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Biomedical subjects

T Onodera

Publications and source records attributed to T Onodera.

At least 73 records · Page 4Linked to original sources
PubMed1997

A new high-molecular-weight glycophorin C variant with a duplication of exon 2 in the glycophorin C gene.

Glycophorin C (GPC) and glycophorin D (GPD) are closely related sialoglycoproteins in the human red blood cell (RBC) membrane. Both are thought to be encoded by the GPC gene (GYPC). We report here the new GPC variant, MAT, with a high-molecular-weight form of GPC and GPD. The murine monoclonal antibody to GPC (CBC-96), which had specificity for the N-terminal region of GPC, gave a stronger reactivity with the MAT RBCs than did normal RBCs in direct agglutination tests. Immunoblotting of the MAT RBC membranes with anti-GPC antibodies showed the apparent molecular weight of GPC.MAT and GPD.MAT was 5000 greater than that of their normal counterparts. cDNA was synthesized from total RNA obtained from three unrelated, heterozygous MAT blood donors and analysed by the polymerase chain reaction with primers that spanned sequences encoded by GYPC. Two fragments were generated: one was 510 bp, the other was 453 bp and corresponded to the normal GPC. Sequencing of the mutant 510-bp fragment showed an insert of 57 nucleotides that corresponds to the entire sequence of exon 2 in GYPC. These results show that MAT is the result of a duplication of exon 2 in GYPC, which probably encodes the two high-molecular weight forms GPC.MAT and GPD.MAT. The MAT mutation is found with a frequency of 0.02% in the Japanese population.

Animals↗
PubMed1997

Application of equine infectious anemia virus core proteins produced in a baculovirus expression system to serological diagnosis.

Equine infectious anemia virus (EIAV) core proteins were obtained from a baculovirus expression system. Recombinant baculoviruses (rBVs) highly expressed the Gag precursor and p26 antigens in an rBV-infected Sf21 cell culture supernatant. Enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) were conducted using the expressed proteins to detect antibodies from experimentally infected horses. The expressed antigens showed low background levels, high specificity and sensitivity in ELISA and AGID. The results of the serological tests using the expressed antigens were identical to those using a manufactured trial antigen. rBVs containing gag and p26 genes were found to express high quality and large quantities of Gag and p26 antigens, respectively. The antigens were quite useful for detecting anti-EIAV antibodies from virus-infected horses.

Animals↗
PubMed1997

c-myc, c-erbB-1 and c-erbB-2 expressions in urothelial carcinoma.

The expression of c-myc, c-erbB-1 and c-erbB-2 in 24 cases of urothelial carcinoma by Southern and northern blot analysis, and immunohistochemistry was examined. The results were compared with the pathological grade and stage. We found elevated mRNA expressions of c-myc and c-erbB-1 in 19 and 11 of 21 cases, respectively, but there was no apparent amplification or rearrangement of these oncogenes in any of the cases examined. By immunohistochemistry using anti-epidermal growth factor receptor antibody, most of the cases showed positive immunoreactivity on the cancer cell membranes, and cancers of higher pathological grade and stage showed more intense staining. By contrast, amplification of c-erbB-2 was detected in four of 24 cases, all of which were assigned to a high pathological grade (G3). Elevated c-erbB-2 mRNA levels appeared to correlate with the pathological grade of the cancers. Positive immunohistochemical reactions to c-erbB-2 were found in the cancer cell membranes in three of 24 cases, which were accompanied by amplification and elevated mRNA levels of c-erbB-2. In conclusion, expressions of c-myc, c-erbB-1 and c-erbB-2 were all elevated in the majority of urothelial carcinomas, but the amplification was not universal.

Blotting, Northern↗
PubMed1997

Curative percutaneous catheter ablation for various supraventricular and ventricular tachyarrhythmias. Results in 187 consecutive patients during the first five years.

Closed-chest transcatheter electrical ablation (catheter ablation) has been applied to various supraventricular and ventricular tachyarrhythmias as a radical therapeutic technique since its introduction in 1982. Currently, it has become a first line therapy for supraventricular tachyarrhythmias except atrial fibrillation and uncommon types of atrial flutter. We first carried out the ablation procedure in 1991 for the treatment of ventricular tachycardia. Up to February 1997, a total of 187 patients underwent catheter ablation in our institution. The aims of this study are to demonstrate our results of catheter ablation in the early 5 years and to show the usefulness of this new curative method. Successful results were obtained in 168 of 187 patients (overall final success rate: 89.8%). The success rates of each category of tachyarrhythmias were 100/105 patients (95%) with WPW syndrome, 41/46 (89%) with atrioventricular nodal reentrant tachycardia, 7/10 (70%) with atrial flutter, 4/4 (100%) with atrial tachycardia, 2/2 (100%) with medically refractory atrial fibrillation, 13/15 (85%) with idiopathic ventricular tachycardia and 3/7 (43%) with sustained ventricular tachycardia associated with structural heart disease, respectively. Complications that required invasive treatments were observed in 3 patients (2 hemopericardium and 1 complete atrioventricular block). Our results indicate that catheter ablation is highly effective in most categories of tachyarrhythmias and can be applied safely without lethal complications.

Adult↗
PubMed1997

Establishment of a sandwich enzyme linked immunosorbent assay for canine interleukin-8.

To estimate canine interleukin-8 (cIL-8) levels in blood plasma samples, a sandwich enzyme linked immunosorbent assay (ELISA) was established. For the development of the sandwich ELISA, polyclonal anti-cIL-8 (capturing), biotinylated anti-cIL-8 (developing) antibodies and glutathione-S-transferase/cIL-8 (GST/cIL-8) fusion protein as an antigen were used. cIL-8 in the fusion protein of GST/cIL-8 was detected in a dose dependent manner. The lowest limit of GST/cIL-8 detectable by this method was 2 ng/ml of GST/cIL-8 (containing; 0.470 ng/ml of cIL-8). IL-8 levels in the plasma samples from apparently healthy dogs were less than 0.470 ng/ ml. Higher levels of IL-8 were detected in the plasma samples of dogs with cystitis, dermatitis, and gastric cancer. These results suggest that the determination of cIL-8 by the sandwich ELISA is useful in diagnosis of inflammatory diseases in dogs.

Animals↗
PubMed1997

Modulation of the blood-aqueous barrier by light exposure in patients with uveitis.

OBJECTIVE: To evaluate the effect of light deprivation on flare measurements in patients with uveitis. DESIGN: Prospective study. SETTING: Eye clinic providing tertiary ophthalmic care in Calgary. PATIENTS: Five consecutive patients with a history of long-standing uveitis. INTERVENTIONS: Flare measurements were obtained with the Kowa FC-1000 flare-cell meter before and after 24 hours of monocular occlusion, and before and after wearing sunglasses during waking hours in a 24-hour period. OUTCOME MEASURE: Aqueous flare. RESULTS: A significant decrease in flare was observed after 24 hours of occlusion in all patients (p < 0.01). One patient showed an interesting change in response when her steroid treatment was tapered and subsequently stopped. Two of three patients showed a decrease in flare after wearing sunglasses. CONCLUSIONS: These preliminary results suggest that sunlight causes breakdown of the blood-ocular barrier in certain cases of uveitis. Further investigation is needed to determine the extent and mechanisms of light-induced breakdown of the blood-aqueous barrier as well as the role of sunglasses and anti-inflammatory medications as potential modulators of light-induced inflammation.

Adult↗
PubMed1996

Identification of a promoter region in the rat prion protein gene.

We have demonstrated the presence of a rat prion protein (RaPrP) gene promoter upstream of multiple initiation sites. A 0.1-kb fragment upstream of the 5'-untranslated region contains specific DNA motifs characteristic of promoter elements including an AP-1 binding site, an inverted CCAAT motif and three inverted Sp-1 binding sites. This fragment directs transcription of a luciferase reporter gene in pheochromocytoma cells (PC12) and rat glioma cells (C6), suggesting that it contains the promoter for the RaPrP gene. To more precisely localize the transcription regulatory elements in this region, a series of 5'-deletion mutants were generated. Deletion analysis showed that an inverted CCAAt and adjoining Sp-1 binding sequences may play an important role in transcription of the RaPrP gene.

Animals↗
PubMed1996

Three-exon structure of the gene encoding the rat prion protein and its expression in tissues.

The prion protein (PrP), encoded by a chromosomal gene, is associated with development of the neurodegeneration of prion-induced diseases. Since determination of the complete structure of the gene encoding PrP is important for understanding gene expression in the central nervous system (CNS), the nucleotide (nt) sequence of the isolated whole gene encoding rat PrP (raPrP) was determined. The rat PrP gene (raPrP) spans 16 kilobases (kb) of the rat genome and contains three exons of 19-47 base pairs (bp), 98 bp, and 2 kb separated by two introns of 2.2 kb and 11 kb. The first and second exons are noncoding, while the third exon contains a short 5' untranslated region, the entire 762-bp open reading frame (ORF), and a 3' untranslated region. The putative raPrP promoter in the 5' flanking region contains putative Sp1, AP-1, and AP-2 binding sites without a consensus TATA box. This TATA box-deficient feature, coupled with the presence of a high G+C content and Sp1-binding sites in the raPrP promoter, characterizes it as a housekeeping gene. Analysis of the raPrP cDNA 5'-end showed that raPrP mRNA transcription was initiated at multiple sites. Northern blot analysis showed that the levels of raPrP mRNA varied among rat tissues, with the highest levels found in the brain and placenta. This determination of raPrP nt sequences, including the introns and the 5' and 3' flanking regions, may make it possible to elucidate cis-acting elements that regulate the expression of this gene in different tissues and cell lines.

Amino Acid Sequence↗
PubMed1996

In vivo administration of serum thymic factor (FTS) prevents EMC-D virus-induced diabetes and myocarditis in BALB/cAJcl mice.

The effect of serum thymic factor (FTS) on the D-variant of encephalomyocarditis (EMC-D) virus-induced diabetes and myocarditis in BALB/cAJcl mice was investigated. Mice pretreated with 50 or 10 micrograms of FTS were infected with 10 or 10(3) PFU of EMC-D virus. In the mice inoculated with 10 PFU of virus, 40% developed diabetes on post-infection day (PID) 14, whereas those treated with FTS (50 micrograms/administration) on day 2 and 1 before infection did not develop diabetes. FTS (10 micrograms)-pretreated mice developed diabetes. In histological observation, FTS non-treated mice which developed diabetes showed severe necrosis and inflammation of mononuclear cells in the islets of Langerhans and myocardia on 19 PID. Mice pretreated with 50 micrograms of FTS, however, manifested mild islet degeneration without any myocardial inflammation. Furthermore, in FTS non-treated mice, immunohistological staining showed a loss of insulin granules. This loss was markedly reversed and insulin granules remained largely intact in FTS-pretreated mice. Viral titers in pancreas of FTS-pretreated mice approximated well to those of non-treated mice on PID 4, 7 and 19. In mice inoculated with higher titer of EMC-D virus (10(3) PFU), however, 50 micrograms of FTS pretreatment did not change the course of these acute pathological developments (diabetes and myocarditis observed from PID 4).

Animals↗
PubMed1996

Myocyte remodeling during the progression to failure in rats with hypertension.

Regional changes in cardiac myocyte shape during the progression to failure with hypertension have not been clearly established. To address this issue, we examined left and right ventricular myocytes from lean, female spontaneously hypertensive/heart failure rats with compensated hypertrophy (approximately 12 months of age) and congestive heart failure (approximately 24 months of age). During this period, body weight did not change, but heart weight increased 59% and lung weight increased 93%. Left ventricular function declined with the onset of failure. Left ventricular myocyte volume increased 27% exclusively because of myocyte lengthening (29% increase). The onset of left ventricular failure resulted in a 72% increase in right ventricular myocyte volume. Right ventricular myocyte growth, however, was proportional, with a 23% increase in myocyte length and 18% increase in myocyte width. Changes in left ventricular myocyte shape were virtually identical to data collected previously from patients with similar disease, suggesting that this is a relevant animal model. Evidence suggests that left ventricular myocyte transverse growth is defective because dilation and failure were associated with cell lengthening, without a change in myocyte diameter. Although severe hypertrophy was present in the right ventricle as a result of left ventricular failure, myocyte growth was proportional, suggesting that cell shape was properly regulated in this chamber.

Animals↗
PubMed1996

Production and characterization of polyclonal anti-canine interleukin-8 antibodies.

Polyclonal anti-canine interleukin-8 (cIL-8) antibodies were raised in rabbits immunized with cIL-8 expressed by E. coli. Polyclonal antibodies were purified by affinity chromatography. In the enzyme linked immunosorbent assay (ELISA), the resulting anti- cIL-8 antibodies showed relatively high reactivities with cIL-8 in the fusion proteins of glutathione-S-transferase/cIL-8 (GST/cIL-8) and maltose binding protein/cIL-8 (MBP/cIL-8), but negligible ones with MBP. Furthermore, Western blotting analysis using these polyclonal antibodies showed distinct bands for cIL-8, GST/cIL-8, and MBP/cIL-8. These antibodies also bound to recombinant human IL-8 (rhIL-8) in ELISA but not in Western immunoblotting. The rHIL-8 (50-800 ng/ml) was chemoattractant for canine neutrophils in a dose dependent manner, but the anti-cIL-8 antibodies did not show the inhibitory effect on the chemotactic activity of rhIL-8 of canine neutrophils, when tested by the chemotaxis assay using Boyden chambers. In addition, GST/cIL-8 and rhIL-8 induced strong and rapid shape change responses of canine neutrophils. However, the anti-cIL-8 antibodies inhibited shape change responses induced by GST/cIL-8 but not by rhIL-8.

ATP-Binding Cassette Transporters↗
PubMed1996

Activation of canine monocytes and polymorphonuclear cells by serum thymic factor (FTS) in vivo.

The effect of serum thymic factor (FTS) was evaluated from the immunoresponse augmented in canine monocytes and polymorphonuclear cells (PMN) using the chemiluminescence technique. FTS did not affect the number of leukocytes and differential count of leukocytes. CL activity of the whole blood was significantly elevated by FTS from 72 hr to 120 hr after administration (p < 0.05), and that at 96 hr after administration was about 3-fold higher than that before the administration. The CL response of PMN was significantly elevated by FTS administration from 24 hr to 96 hr after administration (p < 0.05), and that at 48 hr after administration was about 7-fold higher than prior treatment. FTS also significantly elevated the CL response of monocyte from 24 hr to 96 hr after administration (p < 0.01), and the CL count of monocyte in 24 hr and 48 hr was about 100-fold higher than that before FTS administration. These findings suggested that FTS may be efficacious and useful immuno-potentiator for canine monocytes and PMN.

Animals↗
PubMed1996

Variation in serum creatine phosphokinase activity as indicated in two-phase EMC-D virus-induced myocarditis.

In this study, myocardial damage in the D-variant of encephalomyocarditis (EMC-D) virus-induced myocarditis has been investigated consecutively by measuring serum creatine phosphokinase (CPK) activity. CPK activity in 8 week-old male BALB/cAJcl mice inoculated with EMC-D virus increased to a peak at 4 or 5 days postinoculation (DPI) and then gradually decreased. The CPK activity rose again after 7 DPI until it reached a second peak. In view of the kinetics of CPK activity, two-phase (early and late phase) myocardial damage in EMC virus infection were considered. In the late phase, an increase in cellular infiltration in the myocardium and a decrease in viral titer in the heart were observed. It was therefore suspected that the increase in CPK in the late phase may be caused by cellular infiltration, but not by viral replication. In our results, we suggested that a serial measurement of serum CPK activity might be a useful method for throwing more light on the myocardial damage caused by the autoimmune response. We also used a pathological (TUNEL) method to detect apoptotic cells and some apoptotic myocytes in the myocardium in late phase EMC virus-induced myocarditis.

Animals↗
PubMed1995

Cholecystokinin does not act on the efferent pathway of cholinergic and adrenergic nerves to inhibit ruminal contractions in sheep (Ovis aries).

The effect of exogenous cholecystokinin-octapeptide (CCK-8) on ruminal contractions and the role of efferent pathways of cholinergic and adrenergic nerves on the effect were studied in sheep. Intravenous infusion of CCK-8 at 11.4 and 45.6 pmol/kg/min significantly inhibited the frequency and amplitude of ruminal contractions in conscious sheep. After bilateral cervical vagotomy, intravenous infusion of CCK-8 at 45.6 mol/kg/min had no detectable effect on amplitude of ruminal contractions induced by electric stimulation to the cervical vagus nerve (1 msec, 20 Hz, 5 mA, for 10 sec at 1-min intervals) in anesthetized sheep. The amplitude of contractile responses of ovine ruminal muscle strips to acetylcholine at 5 x 10(-5) M was not inhibited by CCK-8 applied simultaneously at 1 x 10(-9) M. Intravenous infusion of phentolamine at 53.0 nmol/kg/min, propranolol at 101.4 nmol/kg/min, or their combined infusion did not alter the inhibitory action of CCK-8 at either dose on ruminal contractions in conscious sheep. These results suggest that CCK-8, which does not act on the efferent pathway of cholinergic and adrenergic nerves, may reflexively inhibit reticuloruminal contractions via vagal afferent fibers in sheep.

Acetylcholine↗
PubMed1995

Analysis of PrPc mRNA by in situ hybridization in brain, placenta, uterus and testis of rats.

An amyloid-like isoform of a 33- to 34-kD glycoprotein, termed as the scrapie prion protein (PrPsc), plays a critical role in transmissible spongiform encephalopathies of animals and humans. It has even been suggested to present the responsible infectious agent. This protein is a posttranslationally modified form of the cellular isoform of prion protein (PrPc). Hitherto, little has been known about the functions of PrPc. In order to examine the localization of PrPc mRNA in rat tissues, the in situ hybridization technique was performed. In rat brain, PrPc mRNA was predominantly localized within pyramidal cells of the hippocampus, large neurons of the thalamus and neocortex, and Purkinje cells of the cerebellum. In the placenta, not only PrPc mRNA was localized to a subpopulation of decidual cells at the highest levels, it was also expressed in the amnion and mesodermal layer of the yolk sac. Furthermore, PrPc mRNA was also expressed in the myometrium of the uterus and seminiferous tubule in the testis. However, signals were not obtained in the lung, spleen, liver of prenatals and other fetus tissues. The distribution of rat PrPc mRNA portrayed the levels which were different among the various types of cells, suggesting that its expression may be regulated in a tissue-specific manner.

Animals↗
PubMed1995

Detection of the chemotactic factor for canine peripheral blood mononuclear cells and polymorphonuclear cells in the culture supernatant of Cos7 cells transfected with canine interleukin-8 cDNA.

Chemotactic activities in the culture supernatants of Cos7 cells transfected with a cloned canine IL-8 cDNA (pcIL-8SR alpha 14) were evaluated by using mononuclear cells (MNC) and polymorphonuclear cells (PMN) from the peripheral blood of dogs. The culture supernatants of Cos7 cells were collected 66 hr after the transfection of pcIL-8SR alpha 14 (Cos7/cIL-8). Chemotactic activities in the culture supernatants for MNC and PMN were determined as migration distances in Millipore membrane filters in a modified Boyden's chamber method. Peroxidase staining for MNC was effective not only for cells in cytospun smears but also for cells migrated in the filters. PMN in cytospun smears were well stained by peroxidase staining, whereas migrated PMN in the filters were stained weakly. Chemotactic activities in the culture supernatant of Cos7/cIL-8 cells for both MNC and PMN were significantly higher than those of control Cos7 cells. In addition, the culture supernatant of Cos7/cIL-8 cells was chemotactic for peroxidase negative nonadherent MNC (lymphocytes), but not for peroxidase positive adherent MNC (monocytes). This Cos7/cIL-8 supernatant also showed chemotactic activities for neutrophils in a dose dependent manner. These results suggest that the culture supernatant of Cos7/cIL-8 is chemotactic for lymphocytes as well as for neutrophils.

Animals↗