Yasuharu Izumisawa

Systemic atherosclerosis in dogs: histopathological and immunohistochemical studies of atherosclerotic lesions

Histopathological and immunohistochemical studies were carried out on five cases of canine systemic atherosclerosis. The five animals were male, and showed hypercholesterolaemia and hypertriglyceridaemia on biochemical analysis of plasma. Histopathologically, atherosclerotic lesions were seen in the aorta and muscular arteries in many organs, including the heart, spleen, kidneys, lungs, pancreas, alimentary tract, urogenital organs, eyes, prostate and urinary bladder. The lesions were characterized by the deposition of lipids and infiltration of lipid-laden foamy cells in the tunica intima and tunica media, sometimes forming fibrofatty plaques, containing abundant sudanophilic material, cholesterol clefts and mineralized material. The lesions started in the tunica intima and extended to the tunica media and tunica adventitia. Immunohistochemical examination with canine apolipoprotein B-100 (CApoB-100) antibody identified the lipids containing low density lipoprotein. Immunoreactivity to CApoB-100 antibody was recognized in the tunica intima, lipid-laden foamy cell cytoplasm and smooth muscle cells in the tunica media, and fibrofatty plaque. These histopathological and immunohistochemical features were similar to those of human atherosclerotic lesions.

Repair of articular cartilage defects with a novel injectable in situ forming material in a canine model.

We developed an ultra-purified in situ forming gel as an injectable delivery vehicle of bone marrow stromal cells (BMSCs). Our objective was to assess reparative tissues treated with autologous BMSCs implanted using the injectable implantation system into osteochondral defects in a canine model. Forty-eight osteochondral defects in the patella groove of the knee joint were created in 12 adult beagle dogs (two defects in each knee). The defects were divided into a defect group (n = 16), an acellular novel material implantation (material) group (n = 16), and a BMSCs implantation using the current vehicle system (material with BMSCs) group (n = 16). The reparative tissues at 16 weeks postoperatively were assessed through gross, histological, and mechanical analyses. The reparative tissues of the material with BMSCs group were substituted with firm and smooth hyaline-like cartilage tissue that was perfectly integrated into the host tissues. This treatment group obviously enhanced the subchondral bone reconstruction. The compressive modulus of the reparative tissues was significantly higher in the material with BMSCs group than the other groups. This study demonstrated that the implantation of BMSCs using our novel in situ forming material induced a mature hyaline-like cartilage repair of osteochondral defects in a canine model.

Two Cases of Lacaziosis in Bottlenose Dolphins (Tursiops truncatus) in Japan

Lacaziosis, formerly called lobomycosis, caused by Lacazia loboi, is a zoonotic mycosis found in humans and dolphins and is endemic in the countries on the Atlantic Ocean. Although the Japanese coast is not considered an endemic area, photographic records of lacaziosis-like skin lesions were found in bottlenose dolphins (Tursiops truncatus) that were migrating in the Goto Islands (Nagasaki Prefecture, Japan). We diagnosed 2 cases of lacaziosis in bottlenose dolphins captured simultaneously at the same coast within Japanese territory on the basis of clinical characteristics, cytology, histopathology, immunological tests, and detection of partial sequences of a 43 kDa glycoprotein coding gene (gp43) with a nested-PCR system. The granulomatous skin lesions from the present cases were similar to those found in animals from endemic areas, containing multiple budding and chains of round yeast cells and positive in the immune-staining with anti-Paracoccidioides brasiliensis serum which is a fungal species related to L. loboi; however, the gp43 gene sequences derived from the present cases showed 94.1% homology to P. brasiliensis and 84.1% to L. loboi. We confirmed that the causative agent at the present cases was different genotype of L. loboi from Amazon area.

Microstructure and glycosaminoglycan ratio of canine cornea after reconstructive transplantation with glycerin-preserved porcine amniotic membranes.

OBJECTIVE Although amniotic membranes of canine, feline, and equine species have some advantages as corneal transplantation material in many canine ocular diseases, their softness, thinness, and low availability can pose problems. As an alternative, the more abundant porcine amniotic membranes may be used. This paper describes the use of glycerin-preserved porcine amniotic membranes in corneal transplantation in eight normal dogs. METHOD A 0.4-mm deep recipient bed in the axial cornea of the OS of all dogs was created using an 8-mm Barron radial vacuum trephine. The recipient bed was then filled with amnion, and the entire cornea was covered with another piece of the glycerin-preserved membrane. The ocular signs evaluated were corneal opacity and corneal vascularization. The dogs were euthanized on days 5, 10, 20, or 40 after surgery, and samples were collected to evaluate corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content and the glycosaminoglycan (GAG) ratio. RESULTS Corneal opacity was observed immediately after surgery. Restoration of corneal transparency, regression of corneal vascularization, and visualization of the pupil and iris were noted on day 40. CONCLUSIONS The clinical observations were supported histologically by regained corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content, and GAG ratio, suggesting that this technique may be a novel method for the treatment of ocular surface disorders.

Investigation of fellow eye of unilateral retinal detachment in Shih-Tzu.

OBJECTIVE To investigate disease in the fellow eye, and consider the relation to rhegmatogenous retinal detachment (RRD) in Shih-Tzus. ANIMALS STUDIED The fellow eyes of 49 Shih-Tzus (27 male, 22 female; median age: 6.8 years) with unilateral RRD diagnosed by funduscopy or ultrasonography at Rakuno Gakuen University Teaching Animal Hospital were assessed in this study. PROCEDURES Ophthalmic examinations (including menace response, pupillary light reflex, slit-lamp biomicroscopy, and funduscopy) were performed in the subjects. Electroretinography was performed in 12 eyes that developed retinal degeneration. Maximum follow-up period was 42 months. RESULTS Cataracts and vitreous opacity were observed in 26 (53%) and 32 eyes (65%), respectively, by slit-lamp biomicroscopy. Retinal degeneration with various degrees of hyper-reflectivity of the tapetal fundus and/or attenuation of retinal vessels was observed in 35 eyes (71%) on funduscopy. A reduction of amplitude in rod, standard combined and 30 Hz flicker electroretinogram was detected in 5 (42%), 10 (83%), and 6 eyes (50%), respectively. During the follow-up period, RRD was detected in six eyes. CONCLUSION Retinal degeneration was frequently detected by funduscopy and electroretinograms in the fellow eye in Shih-Tzus with RRD. In our subjects, vitreous degeneration was also observed frequently. It has been reported that peripheral retinal degeneration is one of the causes of RRD associated with vitreous degeneration in humans. We assume that primary retinal degeneration with secondary vitreous degeneration is one of the causes of RRD in Shih-Tzus.

Therapeutic effects of intra‐articular ultra‐purified low endotoxin alginate administration on an experimental canine osteoarthritis model

OBJECTIVE This study aimed to elucidate the therapeutic effects of intra-articular administration of ultra-purified low endotoxin alginate (UPLE-alginate) on osteoarthritis (OA) using a canine anterior cruciate ligament transection (ACLT) model. DESIGN We used 20 beagle dogs. ACLT was performed on the left knee of each dog and a sham operation was performed on the right knee as a control. All animals were randomly divided into the control (saline) and therapeutic (UPLE-alginate) groups. Animals in the control and therapeutic groups received weekly injections with 0.7 mL normal saline or 0.7 mL 0.5% UPLE-alginate, respectively, from 0 to 3 weeks after ACLT or sham operation. At 9 weeks after ACLT, the knee joints of all animals were observed using arthroscopy. All animals were euthanized at 14 weeks after ACLT and evaluated using morphologic assessment, histologic assessment, and biomechanical testing. RESULTS Arthroscopic findings showed intact cartilage surface in both groups. Morphologic findings in the therapeutic group showed milder degeneration compared with those of the control group, but there were no significant differences between groups. Histologic scores of the medial femoral condyle (MFC) and lateral femoral condyle (LFC) were better in the therapeutic group than the control group (MFC: p = 0.009, LFC: p = 0.009). Joint lubrication did not differ significantly between groups. CONCLUSION Intra-articular administration of UPLE-alginate in the early stage of OA slowed disease progression in canines. UPLE-alginate may have potential as a therapeutic agent for OA patients and reduce the number of patients who need to undergo total joint arthroplasty.

Hypertrophic Z-Line Observed in Aged One-Humped Camel (Camelus dromedarius)

The ultrastructure of cardiac muscle from 3 aged one-humped camels (Camelus dromedarius) was examined to see if aged camels also develop hypertrophic Z-lines as observed in the heart of aged humans and other animals. The results proved that the aged camel heart develops hypertrophic Z-lines together with other age-associated alterations, including loss of myofibrils and myofilaments, increase of intermyofibrillar spaces and intercellular spaces, and disruption of mitochondrial cristae and intercalated disc structure. Hypertrophic Z-lines are somewhat smaller than those of other species, but occur both in the atria and ventricles. Although the majority of cardiac muscle samples revealed a normal structure, the finding of hypertrophic Z-lines in camel heart is the first such observation in a ruminant animal. The anomalous camel muscle resembles that of other species in terms of cell alterations commonly associated with aged hearts.

Electroretinography recordings using a light emitting diode active corneal electrode in healthy beagle dogs

Electroretinography (ERG) is a well-established diagnostic procedure for objectively evaluating retinal function. In this study, ERG in beagle dogs, which are a popular experimental animal, was performed to determine the normal range of ERG variables and assess differences between the left and right eyes. ERG findings including rod, combined rod-cone, single-flash cone, and 30-Hz flicker responses were recorded with an LED-electrode in 43 sedated beagle dogs. The subjects were divided into young (< 1 year old), adult (1~5 years old), and senile animals (≥ 6 years old). Normal ERG ranges were obtained. Significant differences in b-wave amplitude along with b/a ratio of the combined rod-cone response were found between the young and adult animals as well as young and senile dogs. No significant differences were observed between the left and right eyes. ERG variables in beagle dogs differed by age due to age-related retinal changes. Thus, we propose that normal ERG ranges should be determined according to age in each clinic and laboratory using its own equipment because each institution usually has different systems or protocols for ERG testing.

ANESTHETIC MANAGEMENT OF AN INDO-PACIFIC BOTTLENOSE DOLPHIN (TURSIOPS ADUNCUS) REQUIRING SURGICAL DEBRIDEMENT OF A TAIL ABSCESS

Abstract This report describes the anesthetic management of a 14-yr-old, 160-kg, female Indo-Pacific bottlenose dolphin (Tursiops aduncus) that underwent surgical debridement for a refractory subcutaneous abscess twice within a 6-mo interval. The animal was otherwise in good physical condition at each anesthetic procedure. Following premedication with intramuscular midazolam and butorphanol, anesthesia was induced with propofol and maintained with sevoflurane by intubation. During surgery ventilation was controlled. Blood pressure was indirectly estimated using either oscillometric or pulse oximetry. Presumed hypotension was managed by adjusting the sevoflurane concentration and infusion of dopamine. During recovery, the dolphin regained adequate spontaneous respiration following intravenous administration of flumazenil and doxapram. The dolphin was extubated at 85 min and 53 min after the first and second surgeries, respectively. Successful weaning from the ventilator and initiation of spontaneous respiration was the most important complication encountered. Establishment of a reliable blood pressure measurement technique is critical to success for anesthesia in this species.

Bonding strength evaluation of Cementless Implant by grit blasting

As a method of porous coating for cementless implant, sintering and diffusion bonding are usually used. But the bonding techniques have trouble with decreasing fatigue strength and boundary separation. In this study, we applied grit blasting for porous coating. Polygonal iron particle was sprayed on the titanium base material by compressed-air power.