MAGNETIC RESONANCE IMAGING OF THE CANINE BRAIN AT 7 T

The purpose of this study was to describe relevant canine brain structures as seen on T2-weighted images following magnetic resonance (MR) imaging at 7 T and to compare the results with imaging at 1.5 T. Imaging was performed on five healthy laboratory beagle dogs using 1.5 and 7 T clinical scanners. At 1.5 T, spin echo images were acquired, while gradient echo images were acquired at 3 T. Image quality and conspicuity of anatomic structures were evaluated qualitatively by direct comparison of the images obtained from the two different magnetic fields. The signal-to-nose ratio (SNR) and contrast-to-noise ratio (CNR) were calculated and compared between 1.5 and 7 T. The T2-weighted images at 7 T provided good spatial and contrast resolution for the identification of clinically relevant brain anatomy; these images provided better delineation and conspicuity of the brain stem and cerebellar structures, which were difficult to unequivocally identify at 1.5 T. However, frontal and parietal lobe and the trigeminal nerve were difficult to identify at 7 T due to susceptibility artifact. The SNR and CNR of the images at 7 T were significantly increased up to 318% and 715% compared with the 1.5 T images. If some disadvantages of 7 T imaging, such as susceptibility artifacts, technical difficulties, and high cost, can be improved, 7 T clinical MR imaging could provide a good experimental and diagnostic tool for the evaluation of canine brain disorders.     

Mdivi-1, mitochondrial fission inhibitor,impairs developmental competence and mitochondrial function of embryos and cells in pigs

Mitochondria are highly dynamic organelles that undergo constant fusion/fission as well as activities orchestrated by large dynamin-related GTPases. These dynamic mitochondrial processes influence mitochondrial morphology, size and function. Therefore, this study was conducted to evaluate the effects of mitochondrial fission inhibitor, mdivi-1, on developmental competence and mitochondrial function of porcine embryos and primary cells. Presumptive porcine embryos were cultured in PZM-3 medium supplemented with mdivi-1 (0, 10 and 50 μM) for 6 days. Porcine fibroblast cells were cultured in growth medium with mdivi-1 (0 and 50 μM) for 2 days. Our results showed that the rate of blastocyst production and cell growth in the mdivi-1 (50 μM) treated group was lower than that of the control group (P < 0.05). Moreover, loss of mitochondrial membrane potential in the mdivi-1 (50 μM) treated group was increased relative to the control group (P < 0.05). Subsequent evaluation revealed that the intracellular levels of reactive oxygen species (ROS) and the apoptotic index were increased by mdivi-1 (50 μM) treatment (P < 0.05). Finally, the expression of mitochondrial fission-related protein (Drp 1) was lower in the embryos and cells in the mdivi-1-treated group than the control group. Taken together, these results indicate that mdivi-1 treatment may inhibit developmental competence and mitochondrial function in porcine embryos and primary cells.     

Leiomyosarcoma of urinary bladder in a Shih Tzu dog

A 10-year-old intact male Shih Tzu dog presented with hematuria. Double-contrast cystography revealed a polypoid filling defect at the apex of the urinary bladder. Ultrasonography revealed a heterogeneously hypoechoic intramural mass with minimal vascular flow beneath the submucosal layer. After partial cystectomy, a well-demarcated bladder leiomyosarcoma was diagnosed on histopathology. The patient was alive and well without any clinical signs, recurrence, or metastasis at the 29-month follow-up after the surgical removal of the bladder mass. Leiomyosarcoma should be considered as a differential diagnosis if mass-like lesions are observed in the urinary bladder, although this type of malignancy is rare in canines. Histopathological confirmation is important for predicting prognosis and determining further medical plans.     

A Cathepsin B Inhibitor,E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos

Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.     

Evaluation of left ventricular function with cardiac magnetic resonance imaging and echocardiography after administration of dobutamine and esmolol in healthy beagle dogs

Unlike echocardiography, cardiac magnetic resonance imaging (cardiac MRI) results in a near-exact assessment of cardiac structures and function. However, most veterinary studies have focused on dogs with normal cardiac function. We hypothesized that there would be significant differences in cardiac measurements between cardiac MRI and echocardiography when left ventricular (LV) function was abnormal. This study was undertaken to compare measurements of LV function produced by cardiac MRI and echocardiography in dogs whose LV function was altered by pharmacological agents. This study was conducted with six healthy beagle dogs. We increased left ventricular contractility by administration of dobutamine; we decreased cardiac contractility with esmolol. Stroke volume measurements were made by using both cardiac MRI and echocardiography under seven different conditions with general anesthesia: control, three doses of esmolol (100, 200, and 500 µg/kg/min), and three doses of dobutamine (10, 20, and 50 µg/kg/min). Experiments involving each condition were conducted at least 1 week apart. When LV contractility was normal, ejection fraction (EF) and stroke volume (SV), as measured by echocardiography and cardiac MRI, were not significantly different. However, when contractility was changed by pharmacological agents, EF and SV were overestimated by echocardiography, compared to MRI. Evaluation of cardiac function in patients treated with pharmacological agents should be conducted carefully because EF and SV measured by echocardiography can be overestimated, compared with EF and SV obtained by cardiac MRI.     

Changes in the predicted function of the rumen bacterial community of Japanese Black beef cattle during the fattening stages according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses

We investigated changes in the predicted functions of the rumen bacterial community in Japanese Black beef cattle during fattening. Nine cattle were fed a high-concentrate diet during the early, middle, and late fattening stages consecutively (10–14, 15–22, and 23–30 months of age, respectively). The rumen fluid and solid samples collected at each stage were subjected to sequencing analyses. The sequencing results were clustered and classified into operational taxonomic units (OTUs). Representative sequences and a raw counting table for each OTU were submitted to the Piphillin website. The predicted functions were revealed by the Kyoto Encyclopedia of Genes and Genomes database as the ratio of the total sequence. In the early stage, “Biosynthesis of secondary metabolites” was significantly higher in the fluid fraction than in the solid fraction. “Two-component system” in the middle stage was significantly lower and “Purine metabolism” in the late stage was significantly higher in the fluid fraction than those in the solid fraction. The fluid fraction was significantly correlated with acetic acid, propionic acid, and bacterial metabolism, such as “Biosynthesis of secondary metabolites” and “Sugar metabolism.” Moreover, the solid fraction was correlated with “Purine metabolism” and “Biosynthesis of secondary metabolism”. These results suggest that the rumen bacterial community in Japanese Black beef cattle adapts to changes in rumen conditions by altering their functions in response to a long-term high-grain diet.     

Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming

Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot^TM reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.     

Effect of Omentectomy on Adhesion Formation in Horses

Objective —To determine if omentectomy would decrease the frequency of postoperative intraabdominal adhesions. Study Design —Retrospective study. Animals or Sample Population—44 horses that had either two ventral median celiotomies or a ventral median celiotomy and a necropsy more than 4 days later; 19 of these horses had their omentum removed at the initial surgery. Methods —Data retrieved from the records included location and type of intraabdominal adhesions; location of the surgical lesion; relationship of adhesions to the surgical lesion; surgical procedures; duration of initial surgery; time interval between procedures; age, gender, and breed of the horse; and clinical outcome. Fisher's exact test was used to evaluate the association between categorical explanatory and outcome variables. The effect of potential risk factors on the incidence rate of adhesion formation was estimated using a proportional hazards regression model. Results —Of 25 horses in the nonomentectomy group, 15 (60%) had postoperative adhesions that resulted in the need for a second surgical intervention, whereas of 19 horses that had omentectomy initially, only 4 (21%) had postoperative adhesions that required a second procedure. Rate of adhesion formation was higher in horses that did not have omentectomy initially (incidence ratio rate [IRR], 0.46; 90% confidence interval [CI], 0.18 to 1.19). At initial surgery, 24 horses had a small intestinal lesion, and 20 horses had a large intestinal lesion. Fifteen horses (63%) with small intestinal lesions subsequently developed adhesions compared with four horses (20%) with an initial large intestinal lesion (P= .006). At the second procedure, small intestine lesions were identified in 32 horses and large intestine lesions in 12 horses (1 horse had both small and large intestine lesions), and 1 horse had a gastric lesion. Adhesions were identified as the cause of colic signs in 19 (61%) horses with small intestinal lesions and in none of the horses with large intestine lesions. The frequency of adhesion development leading to colic associated with only the small intestine at the second surgery or necropsy was significantly greater (P= .001) than the frequency only in the large intestine. Conclusions —Omentectomy reduced the rate of postoperative adhesion formation. Adhesions are more likely to occur after small intestinal surgery and if they do occur likely involve the small intestine. Clinical Relevance —Omentectomy is a safe procedure and should be considered prophylactically for reduction of adhesion formation after abdominal surgery in horses.     

Comparison of oxygen consumption patterns between wild and cultured black rockfish Sebastes schlegeli

ABSTRACT:   To compare metabolic activity rhythms between wild and cultured black rockfish Sebastes schlegeli , we measured long-term oxygen consumption rates (OCR) using an automatic intermittent-flow respirometer under constant temperature and darkness. Oxygen consumption rates peaked among wild black rockfish at 12.4 h intervals, which corresponded to a circatidal rhythm. The wild fish were probably exhibiting responses that corresponded to tidal events in their natural environment. However, when captured wild black rockfish were kept under laboratory conditions (12 h light [L] : 12 h dark [D]) for 30 days, the OCR shifted to a circadian rhythm (24.1–24.9 h). The OCR of cultured black rockfish that had been reared in a tank for 9 months peaked at approximately 24 h intervals, corresponding to a circadian rhythm. The results of this study suggest that the differences in OCR patterns between wild and cultured fish were mainly due to differences in the environmental conditions between tidal and non-tidal habitats.