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N Hanada

Publications and source records attributed to N Hanada.

At least 91 records · Page 5Linked to original sources

Immunological properties of the primer-independent glucosyltransferase of Streptococcus mutans serotypes d and g.

Streptococcus mutans serotype g secretes at least three kinds of glucosyltransferase with different enzymological and immunological properties. One of them is a primer-independent enzyme and seems to be the source of primer for the others, both of which are primer-dependent enzymes. Recently, we purified the primer-independent enzyme, the third glucosyltransferase in this group from S. mutans strain AHT-k serotype g. In the present study, we examined the specificity of the antiserum against the primer-independent glucosyltransferase using extracellular culture-conditioned fluids of many strains of the various serotypes of S. mutans. The antiserum cross-reacted with the extracellular culture fluids from strains of serotypes d and a, in addition to serotype g, but not with those of other serotypes, indicating that the primer-independent glucosyltransferase is secreted by the S. sobrinus and S. cricetus, but not by S. mutans and S. rattus. The antiserum did not completely inhibit the activity of the enzyme, even at more than twofold antibody excess, determined by indirect precipitation with immobilized staphylococcal protein A.

Animals↗

Isolation and characterization of the Streptococcus mutans gtfC gene, coding for synthesis of both soluble and insoluble glucans.

The intact gtfC gene from Streptococcus mutans GS-5 was isolated in Escherichia coli in plasmid vector pUC18. The glucosyltransferase activity expressed by the gene synthesized both low-molecular-weight water-soluble glucan and insoluble glucan in a primer-independent manner. Purification of the enzyme by procedures that minimize proteolytic digestion yielded a purified preparation with a molecular weight of 140,000. Insertional inactivation of the gtfC gene with a streptococcal erythromycin resistance gene fragment followed by transformation of strain GS-5 suggested that the gtfC gene product was required for sucrose-dependent colonization in vitro. In addition, evidence for the presence of a third gtf gene coding for soluble glucan synthesis was obtained following the construction of mutants containing deletions of both the gtfB and gtfC genes.

Chromosome Mapping↗

Non-invasive method for early diagnosis of herpes simplex encephalitis.

For the early diagnosis of herpes simplex encephalitis IgG and IgM antibodies to herpes simplex virus in cerebrospinal fluid were measured by an enzyme linked immunosorbent assay (ELISA) and a local production index was calculated. Using these three criteria, 31 cases of various neurological illnesses were analysed. All eight cases of herpes simplex encephalitis were diagnosed correctly in the acute phase, and there were no false positive results.

Adolescent↗

Antigenic characterization of the internal proteins of Newcastle disease virus by monoclonal antibodies.

We have prepared and characterized monoclonal antibodies against the three internal structural proteins, M, P and NP, of Newcastle disease virus. At least two non-overlapping antigenic sites were delineated on the M protein, four on the P, and two on the NP by competitive binding assay. One of the two non-overlapping antigenic sites on the M protein was found to be a cluster of at least three distinct epitopes. Enhancement of antibody binding by the binding of a second antibody was observed with the M protein. The reactivity of these monoclonal antibodies with heterologous strains was studied by enzyme-linked immunosorbent assay. The results indicated that there are both highly conserved antigenic sites and those subject to remarkable change on both M and P proteins. On the other hand, NP appeared to be antigenically more stable.

Antibodies, Monoclonal↗

Purification and characterization of a third glucosyltransferase from Streptococcus mutans serotype g.

Streptococcus mutans strain AHT (serotype g) secretes at least two glucosyltransferases with different pI values. A novel glucosyltransferase with a pI of 5.8 was purified 244-fold from the ammonium sulphate fraction by DEAE-cellulose chromatography, FPLC (Mono Q column, Pharmacia) and hydrophobic chromatography. The enzyme preparation gave a single protein band on analysis by both PAGE and SDS-PAGE, and did not form multiple protein bands detectable by IEF. The Mr was estimated to be about 130,000 by SDS-PAGE and about 135,000 by ultracentrifugal analysis. The apparent Km value and pH optimum of the enzyme were 3.9 +/- 0.2 mM (mean +/- SD) and about 4.7, respectively. The enzyme synthesized water-soluble glucan from sucrose, and the glucan consisted of over 90 mol% 1,6-alpha-D-glucosidic linkages. The enzyme activity was not stimulated by primer dextran. Anti-enzyme serum produced a single precipitin band with the purified enzyme preparation, whereas it did not react with either of the other two known glucosyltransferases.

Chromatography, DEAE-Cellulose↗

Local production of rotavirus specific IgA in breast tissue and transfer to neonates.

Rotavirus specific IgA, secretory component, and IgG were measured by enzyme linked immunosorbent assay in 20 pairs of mothers and babies to estimate antibody transfer from the mother, particularly from breast milk to neonatal faeces. Colostrum contained high titres of specific IgA and secretory component, which decreased gradually. Faeces after breast feeding for three days showed detectable titres of IgA and secretory component, with further increases by seven days. There was a positive correlation between titres of secretory component in breast milk and in faeces. To clarify the mechanism of high anti-rotavirus activity in breast milk, ratios of rotavirus specific IgA in maternal serum samples to breast milk were calculated and compared with those that were herpes simplex virus specific. Significantly higher concentrations were obtained for the herpes simplex virus specific samples, indicating that anti-rotavirus IgA is selectively produced in breast tissue.

Antibodies, Viral↗

Interferon-mediated self-limiting growth of respiratory syncytial virus in mouse embryo cells.

The growth of respiratory syncytial (RS) virus in primary mouse embryo (ME) cells was investigated. The virus yields in ME cells were markedly lower if compared with those in HEp-2 cells, which are fully permissive for RS virus, and a remarkable production of interferon (IFN) was found in the early period of infection of the former cells. The virus yields in ME cells were enhanced when antimouse IFN serum was added to the medium. Indirect immunofluorescent staining of infected ME cells showed that the infection spread in the entire monolayer in the presence of antiserum, whereas it was markedly restricted throughout in the absence of the serum. All the major viral polypeptides were synthesized in the absence of the serum. However, their synthesis rates were greatly enhanced if the antiserum was added. These results suggest that the virus growth in ME cells is self-limiting and that this limited growth is due to autointerference by endogenously produced IFN during the course of infection. Further, this type of growth restriction of RS virus appears to be characteristic of cells of mouse origin by comparative studies that used other cells of different origin.

Animals↗

A long-term follow-up study on the efficacy of further attenuated live measles vaccine, Biken CAM vaccine.

Antibody persistence was measured in 39 children in an open community 12-13 years after immunization against measles with further attenuated live vaccine, Biken CAM. Serum samples of the children taken every two or three years after vaccination had higher, lower, or the same HI antibody titers as those in samples taken 6 weeks after vaccination. These differences reflected a decrease in the titer in some children and subclinical natural reinfection in others. However, all the children still retained detectable antibody in 12 or 13 years after vaccination, indicating long-term persistence of immunity after immunization with Biken CAM vaccine. For evaluation of the protective efficacy of the vaccine, matched controls were studied during the same period. Serological examination revealed that 97.5% of the controls were infected with measles and contracted the disease. In contrast, none of the vaccinees developed clinical infection after close contact with measles patients.

Adolescent↗

pH change in artificial dental plaque formed by glucosyltransferase and some oral bacteria during batch and continuous culture.

Streptococcus mutans alone or cell-free glucosyltransferase (G-Tase)-together with either Streptococcus sanguis, Streptococcus salivarius, Actinomyces viscosus, or Lactobacillus casei cells-formed artificial dental plaque that firmly adhered to glass electrodes in a continuous culture system containing sucrose. The pH in these artificial plaque samples decreased more than did that of the surrounding medium. In the absence of GTase, the bacteria other than S. mutans did not form firmly-adhering plaque on glass electrodes. The pH of the plaque formed with GTase alone did not show the pH decrease seen when the plaque contained bacteria, but, because it catalyzed the synthesis of glucan, it is suggested that the glucan acts as a diffusion barrier to retard acid loss from plaque containing acid-producing bacteria.

Actinomyces↗

Enzymatic and immunological comparison of alanine: glyoxylate aminotransferases from different fish and mammalian livers.

Alanine:glyoxylate aminotransferase was highly purified and characterized from mackerel liver. The purified enzyme had a mol. wt of approx. 200,000 with four identical subunits. It was specific for L-alanine and L-serine with glyoxylate and for L-serine with pyruvate as amino acceptor. The mackerel enzyme was similar to partially purified alanine:glyoxylate aminotransferases from other fish (sardine, gopher gray rock cod and yellow mackerel) liver with respect to mol. wts and substrate specificity. These fish enzymes were similar to mammalian liver alanine:glyoxylate aminotransferases 1 in substrate specificity and to mammalian alanine:glyoxylate aminotransferases 2 in mol. wts. An immunological cross-reactivity of hepatic alanine:glyoxylate aminotransferase was observed between mackerel and other fishes but not between mackerel and mammals.

Alanine Transaminase↗

The relationship between oral health status and biological and psychosocial function in the bedridden elderly.

The present study was conducted in order to determine what item of biological and psychosocial function is related to oral health status in the bedridden elderly. The subjects were 94 elderly individuals (30 males, 64 females) who had been admitted to a nursing home in Chiba city, Japan. We assessed the number of remaining teeth and the number of functional teeth as oral health status variables. Biological and psychosocial function levels were determined using the functional independence measure method developed by the State University of New York at Buffalo. More than 70% of subjects had less than ten remaining teeth. Almost all subjects needed prosthesis treatment. More than 50% of subjects had 27 or less functional teeth. Mann-Whitney U-test and logistic regression models showed that 'expression' was concerned with the number of remaining teeth and 'bladder management', 'locomotion', 'transfers' were related to the number of functional teeth. These data suggest close relation between oral health status and biological and psychosocial function levels in the bedridden elderly.

Journal Article↗

Inhibitory effects of MoAbs against a surface protein antigen in real-time adherence in vitro and recolonization in vivo of Streptococcus mutans.

A surface protein antigen (PAc) of Streptococcus mutans, particularly the A-region of the molecule, has been reported to interact with salivary components on the tooth surface. It might be a candidate antigen inducing the production of antibodies against the adherence of S. mutans to the tooth surface. We investigated the effects of monoclonal antibodies (MoAbs) obtained by immunization of synthetic PAc peptides that completely correspond to the amino acid sequence of part of the A-region. These MoAbs recognize several core B-cell epitopes in the sequence. Two (KH5 and SH2) of these antibodies reacted with both S. mutans and Streptococcus sobrinus, but not with Streptococcus sanguis, Streptococcus salivarius, Porphyromonas gingivalis or Lactobacillus casei. They clearly inhibited the real-time adherence of S. mutans to salivary components in a biosensor. KH5, which showed a real-time inhibition (71%), also significantly prevented the recolonization of S. mutans on the tooth surface in rats. These results suggested that the core B-cell epitope (-Y---L--Y----) recognized by KH5 was the essential sequence in the antigenic epitopes of PAc protein recognized specifically by the inhibitory antibody. Therefore, the amino acid residues were found to be important in the initial attachment of S. mutans to the tooth surface. These results provide for the mechanism of PAc molecule in the initial attachment of S. mutans on the tooth surface and more effective designs for the removal of S. mutans and S. sobrinus from the oral cavity.

Adhesins, Bacterial↗