Aberrantly Expressed tRNA-Val Fragments Can Distinguish Canine Hepatocellular Carcinoma from Canine Hepatocellular Adenoma.

Hepatocellular adenoma (HCA) and hepatocellular carcinoma (HCC) can be difficult to differentiate but must be diagnosed correctly as treatment and prognosis for these tumors differ markedly. Relevant diagnostic biomarkers are thus needed, and those identified in dogs may have utility in human medicine because of the similarities between human and canine HCA and HCC. A tRNA-derived fragment (tRF), tRNA-Val, is a promising potential biomarker for canine mammary gland tumors but has not previously been investigated in hepatic tumors. Accordingly, we aimed to elucidate the potential utility of tRNA-Val as a biomarker for canine HCA and HCC using clinical samples (tumor tissue and plasma extracellular vesicles [EVs]) and tumor cell lines with qRT-PCR assays. We also investigated relevant functions and signaling pathways with bioinformatic analyses (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes). tRNA-Val was markedly downregulated in HCC tumor tissue versus HCA tumor tissue and normal liver tissue, and a similar trend was shown in plasma EVs and HCC cell lines versus healthy controls. Based on areas under the receiver operating characteristic curves (AUCs), tRNA-Val significantly distinguished HCC (AUC = 1.00, p = 0.001) from healthy controls in plasma EVs and HCC from HCA (AUC = 0.950, p = 0.01). Bioinformatics analysis revealed that tRNA-Val may be primarily involved in DNA repair, mRNA processing, and splicing and may be linked to the N-glycan and ubiquitin-mediated proteasome pathways. This is the first report on the expression of tRNA-Val in canine HCC and HCA and its possible functions and signaling pathways. We suggest that tRNA-Val could be a promising novel biomarker to distinguish canine HCC from HCA. This study provides evidence for a greater understanding of the role played by tRNA-Val in the development of canine HCC.

Evaluation of cortical bone strength using a quantitative ultrasound measurement device in dogs.

This study was performed to evaluate cortical bone strength in dogs using a quantitative ultrasound measurement device. In this study, 16 clinically healthy dogs with no lameness underwent measurement of the ultrasound propagation velocity of cortical bone (namely, speed of sound [SOS]) at the radius and tibia. Additionally, computed tomography examination with a calibration phantom was performed in 10 dogs. We calculated the bone mineral density (BMD) and Young's modulus from the computed tomography data using bone strength evaluation software. SOS, BMD, and Young's modulus were statistically compared between the radius and tibia. In addition, we examined the correlation between SOS and BMD and between SOS and Young's modulus. We also examined the correlation between SOS and age in the 13 dogs whose age was known. BMD and Young's modulus were not significantly different between the radius and tibia, but SOS was significantly different (P<0.05). Moreover, SOS and BMD showed a positive correlation in both radius and tibia. Similarly, SOS and Young's modulus showed a positive correlation. In addition, SOS and age showed a strong positive correlation (radius: r=0.77, P<0.05, tibia: r=0.83, P<0.05). Our finding that SOS of the radius and tibia cortical bone was correlated with BMD and Young's modulus indicates that quantitative ultrasound can be useful for evaluating cortical bone strength in dogs.

Common bile duct perforation due to choledocholithiasis in a cat with gallbladder agenesis.

Case summary: An 8-year-old neutered male domestic shorthair cat was presented for further investigation of anorexia, vomiting and lethargy. Abdominal ultrasonography and contrast-enhanced CT revealed choledocholithiasis with suspected bacterial peritonitis and non-visualisation of the gallbladder. During surgery, the common bile duct was noted to be perforated, and a cholelith was found in the abdominal cavity. No gallbladder was confirmed during surgery. Three months postoperatively, the cat underwent CT cholangiography and absence of the gallbladder with a vestigial duplicated gallbladder was diagnosed. Relevance and novel information: Gallbladder agenesis is extremely rare in cats, with only one previous report, but several dogs have been diagnosed based on CT cholangiography and laparoscopy. This report describes gallbladder agenesis concurrent with choledocholithiasis in an adult cat and represents the first report of CT cholangiography in a cat with gallbladder agenesis.

Osteochondral regeneration of the femoral medial condyle by using a scaffold-free 3D construct of synovial membrane-derived mesenchymal stem cells in horses.

BACKGROUND: Medical interventions for subchondral bone cysts in horses have been extensively studied. This study investigated the regeneration of articular cartilage and subchondral bone with scaffold-free three-dimensional (3D) constructs of equine synovial membrane-derived mesenchymal stem cells (SM-MSCs) isolated from three ponies and expanded until over 1.0 × 107 cells at passage 2 (P2). RESULTS: SM-MSCs were strongly positive for CD11a/CD18, CD44, and major histocompatibility complex (MHC) class I; moderately positive for CD90, CD105, and MHC class II; and negative for CD34 and CD45 on flow cytometry and differentiated into osteogenic, chondrogenic, and adipogenic lineages in the tri-lineage differentiation assay. After culturing SM-MSCs until P3, we prepared a construct (diameter, 6.3 mm; height, 5.0 mm) comprising approximately 1920 spheroids containing 3.0 × 104 cells each. This construct was confirmed to be positive for type I collagen and negative for type II collagen, Alcian blue, and Safranin-O upon histological analysis and was subsequently implanted into an osteochondral defect (diameter, 6.8 mm; depth, 5.0 mm) at the right femoral medial condyle. The contralateral (left femoral) defect served as the control. At 3 and 6 months after surgery, the radiolucent volume (RV, mm3) of the defects was calculated based on multiplanar reconstruction of computed tomography (CT) images. Magnetic resonance (MR) images were evaluated using a modified two-dimensional MR observation of cartilage repair tissue (MOCART) grading system, while macroscopic (gross) and microscopic histological characteristics were scored according to the International Cartilage Repair Society (ICRS) scale. Compared to the control sites, the implanted defects showed lower RV percentages, better total MOCART scores, higher average gross scores, and higher average histological scores. CONCLUSIONS: Implantation of a scaffold-free 3D-construct of SM-MSCs into an osteochondral defect could regenerate the original structure of the cartilage and subchondral bone over 6 months post-surgery in horses, indicating the potential of this technique in treating equine subchondral bone cysts.

Relationship between serum estradiol, cathepsin K, and N-telopeptide of type I collagen in female dogs.

Ovariohysterectomized (OHE) female dogs do not develop the osteopenia and osteoporosis associated with decreasing estrogen in post-menopausal women, possibly due to post-OHE bone mineral density retention through a mechanism that remains unclear. In this study, we aimed to elucidate this mechanism by investigating estradiol (E2) and bone markers. Samples were collected from 56 OHE and 43 intact bitches (0.33 to 17.58 years old) and analyzed for serum E2, osteoclast-secreted cysteine protease cathepsin K (CTK), and N-telopeptide of type I collagen (NTx) by ELISA. OHE and intact bitches showed no significant difference in serum E2 or NTx, and there was no correlation between serum E2 and NTx and age and time since OHE. Intact bitches showed a very low correlation between E2 and NTx, but OHE bitches showed no correlation, and serum CTK was generally undetectable in both groups. Our findings suggest the influence of gonadal hormones on bone metabolism does not work effectively in dogs; this is consistent with a shorter duration of exposure to E2 in bitches (through the 4-to-8-month anestrus phase) than women.

Osteochondral regeneration using constructs of mesenchymal stem cells made by bio three-dimensional printing in mini-pigs.

Osteoarthritis is a major joint disease that has been extensively investigated in humans and in model animals. In this study, we examined the regeneration of articular cartilage and subchondral bone using artificial scaffold-free constructs composed of adipose tissue-derived mesenchymal stem cells (AT-MSCs) created using bio three-dimensional (3D) printing with a needle-array. Printed constructs were implanted into osteochondral defects created in the right femoral trochlear groove of six mini-pigs, using femoral defects created in the left femurs as controls. Repair within the defects was evaluated at 3 and 6 months post-implantation using computed tomography (CT) and magnetic resonance (MR) imaging. The radiolucent volume (RV, mm3 ) in the defects was calculated using multi-planar reconstruction of CT images. MR images were evaluated based on a modified 2D- MOCART (magnetic resonance observation of cartilage repair tissue) grading system. Gross and microscopic pathology were scored according to the ICRS (International Cartilage Repair Society) scale at 6 months after implantation. The percentage RV at 3 months postoperation was significantly lower in the implanted defects than in the controls, whereas total scores based on the MOCART system were significantly higher in the implanted defects as compared with the controls. Although there were no statistical differences in the gross scores, the average histological scores were significantly higher in the implanted defects than in the controls. To our knowledge, this is the first report to suggest that artificial scaffold-free 3D-printed constructs of autologous AT-MSCs can be aid in the osteochondral regeneration in pigs. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1398-1408, 2019.

Ganglion cysts arising from a canine stifle joint.

A 10-year-old, neutered male Labrador retriever presented with progressive left hind lameness. Ultrasonography revealed large, subcutaneous, ovoid cysts around the stifle joint. Radiographic and computed tomographic images revealed periosteal reaction of the distal femur. Magnetic resonance (MR) imaging showed a large cyst that was hypointense in T1-weighted images, hyperintense in T2-weighted images and had a thin lining that was enhanced by intravenous gadonium injection. The cyst communicated with the joint cavity and other small cysts around the joint. Histopathology of an excisional biopsy specimen led to diagnosis of ganglion cyst. This report provides MR images of a ganglion cyst in a canine stifle.