Cross-sectional anatomy, computed tomography, and magnetic resonance imaging of the banded houndshark (Triakis scyllium).

Due to their important phylogenetic position among extant vertebrates, sharks are an invaluable group in evolutionary developmental biology studies. A thorough understanding of shark anatomy is essential to facilitate these studies and documentation of this iconic taxon. With the increasing availability of cross-sectional imaging techniques, the complicated anatomy of both cartilaginous and soft tissues can be analyzed non-invasively, quickly, and accurately. The aim of this study is to provide a detailed anatomical description of the normal banded houndshark (Triakis scyllium) using computed tomography (CT) and magnetic resonance imaging (MRI) along with cryosection images. Three banded houndsharks were scanned using a 64-detector row spiral CT scanner and a 3 T MRI scanner. All images were digitally stored and assessed using open-source Digital Imaging and Communications in Medicine viewer software in the transverse, sagittal, and dorsal dimensions. The banded houndshark cadavers were then cryosectioned at approximately 1-cm intervals. Corresponding transverse cryosection images were chosen to identify the best anatomical correlations for transverse CT and MRI images. The resulting images provided excellent detail of the major anatomical structures of the banded houndshark. The illustrations in the present study could be considered as a useful reference for interpretation of normal and pathological imaging studies of sharks.

Successful treatment of a dog with phenobarbital-responsive sialadenosis and an oesophageal stricture.

BACKGROUND: Phenobarbital-responsive sialadenosis (PRS) can cause nausea and vomiting, and is rarely reported in dogs. OBJECTIVES: An 8-year-old neutered, male Pomeranian dog was presented to our teaching hospital with vomiting that began 2 years ago. The clinical signs repeatedly improved and deteriorated despite treatment. METHODS: The only abnormality found on physical examination was salivary gland enlargement, and no specific findings were observed on blood analysis and imaging tests. The results of the fine needle aspirate cytology from the salivary glands revealed possible sialadenosis. Phenobarbital was prescribed, and the patient's symptoms resolved. However, upon discontinuing drug, the patient's clinical signs recurred and did not improve even after re-introduction of phenobarbital and the addition of other anticonvulsant drugs. An oesophageal stricture was observed on an oesophagram, and fibrosis was confirmed endoscopically. A balloon dilation was performed to expand the stenosis. RESULTS: After the first procedure, the patient's clinical signs initially improved, but relapsed 2 weeks later. A total of three oesophageal dilation procedures were performed using a sequentially larger diameter balloon. After the third procedure, the patient's clinical signs were managed without recurrence. The cause of recurrent gastrointestinal signs following the initial successful treatment of phenobarbital-responsive sialadenosis was due to oesophageal stricture formation. CONCLUSIONS: This case report demonstrates the successful management of PRS with subsequent oesophageal stricture formation in a dog.

Primo Bundles Identified by Microcomputed Tomography in Primo Vascular Tissue on the Surface of Rat Abdominal Organs.

BACKGROUND: The primo vascular system (PVS) is a novel network composed of primo nodes (PNs) and primo vessels (PVs). Currently, its anatomy is not fully understood. OBJECTIVES: The aim of this study was to elucidate the three-dimensional PN-PV structure. METHODS: Organ-surface PVS tissue was isolated from healthy and anemic rats. The tissues were analyzed by X-ray microcomputed tomography (CT), hematoxylin and eosin staining, and scanning electron microscopy. RESULTS: From CT images, we identified one or more bundles in a PV. In the PN, the bundles were enlarged and existed in isolation and/or in anastomosis. The transverse CT images revealed four areas of distinct intensities: zero, low, intermediate, and high. The first two were considered to be the sinuses and the subvessels of the PVS and were identified in the hematoxylin and eosin-stained PN sections. The enlargement of the PN from anemic rats was associated with an increase in the intermediate-intensity area. The high-intensity area demarcated the bundle and was overlapped with the mesothelial cells. In scanning electron microscopy, the PV bundles branched out, tapering down to a single bundle at some distance from the PN. Each bundle was composed of several subvessels (∼5 µm). Clustered round microcells (1-25 µm), scattered flat oval cells (∼15 µm), and amorphous extracellular matrix were observed on the surface of the PVS tissue. CONCLUSIONS: The results newly showed that the primo bundle is a structural unit of both PVs and PNs. A bundle was demarcated by high CT intensity and mesothelial cells and consisted of multiple subvessels. The PN bundles contained also sinuses.

Transplantation of hMSCs Genome Edited with LEF1 Improves Cardio-Protective Effects in Myocardial Infarction.

Stem cell-based therapy is one of the most attractive approaches to ischemic heart diseases, such as myocardial infarction (MI). We evaluated the cardio-protective effects of the human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) stably expressing lymphoid enhancer-binding factor 1 (LEF1; LEF1/hUCB-MSCs) in a rat model of MI. LEF1 overexpression in hUCB-MSCs promoted cell-proliferation and anti-apoptotic effects in hypoxic conditions. For the application of its therapeutic effects in vivo, the LEF1 gene was introduced into an adeno-associated virus integration site 1 (AAVS1) locus, known as a safe harbor site on chromosome 19 by CRISPR/Cas9-mediated gene integration in hUCB-MSCs. Transplantation of LEF1/hUCB-MSCs onto the infarction region in the rat model significantly improved overall survival. The cardio-protective effect of LEF1/hUCB-MSCs was proven by echocardiogram parameters, including greatly improved left-ventricle ejection fraction (EF) and fractional shortening (FS). Moreover, histology and immunohistochemistry successfully presented reduced MI region and fibrosis by LEF1/hUCB-MSCs. We found that these overall positive effects of LEF1/hUCB-MSCs are attributed by increased proliferation and survival of stem cells in oxidative stress conditions and by the secretion of various growth factors by LEF1. In conclusion, this study suggests that the stem cell-based therapy, conjugated with genome editing of transcription factor LEF1, which promotes cell survival, could be an effective therapeutic strategy for cardiovascular disease.

Targeted Genome Engineering to Control VEGF Expression in Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells: Potential Implications for the Treatment of Myocardial Infarction.

Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) exhibit potency for the regeneration of infarcted hearts. Vascular endothelial growth factor (VEGF) is capable of inducing angiogenesis and can boost stem cell-based therapeutic effects. However, high levels of VEGF can cause abnormal blood vessel growth and hemangiomas. Thus, a controllable system to induce therapeutic levels of VEGF is required for cell therapy. We generated an inducible VEGF-secreting stem cell (VEGF/hUCB-MSC) that controls the expression of VEGF and tested the therapeutic efficacy in rat myocardial infarction (MI) model to apply functional stem cells to MI. To introduce the inducible VEGF gene cassette into a safe harbor site of the hUCB-MSC chromosome, the transcription activator-like effector nucleases system was used. After confirming the integration of the cassette into the locus, VEGF secretion in physiological concentration from VEGF/hUCB-MSCs after doxycycline (Dox) induction was proved in conditioned media. VEGF secretion was detected in mice implanted with VEGF/hUCB-MSCs grown via a cell sheet system. Vessel formation was induced in mice transplanted with Matrigel containing VEGF/hUCB-MSCs treated with Dox. Moreover, seeding of the VEGF/hUCB-MSCs onto the cardiac patch significantly improved the left ventricle ejection fraction and fractional shortening in a rat MI model upon VEGF induction. Induced VEGF/hUCB-MSC patches significantly decreased the MI size and fibrosis and increased muscle thickness, suggesting improved survival of cardiomyocytes and protection from MI damage. These results suggest that our inducible VEGF-secreting stem cell system is an effective therapeutic approach for the treatment of MI. Stem Cells Translational Medicine 2017;6:1040-1051.

Reduced birth weight, cleft palate and preputial abnormalities in a cloned dog.

The aim of the present study was to report a novel developmental abnormality in a cloned dog. A fibroblast cell line was established from an 8-year-old male German shepherd dog. In vivo matured oocytes were retrieved from a large breed dog, and the nucleus was removed from each oocyte. A donor cell was injected into an enucleated oocyte, and the oocyte-cell couplet was fused electrically. After chemical activation, the resulting embryos were transferred into a naturally estrus-synchronized recipient dog, and two cloned pups were delivered by Cesarean section 60 days later. One cloned pup (Clone 1) was healthy, but the other (Clone 2) had a birth weight of only 320 g and cleft palate, failure of preputial closure at the ventral distal part, and persistent penile frenulum. Clone 2 was raised by stomach feeding until Day 40 after birth, where palatoplasty was performed. The abnormalities in external genitalia in Clone 2 resulted in persistent penile extrusion that was surgically corrected. This complex developmental abnormality has not been reported in dogs previously.

Time course of diurnal rhythm disturbances in autonomic function of rats with myocardial infarction.

The diurnal rhythm of the autonomic function is known to be blunted in heart failure, but the timing of this blunting is not well understood. We examined the time course of the alterations in autonomic function in rats with myocardial infarction (MI) by analyzing heart rate variability (HRV). MI was induced by coronary artery ligation, and HRV was analyzed at 2, 4, 6, and 8 weeks post-MI. Diurnal rhythm in heart rate (HR) was maintained over the study period. However, diurnal rhythm in the standard deviation of averages of normal R-R intervals (SDANN) and the ratio between low and high frequency band powers (LF/HF ratio) were disrupted in MI rats at 2 weeks, which persisted up to 8 weeks, with the exception of 4 weeks. The dark-light differences in the LF/HF ratio changed from negative to positive values between weeks 2 and 4 in the MI rats. We also found decreases in HR, SDANN, and the LF/HF ratio in the dark phase at weeks 6 and 8 and an increase in plasma norepinephrine (NE) level at week 8. Collectively, the results indicate that the timing of the disturbance of diurnal rhythm in SDANN and the LF/HF ratio is different from those in HR and in plasma NE level, suggesting that the mechanisms underlying these changes are different. In addition, there is a transition from the compensatory to the decompensatory phase between 4 and 6 weeks post-MI. These findings may help to understand the progression and pathophysiology of heart failure.

Computed tomographic characteristics of acute thoracolumbar intervertebral disc disease in dogs.

Forty canine patients with a presumptive diagnosis of the intervertebral disc herniation at the thoracolumbar region were imaged. A neurological examination was performed and all patients were classified under four grades by the examination. The degrees of attenuation of the herniated disc material were measured in Housefield units (HU) in each image. The ratio of the area to herniated disc material and the height to disc material were measured. The clinical grade was correlated with the area ratio of the herniated disc material to the spinal cord, but not correlated with the height ratio of that. In the patients with epidural hemorrhage at surgery, HUs of the herniated disc material was lower than those with no epidural hemorrhage at surgery. Noncontrast computed tomography scans of the spine can be useful in diagnosing acute intervertebral disc disease in chondrodystrophoid breeds, evaluating patient status and identifying concurrent epidural hemorrhage.

Enterocutaneous fistula as a result of chronic bite wound repair in a dog.

A 6-year-old castrated male Maltese weighing 4.8 kg was presented with a non-healing wound exhibiting purulent discharge after surgery on scar tissue of a chronic twelve-month-old bite wound on the left caudal abdominal region. The dog had previously undergone four surgeries and had been on continuous antibiotic therapy for eight months. Following radiographic and ultrasonographic examinations, the problem was diagnosed as an enterocutaneous fistula of a herniated bowel loop under the skin. Surgical resection of the fistula involving the bowel loop resolved all symptoms.

Segmental aplasia of the uterine horn with ipsilateral renal agenesis in a cat.

A nine-month-old domestic short haired cat was admitted with the history of acute vomiting, depression and shivering. Abdominal ultrasonography revealed minimum enlargement of the right uterine horn filled with anechoic fluid. On excretory urography, functionally and anatomically normal, enlarged left kidney was found, but right kidney was absent. It was preliminary diagnosed as hydrometra with right renal agenesis. Aiming at the correction of hydrometra, we performed ovariohysterectomy. During spaying, we found a missing segment of distal part of the right uterine horn and absence of ipsilateral kidney and ureter. Compressed uterine structure and segmental aplasia of right uterine horn were found in histopathological investigation. Taken together, it was diagnosed as a segmental aplasia of uterine horn with ipsilateral renal agenesis.

Malignant ocular melanoma in a dog.

A mongrel male dog of three years old was referred to the Seoul National University Veterinary Teaching Hospital following a one month history of glaucoma. On ophthalmic examination, hyphema, glaucoma, uveitis, iridal mass, and loss of vision were noted in the right eye. Ultrasonography and computed tomography revealed a mass with involvement of the entire uvea. Radiographic evaluation did not reveal any evidence of distant metastasis. The right eye was surgically removed because of the high likelihood of neoplasia. A histologic diagnosis of malignant uveal melanoma was made.

Ultrasonographic measurement of optic nerve sheath diameter in normal dogs.

This study was carried out to assess the feasibility of ultrasonographic measurements of the optic nerve sheath diameter (ONSD) in normal dogs and evaluate the effect of breed, sex, body weight and age on biometry of ONSD. The ONSDs were evaluated in 15 dogs (10-50 months old) with normal eye (7 Yorkshire terrier and 8 Maltese). Ultrasonographic measurements of the ONSD were carried out at a constant position located 5 mm behind the optic disc. Eyes were collected immediately after euthanasia, and were used for saline immersion technique and direct measurement by calipers for biometry of ONSD. In this study, there was no significant difference of ONSD between the left and the right eyes, and was no significant difference among ONSD values obtained from ultrasonographical method, saline immersion technique and direct measurement (k=0.95). Also, there was no correlations between ONSD and sex, body weight and age, but was significant between the mean ONSD of Yorkshire terrier and Maltese (p<0.01). The mean ONSD of Yorkshire terrier was 2.10 +/- 0.22 mm and Maltese was 1.63 +/- 0.23 mm. This study suggests that ultrasonographic measurements is useful method for biometry of the ONSD in normal dogs and provides baseline information for the study of evaluating ONSD in various breeds and diagnosing several diseases with the change of the ONSD.

Effect of acupuncture in the treatment of young pigs with induced Escherichia coli diarrhea.

The effect of acupuncture in the treatment of young pigs with induced enteropathogenic Escherichia coli diarrhea was histopathologically evaluated by routine hematoxylin and eosin stain. Thirty two pigs weighed 4-5 kg and aged 21 days old were used in this study. The animals with diarrhea were treated with traditional acupuncture, or enrofloxacin. In the group treated with traditional acupuncture, acupoint GV1 (Jiaochao) was used and in the group treated with antibiotics, enrofloxacin was injected intramuscularly. Ten pigs were inoculated with E. coli, but were not treated and served as nontreated control group. At postinoculation day 6, all pigs of the acupuncture and antibiotic treated groups recovered from diarrhea. In the ascending and descending colons of the nontreated control group, severe infiltration of inflammatory cells in the lamina propria was observed and in the fundic stomach, destruction of the fundic gland architecture and necrotic lesions were observed, however, in the same sites of the acupuncture and antibiotics treated groups, the mucosae of the colon and stomach were relatively similar to those of the normal group. These results indicate that acupuncture treatment is effective in controlling induced E. coli diarrhea in pigs at its early stage.