Search PubMed⌕ Search

Biomedical subjects

N Hirayama

Publications and source records attributed to N Hirayama.

At least 73 records · Page 4Linked to original sources

The biological characterization of field isolates of canine distemper virus from Japan.

Eight isolates of canine distemper virus (CDV) were obtained from seven dogs suffering from distemper by co-cultivation of their mononuclear cells with a marmoset B lymphoblastoid cell line, B95a. Six of the seven dogs had received one or more vaccinations. All of the isolates readily proliferated in B95a cells, but were not completely neutralized by anti-CDV canine plasma, which had high neutralizing activity against the Onderstepoort laboratory strain of CDV. Furthermore, different reactivities of monoclonal antibodies (MAbs) against CDV were observed between the field isolates and laboratory or vaccine strains of CDV in immunofluorescence studies. Immunoprecipitation analysis using MAbs detected the haemagglutinin protein of each new field isolate as 69K, 75K and 155K forms, and the fusion protein as 64K and 65K forms; the corresponding proteins of the Onderstepoort strain were detected as 75K and 61K proteins respectively. It is apparent from these results that the new field isolates of CDV have very different antigenic properties from the Onderstepoort vaccine strain.

Animals↗

Genetic profiles of 12 inbred rat strains for 46 microsatellite loci selected as genetic monitoring markers.

Genetic profiles for 46 microsatellite loci of 12 inbred strains of rats, including 2 congenic strains and a coisogenic strain, have been demonstrated. Rates of loci with different alleles between 2 inbred strains, which are not closely related to each other in origin, were from 71.7% between ACI/N and IS/Kyo strains to 41.3% between F344/N and TM/Kyo. On the other hand, the rates were 0% in both of 2 sets of congenic strains; between F344/N and F344/N-rnu, or between BN/fMaiKyo and BN.IS. When WTC/Kyo and the coisogenic strain TRM/Kyo (WTC/Kyo-tm) were compared for 115 microsatellite loci, no loci with different alleles between the strains were found. The 46 loci should be useful as genetic monitoring markers, since all of the primer pairs generate distinct PCR-products at a fixed annealing temperature of 55 degrees C.

Alleles↗

[The status of extrapulmonary tuberculosis in community hospital].

Fifty patients with extrapulmonary tuberculosis were diagnosed at 4 community hospitals in Tokyo during 1981 to 1992. The percentage of extrapulmonary tuberculosis among the all types of tuberculosis was 10.5% as a whole, and 16% in females and 7.9% in males. The number of patients older than fifth generation was about two times more than that of younger generations. And the patients were more in males than in females under the forties, but were more in females than in males over the sixties. On chest radiogram, there was no pathological findings in 52% of the patients and active pulmonary tuberculosis was detected in 32% of the patients. The majority of the extrapulmonary tuberculosis (30 cases, 60%) was of lymph nodes, especially of cervical lymph nodes, and the remaining were 10 cases of tuberculosis in bone and joint, 4 each in intestine and pericardium, 2 each in liver, peritoneum, kidney, urinary bladder, epididymis and skin. Among 30 cases with tuberculous lymphadenopathy, the disease were observed in cervical lymph nodes by 19 cases, in pulmonary hilar and axillary node by each 4 cases and in abdominal cavity by 3 cases.

Adult↗

Re-evaluation of solid-phase adsorption and desorption techniques for isolation of trace organic pollutants from chlorinated water.

Amberlite XAD resin and activated carbon columns were tested for their abilities to concentrate trace organic pollutants in chlorinated water. Both XAD-2 and XAD-7 resin columns (20 ml) were capable of adsorbing about 30% of total organic halogen (TOX) present in 20 l of drinking water (pH 7) containing about 100 micrograms/l of TOX, whereas the carbon column (10 ml) adsorbed over 90% of TOX. The adsorption capacity of XAD-7 resin was found to be strongly dependent on the solution pH, as compared with those of XAD-2 and carbon adsorbents. Soxhlet and sonication extractions were also evaluated for their abilities to recover the adsorbed organics from the adsorbents, by measurements of TOX, chromatographable compounds and mutagenicity in the eluates. Soxhlet extraction gave higher recoveries than sonication, as measured with the above indices, but these differences were generally small (ca. 20%), with exception of the carbon extracts. The XAD-2 and XAD-7 extracts of drinking water also showed about 3-4 times higher mutagenic activity than the carbon extracts.

Acrylic Resins↗

Structure-activity relationships for mitomycins. Application of the distance and charge analysis method.

Molecular orbital calculations based on coordinates from X-ray analysis have been performed for a set of 24 mitomycins, of which eight compounds have not been isolated so far. To prioritize the order of synthesis of these missing compounds, a new method named DISCA (distance and charge analysis) has been developed. DISCA screens correlations between spatial distribution of charge in molecules and their biological activity. The spatial distribution of charge is represented by several indexes in DISCA. LD50 and ED50 were used as measures of biological activity. DISCA has successfully extracted indexes which have significantly high correlation coefficients. The indexes with the highest correlation coefficient were common to both LD50 and ED50. By use of the correlation functions with high correlation coefficients DISCA has predicted that 9-epi-1a-N-demethylmitomycin D should have the best ED50 and a modest LD50 among the missing mitomycins.

Mitomycins↗

Purification of hemagglutinin from Haemophilus paragallinarum using monoclonal antibody.

The purification of hemagglutinin from Haemophilus paragallinarum was attempted using affinity chromatography with monoclonal antibody. The antigen eluted from the affinity column using potassium thiocyanate buffer agglutinated chicken erythrocytes. In immunoblotting of the eluted antigen, a single band with monoclonal antibody was found as well as the crude antigen. When the chickens were immunized with the eluted antigen, they produced the hemagglutination inhibition (HI) antibody, and they showed protection against challenged exposure with H. paragallinarum strain 221. These results indicated that the HA antigen of H. paragallinarum was a protective antigen.

Animals↗

Antigen-specific proliferative response of peritoneal exudate lymphocytes primed with antigen and bacterial lipopolysaccharide: the roles of Ia+ accessory cells and IL-2.

In vitro antigen-specific proliferation was investigated in a lymphocyte population that had been taken from the peritoneal exudate cells (PEC) of C3H/HeN mice (Iak) primed in vivo with both bacterial lipopolysaccharide (LPS) and horse red blood cells (HRBC) and had been purified by passage through a nylon fiber column (Nfc). The proliferative response of the Nfc-passed lymphocytes primed with HRBC and LPS [T(HRBC+LPS) cells] depended on the dose of antigen in the cultures, and the response was higher than that of cells prepared from mice primed with HRBC alone [T(HRBC) cells]. No response was seen in the cells prepared from the LPS-primed mice [T(LPS) cells] or normal mice [T(N) cells]. The response of the T(HRBC) cells was abolished by previous treatment of the cells with anti-Iak antibody and complement (C), whereas the response of the T(HRBC+LPS) cells was retained after the same treatment, indicating that the Ia- T(HRBC+LPS) cells can proliferate in response to antigen in spite of Ia+ accessory cell-depletion. Supernatants from the cultures of Ia- T(HRBC+LPS) cells in the presence of HRBC showed abundant IL-2 activity, while those of Ia- T(HRBC) cells did not. The IL-2 should be produced by the L3T4 cell population in T(HRBC+LPS) cells in response to antigen, since the previous treatment of the cells with anti-L3T4 antibody and C abrogated the production.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Beta 0-thalassemia trait (IVS-I-1 G-->T) in a Japanese family.

Three subjects in a family with microcytic and hypochromic anemia were studied; red blood cell morphology indicated aniso-poikilocytosis and hypochromasia. Target and tear-drop cells were also noted. In all three cases evaluated, there was an increase in HbA2 levels and a decline in the beta/alpha synthesis ratio. Direct cloning and DNA sequencing identified a point mutation (G-->T) at position 1 of intervening sequence I. The resulting reduction of beta-globin chain synthesis is considered to give rise to beta 0-thalassemia phenotype. This point mutation is to our knowledge, the first case in Japan.

Base Sequence↗

[Two cases of M. avium complex infection presenting as solitary pulmonary nodule].

Case 1 was a 75-year-old woman. Her chest X-ray showed a 3 cm solitary nodule in the right S4, and biopsy obtained by fiberoptic bronchoscopy revealed epithelioid cell granuloma. Four weeks later, Mycobacterium avium complex (MAC) was detected on culture of the bronchial lavage fluid. She was treated with INH, RFP and EB for 12 months and recovered with no relapse. Case 2 was a 75-year-old man with mild diabetes mellitus and hypertension. Chest X-ray showed a 4 cm solitary nodule in the right S9. Percutaneous pulmonary aspiration revealed numerous acid-fast bacilli on smear, and he was treated with INH, RFP and EB. Culture proved M. avium complex. The patient was unable to continue his medications because of gastrointestinal side effects, so right lower lobectomy was performed. Cases of M. avium complex infection presenting as solitary pulmonary nodule are very rare. We found only 12 reported cases in Japan and in other countries. The most usual site of cavitary lesions in M. avium complex infections is both upper lobes, in contrast to the relatively even distribution of solitary pulmonary nodules of the 14 cases (12 reported cases and our 2 cases).

Aged↗

Synthesis and antiallergic activity of 11-(aminoalkylidene)-6,11-dihydrodibenz[b,e]oxepin derivatives.

A new series of 11-substituted 6,11-dihydrodibenz[b,e]oxepin-2-carboxylic acid derivatives was synthesized and demonstrated to be orally active antiallergic agents. These compounds are structurally related to 1 (KW-4994), which we had reported previously to be a new antiallergic agent. Most compounds synthesized exhibited potent inhibitory effects on 48-h homologous passive cutaneous anaphylaxis (PCA) in rats and on IgG1-mediated bronchoconstriction in guinea pigs. Additionally, compounds possessing a terminal carboxyl group at the 2-position of the dibenz[b,e]oxepin ring system exhibited inhibitory effects on specific [3H]pyrilamine binding to guinea pig cerebellum histamine H1 receptors, whereas these demonstrated negligible effects on specific [3H]QNB binding to rat striatum muscarinic acetylcholine M1 receptors. Structure-activity relationship studies revealed that the following key elements were required for enhanced antiallergic activities: (1) a 3-(dimethylamino)propylidene group as the side chain at the 11-position, (2) a terminal carboxyl moiety at the 2-position, and (3) a dibenzoxepin ring system. Among the compounds synthesized, (Z)-11-[3-(dimethylamino)propylidene]-6,11-dihydrodibenz [b,e]oxepin-2-acetic acid hydrochloride (16) was selected for further evaluation. It had an ED50 value of 0.049 mg/kg po in the PCA test in rats and an ID50 value of 0.030 mg/kg po in inhibiting anaphylactic bronchoconstriction in guinea pigs. Furthermore, it had a Ki value of 16 +/- 0.35 nM for the histamine H1 receptor, while it exhibited negligible CNS side effects up to a dose of 600 mg/kg po. Compound 16 is now under clinical evaluation as KW-4679.

Animals↗

Horseshoe kidney and membranous glomerulonephritis with cold activation of complement.

A 48-year-old woman was admitted to our hospital because of proteinuria associated with persistent hypocomplementemia. Intravenous pyelography indicated the presence of horseshoe kidney without other abnormalities. Hypocomplementemia was caused by cold activation of complement. There were some findings suggestive of chronic liver disease (positive HCV antibody, hypergammaglobulinemia, low cholinesterase, etc.). Percutaneous renal biopsy showed the features of multiple evolutional phases of membranous glomerulonephritis.

Cold Temperature↗

Production, characterization and protective effect of monoclonal antibodies to Haemophilus paragallinarum serotype A.

Monoclonal antibodies (mAbs) to Haemophilus paragallinarum serotype A were obtained by fusion of murine myeloma cells (P3-X63-Ag8-U1) and spleen cells from BALB/c mice immunized with whole cells of strain 221. Enzyme linked immunosorbent assay with whole cells was used to show that the monoclonal antibodies are specific for serotype A of H. paragallinarum. Four monoclonal antibodies indicated hemagglutination-inhibition (HAI) activity against serotype A; their titers were 10(4)-10(5). By western blotting, two of these monoclonal antibodies reacted with a protein of molecular weight 39,000. Chickens treated with mAbs possessing HAI activity survived without clinical signs of infection. No challenge strain was isolated from these chickens, indicating that four mAbs with HAI activity suppressed growth of the challenge strain in the nasal cavity, whereas mAbs without HAI activity showed no passive protective effect. These results demonstrated that HI antibodies contributed to protection, and strongly suggest that hemagglutinin (HA) antigen, especially the epitopes which were recognized by these mAbs are important for protective immunity in chickens.

Animals↗

Protective effects of monoclonal antibodies against lethal canine distemper virus infection in mice.

Monoclonal antibodies (MAbs) against the haemagglutinin (H), fusion protein (F) and nucleoprotein of canine distemper virus (CDV) were examined for their ability to protect mice against lethal CDV infection. One MAb against H and two of six MAbs against F protected mice, the protective effect of the anti-H MAb being stronger than that of the anti-F MAbs. The anti-H MAb showed virus neutralizing activity, but the two anti-F MAbs, which recognized the same epitope, did not. Protection by the anti-F MAbs correlated with cell fusion inhibition, but not with complement-dependent neutralization, complement-dependent cytolysis or antibody-dependent cell-mediated cytotoxicity. These results suggest that neutralization by antibody against H and cell fusion inhibition by antibody against F play important roles in the protective mechanism against CDV infection.

Animals↗

Antigenic and functional characterization of rinderpest virus envelope proteins using monoclonal antibodies.

A total of 24 monoclonal antibodies (MAbs) against the haemagglutinin (H) and the fusion protein (F) of rinderpest virus (RPV) were used to characterize their antigenic structure and biological properties, and to analyse natural variation in the envelope proteins of morbilliviruses. The anti-H and anti-F MAbs defined seven and three distinct antigenic sites, respectively. The MAbs to six sites on H were able to neutralize the infectivity of RPV. The addition of guinea-pig complement or anti-mouse immunoglobulin increased the virus-neutralizing antibody titre of most of the anti-H MAbs, including those lacking neutralizing activity. One of the antigenic sites on H was conserved among morbilliviruses and the MAbs to this site had haemagglutination inhibition activity against measles virus (MV). The remaining sites were specific for RPV and varied antigenically between strains of RPV. The anti-F MAbs lacked neutralizing activity, but two of the five MAbs did show activity in the presence of complement or anti-mouse immunoglobulin. On the whole, the antigenic sites on F were conserved in some strains of MV, but not in canine distemper virus. All of the sites on the surface proteins were sensitive to SDS and, although those on F were not affected by 2-mercaptoethanol, five of the seven sites on H were destroyed by it. These results suggest that the epitopes on the envelope proteins are conformation-dependent.

Animals↗

Structure and conformation of a novel inhibitor of angiotensin I converting enzyme--a tripeptide containing phosphonic acid.

A novel phosphotripeptide, (IR)-1-(N-(N-acetyl-L-isoleucyl)-L-tyrosyl)amino-2-(4-hydroxyphenyl)ethy l- phosphonic acid, is a potent inhibitor of angiotensin I converting enzyme (ACE). ACE inhibitory activity in vitro of the peptide is comparable to that of captopril. Its diethylester (C29H42N3O8P, molecular weight, 591.6) crystallizes in the monoclinic space group C2, with cell constants: a = 25.666(9), b = 9.590(8), c = 13.557(2)A, beta = 91.65(2) degrees, Z = 4, Dc = 1.17 g/cm3. The structure was solved by MULTAN 11/82 and refined by full matrix least-squares methods to a final R-factor of 0.063 for 2123 unique reflections (F greater than 3 sigma(F] measured on an Enraf-Nonius CAD-4 diffractometer (CuK alpha, lambda = 1.541 8 A, T = 295 K). The absolute configuration of the alpha-carbon where the phosphonic acid is attached was determined unequivocally by referring to the L-isoleucyl moiety whose absolute configuration is known. The conformation of the molecule is relatively rigid owing to the intramolecular requisites and the resultant relative disposition of hetero atoms, which are necessary to its biological activities, are confined to the corresponding disposition in captopril.

Amino Acid Sequence↗

Survey of infectious coryza of chickens in Indonesia.

A survey of infectious coryza of chickens was performed in West Jawa of Indonesia between 1987 to 1988 by the detection of hemagglutination-inhibition (HI) antibody against Haemophilus paragallinarum (Hpg) in non-vaccinated healthy laying (12 farms, 196 chickens) and native (8 farms, 197 chickens) chickens. HI antibodies against Hpg were detected not only in the native chickens but also in the laying chickens, regardless of the district, and were observed in 70% (14/20) of farms and 19% (73/393) of chickens. HI antibodies against serotype A were detected from 11 farms (55%) and 11% (45/393) in chickens. Those against serotype C were detected from 5 farms (25%) and 8% (30/393) in chickens. Three Hpg strains were isolated from different chickens affected with infectious coryza. Two of them were identified as type A and the other as type C by the rapid plate agglutination test. These results demonstrated that the outbreaks of infectious coryza caused by serotype A and C strains had occurred in Indonesia.

Animals↗

Expression of hemagglutinin of Haemophilus paragallinarum serotype A in Escherichia coli.

The genomic DNA of Haemophilus paragallinarum (Hpg) serotype A strain 221 was cloned into vector plasmid pBR322. The recombinant plasmids were introduced into Escherichia coli (E. coli) strain C600. Subsequently, a total of 277 transformants were obtained. One, designated strain no. 6, expressed hemagglutination activity against chicken erythrocytes. Strain no. 6 contained the recombinant plasmid pNV102, and DNA of about 2.57 kb was inserted into pNV102. When strain no. 6 was cured of pNV102, the strain lost hemagglutination activity. When the cured strain was retransformed with pNV102, hemagglutination activity was restored. E. coli strain no. 6 reacted with monoclonal antibody specific to the hemagglutinin of Hpg serotype A in a dot-blotting analysis. Chickens immunized with the inactivated strain no. 6 produced the hemagglutination inhibition (HI) antibody, and chickens possessing the HI antibody showed protection against challenge exposure by Hpg strain 221.

Animals↗