Histiocytic ulcerative colitis in a French bulldog

A 9-month-old French bulldog was referred for signs of chronic large bowel diarrhea. The dog had an increased frequency of defecation, tenesmus and hematochezia. Flexible colonoscopy showed hyperemia, irregularities and ulcerations with multifocal hemorrhages in the mucosa from the descending colon to the proximal rectum. Multiple colonic biopsies were characterized by infiltrations of PAS positive histiocytes in the lamina propria. A diagnosis of histiocytic ulcerative colitis (HUC) was made, and the animal showed only minimal improvement, although it was treated with nutritional and medical therapies. This is the second case of HUC in French bulldog, a breed which has ancestral relations to Boxer dogs.     

Sodium iodoacetate-induced experimental osteoarthritis and associated pain model in rats

Degenerative lesions were induced in the knee joint of Wistar rats by intraarticular injection of chondrocyte metabolism inhibitor mono-iodoacetate (MIA) at doses of 0, 0.3 or 3 mg/joint. Histopathological examination and the measurement of hind paw weight ratio as an index of joint pain by incapacitance tester were performed. Histological findings that are similar to those observed in human osteoarthritis (OA), such as disorganization of chondrocytes, erosion and fibrillation of cartilage surface, and subchondral bone exposure etc., were observed in a MIA-dose-dependent manner. Saflanin-O fast green staining revealed that marked diffuse reduction of proteoglycan in cartilage tissue of rats treated with MIA. The clinical scores of the joint pain were closely correlated to the grade of histological findings. We conclude that the present experimental model in combination with a novel dual channel weight averager would be very useful for the study of human OA, and could be applied for estimation of therapeutic effect of new anti-OA drugs.     

Use of a rigid-type polyethylene T-cannula,silicone tubing,and adhesion netting for surgical cannulation of ileum of the horse

A rigid-type of polyethylene T-cannula was fitted into the anterior ileum of six horses in order to improve the cannulation techniques. A piece of polyethylene net was fastened onto the intestinal wall around the cannula to prevent dislodgment of the cannula by promoting a secure adhesion between the ileum and the abdominal wall. The cannula barrel sheathed with silicone tubing was exteriorized through a stab incision at the lateral ventral wall on the transverse line of the second lumber vertebra, and a flange was screwed onto the barrel. The feeding regime gradually increased concentrate without roughage prevented any colic signs. The use of these techniques succeeded in the ileal cannulation with no leakage of digesta.     

Successful production of blastocysts following ultrarapid vitrification with step-wise equilibriation of germinal vesicle-stage mouse oocytes

The purpose of this study was to evaluate the viability and subsequent developmental ability of murine germinal vesicle (GV) oocytes ultrarapidly vitrified after step-wise exposure to cryoprotectants (CPAs). Oocytes were transferred to a vitrification solution composed of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 M sucrose in a direct manner (non-preequilibrium) or in a step-wise manner (single-, two-, or ten-step preequilibrium). After ultrarapid vitrification and storage in liquid nitrogen, the oocytes were thawed, washed by diluting the CPAs in five steps, and then subjected to in vitro maturation, fertilization and culture. In the non-preequilibrium group, the rates of post-thawed oocytes surviving, maturing to metaphase-II, developing to blastocysts and to hatching/hatched blastocysts were 91.8, 87.1, 15.9 and 2.3%, respectively. In the single- and two-step groups, the corresponding rates were 97.0-98.2%, 92.2-95.0%, 22.0-29.4% and 8.8-15.6%, whereas in the ten-step group they were 98.2, 91.8, 38.6 and 22.8%, respectively. In the non-vitrified control group, the rates of oocytes maturing to metaphase-II, developing to blastocysts and to hatching/hatched blastocysts were 90.2, 75.2 and 51.5%, respectively. The present study shows that the ultrarapid vitrification of murine GV oocytes by a step-wise manner involving 10 steps preequilibrium may have an advantage in maintaining the viability and subsequent production of blastocysts.     

Distribution of the Consensus Sequences of ABC Transporter Gene among Several Taxonomically Distinct Phytopathogenic Fungi

The distribution of ABC (ATP-binding cassette) transporter genes among several taxonomically distinct phytopathogenic fungi was investigated by Southern blot hybridization, polymerase chain reaction assays and partial sequencing. Consensus sequences of the ABC transporter gene, which might be concerned with multidrug resistance against fungicides and with pathogenicity in phytopathogenic fungi, were found in all of the examined phytopathogenic fungi, which belonged to the Mastigomycotina, Ascomycotina, Basidiomycotina and Deuteromycotina. Received 24 August 2000/ Accepted in revised form 5 December 2000     

Analysis of the Genetic Components of Peridial Tissue of Perithecia in the Blast Fungus

To elucidate perithecial wall development in Magnaporthe grisea, monoconidial isolates were collected from the surfaces of six perithecia in five crosses. Genotypes of these isolates were characterized based on colony color, mating type and repetitive DNA elements. Each set of isolates from four perithecia possessed the genetic markers from only one parent in each cross. However, each set of isolates from the other two perithecia had markers from both parents. Tissues containing both parental genomic components may thus take part in construction of perithecial wall. Received 13 October 2000/ Accepted in revised form 12 March 2001     

Immunohistochemical detection of the enzyme glutamic acid decarboxylase and hormones of the islets of Langerhans in spontaneous insulin-dependent diabetes mellitus in cattle

Immunohistochemical expression of glutamic acid decarboxylase (GAD) enzyme was detected in the pancreatic islets of 12 cattle with spontaneous insulin-dependent diabetes mellitus (IDDM). The most characteristic changes were atrophy and decreased number of pancreatic islets, enlarged islets with vacuolated beta cells, and lymphocytic islet adenitis. Atrophied islets were composed of small islet cells without cytoplasmic insulin-positive granules. Immunohistochemically, GAD was not found in the cytoplasm of atrophied islet cells. Furthermore, enlarged islets consisting of islet cells with vacuolated cytoplasm were frequently observed. The cytoplasm of vacuolated cells contained very few GAD- and insulin-positive granules, indicating beta cell destruction. Enlarged islets with mild lymphocytic infiltrates were frequently observed. These findings suggest that islet cells in cattle with IDDM lose their insulin synthesis function and their ability to regulate hormonal secretion of alpha and delta cells.     

Detection of Babesia microti-like parasite in filter paper-absorbed blood of wild rodents

The first case of human babesiosis was reported in Japan. The epidemiology of this disease in Japanese nature remains unclear. In this study, 97 common field mice captured in Hokkaido, Japan, were examined. Blood specimens absorbed onto filter papers were eluted and tested by nested PCR using specific primers for the B. microti nuclear small subunit rRNA genome. Twenty-three percent (11/47) of Apodemus speciosus and four percent (2/50) of Clethrionomys rufocanus were positive. The 159-bp primary sequences of PCR products tested exhibited 97.5% and 96.8% homology with those of the human isolate in Japan and of U.S. strains of B. microti, respectively.     

No apoptotic cell death of erythroid cells of erythroblastic islands in bone marrow of healthy rats

A possibility of apoptotic cell death in erythropoietic regulation was examined by means of detailed light microscopical histoplanimetry, electron microscopy, the in situ nick-end labeling method, and an immunohistological method in the rat bone marrow. Serum erythropoietin concentrations were shown at normal levels. The erythroid series on a mature process presented several morphological features of apoptosis, i.e. the shrinkage of both nuclei and cytoplasm and the chromatin condensation. In the light microscopical histoplanimetry, however, morphological signs of final apoptotic cell death were never found in any erythroid cell within the erythroblastic islands. This finding was also supported by detailed ultrastructural observation: No erythroid cell bodies were trapped and degraded by the central macrophages of the erythroblastic islands, while the denucleated nuclei with small amount of cytoplasm of late erythroblasts were often trapped and degraded in the macrophages. Nuclear DNA fragmentation was not detected in any erythroblasts, but was detected in the lysosomes of the central macrophages. These findings suggest that erythropoiesis is regulated by other regulatory mechanisms than apoptotic cell death. An additional ultrastructural finding shows that the reticulocytes anchored to the central macrophages are transported into the peripheral blood circulation.     

Characterization of ISPsy2 and ISPsy3, Newly Identified Insertion Sequences in Pseudomonas syringae pv. eriobotryae

Two new active insertion sequences, ISPsy2 and ISPsy3, were isolated from Pseudomonas syringae pv. eriobotryae, the causal agent of stem cankers of loquat trees. ISPsy2 is 1194-bp long, has 16-bp imperfect terminal inverted repeats, and generates a 4-bp target site duplication upon insertion into the selective cartridge of the entrap vector pSHI1063. The nucleotide sequence of ISPsy2 is completely identical with that of the previously identified IS-like element located adjacent to the virulence gene psvA of Pseudomonas syringae pv. eriobotryae NAE6. The single open reading frame of ISPsy2 encodes a 323-amino-acid protein that has similarity to the transposase of the IS5 subgroup of the IS5 family. The ISPsy3 belonging to the IS91 family is 1507 bp in length, does not duplicate its target sequence, GAAC, and presents an 81% sequence homology with IS801 in P. s. pv. phaseolicola. The transposase of ISPsy3 possesses the conserved amino acid motifs found in the rolling-circle replication protein. Southern blot analysis indicated that multiple copies of ISPsy2 and ISPsy3 are present in the genomes of P. s. pv. eriobotryae and some of the other P. s. pathovars tested. Received 16 August 2001/ Accepted in revised form 19 October 2001