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Objective: This study aimed to clarify the association between social mutual aid and psychological stress among residents in a rural district. Materials and Methods: A cross-sectional study based on Andersen's Behavioral Model of Health Care Utilization was conducted on 2,500 residents of City A in Akita Prefecture who were aged 65 years or older. The study was conducted from April 8 to May 15, 2017. Participants were administered a questionnaire containing items on individual characteristics (predisposing, enabling, and need) and contextual characteristics (physical factors). Results: Responses were obtained from 1,236 participants, and data from 974 valid questionnaires were analyzed. Factors related to the high level of psychological stress were "maintenance of confidential relationships that could only be formed in the rural district (low)" and "social support (low)", which are forms of social mutual aid. Use of health services had no association with psychological stress, whereas when psychological stress was high (5 points or higher), the rate of "not participating in community groups (no)" was also high. Conclusion: The findings of this study indicate the need for the objective evaluation of the roles of self-help and mutual help among elderly adults living in a rural district and the mutual help and public help functions represented by health services. It is also important to develop districts that promote the enhancement of social mutual aid so that such help can be fully utilized.

Cell adhesion molecule 1 (CADM1) is aberrantly expressed by T-cell neoplasms such as adult T-cell leukemia/lymphoma (ATLL) and mycosis fungoides (MF). We studied the expression of CADM1 and its splicing variants in Sézary syndrome (SS), MF, other cutaneous T-cell lymphoma (CTCL), and cell lines derived from T- and B-cell lymphomas. Soluble CADM1 was measured in the patients' sera. CADM1+ cells in the blood and skin lesions were examined by flow cytometry and immunostaining, respectively. Soluble CADM1 was measured by ELISA, and the splicing variants of CADM1 transcripts were determined by reverse transcriptase-polymerase chain reaction, followed by sequencing. As a result, circulating CADM1+ cells were significantly increased in seven out of 10 patients with SS, ranging from 7.9% to 74.5% of the CD3+CD4+ fractions (median 33.7%; cut-off value 6.5%). The percentages of CADM1+ cells were usually less than those of circulating Sézary cells. CADM1 was expressed, to various degrees, in six of nine T-cell lines derived from SS, MF, ATLL, and anaplastic large cell lymphoma (ALCL), but negative in B-cell lymphoma-derived cell lines. CADM1+ cells were present in the skin infiltrates of MF, SS, ATLL and ALCL. Serum levels of soluble CADM1 were not significantly elevated in SS/MF. Three major splicing variants of CADM1 expressed by neoplastic T-cells contained different combinations of the exons 7, 8, 9 and 11, including a putative oncogenic variant composed of exons 7-8-9-11. In conclusion, CADM1 is frequently expressed in Sézary cells and cell lines from CTCL.

Mechanical reduction of infectious bacteria by using physical instruments is considered the principal therapeutic strategy for periodontal disease; addition of antibiotics is adjunctive. However, local antibiotic treatment, combined with conventional mechanical debridement, has recently been shown to be more effective in periodontitis subjects with type 2 diabetes. This suggests that some bacteria may invade the inflamed inner gingival epithelium, and mechanical debridement alone will be unable to reduce these bacteria completely. Therefore, we tried to establish infected organ culture models that mimic the inner gingival epithelium and aimed to see the effects of antibiotics in these established models. Mouse dorsal skin epithelia were isolated, and periodontal bacteria were injected into the epithelia. Infected epithelia were incubated with test antibiotics, and colony-forming ability was evaluated. Results indicated that effective antibiotics differed according to injected bacteria and the bacterial combinations tested. Overall, in organ culture model, the combination of amoxicillin or cefdinir and metronidazole compensate for the effects of less effective bacterial combinations on each other. This in vitro study would suggest effective periodontal treatment regimens, especially for severe periodontitis.

The effect on humans of a 6-hour long exposure to cyclic fluctuating air temperature was examined. The tested fluctuating pattern consisted of a gradual sweep-up +2.0°C over 40 minutes and a rather abrupt sweep-down –2.0°C over 20 minutes, within the temperature ranges of 26–28°C and 27–29°C. The experiment on cyclic fluctuating air temperature demonstrates that the thermal dissatisfaction rate for cyclic fluctuating air temperature control between 26°C–28°C is lower than that for constant air temperature control at 26°C, while consuming less energy. Moreover, occupant stress levels measured by salivary alpha-amylase, and subjective fatigue levels under cyclic fluctuating air temperature show that similar or even lower distinct results can be achieved with fluctuating air temperature control compared with constant air temperature control.

The Japanese lifestyle includes activities with a greater range of hip motion, such as squat toilet and sitting on the floor. These activities increase the risk of hip dislocation for patients with total hip arthroplasty. However, research concerning the impact of Japanese lifestyle on quality of life for patients with total hip arthroplasty is limited. This cross-sectional study investigated lifestyle differences between two regions and quality of life related to Japanese lifestyle, controlling age and other covariates. Participants comprised 392 total hip arthroplasty patients with a postoperative period of one to seven years from major hospitals in northern (192 patients) and southern (200 patients) Japan. Participants were administered by the EQ-5D (assessing generic health-related quality of life), Oxford hip score (assessing hip-specific health condition), Japanese lifestyle difficulties (squat toilet, sitting on the floor, etc.) and lifestyle at home. The southern hospital sample retained more of a Japanese lifestyle compared to the northern hospital sample. Nearly 50% of participants reported some level of difficulty related to the Japanese squat toilet, although only a low percentage had it at home. Multivariate linear regression analysis was performed to predict the quality of life, and the final model retained the pain and function of the Oxford hip scores and Japanese squat toilet difficulty. The result indicates that physical environments outside of the house have an impact on quality of life for THA patients in Japan. Some aspects of the Asian lifestyle require a greater range of motion of the hip joint. In non-Western countries, regional differences in lifestyle and physical environments outside of their home need to be taken into consideration in discharge planning and rehabilitation. Further multinational comparison of the Asian lifestyle via prospective research is necessary, controlling pre-operative conditions.

Summary Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are major periodontal pathogens that cause several types of periodontal disease. Our previous study suggested that P. gingivalis gingipains secreted in the subgingival environment are related to the detachment of A.actinomycetemcomitans biofilms. However, it remains unclear whether arginine-specific cysteine proteinase (Rgp) and lysine-specific proteinase (Kgp) play different roles in the detachment of A. actinomycetemcomitans biofilm. The aim of this study was to investigate possible disruptive roles of Kgp and Rgp in the aggregation and attachment of A. actinomycetemcomitans. While P. gingivalis ATCC33277 culture supernatant has an ability to decrease autoaggregation and coaggregation of A. actinomycetemcomitans cells, neither the boiled culture supernatant of ATCC33277 nor the culture supernatant of KDP136 showed this ability. The addition of KYT-1 and KYT-36, specific inhibitors of Rgp and Kgp, respectively, showed no influence on the ability of P. gingivalis culture supernatant. The result of gelatin zymography suggested that other proteases processed by gingipains mediated the decrease of A. actinomycetemcomitans aggregations. We also examined the biofilm-destructive effect of gingipains by assessing the detachment of A. actinomycetemcomitans from polystyrene surfaces. Scanning electron microscope analysis indicated that A. actinomycetemcomitans cells were detached by P. gingivalis Kgp. The quantity of A. actinomycetemcomitans in biofilm was decreased in co-culture with P. gingivalis. However, this was not found after the addition of KYT-36. These findings suggest that Kgp is a critical component for the detachment and decrease of A. actinomycetemcomitans biofilms.

Mechanical reduction of infectious bacteria by using physical instruments is considered the principal therapeutic strategy for periodontal disease; addition of antibiotics is adjunctive. However, local antibiotic treatment, combined with conventional mechanical debridement, has recently been shown to be more effective in periodontitis subjects with type 2 diabetes. This suggests that some bacteria may invade the inflamed inner gingival epithelium, and mechanical debridement alone will be unable to reduce these bacteria completely. Therefore, we tried to establish infected organ culture models that mimic the inner gingival epithelium and aimed to see the effects of antibiotics in these established models. Mouse dorsal skin epithelia were isolated, and periodontal bacteria were injected into the epithelia. Infected epithelia were incubated with test antibiotics, and colony‐forming ability was evaluated. Results indicated that effective antibiotics differed according to injected bacteria and the bacterial combinations tested. Overall, in organ culture model, the combination of amoxicillin or cefdinir and metronidazole compensate for the effects of less effective bacterial combinations on each other. This in vitro study would suggest effective periodontal treatment regimens, especially for severe periodontitis.

Positron emission tomography (PET) is used for staging and response evaluation in primary gastric lymphoma (PGL). However, the implications of [(18)F]-2-fluoro-2-deoxy-D-glucose ((18)F-FDG) uptake in PGL at first diagnosis have not been reported. The relationship between (18)F-FDG uptake and the expression of facilitative glucose transporters (GLUTs), hexokinase II (HK II), and Ki67, as well as malignant potential in PGL, was assessed in this study. We analyzed 23 patients with PGL [nine with diffuse large B-cell lymphoma (DLBCL); seven with high-grade mucosa-associated lymphoid tissue (MALT) lymphoma; and seven with low-grade MALT lymphoma]. The expression levels of GLUT1, GLUT3, HK II, and Ki67 were evaluated according to the percentage of positive area determined by immunohistochemistry. Standardized uptake values correlated significantly with pathological malignant potentials (low-grade/high-grade MALT lymphoma and DLBCL: p = 0.001-0.002), Ki67 (p 0.001), and GLUT1 expression (p = 0.02). We determined that (18)F-FDG uptake is related to GLUT1 expression and tumor histological grade as well as Ki67 in PGL.

Background and Objective Biofilm formation occurs through the events of cooperative growth and competitive survival among multiple species. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are important periodontal pathogens. The aim of this study was to demonstrate competitive or cooperative interactions between these two species in co-cultured biofilm. Material and Methods P. gingivalis strains and gingipain mutants were cultured with or without A. actinomycetemcomitans. Biofilms formed on glass surfaces were analyzed by crystal violet staining and colony counting. Preformed A. actinomycetemcomitans biofilms were treated with P. gingivalis culture supernatants. Growth and proteolytic activities of gingipains were also determined. Results Monocultured P. gingivalis strains exhibited a range of biofilm-formation abilities and proteolytic activities. The ATCC33277 strain, noted for its high biofilm-formation ability and proteolytic activity, was found to be dominant in biofilm co-cultured with A. actinomycetemcomitans. In a time-resolved assay, A. actinomycetemcomitans was primarily the dominant colonizer on a glass surface and subsequently detached in the presence of increasing numbers of ATCC33277. Detachment of preformed A. actinomycetemcomitans biofilm was observed by incubation with culture supernatants from highly proteolytic strains. Conclusion These results suggest that P. gingivalis possesses a competitive advantage over A. actinomycetemcomitans. As the required biofilm-formation abilities and proteolytic activities vary among P. gingivalis strains, the diversity of the competitive advantage is likely to affect disease recurrence during periodontal maintenance.

Above a critical concentration, amphiphilic lipopolysaccharide (LPS) molecules in an aqueous environment form aggregate structures, probably because of interactions involving hydrophobic bonds. Ionic bonds involving divalent cations stabilize these aggregate structures, making them resistant to breakdown by detergents. The aim of this study was to examine expression patterns of stabilized LPS aggregates in Aggregatibacter actinomycetemcomitans, a microorganism that causes periodontitis. A. actinomycetemcomitans strains of various serotypes and truncated LPS mutants were prepared for this study. Following treatment with a two-phase separation system using the detergent Triton X-114, crude LPS extracts of the study strains were separated into detergent-phase LPS (DP-LPS) and aqueous-phase LPS (AP-LPS). Repeated treatment of the aqueous phase with the two-phase separation system produced only a slight decrease in AP-LPS, suggesting that AP-LPS was resistant to the detergent and thus distinguishable from DP-LPS. The presence of divalent cations increased the yield of AP-LPS. AP-LPS expression patterns were serotype-dependent; serotypes b and f showing early expression, and serotypes a and c late expression. In addition, highly truncated LPS from a waaD (rfaD) mutant were unable to generate AP-LPS, suggesting involvement of the LPS structure in the generation of AP-LPS. The two-phase separation was able to distinguish two types of LPS with different physical states at the supramolecular structure level. Hence, AP-LPS likely represents stabilized LPS aggregates, whereas DP-LPS might be derived from non-stabilized aggregates. Furthermore, time-dependent expression of stabilized LPS aggregates was found to be serotype-dependent in A. actinomycetemcomitans.

Tanaka A, Fujise O, Chen C, Miura M, Hamachi T, Maeda K. A novel gene required for natural competence inAggregatibacter actinomycetemcomitans. J Periodont Res 2012; 47: 129–134. © 2011 John Wiley & Sons A/S Background and Objective: Natural competence is the ability of bacteria to take up extracellular DNA and incorporate it into their genomes. Some strains of Aggregatibacter actinomycetemcomitans, a critical periodontal pathogen, are naturally competent for transformation. However, information on natural competence genes is limited for this species. The aim of this study was to confirm the involvement of a novel gene identified near the fimbriae gene cluster in natural competence. Material and Methods: The functions of putative open reading frames (ORFs), designated AA00863–AA00865, in the Oralgen project database for A. actinomycetemcomitans strain HK1651, have not been determined. Using naturally transformable A. actinomycetemcomitans strains D7S-1 and ATCC29523, we created deletion mutants of homologous genes of these ORFs. Natural competence in the study strains was determined using an agar-based transformation frequency assay. Results: Mutation of the AA00865 homolog, which we named urpA in A. actinomycetemcomitans strain D7S-1, resulted in the loss of natural competence, whereas mutations of the AA00864 and AA00863 homologs, located downstream of urpA gene, did not. Similar results were also observed in the mutants of A. actinomycetemcomitans ATCC29523. Complementation of the deleted sequence in the urpA mutant restored natural competence. Conclusion: The urpA gene is a novel gene required for natural competence in A. actinomycetemcomitans and does not exhibit significant homology with any natural competence genes previously identified in other bacterial species.

Sensitization of primary afferent neurons is one of the most important components of pain hypersensitivity after tissue injury. Insulin-like growth factor 1 (IGF-1), involved in wound repair in injured tissue, also plays an important role in maintaining neuronal function. In the present study, we investigated the effect of tissue IGF-1 on nociceptive sensitivity of primary afferent neurons. Local administration of IGF-1 induced thermal and mechanical pain hypersensitivity in a dose-dependent manner, and was attenuated by IGF-1 receptor (IGF1R) inhibition. Tissue but not plasma IGF-1 levels, as determined by enzyme-linked immunosorbent assay, significantly increased after plantar incision. Immunohistochemistry revealed that IGF1R was predominantly expressed in neurons as well as in satellite glial cells in the dorsal root ganglion (DRG). Double-labeling immunohistochemistry showed that IGF1R expression colocalized with peripherin and TRPV1, but not with NF200 in DRG neurons. The IGF1R inhibitor successfully alleviated mechanical allodynia, heat hyperalgesia, and spontaneous pain behavior observed after plantar incision. Expression of phosphorylated Akt in DRG neurons significantly increased after plantar incision and was suppressed by IGF1R inhibition. These results demonstrate that increased tissue IGF-1 production sensitizes primary afferent neurons via the IGF1R/Akt pathway to facilitate pain hypersensitivity after tissue damage.

Porphyromonas gingivalis is considered a critical pathogen of periodontal diseases including recurrent periodontitis. The profound effects of active periodontal treatment (APT) on P. gingivalis elimination were previously demonstrated and revealed that the subsequent P. gingivalis-free or -suppressed status seems to be maintained during early periodontal maintenance (PMT). The aim of the present study was to show the occurrence of microbial recolonization during this early PMT period.In total, 128 sites from 11 generalized chronic periodontitis patients and one generalized aggressive periodontitis patient underwent clinical and microbiologic examination at baseline (Exam-I), after APT (Exam-II), and in PMT (Exam-III). Exam-III was carried out an average of 4.5 +/- 3.5 months after Exam-II. Detection and quantification of putative pathogens were performed using a polymerase chain reaction-based method.The PMT used was effective in maintaining the clinical conditions improved by APT. However, in microbiological examinations, Exam-III showed higher detection frequency and levels of P. gingivalis than Exam-II. This suggests that a P. gingivalis recolonization started in the early PMT period. P. gingivalis-increased sites then showed significantly more severe signs of periodontitis in Exam-I than P. gingivalis-stable sites (bleeding on probing frequency: 76.7% versus 56.5%; suppuration frequency: 41.9% versus 12.9%). On the other hand, in Exam-II, no significant differences of clinical parameters were noted between P. gingivalis-increased and -stable sites.Severe periodontitis sites before APT seemed to place them at risk of P. gingivalis recolonization in the early PMT period, and this microbial restoration could be a cause of recurrent periodontitis.

Porphyromonas gingivalis is considered a critical pathogen in periodontal diseases. It is classified into six genotypes based on diversity of the fimA gene encoding fimbrillin. The present study evaluated the involvement of the fimA genotype in treatment outcome following non-surgical periodontal therapy.Chronic periodontitis patients were enrolled in this study; all received clinical and microbiological examinations at baseline. The detection of subgingival species and identification of P. gingivalis fimA genotypes were performed using polymerase chain reaction based methods. In total, 160 P. gingivalis positive sites with bleeding on probing (BOP) and a probing depth ofor =4 mm were accepted. They were followed up after scaling and root planing.Longitudinal investigation indicated that fimA type I positive sites at baseline were followed by a significantly higher frequency of persistent BOP after treatment than type I negative sites (51.6% versus 27.9%), while types Ib and II were not. Type I positive sites also showed more persistence of Tannerella forsythensis and P. gingivalis after treatment than type I negative sites. In post-treatment investigation, type I positive sites showed higher frequencies of BOP and T. forsythensis detection than type I negative sites (77.8% versus 43.5% and 100% versus 76.1%, respectively).BOP in initially type I positive sites showed little improvement with treatment, and the combined persistence of fimA type I and T. forsythensis seemed to be involved in this poor treatment outcome. The present study demonstrated the potential of P. gingivalis fimA type I as a predictor of persistent BOP after treatment.

Background: The fimA gene, which encodes fimbrillin (FimA), is found in Porphyromonas gingivalis and has been classified into six genotypes based on nucleotide sequence. P. gingivalis that possesses the type II fimA gene is prevalent in adult periodontitis. Objectives: The aim of this study was to investigate the prevalence of P. gingivalis fimA genotypes in Japanese aggressive periodontitis patients, and to examine their virulence. Methods: Subgingival plaque samples were obtained from 223 sites in 18 aggressive periodontitis patients and 95 sites in 22 periodontally healthy young adults. Actinobacillus actinomycetemcomitans, P. gingivalis and Tannerella forsythensis detection, determination of the fimA genotype in P. gingivalis, and the quantification of P. gingivalis were analyzed by polymerase chain reaction (PCR) methods. The proteolytic activities of the P. gingivalis fimA type I and fimA type II were also examined. Results: In aggressive periodontitis patients, the most prevalent fimA genotype was the type II (46.7%), followed by the type Ib and type I, whereas in healthy subjects, the type I fimA was the only genotype detected. The number of P. gingivalis pathogens was the greatest in the type I fimA positive sites, and the frequency of coexisting A. actinomycetemcomitans and T. forsythensis was highest in the type II fimA positive sites in the aggressive periodontitis patients. Both the arginine-specific cysteine proteinase (Arg-gingipain) and lysine-specific cysteine proteinase (Lys-gingipain) activity of the P. gingivalis fimA type I strain were significantly higher than those of the fimA type II strains. Conclusions: These results suggest that differences in virulence exist among different fimA genotypes. Coadherence with other pathogens in P. gingivalis fimA type II-associated aggressive periodontitis and quantitative increases in P. gingivalis in fimA type I-associated aggressive periodontitis are related to this virulence.

Patients suffer considerable pain from fracture of placed implants. In order to reduce the number of fracturing failures in future and to obtain a better understanding of fracturing, a fractured implant has been investigated by means of radiography, scanning electron microscopy and transmission electron microscopy from a metallurgical viewpoint. The blade type implant of Ti-5V alloy has a virgin microstructure of a low density of dislocation with a dispersion of fine precipitates. Further dislocations and twins are introduced into the implant due to repeated loading in use, resulting in possessing a large amount of lattice defects until fracturing. The dislocations are considered to assist calcium penetration and to bring about intergranular fracturing near the surface of the implant. Striations are extensively observed at a center portion of the implant, indicating that the fracturing is caused by fatigue. Microscopic investigations reveal that the fracturing is initiated by some elements such as calcium in saliva in accordance with high density of lattice defects and developed through the fatigue by repeated loading.

Background: Porphyromonas gingivalis is one of the most important periodontopathogens. It produces cysteine proteinases named gingipains. We previously examined the effect of gingipains on abscess formation in a murine model. The rgpA rgpB double and kgp mutants induced smaller abscesses than the wild type. Moreover, the rgpA rgpB kgp triple (gingipain-null) mutant hardly showed lesion formation at all under the experimental conditions used, indicating that genes encoding gingipains are important for P. gingivalis virulence. Objectives: Here, we further report the humoral immune responses induced by P. gingivalis strains. Methods: After the lesions were apparently cured, sera were collected from the mice and immunoglobulin G (IgG) responses against the whole cell antigens of wild-type P. gingivalis were measured. Results: Wild-type strain was found to induce a strong antibody reaction. On the other hand, the rgpA rgpB kgp triple and kgp mutants induced significantly lower antibody responses compared to the wild type. Western blotting analysis confirmed the differences in antibody production. Next, these mice were re-infected with wild-type strain. Mice that were first infected with wild-type strain showed significantly smaller lesion formation than control mice that were first infected with medium only. On the other hand, mice that were first infected with mutant strains devoid of gingipain activities did not show resistance to re-infection and immunoglobulins directed against gingipains may be protective. Conclusions: These results suggest that gingipains play an important role in abscess formation in mice, and humoral immune responses seem to be partly responsible for the resistance to re-infection by P. gingivalis.

Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Bacteroides forsythus are considered major putative periodontal pathogens. However, it remains unclear what combinations or what levels of these bacteria influence treatment outcome. The purpose of the present study was to establish useful pathogenic markers for prediction and assessment of treatment outcome following scaling and root planing (SRP).A total of 1,149 sites in 104 chronic periodontitis patients were clinically examined at baseline. Three months after SRP, 606 sites in 56 of these patients were reexamined. Subgingival plaque samples taken from the examined sites at baseline and 3 months were analyzed for the detection and quantification of A. actinomycetemcomitans, P. gingivalis, and B. forsythus using a colorimetric polymerase chain reaction technique.At baseline, high levels and a combination of P. gingivalis and B. forsythus were frequently detected in diseased sites (74%). SRP reduced the levels and the coexistence of P. gingivalis and B. forsythus (from 75% to 43%). However, in treated sites where there was less reduction of probing depth (2 mm), or where bleeding on probing (BOP) or suppuration was detected, residual coexistence of P. gingivalis and B. forsythus and a high level of P. gingivalis after SRP were significantly more frequent. Furthermore, SRP did not improve BOP at sites exhibiting initially high levels of A. actinomycetemcomitans.These results suggest that the combination of P. gingivalis and B. forsythus, as well as the level of P. gingivalis, is useful in assessing treatment outcome. Furthermore, the high level of A. actinomycetemcomitans before SRP is a possible valuable predictor of treatment outcome.

八ホウ酸二ナトリウム四水和物(DOT, Timbor®)を注入したParaserianthes falcataria(ファルカータ)をコア材としたランバーコア合板の防腐性能を室内標準試験によって評価した。ファルカータに0.12～0.5%DOT水溶液を加圧注入した結果,いずれの濃度においても良好な注入性を示し,各濃度における注入量の平均は607～651kg/m3であった。DOTを加圧注入したファルカータ試験体の中央部付近においてもDOTは浸透していたが,木口端部から内部へ進むにしたがいDOT吸収量(BAE)が減少し,試験体の中央部付近(木口端部から20cm)における吸収量は,木口端部における吸収量の60～70%程度であった。DOTで処理したランバーコア合板の防腐性能を調べた結果,コア材中のDOT吸収量がオオウズラタケに対して2kg/m3 BAE以上,カワラタケに対して1kg/m3 BAE以上で,高い防腐性能を示すことがわかった。また,単板塗布処理あるいは接着剤混入処理にも供試菌の侵入を防止する効果が認められた。

Natural competence is the ability of bacteria to incorporate extracellular DNA into their genomes. This competence is affected by a number of factors, including Ca(2+) utilization and biofilm formation. As bacteria can form thick biofilms in the presence of extracellular Ca(2+) , the additive effects of Ca(2+) -promoted biofilm formation on natural competence should be examined. We evaluated natural competence in Aggregatibacter actinomycetemcomitans, an important periodontal pathogen, in the context of Ca(2+) -promoted biofilms, and examined whether the pga gene cluster, required for bacterial cell aggregation, is necessary for competence development. The A. actinomycetemcomitans cells grown in the presence of 1 mm CaCl2 exhibited enhanced cell aggregation and increased levels of cell-associated Ca(2+) . Biofilm-derived cells grown in the presence of Ca(2+) exhibited the highest levels of natural transformation frequency and enhanced expression of the competence regulator gene, tfoX. Natural competence was enhanced by the additive effects of Ca(2+) -promoted biofilms, in which high levels of pga gene expression were also detected. Mutation of the pga gene cluster disrupted biofilm formation and competence development, suggesting that these genes play a critical role in the ability of A. actinomycetemcomitans to adapt to its natural environment. The Ca(2+) -promoted biofilms may enhance the ability of bacteria to acquire extracellular DNA.

Background Azithromycin is an antibiotic belonging to the macrolides. Previous case reports showed that azithromycin has a regenerative effect on periodontal tissue in addition to improving periodontal gingival inflammation. Recently, we experienced three periodontitis cases, all of which showed severe bone loss. However, their gingival inflammatory signs differed greatly. The present case reports evaluated the regenerative effects of azithromycin on periodontitis sites with different clinical signs of gingival inflammation. Methods In Case 1, generalized chronic periodontitis with severe gingival inflammation was treated with azithromycin before periodontal treatment. In contrast, Case 2 presented with few clinical signs of gingival inflammation, but was treated with azithromycin prescribed within a day of scaling and root planing. In Case 3, teeth with moderate gingival inflammation were treated with azithromycin after a series of scaling and root planing. Results Remarkable alveolar bone growth, regardless of baseline gingival inflammation, was noted in all three cases. Conclusions The use of adjunctive azithromycin in scaling and root planing may be effective for periodontal tissue regeneration. This property may be independent of the degree of baseline gingival inflammation.

Above a critical concentration, amphiphilic lipopolysaccharide (LPS) molecules in an aqueous environment form aggregate structures, probably because of interactions involving hydrophobic bonds. Ionic bonds involving divalent cations stabilize these aggregate structures, making them resistant to breakdown by detergents. The aim of this study was to examine expression patterns of stabilized LPS aggregates in Aggregatibacter actinomycetemcomitans, a microorganism that causes periodontitis. A. actinomycetemcomitans strains of various serotypes and truncated LPS mutants were prepared for this study. Following treatment with a two-phase separation system using the detergent Triton X-114, crude LPS extracts of the study strains were separated into detergent-phase LPS (DP-LPS) and aqueous-phase LPS (AP-LPS). Repeated treatment of the aqueous phase with the two-phase separation system produced only a slight decrease in AP-LPS, suggesting that AP-LPS was resistant to the detergent and thus distinguishable from DP-LPS. The presence of divalent cations increased the yield of AP-LPS. AP-LPS expression patterns were serotype-dependent; serotypes b and f showing early expression, and serotypes a and c late expression. In addition, highly truncated LPS from a waaD (rfaD) mutant were unable to generate AP-LPS, suggesting involvement of the LPS structure in the generation of AP-LPS. The two-phase separation was able to distinguish two types of LPS with different physical states at the supramolecular structure level. Hence, AP-LPS likely represents stabilized LPS aggregates, whereas DP-LPS might be derived from non-stabilized aggregates. Furthermore, time-dependent expression of stabilized LPS aggregates was found to be serotype-dependent in A. actinomycetemcomitans.

Background The phosphorylation of p38 mitogen-activated protein kinase (MAPK) in the dorsal root ganglion (DRG) promotes primary afferent sensitization. The role of p38MAPK signaling in the DRG in the pathogenesis of plantar incision hyperalgesia has not been investigated. Results Levels of phosphorylated p38MAPK (p-p38MAPK) obviously increased in the DRG after plantar incision. Unmyelinated and myelinated DRG neurons that express p-p38MAPK contained small to medium cell bodies, suggesting that p-p38MAPK expression is induced in neurons with C- and Aδ-fibers. The p-p38MAPK inhibitors FR167653 or SB203580 inhibited incision-induced mechanical hypersensitivity and spontaneous pain behavior. The systemic administration of tumor necrosis factor-α (TNF-α) inhibitor prevented subsequent incision-induced activation of p38MAPK in the DRG and alleviated mechanical hypersensitivity after the incision. Conclusions p38MAPK signaling in the DRG plays a crucial role in the development of primary afferent sensitization and pain behavior caused by plantar incision.

The aims of this paper are to survey categorical systems of cleft palate speech that professionals in other countries use in their clinical work and to obtain their opinion of our system usually used in Japan. We sent 217 questionnaires mainly to institutions in the U. S. A. and Canada. Each questionnaire consisted of 3 questions : description of the categorical system used routinely in each institution, types of speech samples used for evaluation, and a request for comments on our system.Fifty-two institutions (24 %) responded. Almost all answered that the speech factors to be evaluated were vocal resonance and articulation. They also used speech in words, sentences and conversation as speech samples. The three most frequently used articulation tests Bzoch Error Pattern Articulation Test, Templin-Darley Test of Articulation and Goldman-Fristoe Test of Articulation. We received various comments from 23 institutions, with 10 answers showing complete agreement with our system.This project gave us meaningful information about American/European clinical work on cleft palate speech, leading us to consider our categorical system to be clinically reasonable.

Pro-inflammatory cytokine high mobility group box-1 (HMGB-1) is involved in inflammation in the central nervous system, but less is known about its biological effects in the peripheral nervous system. In the present study, the role of HMGB-1 in the primary afferent nerve was investigated in the context of the pathophysiology of peripheral nerve injury-induced pain hypersensitivity. Real-time PCR confirmed an increase in HMGB-1 mRNA expression in the dorsal root ganglion (DRG) and spinal nerve at 1 day after spinal nerve ligation (SNL). Induction of HMGB-1 mRNA was observed in both injured L5 and uninjured L4. Immunohistochemistry for HMGB-1 revealed that SNL-induced HMGB-1 expression in the primary afferent neurons and satellite glial cells (SGCs) in the DRG, and in Schwann cells in the spinal nerve. Up-regulation of HMGB-1 was associated with translocation of its signal from the nucleus to the cytoplasm. Injection of HMGB-1 into the sciatic nerve produces transient behavioural hyperalgesia. Neutralizing antibody against HMGB-1 successfully alleviated the mechanical allodynia observed after SNL treatment. Receptor for advanced glycation end products (RAGE), one of the major receptors for HMGB-1, was expressed in the primary afferent neurons and SGCs in the DRG, as well as in Schwann cells in the spinal nerve. These results indicate that HMGB-1 is synthesized and secreted into the DRG and spinal nerve, and contributes to the development of neuropathic pain after nerve injury. Blocking HMGB-1/RAGE signalling might thus be a promising therapeutic strategy for the management of neuropathic pain.

Cladosporium cladosporioides No.9の無細胞抽出液より硫安沈殿,ブチルートヨパール, DEAE-トヨパールおよびトヨパールHW55のクロマトグラフィーにより酢酸エステルを特異的に分解する酢酸エステル分解酵素(Est I, Est II)を単離精製した. 各精製酵素の分子量はゲルクロマトグラフィーによりEst Iは約6.3万, Est II は約6.6万と推定された. また,各酢酸エステル分解酵素の至適反応温度はEst Iは35°C, Est IIは30°Cであった.熱安定性はともに50°C, 30分処理で完全に失活した.至適pHはEst IはpH 9.1, Est IIはpH 9.3であった. pH安定範囲は,前者がpH 7～9,後者がpH 5～6であった.両酵素ともにDFP, PMSFに阻害され,エゼリンで阻害されないことによりセリン残基を有するカルボキシエステラーゼと推定された.SH基阻害剤であるPCMB, DTNBおよび金属イオンキレート剤であるEDTAでは阻害されなかった.また, Ag2+および2価の重金属イオンのCu2+, Hg2+で強く反応を阻害された. エチルエステルに対する基質特異性は,酢酸エステルに比較して最大でも約10～20%の相対活性しかなかった.トリグリセリド,モノグリセリドの長鎖脂肪酸を持つものにはまったく作用せず,短鎖脂肪酸を持つものにわずかに活性を示した.酢酸エステルに対してはEst Iは酢酸イソアミルに対してEst IIは酢酸イソブチルに対して最大の活性を示し,酢酸β-フェネチルに対しても高い分解活性を示した.本酵素はアルコールの酢酸エステルを特異的に分解するカルボキシエステラーゼであった.

Porphyromonas gingivalis is one of the most important periodontopathogens. It produces cysteine proteinases named gingipains. We previously examined the effect of gingipains on abscess formation in a murine model. The rgpA rgpB double and kgp mutants induced smaller abscesses than the wild type. Moreover, the rgpA rgpB kgp triple (gingipain-null) mutant hardly showed lesion formation at all under the experimental conditions used, indicating that genes encoding gingipains are important for P. gingivalis virulence.Here, we further report the humoral immune responses induced by P. gingivalis strains.After the lesions were apparently cured, sera were collected from the mice and immunoglobulin G (IgG) responses against the whole cell antigens of wild-type P. gingivalis were measured.Wild-type strain was found to induce a strong antibody reaction. On the other hand, the rgpA rgpB kgp triple and kgp mutants induced significantly lower antibody responses compared to the wild type. Western blotting analysis confirmed the differences in antibody production. Next, these mice were re-infected with wild-type strain. Mice that were first infected with wild-type strain showed significantly smaller lesion formation than control mice that were first infected with medium only. On the other hand, mice that were first infected with mutant strains devoid of gingipain activities did not show resistance to re-infection and immunoglobulins directed against gingipains may be protective.These results suggest that gingipains play an important role in abscess formation in mice, and humoral immune responses seem to be partly responsible for the resistance to re-infection by P. gingivalis.

Bacteroides forsythus is one of the important periodontopathic bacteria, and this microorganism is known to have an S-layer outside the outer membrane. The S-layer-like antigens were recently isolated from B. forsythus , and they were found to be 270- and 230-kDa proteins in the envelope fraction. In this study, these proteins were confirmed to be specific to B. forsythus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and they were clearly recognized by sera from patients with adult and early-onset periodontitis in Western immmunoblot analysis. We compared the immunoglobulin G (IgG) responses against the purified S-layer-like antigen by enzyme-linked immunosorbent assay. IgG responses against this antigen were low in healthy control subjects, but they were significantly higher in subjects with adult and early-onset periodontitis. Together with the fact that the IgG responses against the crude extract of B. forsythus did not rise significantly in patients with periodontitis, S-layer-like proteins are considered to be specific antigens of B. forsythus and may play an important role in the progression of periodontitis.

A 68-year-old right-handed woman was admitted to our hospital because of difficulty to speak and understand conversation over 10 years. She was able to make herself by writing. No change in her personality or behavioral abnormality was observed so that she could live without help. Although her WAIS score and auditory brain stem response were normal, she could not understand the speech or distinguish the sound. She also spoke plenty of words fluently, resulting in undifferentiated jargon. She did not make any effort in speaking. Her speech was, however, unclear and hard to understand. Brain MRI scan disclosed a moderate atrophy of bilateral temporal lobe and enlargement of Sylvius fissure. A three-dimension reconstructed brain surface image showed enlargement of the perisylvian fissure, and atrophy of the gyrus frontalis inferior, operculum, gyrus temporal superior, bilaterally. Reduced cerebral blood flow was demonstrated on 99mTc-ECD SPECT in the left thalamus and bilateral fronto-temporal lobe. A diagnosis of slowly progressive aphasia with auditory agnosia was made. Our case suggests that bilateral disturbance of neuronal network between the primary auditory area and the secondary auditory area is responsible to the consequence of auditory agnosia.

We reviewed the clinical records of 62 patients with voice disorders without organic diseases of the larynx who were examined in the Department of Otolaryngology, Kurume University Hospital during the 10 years from 1971 to 1980. There were 9 patients with psychogenic dysphonia, 24 with vocal abuse, 4 with spastic dysphonia, 9 with mutational voice disturbance, 8 with virilization of voice, 7 with dysphonia attributed to diseases of other organs than the larynx, and 1 with senile change of voice. The clinical service to the patients with these kinds of dysphonia has been poor in Japan for lack of trained voice pathologists. This has been also the case with our department. Recognition for the necessity of voice pathologists is strongly demanded.