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Biomedical subjects

Nobuhiro Hanada

Publications and source records attributed to Nobuhiro Hanada.

34 records · Page 2Linked to original sources

Establishment of an animal model using recombinant NOD.B10.D2 mice to study initial adhesion of oral streptococci.

An oral biofilm is a community of surface-attached microorganisms that coats the oral cavity, including the teeth, and provides a protective reservoir for oral microbial pathogens, which are the primary cause of persistent and chronic infectious diseases in patients with dry mouth or Sjögren's syndrome (SS). The purpose of this study was to establish an animal model for studying the initial adhesion of oral streptococci that cause biofilm formation in patients with dry mouth and SS in an attempt to decrease the influence of cariogenic organisms and their substrates. In nonobese diabetogenic (NOD) mice that spontaneously develop insulin-dependent diabetes mellitus (IDDM) and SS, we replaced major histocompatibility complex (MHC) class II (A(g7) E(g7)) and class I D(b) with MHC class II (A(d) E(d)) and class I D(d) from nondiabetic B10.D2 mice to produce an animal model that inhibited IDDM without affecting SS. The adhesion of oral streptococci, including Streptococcus mutans, onto tooth surfaces was then investigated and quantified in homologous recombinant N5 (NOD.B10.D2) and N9 (NOD.B10.D2) mice. We found that a higher number of oral streptococci adhered to the tooth surfaces of N5 (NOD.B10.D2) and N9 (NOD.B10.D2) mice than to those of the control C57BL/6 and B10.D2 mice. On the basis of our observation, we concluded that these mouse models might be useful as animal models of dry mouth and SS for in vivo biological studies of oral biofilm formation on the tooth surfaces.

Animals↗

Characteristics and willingness of patients to pay for regular dental check-ups in Japan.

The purpose of this survey was to investigate the relationship between demographic characteristics and willingness of patients to pay for regular dental check-ups in Japan. Questionnaires were distributed at private dental offices and collected via postage-paid return envelopes addressed to the center of the study groups. Questions focused on demographics and willingness to pay for regular check-ups. Five thousand one hundred thirty-two questionnaires were collected (response rate 56.8%). The 3 groups most likely to have regular dental check-ups were found to be the under 20s, 50 to 59 year olds and civil servants. Of these groups, civil servants were found to be the most likely of all to have regular check-ups. More females than males were represented in the sample. More than 60% of the patients responded that they would be willing to pay for regular check-ups if the cost were less than 2,000 yen (about 20 dollars). However, no statistically significant differences were observed in relation to household income. The results suggested that participation in regular dental check-ups might be related to gender and age, but not to household income.

Adolescent↗

Feasibility of eradication of mutans streptococci from oral cavities.

OBJECTIVES: Dental caries prevention programs using chlorhexidine (CHX) have been proposed, but CHX's effect in reducing levels of mutans streptococci (S. mutans and S. sobrinus) appears to last for only a few months. The aim of this study was to attempt to eradicate mutans streptococci from the oral cavity using intensive professional mechanical tooth cleaning (PMTC) and topical application of CHX in custom-made trays. METHODS: Seven adult dentate subjects participated in this study (mean age 53.7+/-5.6, age range 46 to 62, mean DMFT, 9.1+/-4.2). For each subject, PMTC was carried out eight times within ten days. After each PMTC, 1% CHX was applied twice to the tooth surface using custom-made trays. In addition, as home treatment, subjects were required to carry out tooth brushing three times a day, and apply 0.2% CHX in custom trays after brushing in the morning and evening. In addition, subjects rinsed with 0.2% CHX solution after lunch. Salivary levels of mutans streptococci were evaluated using Dentocult-SM at baseline and on days 9, 20, 70, 120. RESULTS: Mutans streptococci were eradicated by day 120 from 4 of the 7 seven subjects participating in this study. Those 3 subjects still harboring mutans streptococci exhibited deep periodontal pocketing. CONCLUSIONS: Eradication of mutans streptococci from the oral cavity is feasible using a combination of CHX application in custom-made trays and intensive PMTC.

Aged↗

Correlation study on oral health and electrocardiogram abnormalities.

The purpose of this study was to investigate the association between periodontal conditions and electrocardiogram test results that were obtained to screen for coronary heart disease risk factors. The present study included a total of 578 subjects who underwent annual medical check-ups at the Total Health Care Center in Otsu, Shiga Prefecture, Japan. To calculate the odds ratios for the electromyography abnormalities, we performed a logistic regression analysis for the oral examination, electrocardiogram, and blood analysis data. The crude odds ratio was obtained by a logistic regression analysis of age, sex, number of missing teeth, number of filled teeth, simplified oral hygiene index, community periodontal index, and blood analysis factors and results indicated there was a statistically significant correlation with the prevalence of electrocardiogram abnormalities. However, electrocardiogram abnormalities have a strong correlation with demographic factors such as sex and age. Therefore the experimental factors representing oral status were reexamined after the odds ratios were adjusted for age and sex. As a result of this adjustment, the new odds ratios that were determined indicated that there were no correlations between the oral factors and the prevalence of electrocardiogram abnormalities.

Adult↗

RGD motif enhances immunogenicity and adjuvanicity of peptide antigens following intranasal immunization.

The use of peptides for various aspects of medical science has been a significant advance. Peptide-based vaccines are promising, but weak immunogenic potency is impeding the clinical application. We have remarkably enhanced the immunogenicity of peptide antigens by addition of motifs that bind to cell attachment proteins, such as arginine-glysine-aspartate (RGD), to the amino acid sequence. The modified peptides induced antigen-specific serum antibodies by intranasal immunization without adjuvants. RGD, an integrin-binding motif was the strongest, among several molecules tested in this experiment, giving an average of 10 times enhancement of antibody titers when incorporated into several peptide antigens. The peptides also acted as an efficient adjuvant following the intranasal immunization with protein antigens. Our data support the feasibility of developing peptide vaccines and peptide adjuvants for intranasal vaccination.

Administration, Intranasal↗

Assessment of oral transmission using cell-free human immunodeficiency virus-1 in mice reconstituted with human peripheral blood leucocyte.

Oral-genital contact is one of the risk factors for the transmission of human immunodeficiency virus (HIV) in adults. In recent reports, oral exposure to simian immunodeficiency virus (SIV) was found to have important implications for the achievement of mucosal transmission of HIV in a rhesus monkey animal model. In the present study, we aimed first to establish a small animal model which did not develop tonsils suitable for HIV oral mucosa transmission, using non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice and NOD/SCID B2m(null) mice grafted with human peripheral blood leucocytes (hu-PBL) and stimulated with interleukin (IL)-4, and second to investigate whether oral exposure to cell-free R5 and X4 HIV-1 could lead to oral transmission of HIV through intact or traumatized mucosal tissues in humanized mice. Both low and high concentrations of cell-free R5 and X4 viruses failed to cause oral transmission with or without trauma in hu-PBL-NOD/SCID and NOD/SCID Beta2m(null) mice, which presented a number of CD4+ cells in gingival tissues and oral cavities with or without tissue injury. The present results show that IL-4-administrated NOD/SCID B2m(null) mice are useful as a small-humanized model for the study of HIV oral infection, which may help to define the window of opportunity for oral transmission by the HIV virus in animal model experiments.

Animals↗

Different frequencies of Streptococcus anginosus infection in oral cancer and esophageal cancer.

Multiple cancers frequently occur in the upper aerodigestive tract. The high incidence rate of multiple carcinomas in this region is often explained in terms of involvement of the same underlying risk factors. It has been reported that the oral bacterium Streptococcus anginosus (S. anginosus) is associated with esophageal, gastric, and pharyngeal cancer tissues. In this study, a highly specific quantification method for S. anginosus DNA using real-time PCR was established. We employed this assay to determine whether S. anginosus is also associated with oral cancer tissues. This precise quantification method revealed different degrees of infection with S. anginosus in esophageal cancer and oral cancer. We assayed 10 ng of genomic DNA from cancer tissues, and found that eight of 18 samples (44%) from the esophagus contained a detectable level (>10 fg) of S. anginosus DNA, whereas this was the case for only five of 38 samples (13%) of oral cancer. The quantity of S. anginosus DNA in the esophageal cancer tissues was significantly higher than in oral cancer. The maximum amount of S. anginosus DNA was approximately ten times higher in esophageal than in oral cancer tissues. In addition, none of the five different oral cancer sites (floor of the mouth, mandibular gingival, maxillary gingival, buccal mucosal, and tongue) showed significant signs of S. anginosus infection. On the other hand, most non-cancerous tissues of the esophagus and tongue showed an undetectable level of S. anginosus. These results suggest that S. anginosus is associated with esophageal cancer, but is not closely related with oral cancer.

Base Sequence↗

Real-time PCR for quantification of Streptococcus mutans.

A real-time polymerase chain reaction (PCR) assay was developed for the quantification of Streptococcus mutans. Primers targeting gtf genes of S. mutans were designed and tested for their specificity using 28 oral streptococcal strains, three other bacterial strains, and human DNA. The primers could amplify specifically the target DNA fragment from a mixture of oral streptococcus genomic DNA containing about 10 fg to 10 ng of S. mutans genome DNA. The real-time PCR produced a linear quantitative detection range over concentrations spanning seven exponential values, with a detection limit of a few copies of S. mutans' genomic DNA per reaction tube. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. mutans in saliva samples. A real-time PCR assay for Streptococcus sobrinus and Streptococcus downei was also established and produced results that corresponded well to those from conventional culture assays for S. sobrinus in saliva samples. These assays will be useful as a new means to assess one of the important risk factors for caries.

Bacterial Typing Techniques↗

Cloning and nucleotide sequence analysis of the Streptococcus sobrinus gtfU gene that produces a highly branched water-soluble glucan.

Streptococcus sobrinus has four gtf genes, gtfI, gtfS, gtfT, and gtfU, on the chromosome. These genes correspond respectively to the enzymes GTF-I, GTF-S1, GTF-S2, and GTF-S3. An Escherichia coli MD66 clone that contained the S. sobrinus gtfU gene was characterized. Immunological properties showed that the protein produced by the E. coli MD66 clone was similar to S. sobrinus GTF-S1. Biological properties and a linkage analysis of the glucans by 13C NMR spectrometry revealed that the protein produced by the E. coli MD66 clone was GTF-S1.

Amino Acid Sequence↗

Survey of dental hygienists and healthcare workers for microorganisms in the oral cavity.

Over 350 bacterial species are known to form communities on tooth surfaces in the oral cavity, and opportunistic pathogens are frequently isolated from samples taken from the oral cavities of immunocompromised hosts, as well as elderly people with poor oral hygiene. To confirm the correlation between the prevalence of opportunistic pathogens in the oral cavity and oral hygiene, dental plaque and saliva samples from 15 dental hygienists and 15 healthcare workers were investigated. No opportunistic pathogens were found in the samples from dental hygienists, however, they were isolated from 10 of the 15 healthcare workers (66.7%). The total amounts of Streptococcus sp. and Lactobacillus sp. were also significantly higher in the healthcare workers. The amount of Streptococcus mutans was also higher, although the difference was not significant. From our results, we concluded that the prevalence of opportunistic pathogens and cariogenic bacteria in the oral cavity may be reduced by proper oral hygiene measures.

Adult↗

Role of stromal-cell derived factor-1 in the development of autoimmune diseases in non-obese diabetic mice.

The chemokine stromal-cell derived factor-1 (SDF-1) controls maturation, trafficking, and homing of certain subsets, lymphoid cells including immunogenic B and T cells, as a ligand of the CXCR4 chemokine receptor. Insulin-dependent diabetes mellitus (IDDM) and Sjögren's syndrome (SS), both highly regulated autoimmune diseases, develop spontaneously in non-obese diabetic (NOD) mice. To investigate the role of SDF-1 in the development of autoimmune diseases, we injected groups of NOD female mice with antibodies to SDF-1 (anti-SDF-1), which resulted in a 30% reduction of diabetes up to 30 weeks of age, delayed average diabetes onset by 10 weeks, and suppressed insulitis. Autoimmune sialoadenitis was evident in anti-SDF-1-injected mice (SDF-1-Ig group) at the same level as in all groups of mice, whether injected with non-specific antibodies or not. In addition, in the SDF-1-Ig group, a greater number of immunoglobulin M (IgM)- IgD- B220(low) CD38+ CD43+ CD23- progenitor B cells and IgM+ IgD+ B220(high) CD43- CD38+ CD24+ CD23+ mature B cells remained in the bone marrow, whereas infiltration of mature IgM+ B cells was less extensive in peripheral tissues. Our results suggested that anti-SDF-1 antibodies injection was effective in inhibiting diabetes and insulitis without affecting autoimmune sialoadenitis or SS in NOD mice. SDF-1 may be an essential chemokine for trafficking and migration of autoreactive B cells in the development of diabetes.

Animals↗

Effects of human interleukin-18 and interleukin-12 treatment on human lymphocyte engraftment in NOD-scid mouse.

NOD/LtSz-prkdc(scid)/prkdc(scid) (non-obese diabetic-severe combine immunodeficiency; NOD-scid) mice grafted with human peripheral blood lymphoid cells have been used as an in vivo humanized mouse model in various studies. However, cytotoxic human T cells are induced in this model during immune responses, which gives misleading results. To assist in grafting of human lymphocytes without the induction of cytotoxic human T cells, we investigated the effects of T helper type 1 (Th1) and Th2 cytokines on human lymphocyte grafting and migration, as well as the production of immunoglobulin deposited in glomeruli and human immunodeficiency virus-1 (HIV-1) infection using NOD-scid mice. Administration of interleukin-18 (IL-18) and IL-12 enhanced the grafting of human CD4+ and CD8+ T cells in the mice, whereas co-administration prevented grafting due to interferon-gamma-dependent apoptosis. Immunoglobulin A (IgA) deposits were observed in mice treated with IL-18 alone, but not in those given phosphate-buffered saline, IL-12 alone, or IL-18 + IL-12. A high rate of HIV infection was also observed in the IL-18-treated group. Together, these results indicate that IL-18 may be effective for the grafting and migration of CD4+ and CD8+ T cells, except for the induction of apoptosis and regulation of class-switching IgA. IL-18-administered NOD-scid mice provide a useful small humanized model for the study of HIV infection and IgA nephropathy.

Animals↗

Relationship between the number of remaining teeth and physical activity in community-dwelling elderly.

The relationship between oral health and general health in the elderly has been much studied. However, further studies focussing on the influence of oral health on the quality of life (QOL) among the elderly are required. The goal of this study was to evaluate the relationship between oral health and physical or cultural activities. Subjects were 101 community-dwelling elderly persons who were functionally independent (mean age 70.3). Oral health status was evaluated according to the number of remaining teeth and the number of functional teeth. Physical and cultural activities were evaluated from self-reported information. The relationship between oral health and physical or cultural activities was examined by logistic regression analysis. About 60% of subjects took part in cultural activities, and less than half actively exercised (leisure sports, 33.6%; travel, 42.6%). Persons with 20 or more remaining teeth were more active in leisure sports (Odds ratio (OR)=4.86, 95% confidence interval (CI)=1.34, 17.38) and travel (OR=5.42, 95% CI=1.63, 18.08) than those with fewer than 20 remaining teeth. These results suggest that the number of remaining teeth is associated with physical activity in elderly persons.

Aged↗

Bone regeneration by recombinant human bone morphogenetic protein-2 around immediate implants: a pilot study in rats.

PURPOSE: Difficulties relating to bone regeneration that complicate immediate implant placement include buccal and/or lingual fenestrations, primary anchorage of the implants, and the need for protection from functional loading during the osseointegration period. The objective of this pilot study was to evaluate bone regeneration by recombinant human bone morphogenetic protein-2 (rhBMP-2) around immediate implants placed in maxillary sockets in rats. MATERIALS AND METHODS: A total of 16 cylindric 0.8 x 1.8-mm commercially pure, solid titanium Implants were placed immediately after gentle extraction of the maxillary first molar teeth of 8 male Wistar rats. The sockets were randomly divided into 3 groups: group 1 (n = 6) received rhBMP-2 with polylactic acid/polyglycolic acid copolymer-coated gelatin sponge carrier; group 2 (n = 5) received only the carrier; and group 3 (n = 5) received no grafting materials following placement The rats were euthanized at 90 days postsurgery for microscopic analysis. RESULTS: In group 1, the implant body remained submerged completely, including the coronal part, which was fully covered by a significant amount (30% of total height) of regenerated cortical bone, even though the implant could easily be pulled out by a tweezer at the time of placement. Close approximation between the implant surface and regenerated bone could also be detected, indicating good bone-to-implant contact. In contrast, only peri-implant bone regeneration occurred in group 2, and an approximate 0.3-mm coronal part of the implant remained exposed. When no grafting materials were used (group 3), almost one third of the total length of the implant was exfoliated out of the socket when no grafting materials were used. DISCUSSION AND CONCLUSIONS: Based on previous study and data from 16 sockets of the present study, it could be concluded that rhBMP-2 facilitated the regeneration of bone around immediate implants. In particular, the bone covering the coronal part could have been regenerated shortly after surgery, which helped to maintain the implant body inside the socket during the integration period in rats.

Animals↗