The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.
Park, So-Young;Choi, Seoung-Hwan;Lee, Su-Jeong;Chang, Moon-Taek;Kim, Hyung-Seop
Journal of Periodontal and Implant Science
/
v.32
no.1
/
pp.41-49
/
2002
The aim of this study was to compare periodontal conditions around mesially tipped molars by a tipping degree. Patients who had been consecutively treated at the Department of Periodontology, Chonbuk National University Hospital from October 1999 to August 2001 were assessed with radiographs taken at their molar regions. Of all molars investigated, 142 molars of 116 patients tipped mesially more than 10 degrees to the line perpendicular to an occlusal plane were selected for the study. The tipped molars were divided into 2 groups with a reference to a tipping degree, i.e., 66 slightly tipped(group 1 : <$30^{\circ}$) and 76 severely tipped molars (group 2 : ${\geq}30^{\circ}$). Probing depth(PD), plaque retention index(PRI) at mesial surfaces of tipped molars and tooth mobility(TM) were recorded at the clinical examination. Tipping degree(TD) and alveolar bony defect(ABD) at the mesial surface of the molars were measured in a radiograph. The results showed that no statistical differences were found between groups in all measured variables. In Pearson correlation analysis performed in the same group, a positive relationship was shown between PRI and PD in the group 1 and, in the group 2, between PRI and PD as well as PRI and ABD(p < 0.01). However, no statistically significant relationship was found between TD and all other variables in both groups. Within limitations of this study, it may be concluded that tipping degree did not seem to influence periodontal conditions, i.e., PD, ABD and TM of mesially tipped molars per se, but plaque presence/absence seemed to mainly affect the periodontal conditions of the tipped molars.
Magan-Fernandez, Antonio;O'Valle, Francisco;Pozo, Elena;Liebana, Jose;Mesa, Francisco
Journal of Periodontal and Implant Science
/
v.45
no.6
/
pp.252-256
/
2015
Purpose: The purpose of this report was to describe the clinical and microbiological characteristics of two rare cases of necrotizing stomatitis, and the outcomes of a non-invasive treatment protocol applied in both cases. Methods: We report two cases of necrotizing stomatitis in a rare location in the hard palate of a 40-year-old woman and a 28-year-old man. Neither had a relevant medical history and both presented with highly painful ulceration in the palate and gingival margin that was accompanied by suppuration and necrosis. 3% hydrogen peroxide was applied to the lesions using sterile swabs, and antibiotic and anti-inflammatory treatment was prescribed to both patients in addition to two daily oral rinses of 0.2% chlorhexidine. Results: In both cases, radiological examination ruled out bone involvement, and exfoliative cytology revealed a large inflammatory component and the presence of forms compatible with fusobacteria and spirochetes. There was a rapid response to treatment and a major improvement was observed after 48 hours, with almost complete resolution of the ulcerated lesions and detachment of necrotic areas with partial decapitation of gingival papillae. Conclusions: Necrotizing periodontal lesions can hinder periodontal probing and the mechanical removal of plaque in some cases due to the extreme pain suffered by the patients. We present a non-invasive treatment approach that can manage these situations effectively.
Background: The role of the interferon consensus sequence binding protein (ICSBP), a member of interferon regulatory factor family, in protecting against a vesicular stomatitis virus (VSV) infection has not been firmly elucidated. Thus, it was investigated utilizing the human promyelocytic leukemia HL-60 cells which do not express ICSBP. Methods: HL-60 cells were stably transfected with plasmid containing cDNA for either ICSBP or DNA binding domain (DBD) and tested for their VSV-susceptibilities. The susceptibility of each transfectant group to a VSV infection was determined by a plaque assay at 1 h, 24 h, and 48 h post-infection in the presence (500 IU/ml) or absence of interferon ${\alpha}$ ($IFN{\alpha}$). Results: In the absence of $IFN{\alpha}$, the three groups showed similar sensitivities to a VSV infection. However, when pre-treated with IFN, the viral titers in both the ICSBP and control clones steadily decreased over 48 h of incubation, indicating the existence of $IFN{\alpha}$-mediated protection against VSV infection. The $IFN{\alpha}$-treated ICSBP clones appeared to be more resistant to infection compared with the control clones, although the difference was not great. On the contrary, the viral titers in the $IFN{\alpha}$-treated DBD clones increased at 24 h then decreased by 48 h. Conclusion: The expression of truncated ICSBP (DBD) does not appear to underlie the impaired protection against a VSV infection in the DBD clones, since even the control clones lacking ICSBP were protected from a VSV infection. This suggests that ICSBP does not play a critical role in the $IFN{\alpha}$- mediated anti-VSV response of HL-60 cells, although it appears to confer some resistance to a VSV infection.
Background: The primary aims of periodontal disease treatment is to remove dental plaque and calculus, the main causes of tooth loss, and restore periodontal tissue destroyed by inflammation. Periodontal disease treatment should also help maintain the alveolar bone, alleviate inflammation, and promote periodontal ligament cell proliferation, which is essential for tissue regeneration. Conventional antibiotics and anti-inflammatories have adverse side effects, especially during long-term use, so there is a need for adjunct treatment agents derived from natural products. The purpose of this study was to investigate whether the herbal flavone baicalein has the osteogenic activity under inflammatory conditions, and assess the involvement of osteoblast immediate early response 3 (IER3) expression. Methods: Human osteoblastic MG-63 cells were cultured with the pro-inflammatory cytokines tumor necrosis factor α and interleukin 1β in the presence and absence of baicalein. Proliferation was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and expression of IER3 mRNA was assessed using real-time polymerase chain reaction. The expression of IER3 protein levels and activation of associated signal transduction pathways were assessed using western blotting. Results: Baicalein increased IER3 mRNA and protein expression synergistically. In addition, baicalein reversed the suppression of cell proliferation, and the downregulation of osteogenic transcription factor runt-related transcription factor 2 and osterix induced by pro-inflammatory cytokines. Baicalein also upregulated the phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK 1/2). The upregulation of IER3 by pro-inflammatory cytokines was blocked by pretreatment with inhibitors of AKT, p38, JNK, and ERK 1/2. Conclusion: Baicalein mitigates the deleterious responses of osteoblasts to pro-inflammatory cytokines. Further, IER3 enhanced the effect of baicalein via activation of AKT, p38, JNK, and ERK pathways.
Many studies reported that the presence of cervical enamel projection (CEP) in cemento-enamel junction(CEJ) is greatly related to periodontal disease. The aim of this study was to investigate the prevalence of enamel projections in buccal, mesial, distal and lingual(palatal) surface of maxillary and mandibular first and second molars on extracted teeth. Among 660 teeth extracted due to the periodontal disease and dental caries in Seoul National University Dental Hospital was examined, 530 teeth which has distinct CEJ were examined with 8 times x electronic magnifier by one examiner. The prevalence of CEP for maxillary teeth (45.49%) was higher than that of mandible (39.62%). The first molar (45.22%) had more CEP than second (39.89%). Furthermore, buccal surface had highest incidence of CEP than other surfaces. The results of this study imply that the clinicians should take good care of the prevalence of CEP when scaling or root planning, plaque control instruction and periodontal surgery.
Bacterial infection and smoking are an important risk factors involved in the development and progression of periodontitis. However, the signaling mechanism underlying the host immune response is not fully understood in periodontal lesions. In this study, we determined the expression of janus kinase (JAK)/signal transducer and activator of transcription (STAT) on Porphyromonas gingivalis lipopolysaccharide (LPS)- and nicotine-induced cytotoxicity and the production of inflammatory mediators, using osteoblasts. The cells were cultured with 5 mM nicotine in the presence of $1{\mu}g/ml$ LPS. Cell viability was determined using MTT assay. The role of JAK on inflammatory mediator expression and production, and the regulatory mechanisms involved were assessed via enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, and Western blot analysis. LPS- and nicotine synergistically induced the production of cyclooxgenase-2 (COX-2) and prostaglandin $E_2$ ($PGE_2$) and increased the protein expression of JAK/STAT. Treatment with an JAK inhibitor blocked the production of COX-2 and $PGE_2$ as well as the expression of pro-inflammatory cytokines, such as tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$ ($IL-1{\beta}$), and IL-6 in LPS- and nicotine-stimulated osteoblasts. These results suggest that JAK/STAT is closely related to the LPS- and nicotine-induced inflammatory effects and is likely to regulate the immune response in periodontal disease associated with dental plaque and smoking.
Journal of the Korean Academy of Esthetic Dentistry
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v.10
no.1
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pp.30-45
/
2001
In recent years, clinicians' and dentists' esthetic demands in dentistry have increased rapidly. The ultimate goal in modern restorative dentistry is to achieve "white" and "pink" esthetics in the esthetically important zones. Therefore, modern esthetic dentistry involves not only the restoration of lost teeth and their associated hard tissues, but increasingly the management and reconstruction of the encasing gingiva with adequate surgical techniques. Interdental space are filled by interdental papilla in the healthy gingiva, preventing plaque deposition and protecting periodontal tissue from infection. This also inhibits impaction of food remnants and whistling through the teeth during speech. These functional aspects are obviously important, but esthetic aspects are important as well. Complete and predictable restoration of lost interdental papillae remains one of the biggest challenges in periodontal reconstructive surgery. One of the most challenging and least predictable problems is the reconstruction of the lost interdental papilla. The interdental papilla, as a structure with minor blood supply, was left more or less untouched by clinicians. Most of the reconstructive techniques to rebuild lost interdental papillae focus on the maxillary anterior region, where esthetic defects appear interproximally as "black triangle". Causes for interdental tissue loss are, for example, commom periodontal diseases, tooth extraction, excessive surgical periodontal treatment, and localized progressive gingiva and periodontal diseases. If an interdental papilla is absent because of a diastema, orthodontic closure is the treatment of choice. "Creeping" papilla formation has been described by closing the interdental space and creating a contact area. In certain cases this formation can also be achieved with appropriate restorative techniques and alteration of the mesial contours of the adjacent teeth. The presence of an interdental papilla depends on the distance between the crest of bone and the interproximal contact point, allowing it to fill interdental spaces with soft tissue by altering the mesial contours of the adjacent teeth and positioning the contact point more apically. The interdental tissue can also be conditioned with the use of provisional crowns prior to the definitive restoration. If all other procedures are contraindicated or fail, prosthetic solutions have to be considered as the last possibility to rebuild lost interdental papillae. Interdental spaces can be filled using pink-colored resin or porcelain, and the use of a removable gingival mask might be the last opportunity to hide severe tissue defects.
The present study has been performed to evaluate the clinical, microbiological, biochemical and immunological parameters associated with the periodontal disease activity in adolescent periodontitis. 21 young adolescents with evidences of periodontal attachment loss participated in the study for upto 3 years of examination. Probing pocket depths and attachment levels of whole dentitions were annually recorded and 4 deepest pockets, with initial probing depth ${\geq}$ 4mm, were selected as the representative experimental sites of a patient. Sites experiencing attachment loss ${\geq}$ 1mm during the 3-year experimental period were designated as the active sites and these sites were examined for the microbiological and biochemical profiles at the time when attachment loss occurred. Microbiological assay included cultural studies and PerioScan for monitoring BANA(+) organisms(e.g. Porphyromonas gingivalis, Treponema denticola, Bacteriodes forsythus). Biochemical assay has been performed for monitoring GCF levels of neutral protease. Serum IgG and IgG2 titers against Porphyromonas gingivalis 381 were determined of a patients at the beginning and the end of the study, respectively for patient-based analysis. The results indicated that the parameters consisting of microbiological cultures and GCF neutral protease exhibited low association with the periodontal disease activity in adolescents. However, the specificity for microbiological culture of the selected periodontopathic organisms(Aa,Pg,Pi) were considerably high. Moreover, the clinical pameters such as bleeding on probing and presence of plaque as well as IgG levels against Pg at the baseline exminations were closely associated with the subsequent evidences of attachment loss during the whole experimental period(3-year).
Purpose: The present study has two aims; firstly, it attempts to verify the presence of oxidative stress by estimating the reactive oxygen species (ROS) levels in periodontal pockets ${\geq}5$ mm as compared to controls. The second aim is to evaluate the effect of lycopene as a locally delivered antioxidant gel on periodontal health and on the gingival crevicular fluid (GCF) levels of 8-hydroxydeoxyguanosine (8-OHdG), a marker of oxidative injury. Methods: Thirty-one subjects participated in this study. In the pretreatment phase, the ROS levels in pockets ${\geq}5$ mm were measured by flow cytometry. Three sites in each subject were randomly assigned into each of the following experimental groups: sham group, only scaling and root planing (SRP) was done; placebo group, local delivery of placebo gel after SRP; and lycopene group, local delivery of lycopene gel after SRP. Clinical parameters included recording site-specific measures of GCF 8-OHdG, plaque, gingivitis, probing depth, and clinical attachment level. Results: The gel, when delivered to the sites with oxidative stress, was effective in increasing clinical attachment and in reducing gingival inflammation, probing depth, and 8-OHdG levels as compared to the placebo and sham sites. Conclusions: From this trial conducted over a period of 6 months, it was found that locally delivered lycopene seems to be effective in reducing the measures of oxidative stress and periodontal disease.
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