Synthesis of Novel Phosphorus-Substituted Stable Isoindoles by a Three-Component Coupling Reaction of ortho-Phthalaldehyde, 9,10-Dihydro-9-oxa-10-phosphaphenanthrene 10-Oxide, and Primary Amines

A three-component coupling reaction of ortho-phthalaldehyde, 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide, and various primary amines readily afforded novel phosphorus-substituted stable isoindoles in good to excellent yields. The importance of the reversible ring-opening of 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide by methanolysis in the three-component coupling reaction became apparent.


Use of ATP Bioluminescence Assay to Evaluate Oral Streptococci

The statistical correlation between the number of oral streptococci and the results of ATP bioluminescence assay was examined and compared with the results from Streptococcus plate counts and an oral bacteria quantification system. Because a significant correlation was found between ATP (RLU) and the number of bacteria in the oral bacteria quantification system for all seven types of oral streptococci examined, ATP would reflect a conditions of oral hygiene. However, using this assay, it was observed it may be difficult to correctly evaluate bacteria that form aggregates. Furthermore, even a small number of bacteria (below 10 CFU/mL) , which cannot be measured by the oral bacteria quantification system, could be estimated by using ATP bioluminescence assay. It was suggested that this assay could be used for quantitative evaluation of the effect of oral cleaning.


Protective effects of resveratrol against 5-fluorouracil-induced oxidative stress and inflammatory responses in human keratinocytes.

-acetylcysteine reduced ROS levels without affecting the expression of pro-inflammatory cytokines. Resveratrol might be useful for preventing 5-FU-induced adverse effects by activating anti-oxidant and anti-inflammatory responses.

Journal of Clinical Biochemistry and Nutrition


Membrane fusion of phospholipid bilayers under high pressure: Spherical and irreversible growth of giant vesicles

Membrane fusion of giant vesicles (GVs) for binary bilayers of unsaturated phospholipids, dioleoylphosphatidylethanolamine (DOPE) having an ability to promote membrane fusion and its homolog dioleoylphosphatidylcholine (DOPC) having an ability to form GV, was investigated under atmospheric and high pressure. While DOPC formed GVs in the presence of inorganic salts with a multivalent metal ion under atmospheric pressure, an equimolar mixture of DOPE and DOPC formed GVs both in the absence and the presence of LaCl3. We examined the change in size and shape of the GVs of this binary mixture in the absence and presence of LaCl3 as a function of time under atmospheric and high pressure. The size and shape of the GVs in the absence of LaCl3 under atmospheric and high pressure and those in the presence of LaCl3 under atmospheric pressure hardly changed with time. By contrast, the GV in the presence of LaCl3 under high pressure gradually changed in the size and shape with time on a time scale of several hours. Namely, the GV became larger than the original GV due to accelerated membrane fusion and its shape became more spherical. This pressure-induced membrane fusion was completely irreversible, and the growth rate was correlated with the applied pressure. The reason for the GV growth by applying pressure was considered on the basis of thermodynamic phase diagrams. We concluded that the growth is attributable to a closer packing of lipid molecules in the bilayer resulting from their preference of smaller volumes under high pressure. Furthermore, the molecular mechanism of the pressure-induced membrane fusion was explored by observing the fusion of two GVs with almost the same size. From their morphological changes, we revealed that the fusion is caused by the actions of Laplace and osmotic pressure.


Applications of a novel biodetection system to saliva using protein fingerprints with data processing.

A fundamental method has been developed focusing on a facile and rapid examination of periodontal disease. Periodontal disease is an oral disease thought to affect 80% of adults, and early detection with treatment is desirable for the improvement of the quality of life. Unfortunately conventional methods are not consistent as the disease is caused by a number of undefined bacteria and detection relies on the skills of the dentist. Thus an objective detection system is required. We have performed an experiment on saliva using a novel biodetection system, designated PepTenChip®. A disease model for saliva was prepared using a specimen from a healthy subject and a mixture of hemoglobin (f-Hb) and lactate dehydrogenase (LDH), which is used as a periodontal disease marker protein with healthy saliva. PepTenChip® is a peptide microarray in which fluorescent labelled structured peptides are immobilized on a novel amorphous carbon substrate. Since the peptides used as capture molecules are fluorescently labelled, labeling of analytes is not necessary. The fluorescence intensity change before and after application of analytes are detected rather than the ON/OFF detection common to conventional microarrays using a set of antigen-antibody. The fluorescence intensity value changes according to the concentration of captured protein allowing the generation of protein fingerprint (PFP) and dendrograms. The present method does not rely on a "one to one" interaction, unlike conventional biodetection, and advantages can be envisaged in the case of an undefined or unknown cause of disease. The statistical analyses, such as multivariate analyses, allow classification of the type of proteins added in saliva as mimetics of disease. PepTenChip® system is useful and convenient for examination of periodontal disease in health care.


Synthesis of Sterically Protected Isoindoles from ortho-Phthalaldehyde

o-Phthalaldehyde (OPA) reacts with O-protected tris(hydroxyalkyl)aminomethanes in the presence of 1-propanethiol to afford a novel class of stable isoindoles. Steric protection provided by the bulkiness of C 3-symmetric primary amines derived from tris(hydroxymethyl)aminomethane could be significant for the stabilization of 1-alkylthio-2-alkyl-substituted isoindoles derived from OPA. A plausible reaction mechanism is proposed to explain the formation of the isoindole and an isoindolin-1-one by-product.


歯肉溝液中の炎症関連バイオマーカーと糖尿病・腎機能マーカーとの関連性

歯肉溝液中の炎症関連バイオマーカーと糖尿病・腎機能マーカーとの関連性


Hydrogen-Rich Water Intake Accelerates Oral Palatal Wound Healing via Activation of the Nrf2/Antioxidant Defense Pathways in a Rat Model.

The wound healing process attempts to restore the integrity and function of the injured tissue. Additionally, proinflammatory cytokines, growth factors, and oxidative stress play important roles in wound healing. The aim of this study was to determine whether hydrogen-rich water intake induces the activation of the Nrf2/antioxidant defense pathway in rat palatal tissue, thereby reducing systemic oxidative stress and proinflammatory cytokine levels and promoting healing-associated genes. A circular excisional wound was created in the oral palatal region, and the wound healing process was observed. The rats were divided into two experimental groups in which either hydrogen-rich water or distilled water was consumed. In the drinking hydrogen-rich water, the palatal wound healing process was accelerated compared to that in the control group. As molecular hydrogen upregulated the Nrf2 pathway, systemic oxidative stresses were decreased by the activation of antioxidant activity. Furthermore, hydrogen-rich water intake reduced proinflammatory cytokine levels and promoted the expression of healing-associated factors in rat palatal tissue. In conclusion, hydrogen-rich water intake exhibited multiple beneficial effects through activation of the Nrf2/antioxidant defense pathway. The results of this study support the hypothesis that oral administration of hydrogen-rich water benefits the wound healing process by decreasing oxidative stress and inflammatory responses.

Oxidative Medicine and Cellular Longevity


Correlation between arteriosclerosis and periodontal condition assessed by lactoferrin and [alpha]1-antitrypsin levels in gingival crevicular fluid.

Patients with periodontal disease exhibit exacerbated atherosclerosis, aortic stiffness, or vascular endothelial dysfunction. However, in a recent scientific statement, the American Heart Association noted that neither has periodontal disease been proven to cause atherosclerotic vascular disease nor has the treatment of periodontal disease been proven to prevent atherosclerotic vascular disease. Therefore, the aim of the present study was to examine the correlation between periodontal condition and arteriosclerosis in patients with coronary artery disease (CAD), which is usually accompanied by systemic arteriosclerosis.We measured levels of gingival crevicular fluid lactoferrin (GCF-Lf) and 1-antitrypsin (GCF-AT) in 72 patients (67 ± 8 years, 56 men) with CAD. Furthermore, we evaluated the maximum intima-media thickness (max IMT) and plaque score of the carotid arteries as well as brachial-ankle pulse wave velocity (baPWV) and flow-mediated dilation (FMD) of the brachial artery, each of which is a parameter for determining arteriosclerosis status. The average level of GCF-Lf was 0.29 ± 0.36 µg/mL and that of GCF-AT was 0.31 ± 0.66 µg/mL, with significant correlation between the two (r = 0.701, P < 0.001). No significant difference in GCF-Lf and GCF-AT levels was observed between patients with single-, double-, and triple-vessel CAD. There were no significant correlations between the arteriosclerosis parameters (ie, max IMT, plaque score, baPWV, and FMD) and GCF-Lf or GCF-AT.No correlation between the GCF biomarkers and the severity of arteriosclerosis was detected. This result may suggest that worsening of the periodontal condition assessed by GCF biomarkers is not a major potential risk factor for arteriosclerosis.


Relationship among salivary antioxidant activity, cytokines and periodontitis: the Nagasaki Island study.

These findings suggest that the evaluation of salivary antioxidant activities, as assessed by electron spin resonance, are associated with periodontitis and various clinical variables in community-dwelling participants (ClinicalTrials. gov number NCT01742728). This article is protected by copyright. All rights reserved.


Biological impacts of resveratrol, quercetin, and N-acetylcysteine on oxidative stress in human gingival fibroblasts.

In periodontitis, production of reactive oxygen species (ROS) by neutrophils induces oxidative stress and deteriorates surrounding tissues. Antioxidants reduce damage caused by ROS and are used to treat diseases involving oxidative stress. This study summarizes the different effects of resveratrol, quercetin, and N-acetylcysteine (NAC) on human gingival fibroblasts (HGFs) under oxidative stress induced by hydrogen peroxide. Real-time cytotoxicity analyses reveals that resveratrol and quercetin enhanced cell proliferation even under oxidative stress. Of the antioxidants tested, resveratrol is the most effective at inhibiting ROS production. HGFs incubated with resveratrol and quercetin up-regulate the transcription of type I collagen gene after 3 h, but only resveratrol sustained this up-regulation for 24 h. A measurement of the oxygen consumption rate (OCR, mitochondrial respiration) shows that resveratrol generates the highest maximal respiratory capacity, followed by quercetin and NAC. Simultaneous measurement of OCR and the extracellular acidification rate (non-mitochondrial respiration) reveals that resveratrol and quercetin induce an increase in mitochondrial respiration when compared with untreated cells. NAC treatment consumes less oxygen and enhances more non-mitochondrial respiration. In conclusion, resveratrol is the most effective antioxidant in terms of real-time cytotoxicity analysis, reduction of ROS production, and enhancement of type I collagen synthesis and mitochondrial respiration in HGFs.


Resveratrol improves oxidative stress and prevents the progression of periodontitis via the activation of the Sirt1/AMPK and the Nrf2/antioxidant defense pathways in a rat periodontitis model.

Oxidative stress is a key factor regulating the systemic pathophysiological effects associated with periodontitis. Resveratrol is a phytochemical with antioxidant and anti-inflammatory properties that can reduce oxidative stress and inflammation. We hypothesized that resveratrol may prevent the progression of periodontitis and reduce systemic oxidative stress through the activation of the Sirtuin1 (Sirt1)/AMP-activated protein kinase (AMPK) and the nuclear factor-E2 related factor 2 (Nrf2)/antioxidant defense pathways. Three groups of male Wistar rats (periodontitis treated with melinjo resveratrol, periodontitis without resveratrol, and control rats with no periodontitis or resveratrol treatment) were examined. A ligature was placed around the maxillary molars for 3 weeks to induce periodontitis, and the rats were then given drinking water with or without melinjo resveratrol. In rats with periodontitis, ligature placement induced alveolar bone resorption, quantified using three-dimensional images taken by micro-CT, and increased proinflammatory cytokine levels in gingival tissue. Melinjo resveratrol intake relieved alveolar bone resorption and activated the Sirt1/AMPK and the Nrf2/antioxidant defense pathways in inflamed gingival tissues. Further, melinjo resveratrol improved the systemic levels of 8-hydroxydeoxyguanosine, dityrosine, nitric oxide metabolism, nitrotyrosine, and proinflammatory cytokines. We concluded that oral administration of melinjo resveratrol may prevent the progression of ligature-induced periodontitis and improve systemic oxidative and nitrosative stress.


Oxidative stress and antibody levels to periodontal bacteria in adults: the Nagasaki Islands study.

OBJECTIVE: Periodontitis is a chronic inflammatory disease of the tissues supporting the teeth and is caused by subgingival plaque. Systemic increases in reactive oxygen species are involved in pathogenesis of periodontitis. This study addressed the relationship between levels of serum oxidative stress and antibodies against putative periodontopathic bacteria and their association with periodontal conditions, in a community-based study. SUBJECTS AND METHODS: Serum samples were measured for reactive oxygen metabolite (ROM) levels and anti-oxidant capacity. The serum levels of immunoglobulin G (IgG) antibodies to Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Eikenella corrodens (Ec) were determined by ELISA. RESULTS: The participants with greater clinical attachment loss had higher serum ROM levels and IgG antibody titers to Pg. Serum ROM levels were positively correlated with antibody titers to Pg, Pi, and Ec. When the participants with greater probing pocket depth and clinical attachment loss were used as the dependent variables, high ROM levels showed a statistically significant associations in multivariate logistic analyses; the adjusted odds ratios were 2.9 (95% confidence interval = 1.0-8.5) and 6.0 (95% confidence interval = 2.0-17.6), respectively. CONCLUSIONS: It was concluded that an increased oxidative stress may be detrimental to periodontitis in Japanese community-dwelling adults.


Association of gingival recession and other factors with the presence of dentin hypersensitivity.

Dentin hypersensitivity (DH) may be present in association with gingival recession. The aim of this study was to determine quantitatively the association of gingival recession and other factors with the presence of DH. One hundred and four Japanese subjects with or without gingival recession were randomly selected. Intact canines and/or first premolars in both maxillary and mandibular quadrants were analyzed. Gingival recession was measured as a vertical length at the buccal site of the teeth. DH was recorded as an ordered categorical variable registering four increasing levels of pain after cold stimulation; from no discomfort to severe pain during and after stimulation (DH1, 2, 3, and 4). Association of DH with periodontal parameters and daily lifestyle was also investigated. Tooth-based analysis of 446 teeth from 104 subjects revealed that DH level was significantly higher in recessive teeth (1, 2, 3, and 4-8 mm) than in non-recessive teeth (0 mm). DH-positive rate in non-recessive teeth was only 18 % (DH1; 14 %, DH2; 3 %, and DH3; 1 %). Highest DH level was observed in teeth with severe recession (4-8 mm), showing DH0; 21 %, DH1; 33 %, DH2; 31 %, and DH3; 15 %. Recession-dependent increase in DH was observed, showing 18, 49, 52, 60, and 79 % DH-positive in teeth with 0, 1, 2, 3, and 4-8 mm recession, respectively. Plaque-free teeth showed a higher DH level than plaque-stained teeth, suggesting that good plaque control may be associated with the presence of DH. There were no significant differences in DH of teeth on the basis of smoking, probing depth, and bleeding on probing. Multiple logistic regression analysis revealed that gingival recession [odds ratio (OR) = 10.2, 95 % confidence interval (CI) = 5.5-18.9] and plaque deposition (OR = 0.3, 95 % CI = 0.2-0.5) were significant contributors to DH. Multilevel modeling analysis revealed that not only gingival recession and plaque deposition but also V-shaped cervical notch and tooth brushing frequency were associated with DH. These results demonstrate that the progression of gingival recession, plaque-free teeth, V-shaped cervical notch, and frequent brushing may be significant predictors of DH in canines and first premolars.


Study on the Subgel-Phase Formation Using an Asymmetric Phospholipid Bilayer Membrane by High-Pressure Fluorometry

The myristoylpalmitoylphosphatidylcholine (MPPC) bilayer membrane shows a complicated temperature-pressure phase diagram. The large portion of the lamellar gel (L(β)'), ripple gel (P(β)'), and pressure-induced gel (L(β)I) phases exist as metastable phases due to the extremely stable subgel (L(c)) phase. The stable L(c) phase enables us to examine the properties of the L(c) phase. The phases of the MPPC bilayers under atmospheric and high pressures were studied by small-angle neutron scattering (SANS) and fluorescence spectroscopy using a polarity-sensitive fluorescent probe Prodan. The SANS measurements clearly demonstrated the existence of the metastable L(β)I phase with the smallest lamellar repeat distance. From a second-derivative analysis of the fluorescence data, the line shape for the L(c) phase under high pressure was characterized by a broad peak with a minimum of ca. 460 nm. The line shapes and the minimum intensity wavelength (λ''(min)) values changed with pressure, indicating that the L(c) phase has highly pressure-sensible structure. The λ''(min) values of the L(c) phase spectra were split into ca. 430 and 500 nm in the L(β)I phase region, which corresponds to the formation of a interdigitated subgel L(c) (L(c)I) phase. Moreover, the phase transitions related to the L(c) phase were reversible transitions under high pressure. Taking into account the fluorescence behavior of Prodan for the L(c) phase, we concluded that the structure of the L(c) phase is highly probably a staggered structure, which can transform into the L(c)I phase easily.


Mucosal immune features to phosphorylcholine by nasal Flt3 ligand cDNA-based vaccination

Phosphorylcholine (PC) is an immunodominant epitope in some pathogens including Streptococcus pneumoniae and it is well-known that PC-specific antibodies (Abs) play a key role in the induction of protective immunity against pneumococcal infection. In this study, we examined whether nasal administration of DNA plasmid encoding Flt3 ligand gene (pFL) as a mucosal adjuvant plus PC-conjugated keyhole limpet hemocyanin (PC-KLH), would elicit PC-specific immune responses, and characterized mucosal immune responses to PC induced by this nasal vaccination. Nasal immunization with pFL plus PC-KLH enhanced induction of PC-specific IgA and IgM Abs in airway secretions when compared with mice given PC-KLH with or without empty plasmid gene (pORF) as controls; in addition to the mucosal immune responses, PC-specific immune responses in serum were also induced. Furthermore, the mucosal and serum IgA and IgM Abs in mice given pFL plus PC-KLH nasally, exhibited high-specificity for the PC molecule. Of interest, the PC-specific Abs bound dose-dependently to anti-T15 idiotype (AB1-2). Thus, the inhibition of S. pneumoniae colonization on the nasal cavity and lungs after nasal challenge with the live organism was significantly elicited in mice immunized with pFL plus PC-KLH compared to that of mice immunized with antigen with pORF. Taken together, these findings show that nasal administration of pFL with PC-KLH elicited T15-like anti-PC IgA and IgM Abs in the respiratory tracts, and further attenuated S. pneumoniae colonization on the respiratory tracts. Nasal administration of Flt3 ligand cDNA with PC may contribute to the development of nasal vaccination for prevention of S. pneumoniae infection.


Nasal Dendritic Cell Targeting Flt3 Ligand As A Safe Adjuvant Elicits Effective Protection Against Fatal Pneumococcal Pneumonia.

We have previously shown that pneumococcal surface protein A (PspA)-based vaccine containing DNA plasmid encoding the Flt3 ligand (FL) gene (pFL) as nasal adjuvant prevented nasal carriage of Streptococcus pneumoniae. In this study, we further investigated the safety and efficacy of this nasal vaccine for the induction of PspA-specific antibody (Ab) responses against lung infection with S. pneumoniae. C57BL/6 mice were nasally immunized with recombinant PspA/Rx1 (rPspA) plus pFL three times at weekly intervals. When dynamic translocation of pFL was initially examined, nasal pFL was taken-up by nasal dendritic cells (DCs) and epithelial cells (nECs), but not in the central nervous systems including olfactory nerve and epithelium. Of importance, nasal pFL induced FL protein synthesis with minimum levels of inflammatory cytokines in the nasal washes (NWs) and bronchoalveolar lavage fluid (BALF). NWs and BALF as well as plasma of mice given nasal rPspA plus pFL contained increased levels of rPspA-specific secretory-IgA and IgG Ab responses that are correlated with elevated numbers of CD8(+) and CD11b(+) DCs and IL-2 and IL-4 producing CD4(+) T cells in the nasopharyngeal-associated lymphoid tisses (NALT) and cervical lymph nodes (CLNs). The in vivo protection by rPspA-specific Abs was evident by reduced numbers of CFU in the lungs, airway secretions, and blood when mice were nasally challenged with Streptococcus pneumoniae WU2. Our findings show that nasal pFL is a safe and effective mucosal adjuvant for the enhancement of bacterial Ag (rPspA)-specific protective immunity through DC-induced Th2-type and IL-2 cytokine responses.


Levels of salivary stress markers in patients with anxiety about halitosis

To investigate the relationship between salivary stress markers and mental stress states in patients complaining of oral malodour. The utility of the salivary stress markers in assessment of mental conditions of those patients was also investigated. The study population included 74 patients, aged 20-59 years, who complained of oral malodour and were referred to the Breath Odor Clinic at Tokushima University Hospital. Patients were classified into two groups, genuine halitosis (GH) and psychosomatic halitosis (PH), according to the results of organoleptic rating measurement. All patients were subjected to examination by the Cornell Medical Index (CMI) Health Questionnaire. Resting saliva was collected and levels of salivary IgA, cortisol and chromogranin A were determined by ELISA. Twenty-three volunteers not complaining of halitosis were included as the control group. Kruskal-Wallis test and Mann-Whitney's U-test were used for statistical analysis. A significant increase was observed in the concentrations of salivary cortisol in the PH group as compared with GH and control groups (p<0.05). Concentrations of IgA and chromogranin A in saliva were not significantly different among the three groups. In addition, higher salivary cortisol concentrations were found in CMI scale III and IV (tendency towards neurosis) than in scale I and II (normal) (p<0.05). Since salivary cortisol reflects a status of chronic stress condition, psychosomatic halitosis might be closely related to this state of chronic stress. Determination of cortisol levels in saliva may provide useful information for evaluating the mental status of patients complaining of halitosis.


Attenuation of inducible respiratory immune responses by oseltamivir treatment in mice infected with influenza A virus.

The antiviral neuraminidase inhibitor oseltamivir (OSV) is widely used to suppress viral replication in the treatment of influenza. Here, we report that OSV administration significantly suppressed respiratory mucosal secretory IgA responses with respect to antigen (Ag)-specific antibody (Ab) production and also the induction of Ag-specific IgA Ab-forming cells, but not systemic IgG responses, in weanling mice as a model of pediatric influenza. Neutralizing activities of the airway fluids in oral OSV-treated mice were significantly less than those of sham-treated mice. Our findings suggest the risk of re-infection in patients showing a low mucosal response following OSV treatment.


Fusobacterium nucleatum envelope protein FomA is immunogenic and binds to the salivary statherin-derived peptide.

We have previously shown that one of the minimal active regions of statherin, a human salivary protein, for binding to Fusobacterium nucleatum is a YQPVPE amino acid sequence. In this study, we identified the FomA protein of F. nucleatum, which is responsible for binding to the statherin-derived YQPVPE peptide. Overlay analysis showed that a 40-kDa protein of the F. nucleatum cell envelope (40-kDa CE) specifically bound to the YQPVPE peptide. The equilibrium association constant between the affinity-purified 40-kDa CE and the YQPVPE peptide was 4.30 x 10(6). Further, the purity and amino acid sequence analyses of the purified 40-kDa CE revealed approximately 98.7% (wt/wt) purity and a high degree of homology with FomA, a major porin protein of F. nucleatum. Thus, a FomA-deficient mutant failed to bind to the YQPVPE peptide. In addition, increased levels of a FomA-specific mucosal IgA antibody (Ab) and plasma IgG and IgA Abs were seen only in mice immunized nasally with cholera toxin (CT) and the purified 40-kDa FomA protein. Interestingly, saliva from mice that received FomA plus CT as a mucosal adjuvant nasally prevented in vitro binding of F. nucleatum to statherin-coated polyvinyl chloride plates. Taken together, these results suggest that induction of specific immunity to the 40-kDa FomA protein of F. nucleatum, which specifically binds to the statherin-derived peptide, may be an effective tool for preventing the formation of F. nucleatum biofilms in the oral cavity.


リアルタイム定量PCR(qPCR)法による唾液中の総細菌数の測定 口腔清潔度の指標としての試み

The state of oral hygiene has been evaluated by employing various indices, such as the Oral Hygiene Index, Plaque Index, Patient Hygiene Performance, and Plaque Control Record (O'Leary's PCR). Measuring the scores of these indices is time- and labor-consuming, requiring manual reading and recording by professionals, standardization of examination methods. In contrast, measuring the total number of bacteria in saliva using quantitative real-time PCR (qPCR) allows the straightforward collection and storage of samples, making it possible to measure many samples at the same time. This technique facilitates the quantification of the number of bacteria within a shorter time than the culture method. In the present study, the total number of bacteria measured by qPCR was found to be significantly decreased (p<0.05) in stimulated whole saliva collected from 5 volunteers by chewing paraffin after tooth cleaning. These findings were in agreement with the decreases shown by Greene's Debris Index and O'Leary's PCR values. There was a significant correlation (r=0.55, p<0.01) between the salivary bacterial counts measured by qPCR from 81 subjects before and after a 4-month interval. These findings suggest that it is possible to measure the total number of bacteria in saliva using qPCR. This technique is convenient and is suggested to be applicable for assessing the oral hygiene status in the future.

口腔清潔度の指標としては,歯面への歯垢の付着度を測定する方法が一般的であるが,診査や記録等の人材的,時間的負担を要することから,集団歯科健診で用いられることは少ない.集団歯科健診においてう蝕や歯周疾患,口臭症のリスクを評価するために,簡便かつ有効な口腔清潔度の評価法の開発が望まれている.リアルタイム定量PCR(qPCR)法による唾液中の総細菌数の測定は,検体の採取や保管が容易で多検体の一括処理が可能であることから,本目的に合致する有望な測定法であると考えられる.本研究では,qPCR法を用いた唾液中の総細菌数の測定が口腔清潔度の評価法として使用できるかどうか検討を行った.5名のボランティアから,ブラッシング中断時と再開後の2回,刺激唾液を採取し,唾液中の総細菌数をqPCR法により測定したところ,Debris Index,O'LearyのPCR値との間に相関は示されなかったものの,ブラッシング再開後では,中断時に比べ,有意に減少した.81名の被験者から4ヵ月の間隔を空けて2回刺激唾液を採取し,唾液中の総細菌数をqPCR法により測定したところ,前後の間に有意な正の相関を認めた.以上の結果より,qPCR法によって唾液中の総細菌数を測定することが可能であることが示された.本法は非常に簡便であることから,今後,口腔清潔度を検討する分野において応用できる可能性が考えられる.


Relationship between Campylobacter rectus and periodontal status during pregnancy.

In a previous study, we showed that the growth of Campylobacter rectus is stimulated by the presence of female sex hormones in the culture medium. In the present study, we examined the relationship between C. rectus levels in the saliva and the periodontal status of pregnant women. Unstimulated whole saliva was collected from 22 pregnant and 15 non-pregnant women. Periodontal pocket depth (PD) and bleeding on probing (BOP) were recorded. A quantitative real-time polymerase chain reaction was performed to determine the concentrations of suspected periodontopathogenic bacteria in the saliva samples. In addition, the concentration of estradiol in the saliva samples was measured by enzyme immunoassay. The average age, number of teeth, and total number of bacteria in the saliva of subjects in both groups were similar. The percentage of sites with a PD = 4 mm and the salivary estradiol concentrations were significantly higher in pregnant women than in non-pregnant women. In addition, the percentage of BOP sites and the C. rectus levels in the saliva of the pregnant women tended to be higher than in non-pregnant women, although these differences were not statistically significant. There were positive correlations between C. rectus levels and estradiol concentrations, and between C. rectus levels and the percentage of sites with PD = 4 mm in the pregnant women. These results indicate that C. rectus levels are higher in the oral flora of pregnant women and that this may be associated with increased salivary estradiol concentrations. This may contribute to periodontal disease progression during pregnancy.


Development of the Japanese version of the Oral Impacts on Daily Performance (OIDP) scale: a pilot study.

The Oral Impacts on Daily Performance (OIDP) is a well-known oral health-related QOL instrument used internationally. The aim of this study was to develop and test a Japanese version of the OIDP for use in interviews. Following an internationally established method, the OIDP scale was translated using standardized methodology that consisted of forward translation, pilot study and backward translation. A pilot study was carried out using the revised version with 47 local residents (range, 40-105 years). In the item analysis of this study, there were no missing values or ambiguous responses. Twenty-eight percent of the participants replied that they 'had discomfort caused by an oral health problem' in the 'eating' items. Cronbach's alpha coefficient was 0.77, indicating the internal consistency reliability of the questionnaire. Denture wearers had significantly higher OIDP scores in covariance analysis with age and sex as adjustment factors (P < 0.05). Based on the results, the questionnaire was re-evaluated. A back translation was produced and approved by the original author. Further assessment and validation of this scale is needed in a study with larger sample size.


Intranasal immunization with phosphorylcholine induces antigen specific mucosal and systemic immune responses in mice.

Phosphorylcholine (PC) is a structural component of a wide variety of pathogens including Streptococcus pneumoniae and Haemophilus influenzae, and anti-PC immune responses are known to protect mice against invasive bacterial diseases. The present study tested the capability of PC as an intranasal plurispecific vaccine against upper airway infections. BALB/c mice immunized with intranasal PC-keyhole limpet hemocyanin (KLH) plus cholera toxin (CT) as a mucosal adjuvant showed increased PC-specific IgM in serum, IgA in nasal wash and saliva, and numbers of PC-specific nasal and splenic antibody producing cells. Enhanced production of IL-4 and IFN-gamma by CD4+ T cells indicated the participation of Th2- and Th1-type cells. Salivary IgA antibodies produced by intranasal immunization with PC-KLH plus CT reacted to most strains of S. pneumoniae and H. influenzae. Further we demonstrated that the clearance of S. pneumoniae and H. influenzae from the nasal tract was significantly enhanced by nasal immunization with PC-KLH and CT. Thus, intranasal vaccination to induce PC-specific immune responses might help to prevent upper airway infections caused by S. pneumoniae and H. influenzae.


Serotype-specific polysaccharide of Streptococcus mutans contributes to infectivity in endocarditis.

Streptococcus mutans and other viridans streptococci have been implicated as major etiological agents of infective endocarditis. The serotype-specific rhamnose-glucose polysaccharide (RGP) of S. mutans has several biological functions that appear to be essential for the induction of infective endocarditis. The aim of this study was to examine the contribution of RGP to the infectivity of S. mutans in infective endocarditis using a rat model. The RGP-defective mutant of S. mutans showed reduced ability to induce infective endocarditis compared to the parental strain. The ability of S. mutans to induce infective endocarditis was not consistent with the binding capacity of the organism to extracellular matrix proteins. The results suggest that S. mutans containing whole RGP is more virulent than the RGP-defective mutant, and the RGP has an important role for the induction of infective endocarditis by S. mutans.


Screening of peptides that inhibit bacterial binding to fibronectin using combinatorial peptide libraries.

International Journal of Peptide Research and Therapeutics


Experimental infective endocarditis induced by human supragingival dental plaque in rats.

Human dental plaque is thought to contribute to disease, not only in the oral cavity but also at other body sites. To investigate the pathogenicity of dental plaque in tissues remote from the mouth, we examined the ability of human supragingival dental plaque to induce infective endocarditis (IE) in rats. In total, 15 out of 27 catheterized rats survived after intravenous injections with human supragingival dental plaque suspensions containing 3 x 10(6) colony-forming units (CFU) of bacterial cells. In surviving rats, infected vegetations were formed in all except one rat. The microbial composition of the infected vegetations was different from that of the respective dental plaque inocula, with Streptococcus oralis comprising the majority of the isolates. In rats affected with endocarditis, the aortic sinus was filled with fibrinous vegetation containing bacteria. Inflammatory cells infiltrated the aortic valve, the aorta adjacent to the valve, and the cardiac muscles. The inoculation of catheterized rats with a cell suspension of S. oralis isolate (5 x 10(6) CFU) was not lethal but capable of inducing endocarditis in all animals. The results suggest that if dental plaque were introduced into the bloodstream, it could serve as a potent source of bacteria causing IE in humans.


Induction of immune response to Streptococcus pneumoniae by administration of oral viridans streptococci via phosphorylcholine determinant.

Expression of the phosphorylcholine (PC) epitope was examined in 48 viridans streptococcal strains, including Streptococcus pneumoniae R36a as the positive control, and their immunogenicity to induce an S. pneumoniae-cross-reactive response was evaluated in mice. Thirteen strains were found to express the PC epitope, while no obvious association was found between the taxonomic categories and PC expression. Serum antibody responses to S. pneumoniae cells were induced in mice by intraperitoneal injection of the PC-positive, but not PC-negative, strains. The cross-reactive antibodies induced by non-pneumococcal oral streptococci were readily inhibited by free hapten PC. IgM was the sole isotype of the anti-pneumococcal and anti-PC antibodies, and the phenomenon of immunological memory was not observed. Since the anti-PC antibody is critically important for resistance against pneumococcal infection in mice, the present results indicate the possibility that PC-expressing oral commensal bacteria have a significant influence on the hosts' responsiveness to S. pneumoniae.

FEMS Immunology and Medical Microbiology


Experimental Abscess Formation Caused by Human Dental Plaque

Human dental plaque consists of a wide variety of microorganisms, some of which are believed to cause systemic infections, including abscesses, at various sites in the body. To confirm this hypothesis experimentally, we examined the abscess-forming ability of native dental plaque in mice, the microbial features of the infectious locus produced by the plaque, and the anti-phagocytic property of microbial isolates. Aliquots of a suspension of supragingival dental plaque containing 6 x 10(6) colony-forming unit of bacteria were injected subcutaneously into the dorsa of mice. Abscess formation was induced in 76 of 85 mice using ten different plaque samples. Thirteen microorganisms were isolated from pus samples aspirated from abscess lesions. The microbial composition of pus, examined in 17 of 76 abscesses, was very simple compared to that of the plaque sample that had induced the abscess. The majority of the isolates belonged to the Streptococcus anginosus group, normally a minor component of plaque samples. S. anginosus was the most frequently detected organism and the most prevalent in seven abscesses, and Streptococcus intermedius and Streptococcus constellatus were predominant in one and three abscess samples, respectively. Each isolate of S. anginosus group produced abscesses in mice, and heat-treated supragingival dental plaque influenced the abscess-forming ability of S. anginosus isolate. These isolates possessed a high antiphagocytic capacity against human polymorphonuclear leukocytes. Our results suggest that human supragingival dental plaque itself is a source of the infectious pathogens that cause abscess formation.


Identification and characterization of bacterial-binding property in the type III repeat domain of fibronectin

To characterize fibronectin binding with Granulicatella adiacens, a causative agent of infective endocarditis, monoclonal antibodies were generated against human fibronectin and selected for their capacity to inhibit the fibronectin binding of the organism. Thermolysin and lysyl-endopeptidase digests of fibronectin were characterized by Western blot. The epitope of inhibitory monoclonal antibody was found in the central portion of fibronectin known as the cell-binding domain, and not in the N-terminal portion known to be the binding region of most microbial species, e.g., Staphylococcus aureus and Streptococcus pyogenes. While these two species could bind to both the N-terminal and central portion, Escherichia coli and G. adiacens bind only to the latter. Excess amounts of free fibronectin in the solution inhibited the bacterial adherence to the N-terminal fibronectin fragment, but not to the central region, thereby suggesting the central region plays a significant role for in vivo bacterial colonization in the presence of high concentrations of soluble fibronectin.

Biochemical and Biophysical Research Communications


本学歯学部における OSCE トライアルの実施方法について

本学歯学部における OSCE トライアルの実施方法について

The Journal of Japanese Dental Education Association

日本歯科医学教育学会雑誌


歯周病罹患状況と臨床検査値および歯周病原性細菌に対する血清抗体価の変化とその関連 --- 同じ被験者を対象とした3年間の調査結果

歯周組織，臨床検査値および血清抗体価に関する3年間の追跡調査

歯周組織，臨床検査値および血清抗体価に関する3年間の追跡調査


Inhibition of fibronectin binding of some bacterial cells by subtle pH increase within the physiological range

The fibronectin (Fn)-binding ability of microorganisms is considered to be involved in their pathogenicities. Granulicatella adiacens, a member of the oral flora and a causative agent of culture-negative infective endocarditis, showed nearly maximum binding to immobilized Fn at pH 7.2 but greatly reduced binding at a slightly higher pH 7.4 and almost no binding at pH 7.6 in the presence of physiological concentration of NaCl (0.15 M). A similar pH-sensitive Fn-binding property was noted with Escherichia coli and Abiotrophia defectiva, but not with Streptococcus pyogenes nor Staphylococcus aureus. In contrast, bindings to laminin and fibrinogen observed for some of these strains were unaffected by the same pH changes. This fastidious pH-dependency of Fn-binding abilities of some bacteria warns that the pH condition must be seriously considered in the in vitro assay of bacterial adherence to fibronectin.


Associations of Periodontal Status with General Health Conditions and Aerum Antibody Titers for Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans

The purpose of the present study was to evaluate associations of periodontal status with general health conditions and with serum antibody titers for Porphyromonas gingivalis (Pg) and Actinobacillus actinomycetemcomitans (Aa) in a Japanese population. Subjects (1,314) aged 40 years or older were recruited for the study. Their periodontal status was evaluated using the community periodontal index of treatment needs (CPITN). Records from general health examinations were used for the present analyses, including data on age, gender, body mass index, systolic and diastolic blood pressure (SBP/DBP); levels of serum alkaline phosphatase, total cholesterol, high-density lipoprotein cholesterol, triglyceride, fasting blood glucose, and C-reactive protein (CRP); counts of white blood cells (WBC) and platelets; smoking habit; and electrocardiogram recordings. Serum IgG antibody titers for Pg fimbrial antigens (IgG.Pg.Fim.), Pg whole-cell antigens (IgG.Pg.whol.), and Aa whole-cell antigens (IgG.Aa.whol.) were determined using enzyme-linked immunosorbent assay. The subjects were divided, according to their maximum CPITN codes, into four periodontal groups: max.CPITN-0/1/2, -3, -4, or -MS (missing sextant). Multivariable logistic regression analyses revealed significant associations of the max.CPITN-4 subjects who were males (odds ratio, OR: 2.03) or smokers (OR: 1.73), or who had high diastolic blood pressure (OR: 1.27), high WBC (OR: 1.24), high serum CRP level (OR: 2.39) and high serum IgG.Pg.Fim. (OR: 1.17), IgG.Pg.whol. (OR:1.54), and IgG.Aa.whol. (OR: 1.19) titers. Significant associations between periodontal status and several health conditions were found in the adult population examined, including gender, smoking habit, diastolic blood pressure, white blood cell counts, C-reactive protein, and serum IgG antibodies to P. gingivalis fimbriae, IgG P. gingivalis whole cell, and IgG A. actinomycetemcomitans whole cell titers.


Immunodominance of conformation-dependent B-cell epitopes of protein antigens

Immunodominance of conformational epitopes over linear ones in four proteins was quantified making use of the B-cell hybridoma technology. The proteins were immunized in their native forms into BALB/c mice, and clonal frequencies of B-cell hybridomas that produce antibodies to the native and denatured forms were determined, using ELISA and immunoblotting. All 16 monoclonal antibodies (mAbs) to Porphyromonas gingivalis fimbria were suggested to recognize conformational epitopes expressed by the oligomer. Ten out of 14 mAbs to Serratia marcescens fimbria and 13 of 15 mAbs to hen lysozyme were also specific to their conformational epitopes. In contrast, all 18 mAbs to a surface protein of Streptococcus mutans, termed PAc, reacted to both the native and denatured forms, thereby indicating the immunodominance of linear epitopes in this protein. The results suggest that B-cell epitopes of proteins possessing stable tertiary or quaternary structures are predominantly expressed by the higher-order structures.


Contribution of conformational stability of hen lysozyme to induction of type 2 T-helper immune responses

It is important to identify characteristics that confer on proteins the potential to induce allergenic sensitization and allergenic disease. Protein allergens carry T-cell epitopes that are capable of inducing a type 2 T helper (Th2) cell response. There is limited information regarding factors that govern the allergenicity of proteins. We previously reported that a decrease in the conformational stability of hen-egg lysozyme (HEL) enhanced its capacity to activate HEL-specific T cells owing to the increased susceptibility to intracellular antigen processing. To determine whether the conformational stability of HEL makes for a critical contribution to allergenic sensitization in vivo, we immunized BALB/c mice with HEL derivatives of different conformational stability, but which retained a similar three-dimensional structure. The magnitude of in vivo T-cell responses, evaluated by ex vivo proliferative responses of lymph node T cells from mice primed with various HEL derivatives, was inversely correlated with conformational stability, as was interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production by splenic T cells in response to HEL. Immunization of the least stable derivative led to a potent IL-4 response and to immunoglobulin E (IgE) antibody production. We propose that the intrinsic allergenicity of proteins can be determined by the degree of conformational stability.


Porphyromonas gingivalisspecific lgG subclass antibody levels as immunological risk indicators of periodontal bone loss

It has been well demonstrated a positive association between the magnitude of host antibody response and periodontal disease status. Previous studies also reported that Porphyromonas gingivalis-specific IgG subclass antibodies were elevated in sera from adult periodontitis patients. However, the rôle and the association of these IgG subclass antibodies to the development of periodontal diseases are poorly understood. The aim of present investigation was to examine the relation of serum IgG subclass antibody levels and alveolar bone loss in treated and untreated periodontitis patients. Serum samples were taken from 20 treated and maintained periodontitis patients (SPT patients), 30 untreated patients and 19 periodontally healthy subjects. We determined the IgG subclass antibody titers to P. gingivalis whole cells using an enzyme-linked immunosorbent assay (ELISA). Subgingival plaque samples were taken from the mesio-buccal surface of 6 randomly selected teeth of SPT patients and evaluated for the presence of P. gingivalis by immunofluorescence microscopy. Clinical measurements were also taken including full mouth intraoral radiographs to measure interproximal alveolar bone loss at baseline (BLS1) and at a 5-year recall visit in the SPT patients (BLS2). Our results indicated that both patient groups had detectable levels of IgG1, IgG2, and IgG4. Significantly higher IgG1 was observed in both patient groups compared to the healthy subjects. The untreated patients also exhibited significantly elevated IgG2 response (p<0.05). The mean IgG4 level of the SPT patients was significantly higher compared to the other subject group (p<0.05). A statistically significant positive correlation between IgG2 levels and changes in bone levels (DeltaBLS: BLS2-BLS1) was seen in the SPT patients (p<0.001). SPT patients with high IgG2 and low IgG4 showed greater bone loss than those with low IgG2 and high IgG4 (p<0.05), although the mean prevalence of P. gingivalis in the 2 groups did not differ significantly. Our data suggest that the prolonged IgG2 response after periodontal treatment may be indicative of recurrent or persistent periodontal destruction.


Periodontal status and serum antibody titers for Porphyromonas gingivalis fimbriae in a rural population in Japan

The present study was undertaken to assess the periodontal status of a rural Japanese population and to study the correlation between the periodontal status and the serum antibody titers for Porphyromonas gingivalis (Pg) fimbriae. A total of 236 individuals were examined for their periodontal conditions by the use of the community periodontal index for treatment needs (CPITN), and serum antibody titers for Pg fimbriae in their peripheral blood samples were evaluated using the enzyme-linked immunosorbent assay. There was a substantially larger proportion of edentulous subjects in the age group older than 60 years. The remaining teeth were 24.1, 23.2, 11.1 and 10.1 per person in the 40-49, 50-59, 60-69 and > or = 70 age groups, respectively. The % of sextants with a CPITN code of missing sextant (MS) increased towards elderly and reached >60% in the age group of > or = 70 years, as the % of the CPITN 2, 1 or 0 sextant decreased. The % of CPITN 4 and 3 sextants did not differ between different age groups and were about 6-8% and 15-20%, respectively. The % of CPITN 1 or 0 sextants was higher in female subjects than in male subjects in the 60-69 and > or = 70 age groups, while the % of CPITN 4 or 3 sextants was higher in male subjects than in female subjects in all age groups. There was no significant difference between various age groups in the mean serum antibody titers for Pg fimbriae. The mean anti-Pg fimbriae antibody titers was significantly higher for the subjects with a maximum CPITN code 4 (max.-CPITN 4 subject) than for the subjects with lower maximum CPITN codes. The antibody titers varied extensively among the max.-CPITN 4 or 3 subjects, but not among the max.-CPITN 2/1/0 or MS subjects. The present study demonstrated that tooth loss is a remarkable event in elderly subjects and that oral prophylaxis and mechanical debridement should be mandatory in the population examined. It was also demonstrated that the serum antibody titers against Pg fimbriae could be useful for screening individuals with moderate to severe periodontitis.


Immunodominant Region of Actinobacillus actinomycetemcomitans 40-kilodalton Heat Shock Protein in Patients with Rheumatoid Arthritis

Bacterial heat shock proteins have been implicated in the pathogenesis of several diseases, and the immunological relationship between rheumatoid arthritis (RA) and Escherichia coli DnaJ has been reported. Since there are similarities in the tissue destruction process of RA and periodontitis, we examined the reactivities of antibodies in sera from RA patients to the DnaJ protein from Actinobacillus actinomycetemcomitans. An enzyme-linked immunosorbent assay showed that IgG titers to the N-terminal conservative region of the DnaJ are significantly higher in RA patients compared with the healthy controls (p < 0.05). Furthermore, we examined IgG titers of disease controls to determine the specificity of the immune responses to this region in RA patients. The difference between RA and infectious disease patients was also significant (p < 0.05). These results suggest that the N-terminal region of DnaJ from A. actinomycetemcomitans may contribute to the etiologic analysis of RA.


Effect of Denture Application on senile patients --- Relationship between mastication and immunological function


Endocardiac infectivity and binding to extracellular matrix proteins of oral Abiotrophia species

Microorganisms of the genus Abiotrophia, formerly known as nutritionally variant streptococci, are members of the oral flora and often isolated from patients with endocarditis, but pathogenicity of oral Abiotrophia species has not been examined yet. In this study, 17 strains isolated from healthy human oral cavities and 7 reference strains (all derived from patients with endocarditis) of Abiotrophia spp. were tested for their abilities to cause infections in damaged heart tissues in catheterized rats and to adhere to extracellular matrix proteins in vitro. The reference strains of A. defectiva and A. adiacens showed high infectivities in the rats. Four oral isolates of these two species showed similarly high infectivities and three had moderate infectivities. Most of 10 oral strains of A. para-adiacens and A. elegans were found to be generally less infective. The highly infective A. adiacens strains showed markedly high fibronectin-binding capacity, suggesting a possible relationship between the fibronectin-binding capacity and damaged heart tissue infectivity of the Abiotrophia species. A. defectiva strains which were also highly infective had moderate levels of binding to fibronectin and other extracellular matrix proteins. Most of A. para-adiacens and A. elegans strains showed low or negligible binding capacities to any extracellular matrix proteins tested.

FEMS Immunology and Medical Microbiology


The molecular weight ratio of monomethoxypolyethylene glycol (mPEG) to protein determines the immunotolerogenicity of mPEG proteins

Immunotolerogenic activity of monomethoxypolyethylene glycol- (mPEG) conjugated proteins is a beneficial property in protein pharmaceutics. However, procedures for the preparation of tolerogenic mPEG proteins have not yet been defined. We prepared mPEG proteins with different mPEG contents using three proteins, hen egg lysozyme, ovalbumin and bovine gamma globulin, and their tolerogenicities to antigen-specific T and B cell responses were examined. We found the most appropriate ratio of tolerance induction to be 1.5-2.0, which is the molecular weight ratio of conjugated total mPEGs to protein. This value may assist in the preparation of tolerogenic mPEG proteins.


Extended blood half-life of monomethoxypolyethylene glycol-conjugated hen lysozyme is a key parameter controlling immunological tolerogenicity

The blood half-life of a protein is prolonged by conjugating a protein with a linear amphiphilic polymer, monomethoxypolyethylene glycol (mPEG). The conjugation gives a protein immunotolerogenicity; hence, it is likely that the long half-life is crucial for the tolerogenicity. We prepared a tolerogenic mPEG conjugate of hen egg lysozyme (mPEG1.5-HEL), which is conjugated 1.5-fold the molecular weight of mPEG against that of HEL, and evaluated the relationship between in vivo stability and the tolerogenicity. mPEG1.5-HEL retained immunogenicity to prime HEL-specific T cell and antibody responses and had a long blood half-life, more than 27 times that of native HEL. The tolerant state was maintained as long as mPEG1.5-HEL was detected in sera. With a decrease in the blood mPEG1.5-HEL level, the tolerant state returned gradually to the responsive state; however, reinjection of mPEG1.5-HEL again restored the tolerance. Thus, the extended blood half-life of HEL by mPEG conjugation is probably vital for establishing and maintaining the tolerant states.


Nasopharyngeal-Associated Lymphoreticular Tissue (NALT) Immunity: Fimbriae-Specific Th1 and Th2 Cell-Regulated IgA Responses for the Inhibition of Bacterial Attachment to Epithelial Cells and Subsequent Inflammatory Cytokine Production


Tolerogenic activity of polyethylene glycol-conjugated lysozyme distinct from that of the native counterpart


A novel approach for detecting an immunodominant antigen of Porphyromonas gingivalis in diagnosis of adult periodontitis

Clinical and Diagnostic Laboratory Immunology


Depression of T-cell Epitope Generation by Stabilizing Hen Lysozyme


Prevention of collagen-induced arthritis (CIA) by treatment with polyethylene glycol-conjugated type II collagen; distinct tolerogenic property of the conjugated collagen from the native one


Hen egg-white lysozyme inhibits biological activities of lipopolysaccharides from periodontopathic bacteria.


Role of the Charged Tail in Localization of a Surface Protein Antigen of Streptococcus mutans


Quaternary structure-dependent idiotope and antigen binding of a monoclonal antibody specific for conformational epitope on type II collagen

We previously generated a monoclonal antibody (mAb) against a putative pathogenic epitope on native type II collagen (CII) for the induction of collagen-induced arthritis in mice (mAb1), and an anti-idiotypic mAb which appears to possess the internal image of the CII epitope (mAb2). In the present study, the structural basis of the antigen/mAb1 and mAb1/mAb2 interactions was examined. When partially SH-reduced mAb1 was analysed on Western blots, only fragments containing both heavy (H) and light (L) chains were recognized by mAb2. When mAb2 was partially SH-reduced, only fragments containing both H and L chains were recognized by mAb1. H and L chains were separated from mAb1 in a reduced, denatured condition, and each chain and a mixture of the two were refolded. mAb2 reacted specifically to the renatured whole IgG molecule of mAb1, but not to the refolded L or to H chains. Recombinant single chain Fv (scFv) generated from mAb1 and mAb2 had properties of the original mAbs, whereas genetically constructed chimeric scFvs, consisting of VH from mAb1 and an irrelevant VL. or VL of mAb1 and an irrelevant VH. did not react either to CII or to mAb2. Thus, interactions among CII, mAb1 and mAb2 appear to depend on quaternary structures containing different protein subunits. These observations support the internal image property of the mAb2. In addition, this dependency on quaternary structure for recognition of proteins may also be relevant to other protein-protein interactions.


Lipopolysaccharides from Porphyromonas gingivalis, prevotella intermedia and Actinobacillus actinomycetemcomitans promote osteoclastic differentiation in vitro


Reduced immunogenicity of monomethoxypolyethylene glycol-modified lysozyme for activation of T cells


抗 Porphyromonas gingivalis 抗体の IgG サブクラスと歯周炎の病態

Journal of the Japanese Association of Periodontology


T cell depletion of human bone marrow using an oxidase-peroxidase enzyme immunotoxin


Enhanced in vitro cytotoxicity of 1,3-bis-(2-chloroethyl)-1-nitrosourea or buthionine sulfoximine combined with a reactive oxygen-generating enzyme immunotoxin


Actinomyces viscosus T14V 株の示す多クローン性B細胞活性化 --- ヒト末梢血リンパ球の応答性


Immunotoxins Containing Glucose Oxidase and Lactoperoxidase with Tumoricidal Properties: in Vitro Killing Effectiveness in a Mouse Plasmacytoma Cell Model


An immunotoxin system intended for bone marrow purging composed of glucose oxidase and lactoperoxidase coupled to monoclonal antibody 097


The Function of Gingival Lymphocytes on the Establishment of Human Periodontitis


Polyclonal induction of IgG antibody forming cells by stimulation with Actinomyces viscosus T14V


Possible role of T cells in the establishment of IgG plasma cell-rich periodontal lesion--Augmentation of IgG synthesis in the polyclonal B cell activation response by autoreactive T cells


T-cell requirement for establishment of the IgG-dominant B-cell lesion in periodontitis


T Cells Modulate T-independent Polyclonal B Cell Activation and Enhance IgG Synthesis --- 特に，多クローン性 IgG 産生の増強について

T細胞による多クローン性B細胞活性化の修飾 --- 特に，多クローン性 IgG 産生の増強について


Utility of Natural Herbs -Focusing on Dental Field Use

天然ハーブの有用性ー歯科領域を中心に