Hee-Chun Lee

Maintenance and characterization of multipotent mesenchymal stem cells isolated from canine umbilical cord matrix by collagenase digestion

Multipotent mesenchymal stem cells have been considered as a novel clinical approach for cell therapy and regenerative medicine. In this study, mesenchymal stem cells (MSCs) were successfully isolated from canine umbilical cord matrix (cUCM; also referred to as Whartons Jelly) by collagenase digestion and further characterized for multipotent properties associated with MSCs. Our cUCM-derived MSCs (cUCM-MSCs) were plastic adherent, spindle-shaped and fibroblast-like cells, maintaining expression of pluripotency markers such as Oct3/4, Nanog, Sox-2 and SSEA-4 as well as normal chromosomal number during a long-term proliferative culture. The cells expressed MSCs-specific surface markers, including CD44, CD90, CD105, and CD184, but did not CD29, CD33, CD34, and CD45. More importantly, cUCM-MSCs could differentiate into mesodermal (adipocyte, osteocyte and chondrocyte) and ectodermal (neuronal cell) cell lineages. These results imply that collagenase digestion would be a highly effective way to isolate multipotent MSCs in abundant amounts.

The rate of Salmonella spp. infection in zoo animals at Seoul Grand Park, Korea

Salmonellosis is an important zoonotic disease that affects both people and animals. The incidence of reptile-associated salmonellosis has increased in Western countries due to the increasing popularity of reptiles as pets. In Korea, where reptiles are not popular as pets, many zoos offer programs in which people have contact with animals, including reptiles. So, we determined the rate of Salmonella spp. infection in animals by taking anal swabs from 294 animals at Seoul Grand Park. Salmonella spp. were isolated from 14 of 46 reptiles (30.4%), 1 of 15 birds (6.7%) and 2 of 233 mammals (0.9%). These findings indicate that vigilance is required for determining the presence of zoonotic pathogen infections in zoo animals and contamination of animal facilities to prevent human infection with zoonotic diseases from zoo facilities and animal exhibitions. In addition, prevention of human infection requires proper education about personal hygiene.

Assessment of glomerular filtration rate with dynamic computed tomography in normal Beagle dogs.

The objective of our study was to determine individual and global glomerular filtration rates (GFRs) using dynamic renal computed tomography (CT) in Beagle dogs. Twenty-four healthy Beagle dogs were included in the experiment. Anesthesia was induced in all dogs by using propofol and isoflurane prior to CT examination. A single slice of the kidney was sequentially scanned after a bolus intravenous injection of contrast material (iohexol, 1 mL/kg, 300 mgI/mL). Time attenuation curves were created and contrast clearance per unit volume was calculated using a Patlak plot analysis. The CT-GFR was then determined based on the conversion of contrast clearance per unit volume to contrast clearance per body weight. At the renal hilum, CT-GFR values per unit renal volume (mL/min/mL) of the right and left kidneys were 0.69 ± 0.04 and 0.57 ± 0.05, respectively. No significant differences were found between the weight-adjusted CT-GFRs in either kidney at the same renal hilum (p = 0.747). The average global GFR was 4.21 ± 0.25 mL/min/kg and the whole kidney GFR was 33.43 ± 9.20 mL/min. CT-GFR techniques could be a practical way to separately measure GFR in each kidney for clinical and research purposes.

Evaluation of partial cranial cruciate ligament rupture with positive contrast computed tomographic arthrography in dogs

Computed tomographic arthrography (CTA) of four cadaveric canine stifles was performed before and after partial cranial cruciate ligament rupture in order to verify the usefulness of CTA examination for the diagnosis of partial cranial cruciate ligament rupture. To obtain the sequential true transverse image of a cranial cruciate ligament, the computed tomography gantry was angled such that the scanning plane was parallel to the fibula. True transverse images of cranial cruciate ligaments were identified on every sequential image, beginning just proximal to the origin of the cranial cruciate ligament distal to the tibial attachment, after the administration of iodinated contrast medium. A significant decrease in the area of the cranial cruciate ligament was identified on CTA imaging after partial surgical rupture of the cranial cruciate ligament. This finding implies that CTA can be used for assessing partial cranial cruciate ligament ruptures in dogs.

Evaluation of glomerular filtration rate by use of dynamic computed tomography and Patlak analysis in clinically normal cats

OBJECTIVE To obtain quantitative variables of the abdominal aorta and both kidneys on the basis of time-attenuation curves (TACs) and to measure glomerular filtration rate (GFR) for each kidney and the global GFR in clinically normal cats by use of dynamic computed tomography (CT) and Patlak analysis. ANIMALS 9 healthy cats. PROCEDURES All the cats were anesthetized with propofol. Anesthesia was maintained by administration of isoflurane, and CT examination was performed in the anesthetized cats. The TACs and renal volume were measured by use of the baseline precontrast and single-slice dynamic scans. The CT-GFR of each kidney and the global CT-GFRs were calculated via Patlak plot analysis. RESULTS CT-GFR results from 7 cats were valid. Peak aortic enhancement was detected between 9.0 and 14.0 seconds after iohexol injection, and the initial peak time of renal parenchymal enhancement was 15 to 24 seconds after iohexol injection. Mean ± SD global GFR was 2.06 ± 0.62 mL/min/kg. Mean ± SD CT-GFR of the right and left kidneys was 0.97 ± 0.32 mL/min/kg and 1.05 ± 0.31 mL/min/kg, respectively. CONCLUSIONS AND CLINICAL RELEVANCE The CT-GFR method can be rapidly and conveniently performed in clinically normal cats. This combined structural-functional approach provided physiologic and morphological information on the kidneys of cats.

Evaluation of the effects of thiopental, propofol, and etomidate on glomerular filtration rate measured by the use of dynamic computed tomography in dogs

OBJECTIVE To evaluate the effects of thiopental, propofol, and etomidate on glomerular filtration rate (GFR) measured by the use of dynamic computed tomography in dogs. ANIMALS 17 healthy Beagles. PROCEDURES Dogs were randomly assigned to receive 2 mg of etomidate/kg (n = 5), 6 mg of propofol/kg (7), or 15 mg of thiopental/kg (5) during induction of anesthesia; anesthesia was subsequently maintained by isoflurane evaporated in 100% oxygen. A 1 mL/kg dosage of a 300 mg/mL solution of iohexol was administered at a rate of 3 mL/s during GFR measurement. Regions of interest of the right kidney were manually drawn to exclude vessels and fatty tissues and highlight the abdominal portion of the aorta. Iohexol clearance per unit volume of the kidney was calculated by use of Patlak plot analysis. RESULTS Mean ± SD weight-adjusted GFR of the right kidney after induction of anesthesia with thiopental, propofol, and etomidate was 2.04 ± 0.36 mL/min/kg, 2.06 ± 0.29 mL/min/kg, and 2.14 ± 0.43 mL/min/kg, respectively. However, no significant differences in weight-adjusted GFR were detected among the treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE Results obtained for the measurement of GFR in anesthetized dogs after anesthetic induction with etomidate, propofol, or thiopental and maintenance with isoflurane did not differ significantly. Therefore, etomidate, propofol, or thiopental can be used in anesthesia-induction protocols that involve the use of isoflurane for maintenance of anesthesia without adversely affecting GFR measurements obtained by the use of dynamic computed tomography in dogs.

Immunologic properties of differentiated and undifferentiated mesenchymal stem cells derived from umbilical cord blood

The expression of immunogenic markers after differentiation of umbilical cord blood (UCB)-derived mesenchymal stem cells (MSC) has been poorly investigated and requires extensive in vitro and in vivo testing for clinical application. The expression of human leukocyte antigen (HLA) classes on UCB-derived MSC was tested by Fluorescence-activated cell sorting analysis and immunocytochemical staining. The undifferentiated MSC were moderately positive for HLA-ABC, but almost completely negative for HLA-DR. The MSC differentiated to chondrocytes expressed neither HLA-ABC nor HLA-DR. The proliferation of MSC was not significantly affected by the allogeneic lymphocytes stimulated with concanavalin A. The responder lymphocytes showed no significant decrease in proliferation in the presence of the MSC, but the apoptosis rate of the lymphocytes was increased in the presence of MSC. Taken together, these findings indicate that UCB-derived MSC differentiated to chondrocytes expressed less HLA class I and no class II antigens. The MSC showed an immunomodulatory effect on the proliferation and apoptosis of allogeneic lymphocytes. These data suggest that the differentiated and undifferentiated allogeneic MSC derived from umbilical cord blood can be a useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

Canine model of ischemic stroke with permanent middle cerebral artery occlusion: clinical features, magnetic resonance imaging, histopathology, and immunohistochemistry.

The purpose of this study was to identify time-related changes in clinical, MRI, histopathologic, and immunohistochemical findings associated with ischemic stroke in dogs. Additionally, the association of cerebrospinal fluid (CSF) and tissue levels of interleukin (IL)-6 with clinical prognosis was assessed. Ischemic stroke was induced by permanent middle cerebral artery occlusion (MCAO) in nine healthy experimental dogs. The dogs were divided into three groups according to survival time and duration of the experimental period: group A (survived only 1 day), group B (1-week experimental period), and group C (2-week experimental period). Neurologic status was evaluated daily. Magnetic resonance imaging (MRI) was performed according to a predetermined schedule. Concentration of IL-6 in CSF was measured serially after ischemic stroke. Postmortem examination was performed for all experimental dogs. During histopathological examination, variable degrees of cavitation and necrosis due to neuronal cytopathic effects, such as pyknotic nuclei and cytoplasmic shrinkage, were observed on the affected side of the cerebral cortex in all dogs. Immunohistochemistry specific for IL-6 showed increased expression in the ischemic lesions. CSF IL-6 concentrations and ischemic lesion volumes 1 day after ischemic stroke were significantly higher in group A compared to groups B and C.

Imatinib Mesylate plus Hydroxyurea Chemotherapy for Cerebellar Meningioma in a Belgian Malinois Dog

ABSTRACT An 8-year-old intact male Belgian Malinois, weighing 37.2 kg, was referred for evaluation due to right side facial paresis, ataxia and a 2-month history of decreased cognitive ability. Physical and neurological examinations revealed mild depression, left-sided head tilt, right-sided facial paresis and ataxia. A well-demarcated, broad-based cerebellar mass and hyperostosis were found on CT imaging of the brain. Based on these CT findings, a cerebellar meningioma was strongly suspected. Hydroxyurea and prednisolone were administered; after 4 weeks, there was reduction in mass size as compared to initial CT results. However, the mass size was found to have grown 6 weeks after hydroxyurea treatment. We then prescribed a combination of imatinib mesylate and hydroxyurea. Two weeks following combination treatment, the mass size had reduced significantly. The mass continuously decreased in size until the patient died during anesthesia. Cerebellar transitional meningioma was confirmed by histopathologic examination. To the author’s knowledge, this is the first reported case of imatinib mesylate plus hydroxyurea therapy for the treatment of meningioma in veterinary medicine.

A Case of Gastric Adenocarcinoma in a Shih-tzu Dog; Successful Treatment of Early Gastric Cancer

ABSTRACT A 9-year-old castrated male Shih Tzu dog was referred to us, because of chronic vomiting. The patient’s hematological, radiographic, ultrasonographic, endoscopic and histological examinations were evaluated for diagnosis. Hematologic analysis indicated moderate anemia and azotemia. Based on the imaging studies, an oval-shaped mass was identified in the gastric pylorus area. A proliferative mass was found on endoscopic examination, and we performed biopsy using grasping forceps. The histopathological findings of the biopsy specimens indicated hypertrophic gastritis, and Y-U pyloroplasty was performed. However, histopathological examination of the surgically resected mass revealed tubular adenocarcinoma of the stomach. Then, carboplatin chemotherapy was performed 4 times for 13 weeks. Clinical signs, such as vomiting, were resolved gradually after surgery and chemotherapy, and the patient’s condition was managed favorably until recently (30 months after surgery). This case report describes clinical features, imaging studies, endoscopic characteristics and histopathological and immunohistochemical features of gastric tubular adenocarcinoma as early gastric cancer in a dog.