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Biomedical subjects

A Shimada

Publications and source records attributed to A Shimada.

At least 91 records · Page 5Linked to original sources

Characterization of monoclonal antibodies generated against Norwalk virus GII capsid protein expressed in Escherichia coli.

The Norwalk virus (NV) causes outbreaks of acute non-bacterial gastroenteritis in humans. The virus capsid is composed of a single 60 kDa protein. The capsid protein of NV36 (genogroup II, Mexico virus type) was expressed in an Escherichia coli system and ten monoclonal antibodies (MAbs) were generated against it. The reactivity of these MAbs was characterized using enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) analysis towards 20 overlapping fragments of the NV36 capsid protein expressed in E. coli. All of the MAbs recognized sequential (continuous) epitopes on the three antigenic regions. Six of the 10 MAbs recognized fragment 2 (equivalent residues 31-70), three MAbs recognized fragment 13 (residues 361-403) and one MAb recognized fragment 7 (residues 181-220), suggesting that the N-terminal domain (residues 1-220) may contain more antigenic epitopes than the C-terminal domain (residues 210-548). Furthermore, two MAbs (1B4 and 1F6) reacted in WB with three purified NV strains (genogroup II) derived from patients' stool samples. It was also found that genogroup I recombinant NV96-908 (genogroup I, KY89 type) could be detected as sensitively as recombinant NV36 (genogroup II) by ELISA with a set of the MAbs produced here.

Amino Acid Sequence↗

Phytotoxicity of indole-3-acetic acid produced by the fungus, Pythium aphanidermatum.

Pythium aphanidermatum causes the serious disease of Pythium red blight on bentgrass. IAA, one of the metabolites that has been isolated from this fungus, showed the same symptom of Pythium red blight on bentgrass at a concentration of 1,000 mg/1. The IAA content in the foliage of bentgrass infected by this fungus was about 200 times that of an untreated control. These results suggest that IAA produced by this fungus was the causal substance of Pythium red blight on bentgrass.

Indoleacetic Acids↗

Connective tissue-type mast cell leukemia in a dog.

An unusual case diagnosed as connective tissue-type mast cell leukemia with marked mastocyte infiltration into visceral organs in a seven-year-old female Curly-Coated retriever is presented. Acute circulatory collapse, emesis, diarrhea, abdominal enlargement, icterus, cyanosis, dyspnea, pulmonary edema, hepatomegary, ascites, and right ventricular enlargement were observed. Hematologic and biochemical examinations revealed mast cell leukemia, mature neutrophilia, monocytosis, thrombocytopenia, hemolytic hyperbilirubinemia, hyperhistaminemia, renal and hepatic injuries. Mast cells were distributed systemically, but predominantly in the diaphragm and liver with a large mass among the serosa of ileum, cecum and colon. Mast cells were stained intensely by both safranin and berberine sulfate.

Animals↗

Neospora caninum infected the alimentary tract of nude mice and was transmitted to other mice by intraperitoneal inoculation with the intestinal contents.

Neospora caninum (BT-2 strain) that originated from the brain of a Holstein calf was serially passaged through 10 generations of BALB/c nude mice by intraperitoneal inoculation. Histological examination of the mice revealed that numerous clusters of tachyzoites appeared in the pancreas, stomach and small intestine as well as in the central nervous system (CNS) and skeletal muscles. Intestinal contents of the infected mice were inoculated intraperitoneally into uninfected nude mice and 3 of the 17 inoculated mice showed clinical signs at post inoculation days 3 to 10. The present experiments demonstrated a proliferation of N. caninum tachyzoites in the mucosa of the alimentary tract and pancreas of the nude mice and the intestinal contents of the mice were infective to other nude mice.

Animals↗

Sequential observation of 2,6-dimethylaniline-induced nasal lesions in a rat two-stage nasal carcinogenesis model after initiation with N-bis(2-hydroxypropyl) nitrosamine.

Male F344 rats received diet containing 3,000 ppm 2,6-dimethylaniline (DMA) after initiation with a single subcutaneous injection of 2,400 mg/kg of N-bis(2-hydroxypropyl)nitrosamine (DHPN), and histological and electron microscopic examinations of the nasal cavity were performed at 4, 13, 26 and 52 weeks to examine sequential changes induced by DMA. Severe atrophy of Bowman's glands and epithelial disarrangement were apparent from week 4, followed by dilatation and/or proliferation of Bowman's glands, degeneration of epithelial cells, and proliferation of undifferentiated epithelial cells from week 13. Focal glandular hyperplasias, dysplastic foci, and adenomas were observed from week 26, and carcinomas at 52 week. These nasal lesions were mostly evident in the olfactory mucosa in the nasal cavity, and their severity and/or incidences, other than atrophy of Bowman's glands, increased with the treatment period. Electron microscopically, carcinoma cells demonstrated desmosomes, dense secretory granules identical to those in normal Bowman's glands, a basement membrane, and microvilli. These results suggest that Bowman's glands are the target of DMA, giving rise to nasal carcinomas after DHPN-initiation.

Adenoma↗

IgG1 is the dominant subclass of antibody against glutamic acid decarboxylase among type 1 diabetes in Japanese.

Autoantibody against glutamic acid decarboxylase (GADA) is a highly sensitive predictor of insulin-dependency in adult diabetic patients as well as young individuals. A considerable number of diabetics who do not reach the insulin-dependent stage have this antibody. Recently, type 1 diabetes has been thought to be caused by T helper 1 (Thl)-type autoimmunity based on studies in non-obese diabetic mice, but it is still difficult to investigate antigen-specific T-cell function in human type 1 diabetes. We therefore assessed an IgG subclass assay for GADA, which should reflect T-helper function against GAD. Sera from 14 type 1 diabetic patients positive for GADA by radioligand binding assay were tested for the IgG subclass of GADA. The assay was based on an enzyme-linked immunosorbent assay, which showed a good correlation with radioligand binding assay. The sera of all but one of the 14 type 1 diabetic patients (93%) were positive for the IgG1 subclass of GADA. The IgG2 and IgG3 subclasses of GADA were also detected in one diabetic patient each who were also positive for IgG1. The IgG4 subclass was not detected in any of the sera we tested. We concluded that IgG1 is the dominant subclass of GADA in Japanese type 1 diabetic patients.

Adolescent↗

Expression of alpha-smooth muscle actin and fibronectin in tubulointerstitial lesions of cats with chronic renal failure.

OBJECTIVE: To examine renal expression of alpha-smooth muscle actin (alpha-SMA) and fibronectin in cats with tubulointerstitial nephritis (TIN) for use in predicting progression to renal fibrosis. ANIMALS: 19 cats with TIN and 9 cats without nephritis. PROCEDURE: Serum creatinine and BUN concentrations were measured. Indices for glomerular extra-cellular matrix (ECM), tubular injury (TI), and fibronectin were determined in renal specimens to quantify the extent of injury and fibrotic lesions. Expression of alpha-SMA in renal tissue was immunohistochemically detected, and correlations were evaluated between the alpha-SMA index and other histologic and clinical variables. RESULTS: The alpha-SMA index in tubulointerstitial areas (1.63 +/- 0.78) was significantly higher in cats with TIN, especially in the periglomerular and peritubular areas, than in cats without nephritis (0.20 +/- 0.14). The alpha-SMA index was significantly associated with the TI index (r2 = 0.70), fibronectin index (r2 = 0.95), BUN concentration (r = 0.64), and serum creatinine concentration () = 0.66). Of special interest was that interstitial alpha-SMA expression appeared evident in the kidneys at an early stage of TIN, prior to the onset of ECM deposition. CONCLUSION AND CLINICAL RELEVANCE: Analysis of results of histologic and clinical examinations revealed that interstitial alpha-SMA expression may have clinical importance and may be a useful early histologic marker for development of chronic renal failure in cats. An immunohistochemical examination for fibrogenic molecules (such as alpha-SMA expression) may provide fundamental information on the pathogenesis of early-stage renal disease and aid clinical management of cats with chronic renal failure, including TIN.

Actins↗

Transurethral holmium laser resection of the prostate.

PURPOSE: We evaluated the efficacy of the holmium:YAG laser for transurethral endoscopic prostatectomy for benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: We treated 103 patients with BPH with holmium:YAG laser resection of the prostate. A high power holmium laser generating a maximum of 60 or 81 W. was used in a pulsed mode, applying energy directly to prostatic tissue via a forward firing 550 micron. fiber transurethrally under direct vision. Treatment outcome was evaluated by the International Prostate Symptom Score, quality of life score, maximum flow rate and post-void residual urine volume. We also compared holmium laser surgery and transurethral resection of the prostate for operative factors, such as surgical duration, bleeding volume and catheterization time. RESULTS: Average symptom score, quality of life score, peak flow rate and post-void residual significantly improved at 1 week, 1 month and 3 months postoperatively, with improvement maintained up to 36 months postoperatively in the holmium:YAG group. Intraoperative bleeding volume was significantly lower and catheterization time was significantly shorter for holmium:YAG than for transurethral prostatic resection. Use of the holmium laser caused no complications. CONCLUSIONS: Because of its effectiveness and safety holmium:YAG resection is a viable potential surgical alternative for symptomatic BPH. The holmium:YAG laser has been verified to be useful for many purposes in urology.

Aged↗

[Long-term result after UPPP for OSAS by evaluation of nocturnal oxygenation].

The nocturnal oxygenation of 45 patients with OSAS who had UPPP at our hospital were evaluated by pulse oximetry before and after surgery and at the time more than one year after UPPP. "Successful case" is defined as the case that showed more than 50% reduced oxygen desaturation index(ODI) after UPPP. Twenty seven cases including 15 severe cases(ODI > or = 40) have showed continuously successful ODI for more than one year. We could not get good result in 8 cases, which kept high ODI. Four of them are now using dental appliance. We have recognized the recurrence of sleep apnea in 10 cases that once got successful results just after UPPP. Our report strongly suggests the importance of long-term follow-up after surgery.

Circadian Rhythm↗

Protamine dissociation before decondensation of sperm nuclei during in vitro fertilization of pig oocytes.

The correlation between morphological changes and the dynamics of protamine in boar sperm chromatin during in vitro fertilization of pig oocytes matured in vitro was assessed. For this purpose, protamine was purified from boar sperm nuclei and an antiserum against protamine was developed. After affinity purification, the antiserum reacted exclusively with boar protamine during western blotting, showing no crossreactivity with core histones. Immunohistochemical evaluation revealed that only fully developed spermatid nuclei in boar testes stained strongly with the antiserum. When pig oocytes matured in vitro were fertilized in vitro, sperm penetration was observed in 37% of oocytes at 2 h after insemination and the penetration rate increased to 99% by 5 h after insemination, accompanied by an increase in polyspermic penetration. Paraffin wax sections of the inseminated oocytes were examined by immunohistochemical analysis with the antiserum. The proportion of condensed sperm nuclei that reacted with the antiserum was 87% of the sperm nuclei that penetrated by 2 h after insemination, and this decreased to 20 and 13% at 3 and 5 h after insemination, respectively. However, none of the decondensing sperm nuclei or male pronuclei reacted with the antiserum during the entire insemination period. These results indicate that a specific antiserum against boar protamine can be raised and, using this serum, it has been demonstrated that protamine is dissociated from boar sperm nuclei before decondensation during in vitro fertilization.

Analysis of Variance↗

Effects of p51/p63 missense mutations on transcriptional activities of p53 downstream gene promoters.

The p51/p63 gene is a homologue of p53, the product of which acts as a transcriptional activator by binding to p53-responsive elements in the promoter regions of several p53 downstream genes. Recently, we identified four distinct mutations in the p51/p63 gene after screening >200 human tumors and cell lines. Because all of the detected p51/p63 mutations were missense mutations, the pathogenic effect of these mutations is difficult to determine without performing a functional analysis. In this study, we examined the transcriptional activity of tumor-derived p51/p63 missense mutations using a yeast-based assay and compared the data with that of artificial p51/p63 missense mutations at residues corresponding to the positions and substituted residues of p53 mutation "hotspots." Although most of the p51/p63 missense mutations at the p53 hotspot residues were unable to transactivate the promoters used in this study, the tumor-derived p51/p63 missense mutations retained their ability to transactivate the MDM2 and/or the BAX promoter but not the p21/WAF1 promoter. These results suggest that the p51/p63 mutation might be involved in an unknown tumor suppression pathway distinct from that of p53.

Amino Acid Sequence↗

Distribution and retention of mercury in metallothionen-null mice after exposure to mercury vapor.

We studied the role of metallothionein (MT) in the distribution and retention of mercury in the brain, lung, liver and kidney of MT-null and wild-type mice after exposure to mercury (Hg0) vapor. Mice were exposed to Hg0 vapor at 5.5-6.7 mg/m3 for 3 h and killed at 1, 24, 72 or 168 h after exposure. One hour after exposure to Hg0 vapor, there were no differences in mercury concentrations in these organs from MT-null and wild-type mice. However, the elimination rate of mercury from the organs, except the brain, were remarkably faster in MT-null mice than in wild-type mice. MT-I and -II levels in the lung and kidney were increased significantly in wild-type mice but not in MT-null mice at 24 h after exposure to Hg0 vapor. At this time point, over 65% of the mercury was retained in the MT fraction of the cytosol of organs from wild-type mice. In contrast, mercury appeared mainly in the high-molecular-weight protein fractions in the cytosol of organs from MT-null mice. In the brain, a large amount of mercury was bound to MT in both strains of mice immediately after exposure. No difference was observed in the elimination rate of mercury from the brain between both strains of mice. Brain MT levels were elevated slightly in wild-type mice at 168 h after exposure but could not be detected in MT-null mice. These data suggest that no detectable MT-I and -II levels were found in the brain of MT-null mice and that mercury was apparently bound to MT-III. Using MT-null mice, we showed also that MT-III may play an important role in the retention of mercury in the brain.

Administration, Inhalation↗

Effect of lycopene on lipid peroxidation and glutathione-dependent enzymes induced by T-2 toxin in vivo.

Lycopene, obtained from fresh tomatoes, was incorporated into the chicks diet. The treatments were: (1) Control, (2) 1.5 mg T-2 toxin/kg body weight/day; (3) 25 mg lycopene/kg body weight/day, (4) 1.5 mg T-2 toxin plus 25 mg lycopene/kg body weight/day. Male broiler chicks, 7-28 days of age, were provided with feed and water ad libitum. Every 7 days, malondialdehyde (MDA) and glutathione (GSH) levels, and enzymatic activities of glutathione-S-transferase (GST), gamma-glutamyltransferase (GGT) and glutathione peroxidase (GP) were evaluated in liver homogenates. Compared to the controls after 7 days of treatment, T-2 toxin increased hepatic MDA concentration (128%). A significant consumption of endogenous antioxidant GSH (45%) was induced as well as a marked increase in hepatic enzymatic activities of GST, GGT, and GP (312, 187, and 324%, respectively). Addition of T-2 plus lycopene, at an approximate ratio of 1:17 in the diet, diminished some parameters measured (P < 0.05). Apparently lycopene participated as an antioxidant agent and also protecting the cellular level of GSH.

Animals↗

Tumor-promoting activity of 2,6-dimethylaniline in a two-stage nasal carcinogenesis model in N-bis(2-hydroxypropyl)nitrosamine-treated rats.

The potential promotion activity on nasal carcinogenesis of 2,6-dimethylaniline (DMA), an alpha2-adrenergic agonist metabolite of xylazine which is used for food-producing animals as a sedative agent, was examined. Male F344 rats received diet containing 0 or 3000 ppm DMA for 52 weeks after initiation with N-bis(2-hydroxypropyl)nitrosamine (DHPN). Histopathological assessment showed the incidence of carcinomas in the DHPN+DMA group (33%) to be significantly elevated as compared with that for the DHPN-alone group (5%). Incidences and/or multiplicity of epithelial hyperplasias and dysplastic foci were also increased in the DHPN+DMA group. These lesions were exclusively observed in the olfactory mucosa. The lowest plasma levels of DMA in tumor- and dysplastic foci-bearing rats were 0.05 and 0.20 microg/ml, respectively. These results indicate that DHPN acts as an appropriate initiator for nasal carcinogenesis and that DMA exerts a tumor-promoting effect on the olfactory mucosa in the rat nasal cavity.

Adenoma↗

The transcriptional activities of p53 and its homologue p51/p63: similarities and differences.

p51/p63 is a novel p53 homologue that has been shown to act as a transcriptional activator through the p53-binding sequence of the p21/WAF1 promoter and to induce apoptosis when it is expressed transiently in a human tumor cell line. We developed transcription assay systems for these two related genes in both Saccharomyces cerevisiae and mammalian cells and used them to investigate the functional similarities and differences of these genes. We found that p51/p63 trans-activated the previously identified p53 target genes, but the degree of the transactivation by p51/p63 differed from that by p53. These results suggest that the cellular signal on p51/p63 cross-talks partially but not completely with that of the p53 pathway.

Cyclin-Dependent Kinase Inhibitor p21↗