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Biomedical subjects

T Nunoya

Publications and source records attributed to T Nunoya.

At least 19 recordsLinked to original sources

Unilateral ocular subalbinism in a laboratory Beagle dog.

Blue discoloration of the iris was found in the left eye of a male laboratory Beagle dog, which had a normal tricolor coat and clinically showed no visual impairment. Ophthalmoscopically, the affected eye revealed red-colored tigroid fundus, in which no tapetum was present. The retinal vasculature and the optic disc showed no noticeable changes. Histopathologically, in the left eye melanocytes had extremely few melanin granules in the anterior segment, including the anterior border layer, in the stroma and sphincter muscle of the iris and in the stroma of the ciliary body and choroid. However, the posterior pigment epithelium of the iris, the pigment epithelium of the ciliary body and the retinal pigment epithelium showed normal pigmentation. The tapetal elements were completely absent. Number and distribution of the S-100 protein-positive melanocytes with or without melanin granules in the iris, ciliary body and choroid of the left eye were similar to those of the normal right eye. Ultrastructurally, melanocytes in the anterior segment of the affected iris possessed no or few melanosomes which were incompletely melanized. In the right eye, no abnormal features were observed. Based on these results, the present case was diagnosed as unilateral ocular subalbinism with tapetal aplasia in a Beagle dog.

Albinism, Ocular↗

Lipoolygosaccharide indirectly enhances inflammatory lesions in lungs as a primary infection site by non-encapsulated and type B Haemophilus influenzae through production of cytokines.

We investigated the role of cytokines in differences in histopathologic changes in the lung between bronchopneumonia caused by non-encapsulated Haemophilus influenzae strain 770235f(0)b(0)and systemic disease caused by type b H. influenzae strain 770235f(0)b(+). Tumour necrosis factor-alpha (TNF-alpha), interleukin-(IL)-6 and IL-1 beta levels in bronchoalveolar lavage fluid (BALF) samples of mice infected with strain 770235f(0)b(0)were higher than in those infected with strain 770235f(0)b(+)until 24 h post-infection. Serum IL-6 rapidly increased in strain 770235f(0)b(0)infection after 72 h post-infection. Serum TNF-alpha level in strain 770235f(0)b(0)infection appeared earlier than in strain 770235f(0)b(+)infection. IL-1 beta production in strain 770235f(0)b(+)infection was later than in strain 770235f(0)b(0)infection. Moreover, a biphasic concentration pattern of TNF-alpha and IL-6 was noted in BALF of mice with strain 770235f(0)b(0)infection.

Animals↗

Anophthalmia and retinal degeneration associated with stenosis of the optic foramen in Fischer 344 rats.

Bilateral anophthalmia was discovered in a male rat (No. 1) and unilateral anophthalmia was found in the left eye of two female rats (Nos. 2 and 3) derived from a Fischer 344 inbred colony. One male rat (No. 4), a littermate of No. 3, had externally normal eyes, but his left eye had severe retinal atrophy, which was detected by ophthalmoscopy. The eyelids in both eyes of No. 1 and in the left eyes of Nos. 2 and 3 were present. Radiography of the skull revealed small optic foramina on both sides of No. 1 and on the affected side of Nos. 2, 3, and 4. Histologically, both globes and optic nerves (ONs) of No. 1 and the left globe and ONs of Nos. 2 and 3 were completely missing. Diffuse retinal degeneration in the left globe and axonal degeneration in the left ON and the right optic tract were observed in No. 4. Dysplasia of the sphenoid bone associated with stenosis of the optic foramen was detected on the affected side of all rats. Thus, ON aplasia in anophthalmic rats and atrophic ON in a rat with retinal degeneration seem to be closely related to stenosis of the optic foramen.

Animals↗

Spontaneous lymphoma in a Japanese White rabbit.

Lymphoma was observed in a 4-month-old female Japanese White rabbit. Grossly, the markedly enlarged mesenteric lymph nodes, prominent Peyer's patches of jejunum, splenomegaly, and enlargement of tracheobronchial lymph nodes, adrenal glands and ovaries were observed. Histologically, neoplastic lymphoid cells proliferated diffusely showing frequent mitotic figures and a characteristic 'starry sky' appearance. Their basophilic cytoplasm contained a few lipid droplets. The mesenteric lymph nodes, Peyer's patches of jejunum, and tracheobronchial lymph nodes were largely replaced by the tumor tissues. The stomach, small intestines, especially the jejunum, liver, spleen, ovaries, and adrenal glands were heavily infiltrated with neoplastic cells. These results suggest that the present lymphoma may have originated from the gastrointestinal lymphoid tissue.

Animals↗

Optic nerve dysplasia associated with meningeal defect in Sprague-Dawley rats.

Unilateral and bilateral dysplasias of the optic nerve (ON) were observed in 20/114 male and 14/110 female Sprague-Dawley rats at 12 weeks of age. Grossly, the intracranial segment of the affected ON had nodular thickening, bifurcation, and curvature. Nodular thickenings were seen in 20 males and 11 females. One female had a bifurcated ON. Curvature was observed in the left ONs of two females. Of 34 ON dysplasias, 12 ONs tapered off into a thin filament at the portion anterior to the dysplastic lesions. The intraorbital segments of the ONs in 33 rats were also reduced in size and were hardly recognizable in the meningeal sheath in 10 rats. Both eyeballs appeared normal in all the animals examined. Histologically, nerve fibers in intracranial and intraorbital segments of the ONs that appeared as slender filaments were markedly reduced in number. Nerve fibers in nodular thickenings were intertwined in haphazard fashion, forming scrollworklike structures. The meningeal sheaths in intracranial segments of the ONs in 15 rats and in intraorbital segments in eight rats were partially missing. The naked portion of the ON protruded into the meningeal spaces or gaps. The data indicate that developmental failures in the ON may have been induced due to insufficient blood supply through the meningeal covering or herniation of growing nerve fibers into the defective meninges. However, etiology and pathogenesis of this condition remain unclear.

Animals↗

New murine model of bronchopneumonia due to cell-bound Haemophilus influenzae.

This murine model of nontypeable (unencapsulated) Haemophilus influenzae (NTHI) bronchopneumonia used organisms bound to mouse fetal lung (MFL) cells as an inoculum. Pretreatment of the mice with 40 microL of 1% formalin 3 days before intranasal instillation of the bacteria was necessary to allow infection. The number of NTHI recovered from the lungs plus trachea on day 7 after instillation was >100 times the number originally inoculated. Later, however, the number of recovered bacteria diminished gradually, and by day 14 it was almost identical to the original inoculum size. Serum IgM also peaked on day 7 after infection, after which IgG increased while IgM decreased. Histologically, bronchoalveolar infiltration of neutrophils was observed on day 3 after inoculation and continued at least for the following 4 days. The present experiment demonstrates that MFL cells can protect bacteria that have invaded the cells from the opsonizing and killing activities of host humoral defense mechanisms.

Animals↗

Establishment and characterization of cell lines, MeMS and MeFS, derived from spontaneous rat mesotheliomas.

New cell lines, designated MeMS and MeFS, were established from the epithelial type of mesotheliomas occurring spontaneously in aged male and female Fischer 344 rats, respectively. Morphologically, MeMS and MeFS expressed the epithelial phenotype. Coexpression of keratin and vimentin in both cell lines was confirmed by the immunocytochemistry, one-dimensional gel electrophoresis and immunoblotting for intermediate filament proteins. Hyaluronic acid synthesis of both MeMS and MeFS was demonstrated by the analysis using high-performance liquid chromatography. MeMS and MeFS seem to be useful models for studying the biological nature of rat mesotheliomas.

Aging↗

Induction of protective immunity in chickens orally immunized with inactivated infectious bursal disease virus.

The present studies were undertaken to examine the effects of oral administration of formalin-inactivated infectious bursal disease virus (IBDV) on the immune responses of chickens. Inactivated IBDV was suspended in phosphate-buffered saline containing sodium bicarbonate. This form of antigen, when administered by oral instillation, induced a serum antibody response against IBDV in chickens and these chickens were protected from subsequent viral challenge. The immunoglobulin class of IBDV-specific antibodies in serum was found to be IgG when determined by enzyme-linked immunosorbent assay. Cholera toxin, which has been reported to have potent mucosal adjuvant properties in mammals, did not enhance the serum antibody response. Oral followed by parenteral administrations of antigen induced an enhanced antibody response in chickens.

Administration, Oral↗

Occurrence of keratoconjunctivitis apparently caused by Mycoplasma gallisepticum in layer chickens.

Natural cases of keratoconjunctivitis, apparently caused by Mycoplasma gallisepticum (MG), in layer chickens are described. The disease occurred in a commercial flock consisting of 36,000 pullets (Babcock), first appearing around 30 days of age. Clinically, affected chickens showed unilateral or bilateral swelling of the facial skin and the eyelids, increased lacrimation, congestion of conjunctival vessels, and respiratory rales. Some of the severely affected chickens closed their eyes. The morbidity reached 27.8%, and it was estimated that approximately 10% died from reduced feed intake due to impaired vision. Ten 70-day-old chickens with clinical diseases were examined for lesions. There was acute to subacute keratoconjunctivitis in all of the chickens, and some exhibited laryngitis. Adherence of mycoplasma organisms to epithelial cells of the conjunctiva, cornea, and larynx was frequently observed. These organisms had an ultrastructure characteristic of MG and showed a positive reaction with rabbit polyclonal antibodies against the S6 strain of MG by immunohistochemical analysis. MG was isolated from tissue homogenates of the eyelids and tracheas of the affected chickens. Many of the chickens had atrophic bursae, and infectious bursal disease virus antigens were detected in necrotic bursal follicles by immunostaining. Therefore, immunosuppression due to infectious bursal disease was implicated in the pathogenesis of keratoconjunctivitis in the present cases.

Animals↗

Establishment and characterization of a cell line (NB-YK) derived from a transplantable rat nephroblastoma.

A new cell line, designated NB-YK, was established from a transplantable rat nephroblastoma (NB-Y) which was derived from a spontaneous nephroblastoma in an aged Fischer 344 rat. NB-YK grew in a piling-up and noncohesive pattern on the plastic surface and formed colonies in a soft agar. The main cell type of NB-YK represented morphology of mesenchymal phenotype and most of the cells contained several secretory granules in their cytoplasm. Immunocytochemically the cells were positive for vimentin, cytokeratin, and laminin. Coexpression of vimentin and cytokeratin in the cells was confirmed by the one-dimensional gel electrophoresis and immunoblotting for intermediate filament proteins. NB-YK cells were tumorigenic and produced fibrosarcoma-like tumor when inoculated subcutaneously or intraperitoneally into syngeneic rats and nude mice. NB-YK seems to be a useful model for studying biological properties of nephroblastoma.

Animals↗

Pathogenesis of highly virulent infectious bursal disease virus infection in intact and bursectomized chickens.

The pathogenesis of highly virulent infectious bursal disease (IBD) virus (IBDV) infection was studied using 6-week-old intact and 5-week-old bursectomized chickens inoculated with highly virulent strain 90-11 or reference strain I. Chickens inoculated with 10(0.7) EID50 of strain 90-11 showed neither clinical signs nor lesions during the 4-day observation period. In contrast, birds inoculated with 10(2.7) or 10(4.7) EID50 developed severe clinical IBD, as well as gross and histologic lesions, typical of IBD, and produced IBDV antigen demonstrable by immunostaining in the bursa of Fabricius (BF), thymus, spleen and bone marrow from day 2 post-inoculation (PI) onwards. The antigen was also detected by the agar-gel precipitation and latex microsphere agglutination tests in a bursal suspension of these birds from day 2 or day 3 PI on. Birds inoculated with 10(6.1) EID50 of strain I developed only slight clinical signs at day 4 PI. Their lesion- and antigen-scores in the BF were almost the same as those in virulent strain-infected chickens, but lesion- and antigen-scores in the other organs were negligible. Bursectomized chickens inoculated with strain 90-11 did not develop clinical IBD despite the presence of infection that was evidenced by histologic lesions in the thymus and spleen as well as IBDV antigen demonstrable by immunostaining in these organs.

Animals↗

An outbreak of porcine reproductive and respiratory syndrome in Japan.

An emerging swine disease principally involving periweaning piglets was examined. The disease was clinically characterized by lethargy, fever, emaciation, coughing, and severe abdominal breathing, hence colloquially named "Heko-heko" disease. The consistent lesions in affected piglets were diffuse interstitial pneumonia with pronounced type II pneumocytic proliferation, meningoencephalitis, and regression of the lymphoid tissues. The causal virus was isolated in primary porcine lung cell (PLC) cultures from various organs of affected piglets and showed serological relatedness to the European porcine reproductive and respiratory syndrome (PRRS) virus. Numerous virus particles, measured about 49 nm in diameter, were detected in the cytoplasm of alveolar epithelial cells and pulmonary macrophages in PLC cultures infected with the isolate. The condition could be experimentally reproduced in conventional piglets by intranasal inoculation with the isolate and the virus was reisolated from the infected animals.

Animals↗

A rapid quantitative method for detecting infectious bursal disease virus using polystyrene latex microspheres.

A monoclonal antibody (mAb) to infectious bursal disease virus (IBDV) was bound to polystyrene latex microspheres. The microspheres agglutinated with extracts of bursae and sera from chickens infected with all strains or isolates of IBDV tested. Agglutination appeared within a 10-min reaction time. The assay could detect a 10(3.7) to 10(4.5) mean embryo infective dose (EID50) of the virus in 0.01 ml and the titer of the assay was 10- to 40-times higher than that of the agar gel precipitin test.

Animals↗

Role of pseudorabies virus glycoprotein II in protection from lethal infection.

A monoclonal antibody (mAb), named 1.21, with complement-dependent neutralizing activity was produced against glycoprotein II (gII) of pseudorabies virus (PRV). By immunoaffinity chromatography using a mAB 1.21 column, gII was purified from Nonidet P40-lysates of PRV infected BHK21/13 cells. When mice and pigs were immunized with purified gII, complement-dependent virus-neutralizing antibodies were produced. The immunized animals survived potentially lethal challenge with PRV. These results indicate that an immunological response against gII plays an important role in the protection from PRV infection.

Animals↗

Isolation and serial propagation of porcine epidemic diarrhea virus in cell cultures and partial characterization of the isolate.

Porcine epidemic diarrhea virus (PEDV) was isolated in Vero cell cultures from the small intestine of a piglet experimentally infected with porcine coronavirus 83P-5, that had been isolated during outbreaks of porcine acute diarrhea and passaged in piglets. The isolation of the PEDV was successful only in Vero cells maintained in the maintenance medium (MM) containing trypsin. Infected Vero cell cultures exhibited CPE characterized by cell-fusion and syncytial formation, as well as cytoplasmic fluorescence when examined by the indirect immunofluorescent test using rabbit anti-83P-5 virus serum. The isolate was adapted to serial propagation in Vero cell cultures by adding trypsin to MM. Vero cell-adapted PEDV was successfully propagated in the MA104, CPK and ESK cell lines in the presence of trypsin in MM. Vero cell-adapted PEDV had morphologic and physicochemical characteristics similar to those of other members of the coronaviridae. The isolate differed serologically from porcine transmissible gastroenteritis (TGE) and porcine hemagglutinating encephalomyelitis viruses, and no antigenic relationship between the isolate and TGE virus could be detected by the indirect immunofluorescent test. Attempts to isolate PEDV in 6 types of primary fetal pig cell cultures and 6 of 10 established cell lines resulted in the failure, probably because these cells were damaged by the action of trypsin.

Animals↗

Primary culture of chicken bursal plical epithelium.

Plical epithelial cells were obtained by trypsin-EDTA treatment of chicken bursa of Fabricius and cultured in the presence of type IV collagen. The culture became confluent six to seven days after seeding. The grown cells showed a positive reaction for cytokeratin by immunostaining and had ultrastructural characteristics of the epithelial cells in vivo. The cell culture will be useful for parasitological and virological studies.

Animals↗