Spinal nephroblastoma in a miniature Dachshund

A 2-year-old castrated miniature Dachshund dog was presented to the Rakuno Gakuen Veterinary Teaching Hospital for diagnosis of progressive hindlimb paresis and ataxia. There was no thoracolumbar intervertebral disk hernia and magnetic resonance imaging revealed an intramedullary spinal cord lesion at the ninth and tenth thoracic vertebrae. Following surgical excision of the neoplasm, there was minor amelioration of neurological signs, but forelimb function was not recovered. The extracted tumor was histopathlogically diagnosed as spinal nephroblastoma.     

Spontaneous Accumulation of Globotriaosylceramide (Gb3) in Proximal Renal Tubules in an ICR Mouse

This report describes spontaneous cytoplasmic vacuolation in the proximal renal tubules of a 7-week-old male ICR [Crlj:CD1(ICR)] mouse. The contents of vacuoles were positively stained with periodic acid-Schiff (PAS) and Sudan black, and the membranes were positive on immunohistochemical staining for lysosomal-associated membrane protein-2 (LAMP-2), a marker of lysosomal membrane. Electron microscopy revealed electron-dense lamellar bodies in the proximal tubular epithelial cells. These histopathological features are similar to those in α-galactosidase A-deficient mice, in which globotriaosylceramide (Gb3), a glycosphingolipid, accumulates in lysosomes. When we performed immunohistochemical staining for Gb3, the contents of vacuoles were positively stained. From these results, spontaneous cytoplasmic vacuolation in the proximal renal tubules in the mouse was identified as lysosomal accumulation of Gb3.     

A new rolling culture‐based in vitro fertilization system capable of reducing polyspermy in porcine oocytes

The high incidence of polyspermy is one of the major obstacles during in vitro fertilization (IVF) in pigs. To overcome this, we developed a novel IVF method, which involves constant rotation. Oocytes matured in vitro were mixed with spermatozoa (0.2 × 10⁵ sperm/mL) in an IVF medium (200 μL) using a 200 μL PCR tube. This tube was then rotated at 1 rpm for 6 h at 38.5°C in a rotation mixer (experimental group). A second PCR tube was simultaneously cultured without rotation (control group). The rate of polyspermy was evaluated 12 h after insemination and was significantly (P < 0.05; 21.0% vs. 48.3%) lower in the experimental group than in the control group. Sperm penetration rate was similar in oocytes from the experimental and control groups (75.2% vs. 83.1%). However, monospermic fertilization rate of the oocytes was significantly (P < 0.05; 44.8% vs. 21.2%) higher in the experimental group than in the control group. Furthermore, the rate of blastocyst formation (30.1% vs. 20.8%) increased in the experimental group, as compared to the control group. This present system will contribute to increase the efficacy of blastocyst production through reduction of polyspermic penetration.     

Seasonal influence on testicular function of male raccoons, Procyon lotor

In order to clarify the breeding capability of male raccoons in Japan, the testes of raccoons, a nuisance animal, collected in Kanagawa Prefecture and Hokkaido were histologically inspected. Furthermore, testosterone concentrations in their blood were measured. The testosterone concentrations increased in winter and the diameter of seminiferous tubules and the spermatogenetic score decreased in summer for the animals captured in Kanagawa Prefecture. For the animals captured in Hokkaido, the diameter of seminiferous tubules did not change and the decrease of the spermatogenetic score in summer was slight. As the above results show, there is a breeding season in male raccoons in Japan, and the reduction of testicular function in summer was greater in animals captured in Kanagawa Prefecture than in animals captured in Hokkaido.     

Association of microalbuminuria and the urine albumin-to-creatinine ratio with systemic disease in cats

OBJECTIVE: To determine the diagnostic usefulness of semiquantitative and quantitative microalbuminuria assays and urine albumin-to-creatinine (UAC) ratio for detecting disease in cats. DESIGN: Prospective study. ANIMALS: 441 cats evaluated at a veterinary teaching hospital. PROCEDURES: Urine samples from cats for which a complete medical record was available were included. Urine dipstick results, urine protein-to-creatinine ratios (cutoffs, 0.1 and 0.4), semiquantitative and quantitative microalbuminuria assay results (cutoff, 1 mg/dL), and UAC ratio values (cutoffs, 100 and 200 mg/g) were determined. Clinical diagnoses determined within 3 months of enrollment were recorded. Sensitivity and specificity were determined with disease status used as the standard. The influences of clinical diagnosis, sex, age, serum urea nitrogen and creatinine concentrations, blood pressure, bacterial urine culture results, rectal temperature, pyuria, hematuria, and bacteriuria were evaluated by means of logistic regression. RESULTS: Of 441 cats that were eligible for inclusion, 40 were healthy and 401 had > or = 1 disease. Results of logistic regression indicated that significant associations existed for age, presence of disease, presence of urinary tract disease, azotemia, hematuria, and pyuria and results of 1 or both of the microalbuminuria assays. CONCLUSIONS and CLINICAL RELEVANCE: Microalbuminuria was associated with underlying disease. Sensitivity and specificity of the microalbuminuria assays for detection of systemic disease were superior to those of other tests. Microalbuminuria testing in conjunction with other screening procedures may increase identification of occult disease. A prospective study evaluating the predictive values of screening tests with and without microalbuminuria determination is needed to validate this recommendation.     

Successful suppression of endogenous α-1,3-galactosyltransferase expression by RNA interference in pig embryos generated in vitro

RNA interference (RNAi) technology using small interfering RNAs (siRNA) has been widely used as a powerful tool to knock down gene expression in various organisms. In pig preimplantation embryos, no attempt to suppress the target gene expression with such technology has been made. The purpose of this study is to demonstrate that the RNAi technology is useful for suppression of endogenous target gene expression at an early stage of development in pigs. Alpha-1,3-Galactosyltransferase (α-GalT) is an enzyme that creates the Galα1-3Gal (α-Gal) epitope on the cell surface in some mammalian species, and removal of the epitope is considered to be a prerequisite for pig-to-human xenotransplantation. We decided to suppress the endogenous α-GalT mRNA expression in pig early embryos, since reduction of α-GalT synthesis is easily monitored by cytochemical staining with Bandeiraea simplicifolia isolectin-B(4), a lectin that specifically binds to the α-Gal epitope, and by RT-PCR analysis. Cytoplasmic microinjection of double-stranded RNA and pronuclear injection of an siRNA expression vector into the embryos generated in vitro resulted in a significant reduction in expression of the α-GalT gene and α-Gal epitope in blastocysts, at which stage the α-Gal epitope is abundantly expressed. Somatic cell nuclear transfer of embryonic fibroblasts stably transfected with an siRNA expression vector also led to a significant reduction in the level of α-GalT mRNA synthesis together with decreased amounts of the α-Gal epitope at the blastocyst stage. These results indicate that the RNAi technology is useful for efficient suppression of a target gene expression during embryogenesis in pigs and suggest the possibility of production of siRNA-expressing pigs for use in xenotransplantation.     

Interpreting gelatinase activity in tumor tissue and serum as a prognostic marker of naturally developing canine tumors

To evaluate the clinical usefulness of tissue and serum gelatinase activity as a prognostic marker of canine tumors, tissue samples from 60 tumors and corresponding serum samples from the same animals were collected at the time of biopsy and surgery. On the basis of histopathology and clinical aggressiveness of metastasis and recurrence (MR), the cases were divided into 6 categories: non-inflammatory (Inf(-)) and inflammatory (Inf(+)) benign, and the Inf(-) MR(-), Inf(-) MR(+), Inf(+) MR(-), and Inf(+) MR(+) malignant. Gelatinase activity was determined semi-quantitatively using gelatin zymogram with a gelatinase standard from cultured canine peripheral blood mononuclear cells stimulated with lipopolysaccharide. No significant difference in gelatinase activities in tissue extracts was evident between the benign and malignant tumors. Inf(+) benign tumors, as well as Inf(-) MR(+), Inf(+) MR(-) and Inf(+) MR(+) malignant tumors, showed significantly higher tissue gelatinase activity than Inf(-) benign. The tissue activity in Inf(-) MR(-) malignant was significantly lower than in Inf(+) MR(-) and Inf(+) MR(+) malignant. The serum activity was significantly higher in the malignant cases than in the control and the benign. Inf(-) MR(+), Inf(+) MR(-) and Inf(+) MR(+) malignant tumors induced significantly higher gelatinase activity in serum than Inf(-) benign tumors. Gelatinase activity in serum was positively correlated with that in tumor extracts. Increased gelatinase in tumor tissue and serum may be correlated with inflammation as well as tumor aggressiveness, and thus should be used in combination with histopathology for predicting tumor metastasis or recurrence.     

Prediction of crude fat content of longissimus muscle of beef using the ratio of fat area calculated from computer image analysis: comparison of regression equations for prediction using different input devices at different stations

Crude fat content of longissimus (ribeye) muscle of beef cattle was predicted from a ratio of fat area (RFA) to area of ribeye muscle calculated from computer image analysis (CIA). Cross sections of 64 ribeyes taken from the 6-7th rib from cattle at experiment station A and cross sections of 94 ribeyes taken from the 6-7th rib from cattle at Experiment Station B were used in this study. Slices (1 to 1.5 cm in thickness) of just the Longissimus dorsi were homogenized and sampled for chemical estimation of crude fat content using petroleum ether. Crude fat content as determined from chemical analysis was used as the true estimate of fat content. A CCD (charge-coupled device) camera was used as the input device at Experiment Station A, and a single-lens reflex camera was used at Experiment Station B to photograph ribeyes for CIA. The contour comparison method, which assigns a threshold value for each marbling particle, was used to obtain accurate binarization in this study. Minimum and maximum of chemical measurements of crude fat were 2.1 and 39.8%, and for CIA calculation of the RFA were 6.1 and 56.8%, respectively. This range covered almost the complete range of the beef marbling standard used in carcass grading in Japan. The equation for the regression of the crude fat content (Y) on RFA (X) calculated from CIA for all of the data was Y = .793X-3.04 with r2 = .96. Regression equations for prediction of crude fat percentage from RFA taking into consideration the effect of experiment station were Y = .741X-2.22 with r2 = .91 for Experiment Station A, and Y = .782X-2.54 with r2 = .91 for Experiment Station B. Analysis of covariance showed that the effects of experiment stations on intercepts and slopes were not significant (P > .10). The ranges of differences between actual and predicted crude fat content from the prediction equation that was calculated without consideration of the effect of station were -6.4 to 4.0%. CIA of cross sections of the ribeye muscle seems to have potential for prediction of crude fat content.