Evaluation of a Collagenase Generated Osteoarthritis Biomarker in Naturally Occurring Canine Cruciate Disease

Objective— To determine the clinical value of a novel osteoarthritis (OA) biomarker in detecting canine cruciate disease.
Study Design— Cross sectional clinical study.
Animals— Dogs (n=22) with cranial cruciate ligament (CCL) rupture and 12 control dogs.
Methods— Concentrations of collagenase-generated cleavage epitope of type II collagen (Col2-3/4C_{long mono}, or C2C) in serum, urine, and joint fluid were compared between a group of dogs with CCL rupture and a control group. Correlation of C2C concentrations to the clinical stage of stifle OA was also evaluated.
Results— There were no significant differences in C2C concentrations in serum, urine, and joint fluid between groups ( P >.05). Subjective scores of lameness, joint effusion, osteophytosis were significantly more severe in the CCL rupture group compared with the control group ( P <.05). There was no significant correlation of C2C concentrations with clinical stage of stifle OA ( P >.05).
Conclusion— This OA biomarker did not detect pathology associated with CCL rupture. Our results suggest that collagenase-specific degradation of type II collagen in articular cartilage may not be involved in the early stage of naturally occurring canine cruciate disease, and that pathology associated with naturally occurring CCL rupture is different from that of experimental OA model.
Clinical Relevance— C2C is not clinically useful in detecting CCL rupture in dogs.     

Study on some functional and compositional properties of yak butter lipid

The present experiment was performed to determine some functional properties of yak butter lipids such as lipid class composition; conjugated linoleic acid (CLA) composition, differential scanning calorimetric (DSC) analysis, tyrosinase inhibition activity and antioxidant property. Yak butter lipids composition contained 98% triacylglycerols, 0.9% free fatty acids, 0.32% free sterols and 0.27% phospholipids. The CLA content in yak butter was 2.5% and the major portion was of cis‐9 and trans‐11 (90%). The DSC analysis of yak butter lipids showed a similar path for transition temperature as cow butter lipids, although the enthalpy of yak lipid was higher (40.0 mJ/mg) compared to cow butter lipids (32.0 mJ/mg). Melting point of yak butter was observed at 41°C. Yak butter with lactic acid, NaCl, citric acid and ascorbic acid showed pronounced tyrosinase inhibition activity. Vegetable oils blended with yak butter have extended the oxidation induction time.     

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 Tibial Plateau Leveling Osteotomy on Femorotibial Contact Mechanics and Stifle Kinematics

Objective— To evaluate the effects of tibial plateau leveling osteotomy (TPLO) on femorotibial contact mechanics and 3-dimensional (3D) kinematics in cranial cruciate ligament (CrCL)-deficient stifles of dogs.
Study Design— In vitro biomechanical study.
Animals— Unpaired pelvic limbs from 8 dogs, weighing 28–35 kg.
Methods— Digital pressure sensors placed subjacent to the menisci were used to measure femorotibial contact force, contact area, peak and mean contact pressure, and peak pressure location with the limb under an axial load of 30% body weight and a stifle angle of 135°. Three-dimensional static poses of the stifle were obtained using a Microscribe digitizing arm. Each specimen was tested under normal, CrCL-deficient, and TPLO-treated conditions. Repeated measures analysis of variance with a Tukey post hoc test ( P <.05) was used for statistical comparison.
Results— Significant disturbances to all measured contact mechanical variables were evident after CrCL transection, which corresponded to marked cranial tibial subluxation and increased internal tibial rotation in the CrCL-deficient stifle. No significant differences in 3D femorotibial alignment were observed between normal and TPLO-treated stifles; however, femorotibial contact area remained significantly smaller and peak contact pressures in both medial and lateral stifle compartments were positioned more caudally on the tibial plateau, when compared with normal.
Conclusion— Whereas TPLO eliminates craniocaudal stifle instability during simulated weight bearing, the procedure fails to concurrently restore femorotibial contact mechanics to normal.
Clinical Relevance— Progression of stifle osteoarthritis in dogs treated with TPLO may be partly the result of abnormal stifle contact mechanics induced by altering the orientation of the proximal tibial articulating surface.     

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.     

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.