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Biomedical subjects

R Wada

Publications and source records attributed to R Wada.

At least 55 records · Page 3Linked to original sources
PubMed2000

In vitro detection of equine arteritis virus from seminal plasma for identification of carrier stallions.

Equine arteritis virus (EAV) was readily isolated in RK-13 cell monolayers by plaque assay from seminal plasma of experimental carrier stallions when they contained high titers of virus regardless of the presence of non-viral cytotoxicity in the seminal plasma. The cytotoxicity interfered with virus isolation from seminal plasma which contained virus at titers less than 10 PFU/ml. However, it was possible to detect the virus in seminal plasma pretreated with PEG (#6000). EAV was consistently identified by RT-PCR from crude seminal plasma which contained virus at titers of more than 10(2.7) PFU/ml. In vitro detection of EAV by virus isolation supplemented with RT-PCR using seminal plasma was proved to be an effective alternative to the standard test mating as a diagnostic method for carrier stallions.

Animals↗
PubMed2000

Macroscopic anatomy of the auditory tube diverticulum (guttural pouch) in the thoroughbred equine--a silicon mold approach.

The characteristics of the equine auditory tube diverticulum (guttural pouches) were studied in detail by the naked eye and silicon mold method. First, we examined the anatomical relationship between the guttural pouches and their associated bones and muscles. Secondly, a silicon mold was fabricated to clarify the three-dimensional aspect of the guttural pouches, paying a special attention to the distribution of major arteries and nerves surrounding it. Thirdly, the volume of the silicon mold was measured by immersing it in water. The guttural pouches are a pair of pouches located dorso-posteriorly to the posterior pharynx. The pouches had a close contact rostrally with the Os sphenoidale, ventrally with the pharynx and the esophagus, and caudally with the Articulatio atlantoaxialis. The left and right guttural pouches had almost the same capacity in each horse. The A. carotis interna, cranial ganglia of cervical nerves and cervical Truncus sympathicus, as well as the N. vagus, N. glossopharyngeus, N. hypoglossus, and Radices spinales of the N. accessorius, were present in the mucosal crease extending from the roof of the guttural pouches towards the middle of the caudal wall. The N. facialis appeared from the Foramen stylomastoideum ran dorsocaudally along the lateral recess. The N. mandibularis appeared from the Foramen lacerum went over the muscular process of an ear bone, ran along the roof of the lateral recess and finally continued with the rostral side of the guttural pouches. Lymph nodes, scattered on the mucosal surface, appeared as tiny indentations with approximately millet seed size on the silicon surface. The capacity of guttural pouches in adult horses (472 +/- 12.4 cm3) was three-fold larger than that in foals (145 +/- 9.4 cm3).

Animals↗
PubMed2000

Histological and morphometrical studies on the mucosa of the equine guttural pouch (auditory tube diverticulum).

The present study attempted to clarify the characteristics of the guttural pouch mucosa in equines and to evaluate its foreign substance clearance ability. The specimens were collected from nine regions (eight in the guttural pouch mucosa, and one in the nasopharynx mucosa). We first examined the pouch mucosa by light and electron microscopy. We then measured the frequency of goblet cells per 200 epithelial cells, the length of the cilia, the thickness of the epithelial cell layer and lamina propria and statistically analyzed the data. The guttural pouch mucosa consisted of stratified columnar epithelia with brush-like cilia, and there were almost no histological differences between adults and foals. The morphometrical study revealed significant differences in goblet cell frequency (p < 0.001) and the thickness of lamina propria (p < 0.05). By contrast, no statistically significant difference was detected in the length of the cilia or the thickness of the epithelial cell layer. These findings suggest that the guttural pouch mucosa provides foreign substance clearance ability, but that its ability varies among different regions of the epithelium.

Age Factors↗
PubMed2000

[A thoracoscopic resection of pulmonary metastasis from breast cancer: a case report with a 27-year disease-free interval].

A 70-year-old woman, who had undergone a right radical mastectomy for breast cancer 27 years previously, was found to have a tumor measuring 15 mm in diameter between the S5 area and the S8 area of the left lung. We suspected it to be either metastatic or primary lung cancer based on preoperatively any conclusive diagnosis. We thus performed a thoracoscopic partial lung resection and pericardial resection. The pathological diagnosis of the resected specimen was pulmonary metastasis from the previous breast cancer, since the pathological findings of the lung lesion were closely similar to those of the previous breast lesion. Using immunohistochemical methods, the tumor cells show positive staining for anti-estrogen receptor antibody. In lung tumor cases in which the patient has undergone a breast cancer resection, even more than 20 years previously, surgeons must not rule out the possibility of recurrence. Thoracoscopic surgery is considered to be most effective method for both making a definitive diagnosis and for performing curative treatment.

Aged↗
PubMed2000

Structure of lamina propria lymphoid follicles and associated epithelium in the gastric mucosa during Helicobacter pylori infection in ulcer-bearing Mongolian gerbils.

To develop a gerbil model of Helicobacter pylori-induced chronic active gastritis comparable in severity to human lesions, we made acetic acid-induced ulcer in the anterior antral wall and concurrently challenged 1 x 10(8) colony-forming units bacteria per os. At 30 and 60 days after inoculation, the number of viable bacteria colonizing on the surface epithelium of the gastric mucosa was larger in ulcer-bearing animals compared to non-bearing ones. Furthermore, in the former animals, neutrophil and mononuclear cell infiltration as well as lymphoid follicle formation in the lamina propria was more prominent. Electron microscopically, lymphoid follicle-associated epithelium displayed specialized structures. Namely, brush cells interposed between mucous epithelial cells and characterized by prominent microfilament bundles and many apical vesicles or caveola specifically embraced the cluster of intraepithelially invading lymphocytes and macrophage-like cells by the attenuated cytoplasm in an analogous manner to M cells in Peyer's patches. The present study has demonstrated that ulcer formation enhances both H. pylori colonization and lamina propria lymphoid follicle formation and suggested that follicle-associated epithelium might play roles in the delivery of intraluminal antigen.

Animals↗
PubMed2000

[Clinicopathological studies of small-sized advanced large bowel cancer less than 2 cm in diameter].

The small sized lesion of the large bowel advanced cancer has been noticed in the development of the large bowel cancer. In this study, 17 lesions of the large bowel advanced cancer within less than 2 cm in diameter were studied clinicopathologically and histologically, according with PG(polypoid growth), NPG(non polypoid growth) classification and the subclassification of NPG, in which NPG-1 was defined that the height of the adjacent mucosa to the carcinomatous lesion was taller than that of the cancerous lesion, and the NPG except for NPG-1 was defined as NPG-2. And all of 17 lesions were NPG type(NPG-1:9 lesions, NPG-2:8 lesions). The results revealed that NPG-1 lesions were significantly smaller in maximum diameter, the invasion depth of its were deeper, these degree of venous invasion were higher and these carcinomatous differentiation were lower, comparing with those of NPG-2 lesions. In conclusion, it was indicated that the NPG-1 lesion was the large bowel advanced cancer which deeply invaded at the small-sized condition.

Adenocarcinoma↗
PubMed1999

Salmonella Abortusequi strains of equine origin harbor a 95kb plasmid responsible for virulence in mice.

Most Salmonella choleraesuis subsp. choleraesuis serovar Abortusequi strains of equine origin harbor a 95kb plasmid, pSA95. Results of PCR and Southern blot analysis suggest that pSA95 contains spv genes. A pSA95-cured strain of S. Abortusequi was 48 times less virulent to mice than its parental strain. Virulence was restored by reintroduction of pSA95. These results provide clear evidence that pSA95 confers virulence on S. Abortusequi in mice. This is the first report describing a virulence plasmid of S. Abortusequi.

Animals↗
PubMed1999

A vital role for glycosphingolipid synthesis during development and differentiation.

Glycosphingolipids (GSLs) are believed to be integral for the dynamics of many cell membrane events, including cellular interactions, signaling, and trafficking. We have investigated their roles in development and differentiation by eliminating the major synthesis pathway of GSLs through targeted disruption of the Ugcg gene encoding glucosylceramide synthase. In the absence of GSL synthesis, embryogenesis proceeded well into gastrulation with differentiation into primitive germ layers and patterning of the embryo but was abruptly halted by a major apoptotic process. In vivo, embryonic stem cells deficient in GSL synthesis were again able to differentiate into endodermal, mesodermal, and ectodermal derivatives but were strikingly deficient in their ability to form well differentiated tissues. In vitro, however, hematopoietic and neuronal differentiation could be induced. The results demonstrate that the synthesis of GSL structures is essential for embryonic development and for the differentiation of some tissues and support the concept that GSLs are involved in crucial cell interactions mediating these processes.

Animals↗
PubMed1999

In vivo pathogenicity and resistance to phagocytosis of Streptococcus equi strains with different levels of capsule expression.

The glossy non-encapsulated strain of Steptococcus equi, NCTC 9682, was compared with the matt strain Hidaka/95/2 which expresses a medium sized capsule and with the mucoid CF32 which expresses a large sized capsule in phagocytosis assays and for virulence in inoculated horses. The three strains, NCTC 9682, Hidaka /95/2 and CF32 produced 2.0, 3.1, and 5.3 mg/g wet cells respectively after 3 h incubation, but similar amounts of M-like proteins, cytotoxin and mitogen. NCTC 9682 showed no resistance to phagocytosis by equine neutrophils regardless of the presence of opsonin while strains Hidaka /95/2 and CF32 showed almost complete resistance to phagocytosis. Furthermore, NCTC 9682 produced no clinical disease although it infected the guttural pouch and caused seroconversion. Typical strangles with guttural pouch invasion was observed in all horses infected with encapsulated strains.

Animals↗
PubMed1999

Streptococcus equi but not Streptococcus zooepidemicus produces potent mitogenic responses from equine peripheral blood mononuclear cells.

Streptococcus equi causes equine strangles. The acute disease has many of the hallmarks of an acute response including high fever, elevated plasma fibrinogen and neutrophilia, affects known to be mediated by proinflammatory cytokines. The objective of this study was to screen-culture supernatants from equine clinical isolates of S. equi and S. zooepidemicus for stimulation of mitogenic responses by horse peripheral blood mononuclear cells. Mitogenicity comparable to that of concanavalin A was detected in culture supernatants of S. equi strains but not in those of S. zooepidemicus. Mitogenicity was neutralised by Proteinase K and a post-strangles convalescent serum, and evidence for the presence of both thermo-resistant and thermo-labile mitogenic factors was obtained. Release of proteinaceous immunogenic mitogens in combination with the antiphagocytic protein SeM unique to S. equi may therefore contribute to some of the severe clinical manifestations of acute strangles in the horse.

Animals↗
PubMed1999

Only limited effects of aminoguanidine treatment on peripheral nerve function, (Na+,K+)-ATPase activity and thrombomodulin expression in streptozotocin-induced diabetic rats.

AIMS/HYPOTHESIS: Aminoguanidine, a potent anti-glycation reagent, is known to be beneficial in experimental diabetic neuropathy. In this study, we explored the mechanisms of how aminoguanidine inhibits neuropathic changes in diabetes and compared its effects with those of insulin treatment. METHODS: Wistar rats, aged 8 weeks, were made diabetic by streptozotocin and given aminoguanidine dissolved in drinking water (1 g/l) for 8 weeks. Effects of daily insulin (protamine-zinc) treatment were also examined for comparison. At the end of the 8 weeks, we examined the peripheral nerve function and (Na+,K+)-ATPase activity and their relation to serum thrombomodulin concentrations that are considered as a marker of endothelial injury. RESULTS: Aminoguanidine treatment reduced the diabetes-induced decrease in tibial nerve conduction velocity by 47% (p < 0.05 vs untreated diabetic rats) and inhibited the loss of sciatic nerve (Na+,K+)-ATPase activity by 54% (p < 0.05 vs untreated diabetic rats). Insulin-treatment of diabetic rats restored these variables by 83% and 75%, respectively (both, p < 0.01 vs untreated diabetic rats). Thrombomodulin concentrations were increased (p < 0.01) in diabetic rats compared with those in non-diabetic controls and unaffected by aminoguanidine treatment. In contrast, the concentrations remained within the normal range in the insulin-treated group. CONCLUSION/INTERPRETATION: Although aminoguanidine treatment improved nerve conduction velocity and (Na+,K+)-ATPase activity, its effects were considerably less than those of insulin and were not apparent in some measures of endothelial cell injury.

Animals↗
PubMed1999

A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder.

Inherited defects in the degradation of glycosphingolipids (GSLs) cause a group of severe diseases known as GSL storage disorders. There are currently no effective treatments for the majority of these disorders. We have explored a new treatment paradigm, substrate deprivation therapy, by constructing a genetic model in mice. Sandhoff's disease mice, which abnormally accumulate GSLs, were bred with mice that were blocked in their synthesis of GSLs. The mice with simultaneous defects in GSL synthesis and degradation no longer accumulated GSLs, had improved neurologic function, and had a much longer life span. However, these mice eventually developed a late-onset neurologic disease because of accumulation of another class of substrate, oligosaccharides. The results support the validity of the substrate deprivation therapy and also highlight some limitations.

Animals↗
PubMed1998

Accelerated loss of islet beta cells in sucrose-fed Goto-Kakizaki rats, a genetic model of non-insulin-dependent diabetes mellitus.

The Goto-Kakizaki (GK) rat is a spontaneously diabetic animal model of non-insulin-dependent diabetes mellitus, which is characterized by progressive loss of beta cells in the pancreatic islets with fibrosis. In the present study, we examined the effects of sucrose feeding on the islet pathology in this model. Six-week-old GK rats were fed with 30% sucrose for 6 weeks to induce severe hyperglycemia, and their condition was compared with that of nontreated rats. Age-matched normal Wistar rats were also given sucrose for the same periods and used for comparison. The sucrose-treated GK rats showed elevated blood glucose levels on oral glucose tolerance tests at 60 minutes and 120 minutes, representing 123% and 127% of values in untreated GK rats, respectively. At the end of the study, the mean beta-cell volume density in GK rats was 50% less than that in untreated Wistar rats. Sucrose feeding further reduced the volume densities of beta cells to only 50% of the levels of age-matched GK rats. Apoptotic cells were found in islet beta cells only in GK rats fed sucrose (mean 0.067%). There appeared to be more islets that immunohistochemically stained strongly positive with 8-hydroxy-deoxyguanosine as a marker of oxidative damage of DNA in GK rats fed sucrose compared with those not given sucrose. GK rats not fed sucrose showed significantly lower proliferative activity of beta cells measured by 5-bromo-2'-deoxyuridine uptake and intensified expression of Bcl-2 immunoreactivities at 6 weeks of age compared with those in age-matched Wistar rats. These two indices were reduced in both GK and Wistar rats with increasing age and were not affected by sucrose feeding in either group. The present study thus indicated that sucrose feeding promoted the apoptosis of beta cells in GK rats through increased oxidative stress without altering their proliferative activity.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1998

Pathogenesis of Babesia caballi infection in experimental horses.

The present study was designed to investigate the role of cytokines in the pathogenesis of Babesia caballi in experimentally infected horses. The expression of cytokine mRNA was determined by using reverse transcription-polymerase chain reaction in two B. caballi-infected horses for 2 weeks after the infection. In one horse, there was up-regulation of interferon-gamma, tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 mRNAs, while in the second horse, expression of only TNF-alpha mRNA was up-regulated. No change was observed in interleukin-4 mRNA in both of the horses. To know the relation between nitric oxide (NO) production and pathogenesis, NO production was assayed in three dexamethasone treated-B. caballi-infected horses. Production of NO in all 3 horses increased significantly before death, although the parasitemia level remained very low. Treatment with NO inhibitor resulted in the suppression of NO production and increased parasitemia level in a horse, which died of the infection. The pathological examination showed that the main cause of the death was dyspnoea and pulmonary edema. Histopathologically, diffuse global mesangial proliferative glomerulonephritis was also observed. These results suggested that NO may be a critical effector molecule of immune defense against parasite. TNF-alpha and NO might be contributing to the pathogenesis in B. caballi infection.

Animals↗
PubMed1997

Pathogenicity and virulence of Rhodococcus equi in foals following intratracheal challenge.

Twelve foals, between 27 and 83 days old, were infected with 2 strains of Rhodococcus equi by intratracheal administration. Ten of the 12 foals were inoculated with 10(4)-10(10) colony forming units (cfu) of ATCC 33701 strain. The other 2 foals were inoculated with 10(9) cfu of a plasmid-cured derivative of the ATCC 33701 strain (ATCC 33701P-). All of the 10 foals challenged with the ATCC 33701 strain showed clinical signs of pulmonary disease within 5-13 days, such as gross lesions associated with acute bronchopneumonia and microscopic lesions associated with granulomatous pneumonia. The two foals challenged with the ATCC 33701P- strain showed neither clinical signs of disease nor gross lesions. Apparently, when lacking plasmid, the virulent Rhodococcus equi lost its pathogenicity.

Actinomycetales Infections↗
PubMed1997

Comparison of tracheal aspiration with other tests for diagnosis of Rhodococcus equi pneumonia in foals.

The diagnostic value of tracheal aspiration was evaluated through comparison with other diagnostic methods using an experimental model of Rhodococcus equi (R. equi) pneumonia in foals. Pneumonia was induced by spraying of the virulent R. equi strain ATCC 33701 into the trachea of foals. All foals developed fever from 11 to 16 days after bacterial inoculation. One foal was euthanized on day 26 due to its poor prognosis, and other foals euthanized on day 43. During the experiment, some tests for diagnosis of Rhodococcus equi pneumonia such as tracheal aspiration, radiography, serodiagnosis and fecal culture were carried out. R. equi was continually isolated from tracheal aspirates collected via a silicone catheter inserted transnasally on day 8 to day 32 after bacterial inoculation. On the other hand, radiography, serodiagnosis and fecal culture were demonstrated to be valuable diagnostic methods, but to be limited compared with tracheal aspiration. Indirect fluorescent antibody technique (IFA) using a monoclonal antibody against the 15- to 17-kDa virulence-associated antigens (VapA) of R. equi and PCR targeting the structural gene of VapA detected bacteria in tracheal aspirates less sensitively than the isolation technique although they were more rapid. Therefore, we conclude that a combination of tracheal aspiration and bacterial isolation was the most valuable method for routine diagnosis of R. equi pneumonia in foals.

Actinomycetales Infections↗