• YAKHAK HOEJI
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• v.52 no.4
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• pp.264-273
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• 2008

The inhibitory effect of omega-3 such as linolenic acid (LNA), docosahexaenoic acid (DNA) and eicosapentaenoic acid(EPA) on the growth of normal cell lines and cancer cell lines was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyItetrazolium bromide (MTT) and 2,3-bis-2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-caboxanilide (XTT) methods. LNA was found to decrease the cell viability of human oral epithelioid carcinoma cells (KB) in the MTT assay, whereas EPA appeared to inhibit the cell adhesion activity of human skin melanoma cells (SK-MEL-3) in the XTT assay analysis. DPPH radical scavenging activity was examined on LNA, DHA and EPA at the concentration of 100 ${\mu}M$, where they showed about 53% scavenging activity. These results suggest that omega-3 unsaturated fatty acid has a potential anticancer activity.

• International Journal of Oral Biology
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• v.34 no.1
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• pp.43-48
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• 2009

Thrombin-induced platelet microbicidal protein (tPMP) is a small cationic peptide that exerts potent in vitro microbicidal activity against a broad spectrum of human pathogens, including Staphylococcus aureus and Streptococcus rattus BHT. Earlier evidence has suggested that tPMP targets and disrupts the bacterial membrane. However, it is not yet clear whether membrane disruption itself is sufficient to kill the bacteria or whether subsequent, presumably intracellular, events are also involved in this process. In this study, we investigated the microbicidal activity of rabbit tPMP toward S. rattus BHT cells in the presence or absence of a pretreatment with antibiotics that differ in their mechanisms of action. The streptocidal effects of tPMP on control cells (no antibiotic pretreatment) were rapid and concentration-dependent. Pretreatment of S. rattus BHT cells with either penicillin or amoxicillin (inhibitors of bacterial cell wall synthesis) significantly enhanced the anti-S. rattus BHT effects of tPMP compared with the effects against the respective control cells over most tPMP concentration ranges tested. On the other hand, pretreatment of S. rattus BHT cells with tetracycline or doxycycline (30S ribosomal subunit inhibitors) significantly decreased the streptocidal effects of tPMP over a wide peptide concentration range. Furthermore, pretreatment with rifampin (an inhibitor of DNA-dependent RNA polymerase) essentially blocked the killing of S. rattus BHT by tPMP at most concentrations compared with the respective control cells. These results suggest that tPMP exerts anti-S. rattus BHT activity through mechanisms involving both the cell membrane and intracellular targets.

• The Journal of Korean Academy of Prosthodontics
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• v.40 no.6
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• pp.560-571
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• 2002

Jaw relations and the recording methods have been controversial aspects of dentistry. The purpose of this study was to quantify the relative muscle activity of the masseter and temporal muscles in relation to different intermaxillary relations recorded by intra-oral tracer during maximal clenching and to decide the optimal mandibular position. Ten volunteers with healthy TMJ were studied. Intra-oral tracer was assembled and bite block was fabricated in the articulator. Intra-oral tracer was placed in the mouth, and four mandibular positions were recorded. EMG activity was recorded on a BIO-PAK system (Bio-Research Associates, Inc. USA.) in masseter and temporal muscle and compared in each mandibular positions. The results were as follows: 1. In comparison with maximum intercuspation, the chewing position was the most similar followed by tapping position, myocentric position and posterior border position. However the differences were not statistically significant. 2. In comparison of bilateral symmetry of masseter muscle, tapping position was the most symmetrical followed by chewing position and maximum intercuspation. Myocentric position and posterior border position were not symmetrical. (P<.05). 3. In comparison of bilateral symmetry of anterior temporal muscle, chewing position was the most symmetrical followed by posterior border position, maximum intercuspation, myocentric position and tapping position. However the differences were not statistically significant. 4. In comparison of proportionality of anterior temporal muscle to masseter muscle activity on left side, posterior border position was the greatest followed by myocentric position, taping position, chewing position and maximum intercuspation. And the proportionality of posterior border position was greater than that of maximum intercuspation. (P<.05). 5. In comparison of proportionality of anterior temporal muscle to masseter muscle activity on right side, myocentric position was the greatest followed by posterior border position, tapping position, maximum intercuspation and chewing position. However the differences were not statistically significant.

• International Journal of Oral Biology
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• v.34 no.4
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• pp.185-190
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• 2009

This study examined the effects of red light generated from a light emitting diode (LED) upon proliferation and mitochondrial stress in human gingival fibroblasts (hGFs). Cells were exposed to LED-generated red light at a clinically relevant intensity and distance with a 610-630 nm wavelength for various times (0-48 min). At different exposure times, cells were processed for the analysis of succinate dehydrogenase (SDH) activity, proliferation, mitochondrial membrane potential (MMP) and cytotoxicity. Cell cycle progression was also investigated by flow cytometry after staining with propidium iodide. Red light exposure was found to inhibit SDH activity and DNA synthesis in hGFs in a time-dependent manner. Light exposure also reduced the MMP levels in these cells and this was closely associated with a $G_0/G_1$ arrest. In contrast, exposure of hGFs to red light for 48 min led to a dramatic loss of MMP with an attendant increase in cytotoxicity. These findings demonstrate that LED-generated red light may cause mitochondrial stress and growth inhibition in hGFs during tooth whitening therapy, depending on the length of the exposure.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7155-7159
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• 2015

Background: Oral cancer is a health problem in Thailand. Cratoxylum formosum subsp. pruniflorum Gogel (Teawdang), normally consumed in northeast Thailand, has proven cytotoxic to cervical cancer cell lines including HeLa, SiHa and C-33A. Recently, Asian oral cancer cell lines, ORL-48 and ORL-136, were established. Therefore, we aimed to study cytotoxicity of Teawdang in these. Total phenolic (TPC) and flavonoid content (TFC), and antioxidant activity of Teawdang were also determined. Materials and Methods: Teawdang was purchased from Khon Kaen market during June-October 2013. Hexane (CHE), ethyl acetate (CEE) and methanol (CME) extracts of its edible part were analyzed for TPC by the folin-ciocalteau method and for TFC by an aluminium colorimetric method. Antioxidant activity and cytotoxicity in normal Vero cells and oral cancer cells were investigated. Cell viability was assessed using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Results: CME and CEE had higher TPC and TFC and antioxidant activity than CHE. Both CME and CEE, at $200{\mu}g$ dry wt/mL, were cytotoxic to the studied oral cancer cell lines. However, CME was cytotoxic to Vero cells whereas CEE was not. Compared to Vero cells, CEE significantly inhibited ORL-48 and ORL-136 growth (p=0.03 and p=0.02, respectively). Conclusions: CEE exhibited cytotoxic effects on the studied oral cancer cell lines but not normal Vero cells. The bioactive compounds in CEE should be further purified and elucidated for their mechanisms of action for development as anticancer agents.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.5
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• pp.470-480
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• 2000

The epithelium of odontogenic cyst seems to be in a specific status of cellular proliferation and cytodifferentiation. With the identification of various genes, which play essential roles in the specific stages of cellular proliferation and differentiation, the cellular conditions of odontogenic cyst epithelium need to be reevaluated. This study aimed to estimate the degree of proliferating, differentiating and apoptotic activities of odontogenic cyst epithelium using antisera of PCNA, Ki-67, MPM-2, transglutaminase C, heat shock protein 70 and $ApopTag$^{(R)}$. method in 19 cases of odontogenic cysts. Cellular changes of the cyst epithelium were measured by intensity of each immunohistochemical staining. Results were as follows: 1. The proliferating activity of the cyst epithelium was slightly lower than that of normal oral mucosal epithelium, with the use of primary antibodies against PCNA, Ki-67, and MPM-2. And the proliferating activity of the epithelium in OKC group was even higher than that of the epithelium in non-OKC group. 2. The odontogenic cysts showed weakly positive reaction with transglutaminase C, but strongly positive reaction with HSP 70. 3. Occasionally, only a few apoptotic cell was observed in the superficial keratin layer of OKC. 4. The hyperplastic cyst epithelium infiltrated with mild inflammatory cells showed diffusely positive reaction with different proliferating factors. From the above results, we presumed that the endogenous proliferating and differentiating activity of the cyst epithelium was slightly lower than that of normal oral mucosal epithelium, and also supposed that the cyst epithelium could be reactivated for the further proliferation by the exogenous factors, such as inflammatory reaction and any chemicophysical irritations.

• International Journal of Oral Biology
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• v.36 no.3
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• pp.155-162
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• 2011

Eugenol (4-allyl-2-methoxyphenol) is a naturally occurring phenolic compound that is widely used in dentistry as a component of zinc oxide eugenol cement that is commonly applied to the mouth environment. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatments with eugenol and cisplatin on human melanoma (G361) cells. To investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment, an MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed. The results indicated that a co-treatment with eugenol and cisplatin induced multiple pathways and processes associated with an apoptotic response in G361 cells including nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, the increase and decrease of Bax and Bcl-2, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of 300 ${\mu}M$ eugenol or 3 ${\mu}M$ cisplatin for 24 h did not induce apoptosis. Our present data thus suggest that a combination therapy of eugenol and cisplatin is a potential treatment strategy for human melanoma.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.2
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• pp.85-99
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• 2004

Purpose: This study observed the changes in the telomerase activity, it's developmental regulation, PCNA expression, and their correlation in rat's upper digestive organs during growth and aging. Materials and Methods: Upper digestive organs(buccal mucosa, gingiva, palate, submandibular and parotid glands, and tongue) were aseptically removed from Sprague-Dawley rats of fetal(gestational 20 days), growing(1, 2, 3, 5, and 7 weeks after birth) and adult(12 week old). Samples for telomerase activity were frozen on liquid nitrogen immediately after sacrifice, and stored until the use at $-75^{\circ}C$ in order to measure it. Telomerase activity was measured by a PCR-based telomeric repeat amplication protoco(TRAP) assay and quantitated with Photometric Telo TAGGG Telomerase PCR ELISA plus(Roche Diagnostics GmbH. Mannheim. Germany). PCNA expression were measured immunohistochemistry with anti PCNA Ab-1, Clone PC10(NeoMark. California. USA). Results: 1. Telomerase activities in buccal mucosa, palate and gingiva were the highest in fetus and decreased gradually or rapidly after birth and then diminished, but In salivary gland and tongue were the highest in fetus and also high at 1 week and then decreased rapidly. 2. PCNA expression in buccal mucosa, gingiva, Tongue and salivary gland was the highest in fetus and decreased gradually and then diminished. but only in palate decreased rapidly after birth and then diminished. Conclusion: The highest telomerase activity of embryonic stage decreased rapidly after birth in rat's upper digestive organs. There may be a developmental regulation of telomerase activity, but not a tissue-specific. This telomerase activity seems correlated closely with PCNA expression in rat's upper digestive system.

• Journal of Oral Medicine and Pain
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• v.32 no.1
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• pp.45-55
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• 2007

Plant extract has attracted considerable interest in oral disease therapy. The present study was performed to observe the antibacterial effect on cariogenic Streptococcus mutans GS5 and Streptococcus sobrinus 6715, and periodontopathic Actinobacillus actinomycetemcomitans Y4 of phytoncide from Chamaecyparis obtusa Sieb. et Zucc employing the measurement of optical density, viable cell counts, and antibiotic sensitivity. The results were as follows: 1. Minimum inhibitory concentration of the phytoncide for S. mutans, S. sobrinus, and A. actinomycetemcomitans was observed to be 0.5%, 1%, and 0.2%, respectively. 2. Minimum bactericidal concentration of the phytoncide for S. mutans, S. sobrinus, and A. actinomycetemcomitans was determined to be 0.5%, 2%, and 0.2%, respectively. 3. The bacteria exposed to the phytoncide become more sensitive to antibiotics. The phytoncide enhanced significantly antibacterial activity of ampicillin against S. mutans and S. sobrinus. It also increased significantly the activity of penicillin and amoxicillin against S. sobrinus. In contrast, the phytoncide augmented the activity of amoxicillin and cefotaxime against A. actinomycetemcomitans but the increase was not statistically significant. The overall results indicate that phytoncide from Chamaecyparis obtusa Sieb. et Zucc used for this study has a strong antibacterial activity against cariogenic and periodontopathic bacteria and that it also has permeabilizing effect on certain antibiotics against these bacteria. Therefore, the phytoncide may be used as a candidate for prevention and therapeutic agent against oral infectious disease including dental caries and periodontal disease.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.587-598
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• 2000

Urea in the oral cavity is hydrolyzed mainly by bacterial ureases to ammonia, which in turn, raises pH of the oral environment, maintaining oral pH homeostasis, thereby inhibiting dental caries. Streptococcus salivarius has been shown to be a major contribution to oral ureolysis. Synthesis of urease by S. salivarius appears to be constitutive, but can be greatly enhanced in the acidic environment. It has been presumed that ureolytic activity of S. salivarius strains isolated from caries-active site is greater than that of strains from caries-free site. However, no in vivo study has supported the presumption. The present study was performed to observe the ureolytic activity of S. salivarius strains isolated from different environments in the same individual, finding out whether the ureolytic activity is related to dental caries. For the purpose, S. salivarius strains were isolated from caries-active site (>C2), a caries-free site of the tooth, and the dorsum of the tongue of each of 50 patients having decayed teeth. The strains isolated from the patients who harbored S. salivarius in more than two sites were selected and then their ureolytic activities were measured. In order to examine clonal diversity of the strains, their ureC genes were amplified by polymerase chain reaction (PCR) and then restricted with EcoRV, and the protein profiles of the strains were compared by SDS-PAGE. The results were as follows: 1. Of 50 patients, 13 patients harbored S. salivarius in more than two sites; a total of 61 S. salivarius strain were isolated from the patients and selected for the study. 2. Of 17 isolates from the caries-active site of 9 patients harboring S. salivarius in more than two sites including carious lesion, 10 (58.8%) showed a high ureolytic activity (> 200 ${\mu}mol/min/mg$). While, 19 out of 44 isolates (43.2%) from the caries-free site of the teeth and the dorsum of the tongues of 13 patients were the strains with a high ureolytic activity. 3. Of 9 patients harboring S. salivarius in more than two sites including caries-active site. 6 patients were found to have the strains in the caries-active site showing a lower ureolytic activity than the strains in the other sites. 4. Of 34 isolates with ureolytic activity higher than 40 ${\mu}mol/min/mg$, 32 isolates produced 0.54-Kbp PCR products regardless of the sites of bacterial collection. In contrast, of 27 isolates with ureolytic activity lower than 40${\mu}mol/min/mg$, 26 isolates yielded 1.3-Kbp PCR products or none regardless of the sites. 5. Different clonal types of S. salivarius with relatively higher and lower ureolytic activities were found in the same individuals and even in the same sites. 6. None of strains showing different ureolytic activity appeared to be the same clonal type. The overall results suggest that ureolytic activity of the isolates does not appear to be related to differences of the environments but related to their own genetic traits.