France). The number of reported outbreaks has increased steadily during the last decade. Serodiagnosis is hampered by the considerable number of false‐positives and ‐negatives of the internationally prescribed tests. The major problem leading to low sensitivity and specificity of complement fixation test (CFT) and enzyme‐linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e. crude preparations of whole cells. Future perspectives for the development and evaluation of serological test kits using well‐characterized single antigens are discussed in the light of recent molecular research on B. mallei and the closely related saprozoonotic agent B. pseudomallei.     

Growth response of Ficus benjamina to limited soil volume and soil dilution in a skeletal soil container study

The interactive effects of rooting volume and nutrient availability in a skeletal soil medium designed to meet street tree and pavement needs were observed in a containerized experiment. Benjamin fig (Ficus benjamina L.) was grown in a stone-soil blended skeletal soil material (CU-Soil™) and compared to a loam soil. The same topsoil used as the soil component in the skeletal soil material was used as the sole component in the comparison soil-only treatment.Plants grown in the skeletal soil material had reduced leaf tissue N content and depressed growth compared with plants grown in non-diluted soil. No other mineral deficiencies were found. Leaf number, chlorophyll concentration, shoot weight, and root characteristics were all affected.Reduced growth from soil dilution could be offset by the provision of an enlarged rooting volume for root development. Large containers of skeletal soil were observed to have smaller root systems compared to equivalent net volumes of loam soil at the first two harvest dates of the study. By the end of the study, the large containers of skeletal soil were observed to have developed larger root systems compared to equivalent net volumes of loam soil; resulting in comparable leaf N levels and total plant dry matter. Plants in skeletal soil had lower shoot: root ratios at the end of the study. Investing resources to further root growth in times of nutrient shortages is a probable plant reaction as evidenced by differences in specific root length between treatments. The study allowed a method for directly partitioning the containerization effect by having equivalent amounts of soil over two volumes.