A sensitive enzymatic method (SK-013) for detection of Treponema denticola, Porphyromonas gingivals and Bacteroides forsythus in subgingival plaque samples

An enzymatic method, SK-013, was developed for rapid detection of the peptidase activity in subgingival plaque samples. This method was found to have specificity for Porphyromonas gingivalis, Treponema denticola, Bacteroides forsythus, and some Capnocytophaga strains. The purpose of this study was to determine whether SK-013 could indicate the presence of periodontopathic bacteria, including T. denticola, P. gingivalis and B. forsythus, which produce trypsin-like enzymes. Subgingival plaque samples were taken from 10 clinically healthy sites and 30 periodontally diseased sites with 3 paper points. SK-013 activity of plaque samples was assayed, and the numbers of T. denticola, P. gingivalis and B. forsythus in the sample were counted by immunofluorescence technique. In diseased sites, the SK-013 activity was significantly correlated with clinical parameters such as Gingival Index, Plaque Index, probing depth and bleeding on probing. A significant correlation was found between the presence of these organisms and SK-013 activity. Correlation coefficients between the presence of T. denticola and SK-013 activity were higher than those with other organisms. These findings indicate that the SK-013 is useful as an indicator of cell population of T. denticola, P. gingivalis and B. forsythus in subgingival plaque.     

Effect of ENU (ethylnitrosourea) mutagenesis in cleft lip and/or palate pathogenesis in mice

Cleft lip and/or palate (CL/P) are caused by many factors. The aim of this study was to investigate the effects of genetic point mutations in CL/P pathogenesis. ICR and AJ strain mice were used. Ethylnitrosourea (ENU) was injected into 10-week-old male mice (G0) intraperitoneally at a dose of 250 mg/kg. The males were bred with two untreated virgin females of the same strain on day 100 after injection. The uterine contents (G1) of one female were examined on day 18.5 of pregnancy. From the other female, the offspring were delivered naturally, and F3 mice (G3) were also examined in the same way. In ICR strain mice, cleft palate only (CPO) was increased in both the G1 and G3. The frequency was significantly higher in the G3 than the G1 generation. Cleft lip was not observed. In AJ strain mice, CL/P increased in both the G1 and G3. In the G3, the frequency of CPO was increased significantly. Genes related to CPO may be recessive in phenotype. CPO and cleft lip differ from a genetic viewpoint. Point mutations play a significant role in cleft lip and palate.     

P53 and MDM2 co-expression in tobacco and betel chewing-associated oral squamous cell carcinomas

Oral cancers of tobacco and betel chewers represents a unique in-vivo model to understand the genotoxic effect of tobacco and betel carcinogens on oncogenes and tumor suppressor genes. Coordinated interactions of p53 and MDM2 play an important role in regulation of critical growth control gene following exposure to DNA damaging agents. The purpose of this study is to determine if the tumor suppressor function of p53 is inactivated by mutation or other alternative mechanisms in carcinogen-induced oral squamous cell carcinoma (SCC), and to investigate the clinicopathological significance of p53 and MDM2 expression. The p53 mutation in oral SCC of tobacco and betel chewers (n=40) was detected by polymerase chain reaction - single strand conformation polymorphism (PCR-SSCP) analysis and immunohistochemistry (IHC) was done to investigate p53 and MDM2 proteins overexpression. The incidence of p53 mutation was relatively low (17.5%), but there was a high prevalence of MDM2 overexpression (72.5%). In the total of 40 cases, IHC phenotype showed p53 positive immunostaining with MDM2 positive immunostaining (p53+/MDM2+) 62.5%, p53 negative immunostaining with MDM2 negative immunostaining (p53-/MDM2-) 15%, p53 positive immunostaining with MDM2 negative immunostaining (p53+/MDM2-) 12.5%, and p53 negative immunostaining with MDM2 positive immunostaining (p53-/MDM2+) 10%. A significant correlation was found between MDM2 and p53 overexpression (p=0.0289). Moreover, p53+/MDM2+ phenotype was significantly associated with poorly differentiated tumors (p= 0.0007). These results conclude that other factors than p53 mutation is likely to be the targets of tobacco/betel carcinogens and MDM2 may play an important role in tobacco/betel chewing-related oral SCCs. Overexpression of MDM2 protein may constitute an alternative mechanism for p53 inactivation.     

Coptidis rhizoma inhibits growth and proteases of oral bacteria

OBJECTIVE: The aim of this study was to investigate the antibacterial effect of Coptidis Rhizoma (CR), a traditional medicinal plant, on oral bacteria. MATERIALS AND METHODS: CR extract was prepared by boiling CR in water for 2 h. Alkaloids contained in CR extract were assayed by high performance liquid chromatography (HPLC). Antibacterial activity of CR extract was estimated from the lowest concentration that did not permit bacterial growth (minimum inhibitory concentration, MIC) and the concentrations that inhibited 50% of bacterial proteolytic activity (IC50). RESULTS: CR extract inhibited the growth of Actinomyces naeslundii, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Actinobacillus actinomycetemcomitans at MIC of 0.031-0.25 mg ml(-1), whereas it had less inhibitory effect (MIC: 0.5-2 mg ml(-1)) on the growth of Streptococcus and Lactobacillus. The major active component of CR extract was berberine (Ber), an alkaloid, and its inhibiting specificity to bacterial growth was similar to that of CR extract. CR extract and Ber were bacteriostatic at the MICs against most of the bacteria, and bacteriocidal at the concentrations higher than the MICs. Ber inhibited the activities of collagenase from P. gingivalis and A. actinomycetemcomitans. CONCLUSION: CR extract and Ber had an inhibitory effect on periodontopathogenic bacteria. These results suggest the possibility of their clinical application for the treatment of periodontal diseases.     

18q-syndrome with cleft lip and palate. A clinically diagnosed case

A case of a child with 18q-syndrome was encountered. The patient presented cleft lip and palate, narrow auditory canals, overlapping fingers, undescended testes and micropenis. Chromosome analysis demonstrated partial deletion of the long arm of one chromosome 18 in the patient and normal chromosomes in the parents. A cheiloplasty for repair of bilateral complete cleft lip was performed.     

Treatment of intrabony periodontal defects with enamel matrix derivative in private practice: a long-term retrospective study

The present study describes treatment of intrabony periodontal defects with enamel matrix derivative (EMD) in private practice. Ten patients with clinical diagnosis of chronic periodontitis were subjected to data analysis. A total of 18 teeth with various osseous defects received regenerative therapy with EMD, and were followed for a minimum of 2 years. Treatment of the intrabony defects with EMD led to a statistically significant improvement in the mean value of probing depth at 1-year when compared with at the baseline (p<0.01). Reduction in probing depth was achieved with minimal recession of the gingival margin, and was maintained over the 2-year observation period with no significant change. Mean values of attachment gain at 1 and 2 years were of clinical significance: 3.39+/-1.46 mm and 3.22+/-1.40 mm, respectively. Although one tooth was extracted because of subsequent loss of attachment and bone, most teeth treated have been successfully maintained for 2 to 7 years with no significant signs of disease progression. In conclusion, EMD treatment of intrabony osseous defects yielded clinically favorable responses. The gain in clinical attachment can be longitudinally maintained in a private practice setting. Further controlled studies are needed to elucidate the clinical significance of EMD treatment.     

Influences of interferon-gamma on cell proliferation and interleukin-6 production in Down syndrome derived fibroblasts

Objective

Down syndrome, a frequently encountered genetic disorder, is usually associated with medical problems related to infectious disease, such as periodontal diseases and prolonged wound healing. Although affected individuals are considered to have clinical problems related to high interferon (IFN) sensitivity, the molecular mechanisms of IFN activities are not completely understood.

Design

Down syndrome derived fibroblasts, Detroit 539 (D1) and Hs 52.Sk (D2) cells, were used. To analyse the expressions of interferon (IFN) receptors and downstream of IFN-γ, western blotting was performed. Cell proliferation was determined by counting cells following trypan blue staining. Media levels of IL-1β, TNF-α, and IL-6 were quantified using ELISA.

Results

IFN-γ receptor 2 and IFN-α receptor 1, but not IFN-γ receptor 1, were highly expressed in D1 and D2 cells, as compared to the control fibroblast cells. Cell proliferation by D1 and D2 cells was lower than that by the control fibroblasts, further, IFN-γ had a greater effect to inhibit cell proliferation by D1 and D2 cells. In addition, IFN-γ treatment increased the phosphorylation of STAT1 and MAPK in D1 cells as compared to normal fibroblasts. Also, the presence of exogenous IFN-γ in the growth medium significantly induced IL-6, but not IL-1β or TNF-α, in D1 and D2 cells.

Conclusion

Taken together, our results are consistent with hypersensitive reactions to IFN-γ seen in patients with Down syndrome and may provide useful information to elucidate the mechanisms of IFN-γ activities in those individuals.     

Cloning, characterization and immunolocalization of human ameloblastin

Amelogenesis imperfecta is a broad classification of hereditary enamel defects, exhibiting both genetic and clinical diversity. Most amelogenesis imperfecta cases are autosomal dominant disorders, yet only the local hypoplastic form has been mapped to human chromosome 4q between D4S242 1 and the albumin gene. An enamel protein cDNA, termed ameloblastin (also known as amelin and sheathlin), has been isolated from rat, mouse and pig. Its human homolog has been mapped to chromosome 4q21 between markers D4S409 and D4S400, flanking the local hypoplastic amelogenesis imperfecta critical region. Therefore, ameloblastin is a strong candidate gene for this form of amelogenesis imperfecta. To facilitate genetic studies related to this dental disease, we isolated and characterized a human ameloblastin cDNA. A human third molar cDNA library was screened and two ameloblastin clones identified. Nucleotide sequencing of these cDNAs indicated alternative splicing of the putative open reading frame, use of different polyadenylation signals, and a high degree of similarity to reported rat, mouse and porcine cDNAs. Immunohistochemistry studies on embryonic human teeth using an antibody to recombinant ameloblastin indicated ameloblastin expression by ameloblasts with localization in the enamel matrix associated with the sheath structures.     

Influence of firing times and temperature on color stability of low-fusing porcelains for high-functional gold alloys

The purpose of this study was to investigate the effect of firing conditions on color stability. Three commercially available porcelains for high functional gold alloys, Carrara (CA), Deguceram Gold (DE) and Duceragold (DU) were used. In this study color stability was investigated under fire treatment of 1st, 3rd, 5th times and firing temperatures of 0, 20, and 40 degrees C higher than the manufactures standard temperature (CA: 845 degrees C, DE: 780 degrees C, DU: 780 degrees C). L* was degreased in CA 5 times, DE 3 times, DU by repeated firing. No difference was observed among L* of firing temperatures DE and DU, but L* was degreased in CA with firing temperatures of 40 degrees C higher. a* was degreased in CA, DU, DE with repeated firing, a* was degreased 5 times at firing temperatures of higher. b* was degreased in DU, but b* was increased in CA and DE with 5 firings. b* was increased with 5 findings at firing temperatures of 40 degrees C higher.