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The aim of this study was to examine the effect of adjunct local minocycline administration on the microbiological parameters of subgingival plaque samples in the residual periodontal pockets. Ten chronic periodontitis patients under a supportive periodontal therapy regimen were recruited. After subgingival debridement, either 2% minocycline gel, Periocline™, (Test Group) or a placebo (Control Group) was administered to the selected sites once a week for three weeks. Subgingival plaque was collected at baseline, and at four weeks and eight weeks. The microbiological composition was analyzed by 16S ribosomal RNA sequencing. In the Test Group, α-diversity (evenness) decreased compared to the baseline (p = 0.005) and was lower compared to the control group at four weeks (p = 0.003). The microbial community composition between the two groups was significantly different at four weeks (p = 0.029). These changes were attributable to a decrease in the bacteria associated with periodontitis and an increase in the bacteria associated with periodontal health. Additionally, the improvement in bleeding on probing continued at eight weeks; however, there were little microbial effects of 2% minocycline gel observed at eight weeks. The control group demonstrated no change throughout the eight-week experimental period. Thus, local minocycline administration can change the subgingival microbial community of residual periodontal pockets.

DOI： 10.3390/dj8040123

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Language：Japanese   Publisher：(NPO)日本歯科保存学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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BACKGROUND AND PURPOSES: Osteoporosis and osteopenia are multifactorial diseases characterized by low bone mineral density (BMD) and are susceptible to genetic and environmental risk factors. The macrophage erythroblast attacher (MAEA) was discovered as a protein to mediate the attachment of erythroid cells to macrophages and is essential for bone marrow hematopoiesis. MAEA is expressed in a wide range of cells and tissues including osteoblasts and osteoclasts. Recent studies have shown that a single nucleotide polymorphism (SNP) rs6815464 (C/G) in the MAEA gene increases the susceptibility of type 2 diabetes mellitus. However, the contribution of MAEA to bone metabolism remains unknown. Therefore, we performed this study to evaluate the association between MAEA polymorphism and low BMD. METHODS: In a cross-sectional study with postmenopausal Japanese women living in the Yokogoshi area, Niigata City, we evaluated whether rs6815464 was associated with low BMD. Blood samples were collected from 353 subjects (age 63.8 ± 5.4 years). The MAEA genotype was determined by TaqMan assay. BMD was assessed by dual-energy X-ray absorptiometry at the lumbar spine (L2-L4), hip and femoral neck. Low BMD was defined as a T-score <-1. RESULTS: The percentage of subjects with low BMD in the lumbar spine, total hip and femoral neck were 71%, 75% and 84% respectively. After adjusting age, BMI, HbA1c, smoking and alcohol consumption, the G-allele carriage was found to be associated with low BMD of total hip (odds ratio = 2.11, 95% CI: 1.14-3.91, P = 0.018), but not of the lumbar spine or femoral neck. CONCLUSION: The MAEA gene polymorphism rs6815464 was associated with low hip BMD in postmenopausal Japanese women.

DOI： 10.1016/j.gene.2019.03.027

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OBJECTIVES: Macrophage erythroblast attacher (MAEA) is a membrane protein that regulates the development of mature macrophages by mediating attachment with erythroblasts. A polymorphism rs6815464 (C/G) in MAEA gene was reported to be associated with type II diabetes. Along with diabetes, osteoporosis shows an increased prevalence in postmenopausal females, and both diseases have been reported to be associated with periodontitis. Therefore, we explored the relevance of the MAEA polymorphism to periodontitis, bone mineral density (BMD) and haemoglobin A1c (HbA1c). DESIGN: This was a cross-sectional study with the final sample comprised of 344 postmenopausal Japanese females. Probing pocket depth (PPD) and clinical attachment level (CAL) were measured. Genotype was determined by TaqMan assay. Blood biochemical parameters and BMD of the lumbar spine were evaluated. RESULTS: No differences were found in age, body mass index, HbA1c, BMD, number of teeth, bone metabolism parameters between the genotypes. Mean CAL and percentage of sites with PPD or CAL ≥ 5 mm were higher in the G-allele carriers than in the non-carriers. Multiple logistic regression analyses revealed that G-allele carriage was associated with severe periodontitis (odds ratio = 3.73, 95% CI = 1.36-10.19). CONCLUSION: Our results suggested that the MAEA gene polymorphism was independently associated with severe periodontitis.

DOI： 10.1016/j.archoralbio.2019.04.008

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Most studies that have demonstrated an association between number of remaining teeth and cognitive impairment have treated teeth as a continuous variable, although the relationship is nonlinear. The aim of this cross-sectional study was to determine the critical number of remaining teeth in hospital outpatients at which the association with cognitive impairment becomes apparent. Japanese adults living on Sado Island who visited Sado General Hospital were invited to participate in Project in Sado for Total Health. In total, 2,530 adults were interviewed and had their teeth counted; 1,476 of these individuals also completed the Mini-Mental State Examination (MMSE) and underwent measurement of their serum high-sensitivity C-reactive protein (hsCRP) levels. Patients on dialysis and those with hsCRP ≥ 10 mg/L were excluded. The final study group consisted of 565 adults (290 men and 275 women) of mean age 69.8 (range 29-91) years. An MMSE score < 24 was considered to indicate cognitive impairment. The subjects were categorized according to whether they had an edentulous jaw or one to 10, 11-20, 21-27, or ≥28 remaining teeth. One hundred twenty-eight of the 565 study participants were diagnosed to have cognitive impairment. Multiple logistic regression analysis revealed associations of cognitive impairment with older age, ischemic heart disease, smoking, and alcohol consumption. After adjustment for covariates, having one to 10 remaining teeth was significantly associated with cognitive impairment. There is a significant association between having only one to 10 remaining teeth and cognitive impairment in hospital outpatients.

DOI： 10.1002/cre2.147

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Blackwell Munksgaard  

Background and Objectives: Previous reports suggest that several serum biomarkers play roles in the pathogenesis, inflammatory response, and oxidative stress in periodontitis caused by bacterial infections, linking chronic periodontitis to atherosclerotic vascular disease (ASVD). The aim of this preliminary study was to investigate, in a Japanese cross-sectional community survey, potential serum biomarkers of periodontitis that are associated with ASVD and chronic periodontitis. Material and Methods: The study cohort included a total of 108 male subjects who underwent annual health examinations. Serum biomarkers (high-sensitivity C-reactive protein [hs-CRP], proprotein convertase subtilisin/kexin type 9 [PCSK9], interleukin-6, tumor necrosis factor-α, soluble CD14, myeloperoxidase, matrix metalloproteinase-3, adiponectin, total bilirubin [TBIL], and serum lipids) were analyzed to determine their association (if any) with periodontal parameters. Aortic stiffness was evaluated using the brachial-ankle aortic pulse wave velocity (PWV) index and the cardio-ankle vascular index (CAVI). Results: The concentrations of PCSK9 and hs-CRP were increased (P =.001 and.042, respectively), and the concentration of TBIL was decreased (P =.046), in subjects with periodontal disease (determined as a probing depth of ≥4 mm in at least one site) compared with periodontally healthy subjects. The ratio of low-density lipoprotein cholesterol (LDL-C) to high-density lipoprotein cholesterol and the concentrations of triglycerides, remnant-like particles-cholesterol, and oxidized LDL were elevated in subjects with periodontal disease compared with periodontally healthy subjects (P =.038,.007,.002, and.049, respectively). Multivariate regression analyses indicated that the number of sites with a pocket depth of ≥4 mm was associated with the concentration of PCSK9 and inversely associated with the concentration of TBIL independently (standardized β =.243, P =.040
standardized β = −.443, P =.0002
respectively). Analysis of receiver operating characteristic curves of PCSK9 indicated moderate accuracy for predicting the presence of disease sites (probing depth ≥ 4 mm) (area under the curve = 0.740). No significance in the values of PWV and CAVI was observed between subjects with periodontal disease and periodontally healthy subjects. Conclusion: In Japanese male subjects, the concentrations of serum PCSK9 and TBIL were correlated with periodontal parameters. Moreover, PCSK9 could be a candidate biomarker for diagnosing chronic periodontitis, and may also have potential to evaluate the risk for periodontitis to cause ASVD. Longitudinal studies of larger populations are necessary to confirm the exact association of periodontitis with increased serum PCSK9 and decreased TBIL.

DOI： 10.1111/jre.12533

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Background: The interaction of periodontopathic bacteria with host immune system induces the production of inflammatory mediators which leads to alveolar bone loss (ABL), the essential feature of periodontitis. Concurrently, periodontal diseases cause the elevation of blood cytokine levels, the alteration of gut microbiota and the dissemination of enterobacteria to the liver. Owing to these mechanisms, periodontal disease might be a risk for liver dysfunction. Several epidemiological studies have reported associations between periodontal diseases and liver dysfunction, although the association between ABL and liver dysfunction has not been investigated. This cross-sectional study determined if elevated serum liver enzyme levels were associated with ABL in Japanese adults. Material and Methods: Japanese adults living on Sado Island who visited Sado General Hospital were invited to participate in the study. Participants over 40 years of age who underwent dental panoramic radiography and blood tests were included. Drinking and smoking habits were self-administered. After excluding patients with edentulous jaw, diagnosed liver diseases, and those on dialysis, data from 44 men and 66 women with a mean age of 73 years were analyzed. The average percentage of ABL for each participant was calculated for mesial and distal sites of all remaining teeth. The levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) were determined. Univariate analyses were performed to select covariates to be put in multivariate analyses. The association between elevated serum liver enzyme levels and the highest quartile of ABL were assessed by multiple logistic regression analysis. Results: After adjusting for covariates, no significant association was found between elevated serum AST, ALT, or GGT levels as dependent variables and the highest quartile of ABL as an explanatory variable. Conclusions: There was no significant association between the elevation of serum liver enzyme levels and ABL in Japanese adults. Key words:Liver enzymes, dental panoramic radiography, alveolar bone loss, Japanese adults.

DOI： 10.4317/jced.54555

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OBJECTIVES: An interrelationship between rheumatoid arthritis (RA) and periodontitis has been suggested due to their common pathogenic mechanisms. Protein carbamylation and neutrophil extracellular traps (NETs) formation have been shown to be related to autoimmune conditions, including RA, but their association with periodontitis has not been elucidated. Therefore, we assessed whether or not circulating levels of carbamylated protein (CarP) and NETs are associated with periodontitis severity and influenced by periodontal treatment. METHODS: We conducted a retrospective case-control study that included 40 patients with RA and periodontitis, 30 patients with periodontitis, and 43 systemically and periodontally healthy controls to assess the circulating levels of CarP and NETs and rheumatologic and periodontal conditions. The same assessments were also performed in 22 patients with RA and periodontitis after 2 months of periodontal treatment, including oral hygiene instruction and full-mouth supragingival scaling. RESULTS: Patients with RA and periodontitis showed significantly higher serum levels of CarP and NETs than the control group (P = 0.04 and P < 0.001, respectively). The serum levels of CarP and NETs were significantly correlated positively with the mean values of probing depth (P = 0.01 and P = 0.007, respectively) and clinical attachment level (P = 0.007 and P = 0.001, respectively) in the 40 patients with RA and periodontitis. Multiple logistic regression analyses also revealed significantly positive associations between the serum levels of CarP and NETs and moderate to severe periodontitis (P = 0.03 and P = 0.001, respectively). Furthermore, periodontal treatment significantly decreased the serum levels of CarP and NETs in patients with RA and periodontitis (P = 0.03 and P = 0.02). CONCLUSION: The circulating levels of CarP and NETs are associated with periodontitis severity and influenced by periodontal treatment in patients with RA.

DOI： 10.1371/journal.pone.0192365

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

OBJECTIVES: The aim of this study was to examine the association between periodontal disease and renal function in elderly women with different genotypes. MATERIAL AND METHODS: A total of 332 postmenopausal never-smoking women were analysed. Poor renal function was defined as serum cystatin C > 0.91 mg/l. Periodontal disease markers such as periodontal inflamed surface area (PISA) were evaluated. Selected variables, including PISA quartile, body mass index (BMI), HbA1C and age in Arg allele carriers and non-carriers based on the beta-3 adrenergic receptor, or between Ala allele carriers and non-carriers based on peroxisome proliferator-activated receptor gamma, were analysed using multiple logistic regression analysis. RESULTS: The odds ratios of serum cystatin C level and PISA (fourth quartile) were significantly positive for both Arg (2.52; p = 0.035) and Ala allele non-carriers (2.36; p = 0.021). A significant association was also found between serum cystatin C level and BMI for both Arg (1.18; p = 0.001) and Ala allele non-carriers (1.12; p = 0.003). CONCLUSION: The results of this study suggest that periodontal inflammation might be associated with renal function. Furthermore, in both the Arg and Ala allele non-carriers, the associations between BMI and PISA for renal function became stronger.

DOI： 10.1111/jcpe.12708

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BACKGROUND: Several studies have reported inconsistent results regarding the association between the PPARγPro12Ala polymorphism and obesity. Obese individuals had higher C-reactive protein (CRP) levels compared with those of normal weight, and PPARγ activation could significantly reduce serum high-sensitive CRP level. We have previously suggested that the Pro12Ala polymorphism represents a susceptibility factor for periodontitis, which is a known risk factor for increased CRP level. OBJECTIVES: The aim was to investigate associations between PPARγ gene polymorphism, serum CRP level, BMI and/or periodontitis among post-menopausal Japanese women. MATERIALS AND METHODS: The final sample in this study comprised 359 post-menopausal Japanese women. Periodontal parameters, including PD, CAL and BOP, were measured per tooth. PPARγPro12Ala genotype was determined by PCR-RFLP. Hs-CRP value was measured by a latex nephelometry assay. RESULTS: No significant differences in age, BMI or periodontal parameters were found between the genotypes. The percentages of sites with PD ≥ 4 mm were significantly higher among the hsCRP ≥ 1 mg/l group than the hsCRP < 1 mg/l group (p = 0.003). Positive correlations were found between serum hsCRP levels and the percentages of sites with PD ≥ 4 mm (p = 0.043) in PPARγ Ala allele carriers, and BMI (p = 0.033) in non-carriers. After adjustment for model covariates, BMI was significantly associated with serum hsCRP level. CONCLUSION: The PPARγPro12Ala polymorphism was not independently associated with periodontitis, serum CRP level or BMI in post-menopausal Japanese women. However, serum hsCRP level correlated with periodontitis in Ala allele carriers, and with BMI in non-carriers.

DOI： 10.1111/ger.12110

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BACKGROUND: It has been hypothesized that β-3 adrenergic receptor and peroxisome proliferator-activated receptor gamma (PPARγ) might have gene-environmental and gene-gene interactions in periodontal disease. The purpose of this study is to elucidate the interaction between β-3 adrenergic receptor and PPARγ gene polymorphism with periodontal disease. METHODS: Three hundred thirty-two postmenopausal females were enrolled, and their serum high-sensitivity C-reactive protein (hsCRP) and hemoglobin A1c (HbA1c) were examined. β-3 adrenergic receptor and PPARγ genotypes were then determined. An oral examination was performed. The number of remaining teeth was counted, and the probing depth (PD) and clinical attachment level (CAL) were measured. Prevalence-rate ratios (PRRs) were calculated by multiple Poisson regression analyses to evaluate the relationship among periodontal disease markers, such as the number of sites with CAL 4 to 5 or ≥6 mm or PD 4 to 5 or ≥6 mm, and β-3 adrenergic receptor polymorphisms, PPARγ polymorphisms, and the interaction term adjusted by age, hsCRP, and HbA1c, after converting the number of remaining teeth (n) to an offset variable. RESULTS: In the participants with body mass index (BMI) ≥25, PRRs of β-3 adrenergic receptor genotype (Trp/Arg and Arg/Arg) for periodontal disease markers were 0.13 to 0.70 (P <0.0001 to 0.74), those of PPARγ genotype (Pro/Pro) were 0.66 to 3.14 (P = 0.01 to 0.68), and those of the interaction term for the two genotypes were 1.69 to 12.61 (P <0.0001 to 0.33). However, in the participants with BMI <25, a constant tendency was not observed. CONCLUSION: The results confirmed a positive relationship between the interaction term for β-3 adrenergic receptor genotype and PPARγ genotype and various periodontal markers in obese elderly females.

DOI： 10.1902/jop.2015.140472

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier Ltd  

Objective IL-6 plays critical roles in bone resorption and the pathogenesis of periodontitis in both inflammation and alveolar bone loss. A negative correlation was observed between periodontitis and truncal bone mineral density (BMD) in postmenopausal women. The C allele carriers of a genetic polymorphism IL-6-572G/C have higher levels of serum IL-6 compared to G allele carriers. We investigated the possible effect of IL-6-572G/C polymorphism on the relationship between low BMD and periodontitis in postmenopausal women. Subjects and methods A total of 300 postmenopausal Japanese women who lived in Yokogoshi area of Niigata City, Japan, participated in this study. Genomic DNA was extracted from peripheral blood. The IL-6-572G/C genotypes were determined by the restriction fragment length polymorphism method. Bone mineral density (BMD) of right femoral neck and serum bone metabolism markers were measured. Low BMD was defined to have the BMD &lt
80% of the mean for young adults. Periodontal parameters at two sites per tooth were measured. Results Serum osteocalcin levels were significantly lower in the IL-6-572G/G genotype (p = 0.025). In the -572G allele non-carriers, percentages of PPD ≥ 4 mm sites were significantly higher in low BMD group compared with the healthy control group (p = 0.021). Logistic regression analysis revealed low BMD to be associated with periodontitis (Odds ratio = 1.736, p = 0.027) after adjusted with IL-6-572G carriage, age, serum albumin level. Conclusions IL-6-572G/C polymorphism was not an independent risk factor of low BMD or periodontitis, but may affect the relationship between the two diseases in postmenopausal Japanese women.

DOI： 10.1016/j.archoralbio.2014.12.005

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Language：Japanese   Publisher：The Japanese Society of Conservative Dentistry  

Purpose: Mouth rinse with chlorhexidine gluconate (CHG) has been reported to be effective as a chemical plaque control. However, undesirable side effects such as allergy are known problems. Therefore, we developed a new mouth rinse formulated with both chlorhexidine hydrochloride (CHH) and cetylpyridinium chloride (CPC) as anti-microbial constituents and tested its effects on oral bacteria in patients with chronic periodontitis.<br> Methods: This was a short-term, randomized, controlled clinical trial. Systemically healthy 30 patients with ≥20 teeth were enrolled if they had 1 to 7 sites of residual pockets ≥5 mm after initial periodontal therapy with scaling and root planing. Patients were assigned to three groups and used placebo (control group), or rinse with 0.05% CHG (CHG group), or rinse with 0.05% CHH and 0.05% CPC (CHH+CPC group), for 4 weeks. In each patient, saliva, tongue coating and supragingival plaque were collected and microbiological parameters were measured with real-time PCR. Terminal restriction fragment length polymorphism (T-RFLP) analyses were also performed.<br> Results: No adverse event was observed. At the baseline, no difference was found in age, sex, number of teeth, or clinical periodontal parameters between the groups. Comparing the data at baseline and after 4 weeks, total bacterial counts and <i>S. mutans</i> in saliva, total bacterial counts in tongue coating and total streptococci in supragingival plaque were significantly decreased in the CHH+CPC group. In the CHG group, total bacterial counts in supragingival plaque were significantly decreased. Significant decreases of total bacterial counts and streptococci in tongue coating were detected in the control group. As the results from T-RFLP analyses, saliva from the CHH+CPC group showed significant reductions of presumed numbers of Streptococcus group in the digested fragments by <i>Msp I</i>, and Streptococcus, Eubacterium group, Parvimonas group and Porphyromonas, Prevotella group in the digested fragments by <i>Hha I</i>. Additionally, in supragingival plaque obtained from the CHG group after 4 weeks, presumed Streptococcus, Veillonella group from <i>Hha I</i> digests were significantly reduced. No change was observed between baseline and after 4 weeks in the other bacterial data.<br> Conclusion: The new mouth rinse formulated with CHH and CPC might be used safely and have equal to greater effects reducing oral bacteria than existing mouth rinse formulated with CHG alone.

DOI： 10.11471/shikahozon.57.219

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OBJECTIVES: The purpose of this study was to elucidate whether the association between beta-3 adrenergic receptor polymorphism and periodontal disease is modified by body weight. MATERIAL AND METHODS: We enrolled 332 postmenopausal women and determined their HbA1C levels (%) and beta-3 adrenergic receptor (rs4994) genotypes. Periodontal parameters including clinical attachment level (CAL) were measured. After selecting subjects for each body mass index (BMI) level, the prevalence rate ratio (PRR) by multiple Poisson regression analysis was calculated to evaluate the relationship between periodontal disease and beta-3 adrenergic receptor polymorphism. The number of sites with CAL≥6 mm was used as a dependent variable, and beta-3 adrenergic receptor genotype [categorized as Arg non-carriers (reference) or Arg carriers], age (y) and HbA1C (%) were adopted as independent variables. We converted the number of probing sites (n) to an offset variable. RESULTS: The PRR of the beta-3 adrenergic receptor genotype for the number of sites of CAL≥6 mm showed a positive association in subjects with BMI≥25.0 and increased markedly with BMI. The PRR in subjects with BMI≥30 was 3.10 (p < 0.0001). CONCLUSION: This study indicates a positive association between periodontal disease and the beta-3 adrenergic receptor genotype in obese individuals.

DOI： 10.1111/jcpe.12235

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PERGAMON-ELSEVIER SCIENCE LTD  

OBJECTIVE: Biomarkers in gingival crevicular fluid (GCF) have been investigated; however, measurements were limited by the small sample volume available. The aim of this study was to determine the levels of 40 different cytokines and chemokines in GCF samples. DESIGN: Eleven patients with generalised chronic periodontitis participating in a supportive periodontal therapy programme with remaining probing pocket depths (PDs) of >5mm were enrolled. One healthy and two diseased sites were sampled in each subject. Forty biomarkers in GCF were examined using a multiplex bead immunoassay. Porphyromonas gingivalis from the diseased sites was quantified by real-time polymerase chain reaction. RESULTS: Twenty-six biomarkers were detected in the GCF samples using the multiplex bead immunoassay. The levels of nine biomarkers were significantly different between the diseased and healthy sites after adjustment with Bonferroni's correction. The level of 26 biomarkers in diseased sites was compared between bleeding on probing (BOP)-positive and BOP-negative sites. Interleukin (IL)-1β and interferon-inducible protein (IP)-10 levels were significantly higher in BOP-positive diseased sites than BOP-negative diseased sites after adjustment for multiple comparisons (IL-1β, p=0.0007, IP-10; p=0.0009). In addition, the levels of IL-1β in GCF were found to be strongly correlated with the P. gingivalis ratio (r=0.646, p=0.0012). CONCLUSION: IL-1β levels in GCF correlate with the PDs, BOP and the presence of P. gingivalis in subgingival plaque. Multiplex bead assays can be useful in GCF studies. These findings can help in identifying new diagnostic methods in the diagnosis of periodontal disease.

DOI： 10.1016/j.archoralbio.2012.11.012

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OBJECTIVES: PPARg regulates bone metabolism and inflammation. Our previous study suggested PPARg Pro12Ala polymorphism to represent a susceptibility factor for periodontitis in pregnant Japanese women. Several recent papers have drawn attention to a possible link between low bone mineral density (BMD) and periodontitis in postmenopausal women. Since the pathogenesis for both involve bone remodeling, they might share common risk factors such as gene polymorphisms and vitamin D level. The present study investigated possible associations between the PPARgPro12Ala polymorphism, periodontitis, BMD and serum 25(OH)D in postmenopausal Japanese women. MATERIALS AND METHODS: PPARgPro12Ala genotypes of 359 women were determined by PCR-RFLP. BMD and periodontal parameters of each woman were measured. Serum 25(OH)D levels were determined by radioimmunoassay. RESULTS: PPARgPro12Ala polymorphism was not associated with periodontitis or BMD as an independent factor. Serum 25(OH)D was significantly higher in Ala allele carriers compared to non-carriers. Only in the Ala allele carriers, positive correlations were found between mean clinical attachment level and BMD, between BMD and 25(OH)D, and between percentage of sites with probing depth ≥4 mm and 25(OH)D. CONCLUSIONS: PPARgPro12Ala polymorphism was not independently associated with periodontitis or BMD. However, the polymorphism might be a modulator of the relationship between the two conditions in postmenopausal Japanese women.© 2012 John Wiley &amp
Sons A/S.

DOI： 10.1111/odi.12032

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Language：Japanese   Publisher：The Japanese Society of Conservative Dentistry  

Purpose: Periodontopathic bacteria and inflammation in a residual periodontal pocket after periodontal therapy increase the risk for further progression of periodontitis. A tooth cleaning gel containing antibacterial and anti-inflammatory agents would be effective to inhibit such progression. The tooth cleaning gel 'Jellcoat F' contains 0.05% chlorhexidine hydrochloride as an antibacterial and β-glycyrrhetinic acid as an anti-inflammatory. We evaluated the effects of Jellcoat F on residual periodontal pockets with clinical, microbiological and biochemical analyses. Methods: The present study had a randomized, controlled, double-blinded design. Systemically healthy patients with &ge;20 teeth were enrolled if they had at least two teeth with residual pockets of 6-7 mm depth after at least a month from the completion of active periodontal therapy containing scaling and root planing. Twenty patients were randomly assigned to the test group who used Jellcoat F or the control group who used placebo. Gingival index (GI), plaque index (Pl I), probing pocket depth and bleeding on probing were recorded. In each patient, gingival crevicular fluid (GCF) was collected from one of the teeth with residual pockets, and subgingival plaque from the other. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in GCF and Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Treponema denticola in subgingival plaque were determined. After the collections, residual pockets were filled with Jellcoat F or placebo. Patients were instructed to brush their teeth with Jellcoat F or placebo and also apply the gel with a retainer for 10 minutes before sleeping. After 4 weeks, the same examinations were performed. Results: No adverse event was observed. At the baseline, no difference was observed between the test and control groups in age, sex, number of teeth, levels of periodontopathic bacteria, AST and ALT. Comparing the baseline and 4 weeks, only GI and Pl I were significantly decreased in the test group. No other change was found in both test and control groups. Changes of measurements during the 4 weeks were not statistically different between the groups. The effect of Jellcoat F to improve GI remained significant after being adjusted for age and sex. Conclusion: Application of Jellcoat F for 4 weeks on residual pockets after periodontal therapy with teeth brushing and a retainer did not significantly change the levels of periodontopathic bacteria in subgingival plaque, AST and ALT in GCF However, it might suggest the reduction of clinically evaluated supragingival plaque and gingival inflammation.

DOI： 10.11471/shikahozon.56.344

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FcγRIIB contains a unique immunoreceptor tyrosine-based inhibition motif (ITIM) and functions as a negative feedback regulator of leucocyte activation and antibody production. We have previously reported FcγRIIB-nt645+25A/G gene polymorphism to be associated with prevalence and severity of periodontitis, FcγRIIB expression level on peripheral B lymphocytes and the serum IgG level against periodontopathic bacteria. Previous studies have reported maternal periodontal disease to be associated with an increased risk for preeclampsia. Therefore, FcγRIIB-nt645+25A/G gene polymorphism may be associated with preeclampsia by affecting immune response to periodontopathic bacteria in pregnant women. To elucidate whether FcγRIIB-nt645+25A/G gene polymorphism has associations with preeclampsia and/or periodontitis in pregnant Japanese women, a case-control study was carried out on women with preeclampsia (n=13) and without preeclampsia (n=106). Maternal periodontal parameters and bacterial data of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque were collected within 5days of delivery. FcγR genotypes of each woman were determined using the genomic DNA isolated from peripheral blood. Serum IgG levels specific for each bacteria were determined. There was a significant association between FcγRIIB-nt645+25A/G polymorphism and preeclampsia (P=0.013). The frequency of the FcγRIIB-nt645+25AA genotype was higher in the preeclampsia group compared with the nonpreeclampsia group (P=0.007). The DNA level of A. actinomycetemcomitans from subgingival plaque was shown to be higher in the preeclampsia group (P=0.017). In conclusion, maternal FcγRIIB-nt645+25A/G polymorphism and subgingival DNA level of A. actinomycetemcomitans were significantly associated with the prevalence of preeclampsia in a limited number of Japanese women independently with periodontal infection. Further investigations should be performed to confirm this association in a larger population and to determine the biological process of the association. © 2012 Blackwell Publishing Ltd.

DOI： 10.1111/j.1744-313X.2012.01124.x

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Many studies have reported an association between periodontal disease and preterm birth, although this remains controversial. Cytokines and antibodies produced to give resistance to infection can enter the bloodstream and cause preterm labor. We analyzed maternal genetic polymorphisms in various immunoregulatory genes that could affect both preterm birth and periodontitis. A total of 1099 women referred to the Department of Obstetrics and Gynecology, Niigata University Medical and Dental Hospital were candidates for participation, 424 of whom refused, and 553 were excluded. The final number of subjects was 122 (51 with preterm birth, 71 with term birth). Genomic DNA was isolated from venous blood, and 22 polymorphisms were determined: IL-1A, IL-1B, IL-1RN, IL-2, IL-4, IL-6, IL-10, TNFA, TNFRI, TNFRII, FcγRIIA, FcγRIIB, FcγRIIIA, FcγRIIIB, and FcαR. Within five days of labor, periodontal parameters were evaluated, and bacteria from subgingival plaque were detected using real-time PCR. There was no difference in the prevalence and degree of periodontitis between term and preterm births. Chi-squared tests showed that an age <33 years and FcαR(+56)T/C alleles were associated with preterm birth. Multiple logistic regression analysis represented a model with significant fitness in which four variables were associated with preterm birth: maternal age, number of Aggregatibacter actinomycetemcomitans, IL-6(-572)G/C, and FcαR(+56)T/C. In conclusion, there was no association between preterm birth and periodontitis in this study. A. actinomycetemcomitans, IL-6, and FcαR were suggested to be associated with preterm birth. Multiple logistic regression models with both genetic and environmental factors would be useful for evaluating susceptibility to preterm birth.

DOI： 10.1016/j.jri.2012.01.005

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AIM: To determine whether periodontitis and three prominent members of the periodontal flora are associated with the development of preeclampsia (hypertension plus proteinuria) Materials and Methods: The samples were composed of 127 systemically healthy women. Within 5 days after labour, clinical periodontal parameters and Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque were evaluated. Maternal serum IgG antibody specific for each bacteria was determined by enzyme-linked immunosorbent assay. Multivariate logistic regression analysis was used to control for confounders (maternal age, body mass index before pregnancy, parity, and smoking). RESULTS: Eighteen women were affected with preeclampsia. The number of A.actinomycetemcomitans was shown to be significantly associated with preeclampsia in the logistic regression model (odds ratio; 1.7, 95% confidence interval; 1.1–2.7). There were statistically significant differences between the preeclamptic and control groups in body mass index before pregnancy, pre-term birth and low birthweight (respectively, p = 0.014, p = 0.010 and p < 0.0001). We found no statistically significant association between preeclampsia and periodontal clinical parameters or the presence of periodontitis. CONCLUSION: In systemically healthy pregnant women, our findings suggested that the levels of maternal subgingival A. actinomycetemcomitans DNA were elevated in preeclamptic women.

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Sugita N, Iwanaga R, Kobayashi T, Yoshie H. Association of the Fc RIIB-nt645+25A/G polymorphism with the expression level of the Fc RIIb receptor, the antibody response to Porphyromonas gingivalis and the severity of periodontitis. J Periodont Res 2012; 47: 105113. (C) 2011 John Wiley & Sons A/S

DOI： 10.1111/j.1600-0765.2011.01411.x

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Background and Objective: Recently, numerous studies have investigated the association of preterm birth with periodontitis. FcγRIIb is a human low-affinity receptor for immunoglobulin G (IgG). We have previously demonstrated single nucleotide polymorphisms (SNPs) of FcγRIIb to be associated with periodontitis and the serum-specific IgG level against periodontopathic bacteria. In this study, we investigated whether FcγRIIB gene polymorphisms were associated with periodontitis and/or pregnancy outcome. Material and Methods: We assessed the periodontal conditions of 122 Japanese pregnant women within 5d of delivery, and polymorphisms in FcγRIIB and in other Fcγ receptors were detected from the genomic DNA. Using clinical and genomic data, we analyzed the relationship between periodontitis, preterm birth and Fcγ receptor polymorphisms. Results: A significant difference was observed in the distribution of FcγRIIB-nt645+25A/G (rs2125685) between preterm and term birth groups, with a higher prevalence of nt645+25AA in the preterm birth group (p=0.032). Additionally, the FcγRIIB-nt645+25GG carrier showed significantly higher results for the prevalence of periodontitis (p=0.048), mean pocket depth (p=0.021), mean clinical attachment level (p=0.010), percentage of sites with pocket depth ≥4mm (p=0.005) and percentage of sites with clinical attachment level ≥3mm (p=0.007) than the AA carrier. An association between preterm birth and periodontitis was not observed in this study. Conclusion: These findings suggest that FcγRIIB-nt645+25AA carriers are more likely to experience preterm birth than FcγRIIB-nt645+25AG and GG carriers. Also, women with FcγRIIB-nt645+25G exhibited a greater tendency to have periodontitis than those with nt645+25A. © 2011 John Wiley &amp
Sons A/S.

DOI： 10.1111/j.1600-0765.2010.01338.x

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BACKGROUND: Previous studies suggest that periodontitis is closely related to obesity and metabolic syndrome. Leptin, a pleiotrophic hormone produced by adipose tissue, has been reported to be related to periodontitis. This study investigates the effects of periodontal treatment on the serum levels of leptin and other cytokines in patients with chronic periodontitis (CP). METHODS: Serum samples were taken from 33 CP patients (22 non-smokers, 11 smokers) and 18 healthy subjects. The serum leptin, adiponectin, tumor necrosis factor-alpha, interleukin (IL)-6, and C-reactive protein (CRP) levels were measured before and after non-surgical periodontal treatment. RESULTS: Significant differences between healthy and CP patients were found in serum leptin, IL-6, and CRP levels (P = 0.0018, P = 0.0064, and P = 0.0095, respectively). The serum leptin level was associated with mean probing depth, mean clinical attachment level, mean alveolar bone loss, and body mass index. There were significant associations between serum leptin levels and IL-6 and CRP levels. After non-surgical periodontal treatment, serum leptin, IL-6, and CRP levels were significantly decreased (mean +/- SD before and after, P value, respectively: leptin, 8.02 +/- 5.5, 7.10 +/- 4.4, P = 0.015; IL-6, 1.73 +/- 1.02, 1.36 +/- 0.73, P = 0.048; and CRP, 802.0 +/- 1065, 491.2 +/- 479.3, P = 0.047). CONCLUSIONS: Periodontal treatment is effective in reducing serum leptin, IL-6, and CRP levels. The results suggest that leptin, IL-6, and CRP could be mediating factors that connect metabolic syndrome and periodontitis.

DOI： 10.1902/jop.2010.090741

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BACKGROUND: Recent studies suggest an association between maternal periodontitis and preterm birth, although the association remains controversial. It was suggested that mechanisms such as a genetic predisposition for a hyperinflammatory response cause periodontitis and preterm births. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a nuclear hormone receptor and ligand-dependent transcription factor. PPARgamma inhibits the transcriptional activity of the genes that produce proinflammatory mediators and repress periodontitis. Recently, a common polymorphism, proline(PRO)-to-alanine(ALA) mutation at codon12 in exonB (Pro12Ala: rs 1801282) PPARgamma, was reported to reduce the ability to transactivate responsive promoters. In this study, we tested whether the PPARgammaPro12Ala polymorphism was associated with maternal periodontitis and/or preterm birth. METHODS: Genomic DNA was isolated from the venous blood of pregnant Japanese women (term birth: n = 72; preterm birth: n = 58). The PPARgammaPro12Ala genotype was determined by polymerase chain reaction (PCR)-restriction fragment length polymorphism. Within 5 days after labor, clinical periodontal parameters were evaluated, and periodontopathic bacteria from the subgingival plaque were detected by species-specific PCR. RESULTS: The mean clinical attachment level (P = 0.012), mean probing depth (P = 0.031), mean gingival index (P = 0.037), and percentages of sites with bleeding on probing (P = 0.041) in women with the PPARgammaPro12Ala genotype were significantly higher than in women with the PPARgammaPro12Pro genotype. However, there was no association between preterm birth and periodontitis. CONCLUSION: We suggest that the PPARgammaPro12Ala polymorphism may represent a genetic susceptibility factor for the clinical measurements of periodontitis in a limited number of pregnant Japanese women, but it probably cannot influence the relationship between periodontitis and preterm birth.

DOI： 10.1902/jop.2010.090669

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BACKGROUND: Porphyromonas gingivalis (Pg) is one of the most harmful periodontal pathogens and it has been reported that Pg is associated with preterm birth (PTB), intrauterine growth retardation (IUGR) and pregnancy-induced hypertension (PIH), discovered by animal experiments and clinical research. The relationship between adverse pregnancy outcomes and maternal antibody response to Pg is controversial. On the other hand, the serum C-reactive protein (CRP) has been recognised as a reliable serum marker of periodontal disease. AIMS: To determine the significance of antibody responses to Pg affecting pregnancy outcomes in the first trimester. METHODS: A case-control study was carried out on women with PTB (n = 58), IUGR (n = 91), PIH (n = 32) and without any complications (control, n = 98). The serum level of the CRP and IgG1 against 40-kDa outer membrane protein of Pg (anti-40-kDa OMP Pg-IgG1) in the first trimester was measured. RESULTS: The IUGR group, and PTB patients whose placentas were diagnosed as chorioamnionitis or whose vaginal flora included Lactobacilli, showed a lower level of anti-40-kDa OMP Pg-IgG1 than the control group. There was no difference in the serum CRP level between each case group and control group. CONCLUSIONS: These results suggest that a lack of humoral immunity against Pg in early pregnancy is associated with IUGR and some PTB.

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Language：Japanese   Publisher：JAPANESE SOCIETY OF PERIODONTOLOGY  

DOI： 10.14833/amjsp.2009f.0.28.0

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Human Fc gamma RIIB is one of the receptors for immunoglobulin G (IgG) and suppresses the activation of B lymphocytes through cross-linking with the B cell receptor via immune complexes. This function of Fc gamma RIIB is essential for the negative regulation of antibody production. Our previous study has demonstrated the gene polymorphism Fc gamma RIIB-I232T to be associated with periodontitis. The polymorphism Fc gamma RIIB-232T has been reported to inhibit B-cell antigen receptor signaling more effectively compared to Fc gamma RIIB-232I, while other groups concluded that Fc gamma RIIB-232T had no ability to inhibit activatory receptors. In this study, we examined whether Fc gamma RIIB-I232T polymorphism would change the IgG antibody response to the periodontopathic bacteria Porphyromonas gingivalis.
Forty-seven patients with periodontitis were genotyped with the direct sequencing of genome DNA. Serum IgG and specific IgG subclass levels for the sonicate of P. gingivalis and the recombinant 40 kDa outer membrane protein (OMP) were determined.
No significant difference in the total IgG level and IgG response to P. gingivalis sonicate were observed between sera from Fc gamma RIIB-232T carriers and non-carriers. The Fc gamma RIIB-232T carriers revealed a significantly lower IgG(2) response to P. gingivalis 40 kDa OMP compared to non-carriers (p = 0.04, Mann-Whitney U-test). Lower responses of Fc gamma RIIB-232T carriers were also observed in specific IgG and IgG(1) levels. The Fc gamma RIIB-232T carriers revealed a low level of IgG(2) response to P. gingivalis 40 kDa OMP, even with a high average probing pocket depth.
These results suggest that association of the Fc gamma RIIB-232T allele with periodontitis might be related to the lower levels of antibody response to P. gingivalis.

DOI： 10.1111/j.1600-0765.2007.01078.x

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BACKGROUND: The pathobiology of rheumatoid arthritis (RA) is similar to that of periodontitis in that proinflammatory cytokines and immunoglobulin G Fc receptor (FcgammaR) play an important role. Functional polymorphisms of interleukin (IL)-1 and FcgammaR were shown to be associated with susceptibility to both diseases. Therefore, we evaluated whether the IL-1 and FcgammaR gene polymorphisms represent a common risk factor for RA and periodontitis. METHODS: The study population consisted of Japanese adults with RA (RA group; N = 100), periodontitis only (P group; N = 100), and healthy individuals with no systemic or oral disease (H group; N = 100). Clinical periodontal condition was defined by measurements of probing depth, clinical attachment level, and bleeding on probing. Genomic DNA was isolated from peripheral blood and analyzed for determination of IL-1 genotypes (IL-1A+4845, IL-1B+3954, and IL-1RN+2028) and FcgammaR genotypes (FcgammaRIIA, FcgammaRIIIA, and FcgammaRIIIB) by allele-specific polymerase chain reactions. RESULTS: Among 100 patients with RA, 86% showed periodontal tissue destruction. However, the RA group exhibited milder levels of periodontal tissue destruction than the P group (P <0.01). There was a significant difference in the distribution of IL-1B+3954 C/T genotypes between the RA and P groups and between the RA and H groups (P = 0.03 for both comparisons), with enrichment of the T allele in the RA group (P = 0.04; odds ratio, 2.9 for both comparisons). The combination of IL-1A+4845 T and IL-1+3954 T alleles yielded a strong association with RA and periodontitis (RA versus P group: P = 0.00001; RA versus H group: P = 0.00001). CONCLUSIONS: These results failed to show that IL-1 and FcgammaR gene polymorphisms constitute a common risk factor for RA and periodontitis. However, it was suggested that the distributions of IL-1B+3954 genotypes and IL-1A+4845 and IL-1B+3954 haplotypes were unique to the patients with RA and periodontitis.

DOI： 10.1902/jop.2007.070136

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The functional bi-allelic polymorphism of immunoglobulin G (IgG) Fc receptor (Fc gamma R) IIa influences the efficiency of human IgG2 binding. Our previous study showed that the high affinity Fc gamma RIIa genotype (-H/H131) was associated with periodontitis risk. As interleukin-1 (IL-1) is one of the major causes of periodontal tissue destruction, it is hypothesized that the Fc gamma RIIa-H/H131cross-linking could induce an increased IL-1 release by mononuclear cells. In this study, we evaluated the intracellular expressions of IL-1 beta in CD14 positive cells upon stimulation with human IgG2 by flow cytometry. Fc gamma RIIa-H/H131 subjects exhibited a higher percentage of IL-1 beta-producing cells than Fc gamma RIIa-R/H131 and -R/R131 subjects (P &lt; 0.05). These results support the concept that Fc gamma RIIa genotype may affect IL-1 beta production, possibly leading to interindividual differences in periodontitis risk.

DOI： 10.1111/j.1744-313X.2007.00701.x

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BACKGROUND: The pathobiology of systemic lupus erythematosus (SLE) is similar to that of periodontitis in that the immunoglobulin G Fc receptor (FcgammaR) and proinflammatory cytokines play an important role. Genetic variations of FcgammaR and interleukin (IL)-1 are associated with susceptibility to both diseases. Therefore, we evaluated whether the combination of FcgammaR or IL-1 polymorphic genes represents a common risk factor for SLE and periodontitis. METHODS: The study population consisted of Japanese adults with SLE and periodontitis (SLE+P group; n = 46), SLE only (SLE group; n = 25), periodontitis only (P group; n = 58), and healthy individuals with no systemic or oral disease (H group; n = 44). Clinical periodontal condition was evaluated by measurement of probing depth, clinical attachment level, and alveolar bone loss. Genomic DNA was isolated from peripheral blood and analyzed for determination of FcgammaR genotypes (FcgammaRIIA, FcgammaRIIB, FcgammaRIIIA, and FcgammaRIIIB) and IL-1 genotypes (IL-1A +4845 and IL-1B +3954) by allele-specific polymerase chain reactions or DNA sequencing. RESULTS: A significant overrepresentation of the R131 allele of stimulatory FcgammaRIIA and the 232T allele of inhibitory FcgammaRIIB was found in the SLE+P group compared to the H group (P = 0.01 and P = 0.0009, respectively). The combination of FcgammaRIIA-R131 and FcgammaRIIB-232T alleles yielded a strong association with SLE and periodontitis (SLE+P group versus P group: P = 0.01, odds ratio: 3.3; SLE+P group versus H group: P = 0.0009, odds ratio: 11.2). Furthermore, SLE patients with the combined FcgammaR risk alleles exhibited more severe periodontal tissue destruction compared to other SLE patients. The frequencies of IL-1 polymorphic alleles were too low to assess the association with SLE or periodontitis. CONCLUSION: The combination of stimulatory FcgammaRIIA and inhibitory FcgammaRIIB genotypes may increase susceptibility to SLE and periodontitis in the Japanese population.

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We describe the cloning and characterization of Siglec-H, a novel murine CD33-related siglec-like molecule with 2 immunoglobulin domains. Unlike other CD33-related siglecs, Siglec-H lacks tyrosine-based signaling motifs in its cytoplasmic tail. Although Siglec-H has the typical structural features required for sialic acid binding, no evidence for carbohydrate recognition was obtained. Specific monoclonal and polyclonal antibodies (Abs) were raised to Siglec-H and used to define its cellular expression pattern and functional properties. By flow cytometry, Siglec-H was expressed specifically on plasmacytoid dendritic cell (pDC) precursors in bone marrow, spleen, blood, and lymph nodes. Staining of tissue sections showed that Siglec-H was also expressed in a subset of marginal zone macrophages in the spleen and in medullary macrophages in lymph nodes. Using bone marrow-derived pDC precursors that express Siglec-H, addition of Abs did not influence cytokine production, either in the presence or absence of synthetic oligodeoxynucleotides containing unmethylated cytosine-guanine motifs (CpG). In comparison, Siglec-H functioned as an endocytic receptor and mediated efficient internalization of anti-Siglec-H Abs. By immunizing mice with ovalbumin-conjugated anti-Siglec-H Ab in the presence of CpG, we demonstrate generation of antigen-specific CD8 T cells in vivo. Targeting Siglec-H may therefore be a useful way of delivering antigens to pDC precursors for cross-presentation.

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BACKGROUND: FcgammaRIIIb genotypes and smoking are risk factors for periodontal disease. However, the interaction of FcgammaRIIIb- NA1-NA2 polymorphism with smoking remains unclear. The purpose of this study was to determine if FcgammaRIIIb-NA1-NA2 polymorphism and smoking are associated with periodontal disease progression among elderly people. METHODS: Among 70-year-old subjects, 164 with neither diabetes mellitus nor blood sugar > or =140 mg/dl, who had more than 20 teeth and who could participate in both the baseline and the follow-up examinations were included in the study. The NA1 group comprised subjects with FcgammaRIIIb-NA1NA1 genotype (N = 53), while the NA2 group included subjects with FcgammaRIIIb-NA1NA2 or NA2NA2 genotype (N = 111). We examined the progression of periodontitis by measuring attachment loss during 3 years. RESULTS: The frequency of subjects who showed > or =4 mm additional attachment loss at one or more sites was 55.6% for smokers and 37.2% for non-smokers. The odds ratio (OR) was 2.13 (confidence interval [CI]: 0.92 to 4.76). We found a better association between periodontal progression and smoking in the NA2 group. The OR for smokers was 3.03 (CI:1.12 to 8.33, P = 0.028). Additionally, the mean number of sites with > or =4 mm additional attachment loss per person between smokers and non-smokers in the NA2 group or between smokers and non-smokers in the NA1 group was 2.90 3.42 and 0.74 1.53 or 0.57 0.79 and 0.68 1.03, respectively (P <0.001; analysis of variance [ANOVA]). CONCLUSION: Our results may suggest an association between smoking and periodontal disease progression in elderly people with FcgammaRIIIb-NA2 polymorphism.

DOI： 10.1902/jop.2005.76.2.250

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OBJECTIVE: The purpose of the present study was to investigate the effect of smoking cessation on the peripheral neutrophil mRNA expression levels for inflammatory cytokines, chemokine, growth factor and matrix metalloproteinase (MMP). MATERIAL AND METHODS: Sixteen male smokers (aged 22-39 [25.3+/-4.0] years), with no clinical signs of periodontal and systemic diseases, were recruited. The experiment was performed before (baseline) and at 1, 4 and 8 weeks after smoking cessation. The status of smoking and smoking cessation was verified by exhaled carbon monoxide (CO) concentration and serum cotinine concentration. Neutrophils were isolated from each subjects' peripheral blood, then the cell was stimulated with N-formyl-methionyl-leucyl-phenylalanine (FMLP). The mRNA expression levels for interleukin (IL)-1 beta, IL-8, tumor necrosis factor (TNF)-alpha, vascular endothelial growth factor (VEGF) and MMP-8 were analyzed by semiquantitative reverse transcription-polymerase chain reactions. The same experiment was performed on 11 non-smoking controls (four female and seven male), aged 23-27 (24.4+/-1.2) years. RESULTS: Eleven of 16 smokers successfully completed smoking cessation for 8 weeks. At 1 day after smoking cessation, there was a statistically significantly lower CO concentration than at baseline (p<0.01). Also, cotinine concentration markedly decreased at the second measurement, which was taken at 1 week. All of the analyzed mRNA expression levels of neutrophils from smokers were statistically significantly lower than that in non-smokers (p<0.01: IL-1 beta, IL-8, VEGF; p<0.05: TNF-alpha, MMP-8). The MMP-8 mRNA levels were statistically significantly increased at 8 weeks after smoking cessation compared with the baseline (p<0.05). Although the other mRNA expression levels were also elevated gradually from the baseline, they did not reach the statistically significant levels at 8 weeks after smoking cessation. CONCLUSION: The results showed that the neutrophil transcript levels in smokers were generally lower than those in non-smokers, which could be related to an impairment of neutrophils by smoking effects. The significant increase of MMP-8 mRNA levels were associated with the effects of smoking cessation, while recovery of the other mRNA levels seemed to require a bit longer period beyond 8 weeks after smoking cessation.

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BACKGROUND: Leukocyte Fc receptors for immunoglobulin G (FcgammaR) play a major role in the handling of immune complexes and pathogens in systemic lupus erythematosus (SLE) and periodontitis. Both diseases have been shown to be partly influenced by genetic components including FcgammaR genotype. The aim of this study was therefore to evaluate whether FcgammaR gene polymorphisms are associated with periodontitis risk in SLE patients. METHODS: The study subjects consisted of 42 SLE patients with periodontitis (SLE/P), 18 SLE patients without periodontitis (SLE/H), 42 healthy subjects with periodontitis (H/P), and 42 healthy subjects without periodontitis (H/H), who were all unrelated Japanese non-smokers. Genomic DNA was isolated from peripheral blood, and FcgammaR genotypes for 3 biallelic polymorphisms (FcgammaRIIa-R131/H131, FcgammaRIIIa-158V/158F, FcgammaRIIIb-NA1/NA2) were determined by allele-specific polymerase chain reactions. RESULTS: The SLE/P group was found to have more mild levels of periodontal destruction than the H/P group (P < 0.01). There was a significant difference in the distribution of FcgammaRIIa genotypes between SLE/P and H/H groups (P = 0.004). A significant overrepresentation of the FcgammaRIIa-R131 allele was found in the SLE/P group compared to the H/H group (SLE/P versus H/H: odds ratio [OR] 3.13, 95% confidence interval [CI] 1.46-6.77, P = 0.0013). Furthermore, the prevalence of periodontitis was found to be 70% in SLE patients. The FcgammaRIIa-R131 allele was also found to be overrepresented in the SLE/P group compared to the SLE/H group (SLE/P versus SLE/H: OR 3.40, 95% CI 1.18-10.25, P = 0.011). CONCLUSION: These results show the FcgammaRIIa-R131 allele to be associated with periodontitis risk in SLE patients.

DOI： 10.1902/jop.2003.74.3.378

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Differential gene expression was investigated in neutrophils stimulated with N-formyl-methionyl-leucyl-phenylalanine using RNA fingerprinting by arbitrarily pruned polymerase chain reaction (RAP-PCR). The cells were isolated from 3 groups of subjects: patients with generalized aggressive periodontitis (Aggressive-P, n=6), generalized chronic periodontitis (Chronic-P, n=6) and healthy controls (H, n=8). Our results show that 37 genes were upregulated, while 27 genes were down-regulated in all Aggressive-P neutrophils by using RAP-PCR with 45 primer pairs. Reverse transcript ion-PCR analyses revealed that mRNA levels were significantly different (p &lt; 0.05) for heat shock transcription factor 4b (HSF4b) gene, Kruppel-like zinc finger transcription factor 9 (Zf9) and muskelin genes. HSF4b was greater in neutrophils from Aggressive-P compared to groups I-I and Chronic-P. Zf9 and muskelin genes were lower in Aggressive-P compared to the H groups, but no significant difference was noted compared to the Chronic-P group. The control genes. IL-1&lt;beta&gt; and VEGF genes. were expressed at a significantly higher level in Aggressive-P and Chronic-P than H (p &lt; 0.01, p &lt; 0.05). In conclusion, the RAP-PCR technique used in this study enabled us to identify 3 Aggressive-P related genes, which had not been reported previously. Neutrophil functions in Aggressive-P patients are suggested to be altered by regulatory factors of the immune system including HSF4b (transcription factor), Zf9 (activator of TGF-beta) and muskelin (cellular adhesion).

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BACKGROUND: Functional polymorphisms of immunoglobulin G (IgG) Fc receptors (FcγR) have been shown to be associated with generalized aggressive periodontitis (GAgP) or recurrence of chronic periodontitis (CP) in Japanese patients. The purpose of this study was to evaluate whether FcγR polymorphisms are also associated with severity of CP. METHODS: Fifty Japanese non-smoking patients with severe CP and 39 Japanese non-smoking patients with moderate CP were identified according to established clinical criteria, including measurements of probing depth (PD), clinical attachment level (CAL), and alveolar bone loss (BL). FcγR genotypes for 3 bi-allelic polymorphisms (FcγRIIa-R/H131, FcγRIIIa-158V/F, FcγRIIIb-NA1/NA2) were determined in these CP patients and 64 race-matched, non-smoking healthy controls by means of allele-specific polymerase chain reactions. RESULTS: There was a significant over-representation of FcγRIIIa-158V allele in severe CP patients compared to moderate CP patients (odds ratio 2.03, 95% confidence interval [CI] 1.03-4.01, χ 2 = 4.86, P = 0.028). In addition, we found a strong association between CP severity and FcγR composite genotype comprising FcγRIIIa-158V plus FcγRIIIb-NA2 (severe CP versus moderate CP: odds ratio 4.69, 95% CI 1.52-15.10, χ 2 = 9.35, P = 0.002; severe CP versus healthy controls: odds ratio 4.10, 95% CI 1.62-10.59, χ 2 = 11.13, P = 0.0009). Moreover, CP patients positive for the composite genotype exhibited more severe signs of periodontitis than composite genotype-negative individuals (positive versus negative; mean PD: 3.8 mm versus 3.2 mm, P = 0.005; mean CAL: 4.5 mm versus 3.7 mm, P = 0.005; mean % BL: 37.6% versus 29.9%, P = 0.008). CONCLUSION: Our results document the FcγRIIIa-158V allele and possibly FcγRIIIb-NA2 to be associated with severity of CP in Japanese patients. J Periodontol 2001;72:1324-1331.

DOI： 10.1902/jop.2001.72.10.1324

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Early-onset periodontitis (EOP) is considered to have a genetic basis which has not been clearly defined. Genetic polymorphisms of the vitamin D receptor (VDR-B-b) and the immunoglobulin-Fcγ receptor IIIb (FcγRIIIb-NA1-NA2) are associated with bone metabolism and infectious diseases, respectively. The purpose of this study was to investigate the associations of EOP with VDR and FcγRIIIb polymorphisms. Subjects were comprised of those with generalized EOP (G-EOP, n = 42), adult periodontitis (AP, n = 52), and healthy control (HC, n = 55). VDR and FcγRIIIb genotypes were determined by allele-specific polymerase chain-reactions. Our results indicated that frequencies of the VDR-B non-carrier and the FcγRIIIb-NA2 carrier were lower in the G-EOP compared with the AP and HC groups. Furthermore, we found a strong association between G-EOP and the VDR-FcγRIIIb composite genotype (G-EOP vs. AP - OR = 5.09, p = 0.009
G-EOP vs. HC - OR = 5.93, p = 0.004). In conclusion, no correlation was found between the VDR genotype and G-EOP. However, the VDR and FcγRIIIb genotype combination may be associated with susceptibility to G-EOP.

DOI： 10.1177/00220345010800120501

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Background: Functional polymorphisms of immunoglobulin G (IgG) Fc receptors (FcγR) have been shown to be associated with generalized aggressive periodontitis (GAgP) or recurrence of chronic periodontitis (CP) in Japanese patients. The purpose of this study was to evaluate whether FcγR polymorphisms are also associated with severity of CP. Methods: Fifty Japanese non-smoking patients with severe CP and 39 Japanese non-smoking patients with moderate CP were identified according to established clinical criteria, including measurements of probing depth (PD), clinical attachment level (CAL), and alveolar bone loss (BL). FcγR genotypes for 3 bi-allelic polymorphisms (FcγRIIa-R/H131, FcγRIIIa-158V/F, FcγRIIIb-NA1/NA2) were determined in these CP patients and 64 race-matched, non-smoking healthy controls by means of allele-specific polymerase chain reactions. Results: There was a significant over-representation of FcγRIIIa-158V allele in severe CP patients compared to moderate CP patients (odds ratio 2.03, 95% confidence interval [CI] 1.03-4.01, X2 = 4.86, P = 0.028). In addition, we found a strong association between CP severity and FcγR composite genotype comprising FcγRIIIa-158V plus FcγRIIIb-NA2 (severe CP versus moderate CP: odds ratio 4.69, 95% CI 1.52-15.10, X2 = 9.35, P = 0.002
severe CP versus healthy controls: odds ratio 4.10, 95% CI 1.62-10.59, X2 = 11.13, P = 0.0009). Moreover, CP patients positive for the composite genotype exhibited more severe signs of periodontitis than composite genotype-negative individuals (positive versus negative
mean PD: 3.8 mm versus 3.2 mm, P = 0.005
mean CAL: 4.5 mm versus 3.7 mm, P = 0.005
mean % BL: 37.6% versus 29.9%, P = 0.008). Conclusion: Our results document the FcγRIIIa-158V allele and possibly FcγRIIIb-NA2 to be associated with severity of CP in Japanese patients.

DOI： 10.1902/jop.2001.72.10.1324

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：American Academy of Periodontology  

Background: Genetic polymorphisms of immunoglobulin G (IgG) Fc receptors (FcγR) were recently shown to be associated with recurrence rates of adult periodontitis (AP). The purpose of this study was to evaluate whether FcγR polymorphisms are also associated with generalized early-onset periodontitis (G-EOP) in Japanese patients. Methods: Thirty-eight Japanese patients with G-EOP and 83 Japanese patients with AP were identified according to established clinical criteria, including measurements of probing depth, clinical attachment level, and alveolar bone level. FcγR genotypes for 3 bi-allelic polymorphisms were determined in these G-EOP and AP patients and 104 race-matched healthy controls by means of allele-specific polymerase chain reactions. Results: There was a significant difference in the distribution of FcγRIIIb genotypes between G-EOP patients and healthy controls (P = 0.02). Additionally, a significant over-representation of FcγRIIIb-NA2 allele was observed in G-EOP patients as compared to AP patients and controls (P = 0.02, P= 0.009, respectively). Moreover, we found a strong association between G-EOP and the composite genotype comprising FcγRIIIb-NA2 and FcγRIIIa-158F (G-EOP versus controls: odds ratio 2.4, 95% CI 1.0-6.0, χ2 = 4.13, P = 0.04). Conclusions: This study indicates that the FcγRIIIb-NA2 allele and possibly FcγRIIIa-158F could be associated with susceptibility to G-EOP in Japanese patients.

DOI： 10.1902/jop.2000.71.9.1425

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DOI： 10.2177/jsci.21.supplement_225

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Our previous studies showed that the expression of CD23 on polymorphonuclear leukocytes (PMNLs) in gingival crevicular fluid (GCF) from adult periodontitis (AP) patients was higher than in autologous peripheral blood (PB). Percentages of eosinophils in GCF PMNLs ranged between 6 and 14%. The purpose of the present studies was to increase understanding of the potential role of eosinophils and their products, including CD23, in periodontal disease. We analysed the eosinophil fraction in GCF and PB by flow cytometry using monoclonal antibodies to CD23b (BB10), eosinophil cationic protein (ECP) in stored and secretory forms (EG1 and EG2), and CD67 (80H3). Simultaneously, we measured IgE and soluble CD23 titer and GCF and serum by ELISA. Flow cytometric analysis of BB10, EG2 and 80H3 binding showed that GCF eosinophils from AP were activated. A large BB10(+) EG2(+) cellular fraction was detected in GCF from AP whereas it was very low in autologous serum (9.30+/-2.460 vs 0.16+/-0.10, p&lt;0.001). GCF from gingivitis patients exhibited no flow cytometric evidence for the presence of BB10(+) EG2(+) cells. BB10(+) EG1(+) cells, or inactivated eosinophils rated lower in GCF than in PB both in gingivitis and periodontitis patients (0.45+/-0.63 vs 1.83+/-0.96 and 0.15+/-0.30 vs 1.30+/-0.20, p&lt;0.05, respectively). IEE titer in AP patients reached 1208.1+/-421.2 IU/ml in GCF while only 49.1+/-50.4 in sera. Soluble CD23 in GCF reached 236.1+/-81.3 ng/ml in GCF and 5.6 +/- 1.8 ng/ml in sera. GCF of gingivitis patients, however, contained no detectable sCD23. Thus, GCF from AP patients displayed a high rate of activated eosinophils secreting ECP, while GCF of gingivitis patients did not. These results suggest that ECP-secreting activated eosinophils are relevant to the pathology of adult periodontitis.

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Language：Japanese   Publisher：The Japanese Society of Periodontology  

Various laboratory techniques have been applied to the microbiological diagnosis of periodontal disease. However, certain disadvantages persist. In this study we examined the specific peptidase activity in periodontal pockets by using the chair-side test system, SK-013 as a microbiological diagnostics technique. SK-013 can rapidly (within 15 min) evaluate the peptidase activities derived from periodontopathic bacteria such as Treponema denticola and Bacteroides species. We applied SK-013 to patients with periodontitis, gingivitis, dental caries, other types of periodontal infectious disease and periodontally healthy controls and then evaluated the clinical specificity of SK-013, assessing its cut-off level. SK-013 activity was determined both spectrophotometrically and by using the color standard. Phase contrast microscopy was also utilized for determination of subgingival microorganisms. SK-013 activity was specifically detected in the periodontitis group. Setting its cut-off level at 0.2 Try U/ml based on the number of spirochetes in the subgingival pocket, we got good sensitivity, specificity, predictive value and efficacy in terms of the diagnostic value of SK-013. Using the same cut-off level, a good coincidence was also shown in both spectrophotometrically and by using the color standard. These results indicate that SK-013 is a useful chair-side diagnostic method for periodontal disease.

DOI： 10.2329/perio.33.154

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Language：Japanese   Publisher：The Japanese Society of Periodontology  

The purpose of the present study was to examine the diagnostic value of SK-013 (Sunstar, Osaka) in evaluating the efficacy of periodontal pocket curettage. SK-013 is a highly sensitive reagent which is used to measure the specific peptidase activities produced by Treponema denticola, Bacteroides (Porphyromonas) gingivalis and Bacteroides forsythus. Sixty-four periodontitis patients were selected for the study. Of the 64 patients, 33 were treated by plaque control instruction and periodontal pocket curettage (T group), and the other 31 patients received no treatment for periodontal disease (NT group). One site was monitored in each patient. The clinical status of each site was assessed using clinical indices such as the gingival index (GI), plaque index (PlI), probing depth (PD), attachment level (AL), bleeding on probing (BOP) and tooth mobility. In the T group, the clinical and enzymatic examinations were made at five separate appointments as follows: (1) at baseline prior to plaque control instruction; (2) 4 weeks after plaque control instruction; and (3) 2, 4, and 8 weeks after periodontal pocket curettage. The same examinations were performed but, only at baseline and after 12 weeks, in the NT group. As a result, periodontal pockets were reduced in depth following plaque control instruction, while the enzymatic activity showed no significant change before and after plaque control. Two weeks after periodontal pocket curettage, both clinical indices (GI, PD, AL, BOP) and SK-013 activity were significantly reduced and these improvements were maintained for 4 weeks. No significant discrepancy was observed in the NT group between clinical assessment and enzymatic activity during the experimental period. This finding suggested that SK-013 is a clinically useful diagnostic method for assessing the effect of initial periodontal therapy.

DOI： 10.2329/perio.33.164

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Other Link： http://search.jamas.or.jp/link/ui/1993025542

Language：English   Publishing type：Research paper (scientific journal)  

Ten periodontally affected subjects and 10 healthy subjects were selected in order to examine the duration patterns of percussion sounds in all upper teeth, excluding the 8′s. Probing depth, loss of attachment, tooth mobility by ‘Periotest’, bone loss and percussion sound duration via occlusal sound analyser were recorded during the pretreatment period. The following results were obtained from the study: the normal duration of percussion sounds ranged from 4.40–5.33 ms
percussion sound values of periodontal patients were of longer duration than those of healthy subjects
a close correlation between duration and other individual parameters was found in all upper teeth, with the exception of molars. Copyright © 1990, Wiley Blackwell. All rights reserved

DOI： 10.1111/j.1365-2842.1990.tb01429.x

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Language：Japanese   Publisher：JAPANESE SOCIETY OF PERIODONTOLOGY  

There have been various laboratory methods for the microbiological diagnosis of periodontal disease. However, there have been some disadvantages in these methods.<BR>In this study of the application of a chair-side test for microbiological diagnosis, the activity of peptidases in periodontal pockets of patients was examined by using the assay system SK-013. SK-013 consists of synthetic substrates and is capable of rapidly (in 15 min) evaluating the activity of specific peptidase from <I>Treponema denticola</I> and <I>Bacteroides species</I>. Using SK-013, we evaluate the correlation betweenthe enzymatic activity, clinical periodontal parameters and subgingival level of microorganisms, including phase contrast microscopy. We calculated the sensitivity and efficacy of SK-013 as a diagnostic indicator in the presence of Spirochetes and in periodontitis. A positive correlation were demonstrated between enzymatic activity, clinical periodontal parameters, the numbers of total cell count, Spirochetes, and M & S ratio. SK-013 was highly sensitive and efficacous (sensitivity: 92%, efficacy: 96%).<BR>We concluded that the assay system SK-013 is a useful chair-side method for diagnosing periodontal disease.

DOI： 10.2329/perio.32.241

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Language：Japanese   Publisher：JAPANESE SOCIETY OF PERIODONTOLOGY  

The purpose of the present study was to examine the diagnostic value of the SK-013 (Sunstar, Osaka) in evaluating the effect of initial preparation. The SK-013 is a highly sensitive reagent to measure peptidase activity specifically produced by <I>Bacteroides gingivalis</I>, <I>Bacteroides forsythus</I>, and <I>Treponema denticola</I>. Thirty-six sites in 16 periodontitis patients were monitored in this study. The clinical status of each site was assessed using the clinical indices such as gingival index (GI), plaque index (PlI), probing depth (PD), and bleeding on probing (BOP). The composition of subgingival microflora in samples was determined using phase contrast microscopy. Clinical, microbiological, and enzymatic examinations were made at five separate appointments, as follows: (1) at baseline prior to plaque control instruction; (2) 4 weeks following plaque control instruction; and (3) 2, 4, 8 weeks following scaling and root planing of periodontal sites. The results of these examinations were compared statistically. The correlation was positive between the total counts of bacteria including spirochetes and motile rods and the enzymatic activity identified using the SK-013. Also the enzymatic profile was highly correlated with microbiological findings other than clinical indices. These results show that the SK-013 is a clinically useful diagnostic method for assessing the effect of initial periodontal therapy.

DOI： 10.2329/perio.32.249

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DOI： 10.11471/shikahozon.58.109

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Other Link： http://search.jamas.or.jp/link/ui/2005108491

Language：English   Publisher：NATURE PUBLISHING GROUP  

Human type II low-affinity receptor for immunoglobulin G (FcgammaRII) constitutes a clustered gene family consisting of FcgammaRIIA, IIB and IIC genes. FcgammaRIIB is unique in its ability to transmit inhibitory signals in B cells via immunoreceptor tyrosine-based inhibitory motif (ITIM). B-cell activation and subsequent elevated production of IgG are the immunopathological features of inflammatory disease such as periodontitis. To determine whether an association with periodontitis susceptibility exists, genetic polymorphisms of FcgammaRIIB were examined in Japanese patients with aggressive periodontitis (AGP) and chronic periodontitis (CP), and in the race-matched healthy controls (HCs). A significant difference was observed in the distribution of FcgammaRIIB - 232I/T allele (exon 5) between the AGP and HC groups, with enrichment of the 232T in the AGP group (P = 0.006). In addition, the FcgammaRIIB- nt 646 - 184A/G allele (intron 4) distribution was significantly different between the CP and HC groups, with enrichment of the nt 646 - 184A in the CP group (P = 0.011). These results document the association of FcgRIIB gene polymorphisms with susceptibility to periodontitis in the Japanese.

DOI： 10.1038/sj.gene.6364021

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Language：English   Publisher：BLACKWELL MUNKSGAARD  

Variation screening for the immunoglobulin G Fc receptor IIB (FcgammaRIIB) gene was performed with the genomic DNA from 100 healthy Japanese subjects. We identified 3 non-synonymous and 2 synonymous substitutions and 2 single-nucleotide polymorphisms in an intron region. These substitutions were found to be located in the ligand-binding domain and the intron, which might alter the function of FcgammaRIIb.

DOI： 10.1034/j.1399-0039.2001.580509.x

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Language：English   Publisher：AMER SOC MICROBIOLOGY  

Porphyromonas gingivalis has been implicated as a causative pathogen in periodontitis. Immunotherapeutic approaches have recently been suggested to aid in the clearance of P, gingivalis from disease sites. Because antibody-Fe receptor (FeR) interactions play a role in the effector functions of polymorphonuclear neutrophils (PMN), we evaluated which FeR on PMN from gingival crevicular fluid (GCF) serves as an optimal target molecule for FcR-directed immunotherapy. GCF PMN and peripheral blood (PB) PMN from adult periodontitis patients were analyzed for their immunoglobulin G (IgG) and IgA FcR (Fc gammaR and Fc alphaR, respectively) expression and function by studying IgG- and IgA-mediated elimination of P. gingivalis. GCF PMN exhibited higher Fc alpha RI and Fc gamma RI levels and lower Fc gamma RIIa and Fc gamma RIIIb levels than PB PMN. Functional studies revealed that GCF PMN exhibited less of a capacity to phagocytose and kill IgG1-opsonized P. gingivalis than PB PMN. IgA1-mediated phagocytosis and killing capacity was, however, comparable between GCF PMN and PB PMN. In summary, these in vitro results document that Fc alpha RI represents a candidate target for FcR-directed immunotherapy for the clearance of P. gingivalis.

DOI： 10.1128/IAI.69.5.2935-2942.2001

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Immunoglobulin G Fc receptor mb (Fc gamma RIIIb) is constitutively expressed on neutrophils, and has three allelic forms: Fc gamma RIIIb-NA1, Fc gamma RIIIb-NA2, and Fc gamma RIIIb-SH. We identified two Japanese subjects in whom an A to G substitution at nt 221 changes asparagine (N) to serine (S) at amino acid position 45 in the Fc gamma RIIIb-NA2 gene. Fc gamma RIIIb-NA2-specific mono-clonal antibodies (GRM1 and PEN1) did not bind to mutant neutrophils, which lack an N-linked glycosylation site. Furthermore, IgG3-mediated neutrophil phagocytosis by mutant was slightly increased as compared to wildtype donors.

DOI： 10.1034/j.1399-0039.2001.057004363.x

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Many elderly people show minimum periodontal tissue destruction, which might be partly due to genetic advantages in host immune response against periodontopathic bacteria. The human IgG Fc receptor IIIb on neutrophils bears a NA1-NA2 polymorphism. The Fc gamma RIIIb-NA1 displays a more efficient interaction with IgG1- and IgG3-opsonized bacteria, compared with the Fc gamma RIIIb-NA2. We investigated a 70-year-old Japanese population (n = 599) to determine whether the Fc gamma RIIIb polymorphism was associated with resistance to periodontitis. Among subjects with greater than or equal to 20 teeth present, periodontitis-resistant (n = 46) and periodontitis-susceptible groups (n = 73) were selected based on the percentage of sites with greater than or equal to 4 nim probing attachment loss in the entire dentition. The Fc gamma RIIIb-NA1 allotype was overrepresented in the periodontitis-resistant group, compared with the periodontitis-susceptible group (chi (2) = 4.89, p = 0.03, odds ratio = 1.87, 95% CI, 1.07 to 3.28). This suggests that Fc gamma RIIIb-NA1 may be associated with resistance to periodontitis.

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Differential gene expression was investigated in neutrophils stimulated with N-formyl-methionyl-leucyl-phenylalanine using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). The cells were isolated from 3 groups of subjects: patients with generalized aggressive periodontitis (Aggressive-P, n = 6), generalized chronic periodontitis (Chronic-P. n = 6) and healthy controls (H, n = 8). Our results show that 37 genes were upregulated, while 27 genes were down-regulated in all Aggressive-P neutrophils by using RAP-PCR with 45 primer pairs. Reverse transcription-PCR analyses revealed that mRNA levels were significantly different (p &lt
0.05) lor heat shock transcription factor 4b (HSF4b) gene, Kruppel-like zinc finger transcription factor 9 (Zf9) and muskelin genes. HSF4b was greater in neutrophils from Aggressive-P compared to groups H and Chronic-P. Zf9 and muskelin genes were lower in Aggressive-P compared to the H groups, but no significant difference was noted compared to the Chronic-P group. The control genes, IL-1β and VEGF genes, were expressed at a significantly higher level in Aggressive-P and Chronic-P than H (p &lt
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0.05). In conclusion, the RAP-PCR technique used in this study enabled us to identify 3 Aggressive-P related genes, which had not been reported previously. Neutrophil functions in Aggressive-P patients are suggested to be altered by regulatory factors of the immune system including HSF4b (transcription factor), Zf9 (activator of TGF-β) and muskelin (cellular adhesion).

DOI： 10.1034/j.1600-0765.2001.360607.x

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Differential gene expression was examined in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis (P. gingivalis-LPS) using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). LPS from Escherichia coli (E. coli-LPS) was used as control. More than 200 differently expressed transcripts were found in 8 subjects showing differential regulation at the transcriptional level. Densitometric analyses revealed that 42-100 genes were upregulated, while 53-116 genes were downregulated by P. gingivalis-LPS compared with E. coli-LPS. The results of sequencing identification showed the presence of supervillin (SVIL) and vascular endothelial growth factor (VEGF) genes in the clones which were upregulated by P. gingivalis-LPS. Consequently, semiquantitative analyses proved that the level of mRNA for SVIL and VEGF were significantly higher in P. gingivalis-LPS-stimulated neutrophils than in other bacterial (E. coli, Actinobacillus actinomycetemcomitans, Prevotella intermedia) LPS- or synthetic lipid A-stimulated neutrophils. Our findings suggest that an elevated mRNA expression for SVIL could be associated with impaired function of neutrophils when stimulated by P. gingivalis-LPS. Further, the VEGF mRNA over-expression might be related to the pathogenesis of P. gingivalis-associated periodontitis. The RAP-PCR technique used in this study enabled us to identify a number of P. gingivalis-LPS regulated genes which hitherto have not been reported.

DOI： 10.1034/j.1600-0765.2001.360304.x

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BACKGROUND: Genetic polymorphisms of immunoglobulin G (IgG) Fc receptors (FcγR) were recently shown to be associated with recurrence rates of adult periodontitis (AP). The purpose of this study was to evaluate whether FcγR polymorphisms are also associated with generalized early-onset periodontitis (G-EOP) in Japanese patients. METHODS: Thirty-eight Japanese patients with G-EOP and 83 Japanese patients with AP were identified according to established clinical criteria, including measurements of probing depth, clinical attachment level, and alveolar bone level. FcγR genotypes for 3 bi-allelic polymorphisms were determined in these G-EOP and AP patients and 104 race-matched healthy controls by means of allele-specific polymerase chain reactions. RESULTS: There was a significant difference in the distribution of FcγRIIIb genotypes between G-EOP patients and healthy controls (P = 0.02). Additionally, a significant over-representation of FcγRIIIb-NA2 allele was observed in G-EOP patients as compared to AP patients and controls (P = 0.02, P = 0.009, respectively). Moreover, we found a strong association between GEOP and the composite genotype comprising FcγRIIIb-NA2 and FcγRIIIa-158F (G-EOP versus controls: odds ratio 2.4, 95% CI 1.0-6.0, X2 = 4.13, P = 0.04). CONCLUSIONS: This study indicates that the FcγRIIIb-NA2 allele and possibly FcγRIIIa-158F could be associated with susceptibility to G-EOP in Japanese patients. J Periodontol 2000;71:1425-1432.

DOI： 10.1902/jop.2000.71.9.1425

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Language：English   Publisher：ELSEVIER SCIENCE BV  

Leukocyte IgG receptors (Fc gamma R) are important immune-response modulating molecules. Fc gamma RIIIa is expressed on macrophages, NK-cells and gamma delta-T cells and exhibits a genetically determined, functional polymorphism at nucleotide 559. This allelic difference predicts either a phenylalanine (F158) or valine (V158) at amino acid 158 in the membrane-proximal extracellular domain, and has been shown to be associated with autoimmune and infectious diseases. Published methods to determine Fc gamma RIIIa genotypes are cumbersome. Therefore, we developed a novel, rapid and reliable PCR-based method to determine Fc gamma RIIIa genotypes. Comparison of genotyping results with direct Fc gamma RIIIa sequencing of 60 blood donors showed 100% accuracy of this new method. Since genotype frequencies of Fc gamma R polymorphisms depend strongly on race and ethnicity, we compared Fc gamma RIIIa genotype frequencies of 176 Caucasian Dutch and 104 Japanese blood donors. Interestingly, these frequencies were not significantly different (P&gt;0.1), in contrast to the Fc gamma RIIa and Fc gamma RIIIb genotype frequencies (P&lt;0.001). (C) 2000 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S0022-1759(00)00240-4

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Polymorphonuclear neutrophils (PMNs) are essential in host defense against periodontopathic bacteria such as Porphyromonas gingivalis. The uptake of immunoglobulin G (IgG)-opsonized bacteria via IgG Fc receptors (Fc gamma R) on PMN constitutes a central defense mechanism in periodontium. Fc gamma RIIIb is the most abundantly expressed Fc gamma R on PMN and is functionally polymorphic. The Fc gamma RIIIb-NA1 and IIIb-NA2 allotypes interact differently with IgG1- and IgG3-opsonized particles. We recently showed recurrence rates of adult periodontitis (AP) to be higher in patients carrying at least 1 Fc gamma RIIIb-NA2 allele. In this study we evaluated the functional relevance of the Fc gamma RIIIb polymorphism to anti-P. gingivalis PMN effector functions. Our results showed Fc gamma RIIIb-NA2-carrying PMN from both patients with AP and healthy controls to be less efficient in phagocytosis and induction of oxidative burst upon interaction with IgG1- and IgG3-opsonized P. gingivalis. These functional differences between Fc gamma RIIIb-NA1 and IIIb-NA2 were observed in the presence of CD32-blocking antibody fragments, but not upon blocking CD16. Moreover, PMNs from AP patients exhibited increased Fc gamma RIIIb-allelic differences in IgG3-induced oxidative burst compared to control PMNs. These results support the concept that Fc gamma RIIIb heterogeneity may influence the clinical course of AP.

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Language：English   Publisher：BLACKWELL SCIENCE LTD  

The immunoglobulin receptor Fc gamma RIIIa (CD16) is distributed on natural killer (NK) cells, macrophages, and gamma delta T cells, and is polymorphic. Fc gamma RIIIa-158V has a higher affinity for both monomeric and immune complexed IgG1, IgG3, and IgG4 than IIIa-158F. We determined Fc gamma RIIIa-158V/F genotypes of Japanese patients with adult periodontitis. A significant over-representation of Fc gamma RIIIa-158F was found in patients with recurrence, compared with patients without recurrence, making Fc gamma RIIIA a candidate gene for recurrence risk of adult periodontitis.

DOI： 10.1046/j.1365-2249.1999.00984.x

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DNA binding activity of NF-kappa B and AP-1 were examined in neutrophils stimulated with LPS purified from P. gingivalis, a major pathogenic bacteria of periodontitis lesion. Porphyromonas gingivalis LPS enhanced the activity reaching a peak at a concentration of 500 ng/ml in the absence of serum. The NF-kappa B activation stimulated with 10 ng/ml of P. gingivalis LPS was suppressed approximately 44% by treatment of neutrophils with anti-CD14 antibody under the presence of serum. Increase in the steady-state IL-8 mRNA level was concomitantly observed by stimulation of neutrophils with 500 ng/ml of P. gingivalis LPS under the absence of serum. These results indicate that P. gingivalis LPS activates NF-kappa B and AP-1 in both serum-dependent and -independent manners, followed by increased IL-8 transcription in neutrophils, and suggested a role for P. gingivalis LPS in IL-8 synthesis by neutrophils in inflamed gingiva and GCF.

DOI： 10.1023/A:1022344031223

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Language：English   Publisher：PERGAMON-ELSEVIER SCIENCE LTD  

A previous study has demonstrated that complement receptors on the surface of polymerphonuclear leucocytes (neutrophils) in gingival crevicular fluid significantly increased compared with those in autologous peripheral blood obtained from periodontitis-affected subjects. The present study attempted to determine the mRNA levels of complement receptor types 1 and 3 (CR1, CR3) on neutrophils in gingival crevicular fluid using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. Gingival crevicular fluid samples were obtained from 11 adult periodontitis patients by gingival crevicular washing, and venipunctured peripheral blood was used as a control. RT-PCR analysis was performed using the primer sets for CR1, CR3 and beta-actin. Digoxigenin-labelled RNA probes were synthesized from RT-PCR products for in situ hybridization. Both CR1 and CR3 mRNA levels relative to beta-actin were significantly lower in crevicular fluid neutrophils than in peripheral blood neutrophils (crevicular fluid-CR1: 32.75+/-22.93%, peripheral blood-CR1: 65.30+/-43.25%, p &lt;0.005; crevicular fluid-CR3: 9.09+/-5.34%, peripheral blood-CR3: 30.14+/-18.80%, p &lt;0.005). In in situ hybridization, a greater majority of neutrophils showed positive CR1 and CR3 mRNA expression, while only a few neutrophils showed positive signals in gingival crevicular fluid. Data in the present study suggest that increased expression of complement receptors on the neutrophil cell surface appears to be unrelated to de novo synthesis. (C) 1997 Elsevier Science Ltd.

DOI： 10.1016/S0003-9969(96)00110-0

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DOI： 10.2329/perio.38.243

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DOI： 10.2329/perio.38.243

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Polymorphonuclear leukocytes (PMNLs) are the most numerous cell population among the cellular infiltrates in gingival crevicular fluid (GCF) and play important roles in the host-defensive system in the gingival crevices. We determined the percentage of neutrophils, eosinophils and basophils in total PMNLs by light microscopic observation using Randolph-methylene blue staining, then assessed flow cytometric differences in the expression of CR3, FcgammaIII, FcepsilonRII, LFA-1alpha, and LFA-1beta on PMNL in GCF and peripheral blood (PB) from 21 patients with adult periodontitis (AP) and 13 healthy donors. Percentages of basophils and eosinophils were higher in GCF than in PB. In both AP patients and healthy subjects, expression of CR3 and FcepsilonRII was higher while FcgammaRIII was lower in GCF than in PB. The statistical analysis showed that the expressions of FcgammaRIII and FcepsilonRII on GCF PMNLs were lower in AP patients than in healthy subjects. Expressions of LFA-1alpha and beta on GCF were similar to those on PB PMNLs. PB PMNLs stimulated in vitro with Porphyromonas gingivalis culture supernatant and fMLP displayed an expression pattern of CR3, FcgammaRIII and FcepsilonRII on GCF PMNLs. However, C5a and IL-1 failed to induce changes in FcgammaRIII and FcepsilonRII. The results indicate that GCF neutrophils are activated, present enhanced adhesion and a decreased IgG-binding ability which would reflect that they are at the terminal stage of activation, and that GCF contains a larger eosinophil-fraction than in PB. Moreover, these GCF eosinophils appear to be activated.

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Language：Japanese   Publisher：Japanese Society of Stomatognathic Function  

CiNii Article

CiNii Books

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