• 보존과학연구
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• s.29
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• pp.137-148
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• 2008

Analyses of ancient DNA (aDNA) from archaeological and historical skeletal material are characterized by low quality. Many soil contaminants such as humic acid, fulvic acid, and bone collagen are often co-extracted with aDNA and inhibit amplification by polymerase chain reaction (PCR). In this study, we compared with two methods of DNA extraction by phenolchloroform extraction and silica-bead extraction. In addition, we applied new protocol, ultra sonication based silica-bead extraction method to extract aDNA from some ancient human skeletal remains. This method was more effective by both mitochondrial DNA (mtDNA) and amelogenin gene amplification.

• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.497-513
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• 1995

Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

• Journal of Oral Medicine and Pain
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• v.19 no.2
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• pp.205-219
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• 1994

Cigarette butts from 5 smokers were gathered and then, placed in room temperature for 1, 3, 5, 7, 15 days. The possible use of the cigarette butts for individual identification was evaluated in sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene from the extracted DNA. 1. DNA extraction was possible in cigarette butts weree left in room temperature for 15days, so it can be applicatable to individual identification by polymerase chain reaction(PCR). 2. Amplification of X-Y homologous amelogenin gene by PCR made it possible to identify the sex in saliva stains (cigarette butts). 3. Amplification of D1S80 locus can be acquired from adding the boving serum albumin and hot start PCR procedures from forensic samples such as saliva stains (cigarette butts), so the AMP-FLPs examining is possible. 4. Genotype could be determined simply and rapidly using Amplitype$TM$ HLA-DQ$\alpha$ forensic kit in examining the HLA-DQA1 gene. From the investigation, DNA extraction, sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene was successfully done even though the cigarette butts were left for 15 days at room temperature. Therefore cigarette butts are highly reliable and applicatable as molecular biologic samples for individual identification.

• International Journal of Oral Biology
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• v.34 no.3
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• pp.143-150
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• 2009

Amino acid transporters play important roles in supplying nutrients to cells. In our current study, we investigated the expression of LAT1 and measured the amino acid uptake in ameloblast cultures to further elucidate the roles of this transporter during the differentiation of these cells. RT-PCR, observations of cell morphology, Alizaline red-S staining, and uptake analyses were performed following the experimental induction of differentiation in the cultures. LAT1 mRNA was detectable and found to gradually increase over time whereas LAT2 mRNA was not evident in the ameloblast cultures. Transcripts of 4F2hc, a cofactor of LAT1 and LAT2, were also found to be expressed in ameloblast cultures and increase with time. Amelogenin mRNA was expressed in the early stage ameloblast cultures. L-leucine uptake was observed to increase over 14 days of growth in culture. Our data suggest that LAT1 has a key role in the differentiation of ameloblasts and in providing these cells with neutral amino acids, including several essential amino acids.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.177-183
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• 2018

The objective of this study was to examine the expression pattern of Kelch-like ECH-associated protein 1 (Keap1) in the maxillary $2^{nd}$ molar germs of rats. We used the maxillary $2^{nd}$ molar germs in rats' pup at postnatal day 3 (bell stage), 6 (crown formation stage) and 9 (root formation stage). The investigation on mRNA and protein levels were done using reverse transcription - polymerase chain reaction and western blot. Localization of Keap 1 in the maxillary $2^{nd}$ molar germs were revealed through immunofluorescence staining. Keap1 from the maxillary 2nd molar germs were mostly manifested on postnatal day 3 and dramatically decreased on postnatal day 6 and 9 at mRNA and protein levels, while amelogenin and ameloblastin increased during the development of maxillary 2nd molar germs. During immunofluorescence analysis, the strong immunoreactivity against Keap1 was detected in the apical side of ameloblasts at the presecretory and secretory stages. However, Keap1 expression was hardly observed in the ameloblasts at the maturation stage. These results shows that Keap1 is strongly expressed in the presecretory and secretory ameloblasts of amelogenesis, and suggest that Keap1 may be a crucial molecule for the regulatory mechanisms tasked with the formation of enamel layer.

• Journal of Oral Medicine and Pain
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• v.21 no.2
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• pp.225-242
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• 1996

The author studied to determine the sex and age of 8 ancient human skeletal remains, which had excavated from ancient tombs located in Yeanri, Kimhae. Some kinds of personal identification methods their skulls and teeth were used for this study and the results were obtained as follows : 1. Sex determination was possible in ancient teeth from 4th to 5th century, using detection of X-Y homologous amelogenin gene by polymerase chain reaction. 2. DNA analysis proved that the materials examined were all male, but which always did not coincide with the results from other methods for sex determination including comparison of sexual differentiation of cranium and teeth and use of discriminant functions in the dental measurement. 3. There was little difference of the estimated ages between the methods by regression of pulp cavity and attrition in teeth. The ages from these two methods always did not coincide with ones from evaluation closure of cranial and palatal sutures. 4. Sex and age of the materials were determined as follows ; $\cdot$ Y9 was estimated to be male in early sixties. $\cdot$ Yl2 was estimated to be male in late twenties to early thirties. $\cdot$ Y37 was estimated to be male in early forties. $\cdot$ Y70 was estimated to be male in early sixties. $\cdot$ Y87 was estimated to be male in late forties. $\cdot$ Y109 was estimated to be male in early forties. $\cdot$ Yl29 was estimated to be male in late thirties to early forties. $\cdot$ Yl42 was estimated to be male in late fifties to early sixties.

• Journal of Sasang Constitutional Medicine
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• v.21 no.1
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• pp.227-236
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• 2009

1. Objectives As a basic trial for identification of Sasang constitutional gene marker, we genotyped and analysed statistical relationships of STR(short tandem repeat) alleles and its distribution in each constitution. 2. Methods After obtaining basic constitutional data with questionnaire (QSCC II), decision of constitution was made by 3 different constitution specialists' diagnosis, and only the samples of specialists' agreement of each constitution by discussion were taken into this research. Using multiplex PCR kit, total 146 constitutional samples were amplified in 16 autosomal STR marker, genotyped, and analysed statistically. Among 16 markers, 15 were analysed in this study excluding the amelogenin marker is used for in gender identification. 3. Results and Conclusions It is difficult to determine the relationship between constitution and STR marker as the sample size is small, however, Penta D and vWA were shown to be related statistically with constitution. It has been know that STRs has no genetic informations, however there are some recent research results showing STRs as a regulatory element, relationship between microsatellite instability and repeat number and size, and post-transcriptional sigualing. STRs which is not known about its function currently, are proposed to have function and/or regulatory activities anyhow with Sasang constitution. It is believed that the results of this study can halp determine and deatify the markers related to Sasang Constitutional Medicien.

• International Journal of Oral Biology
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• v.45 no.4
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• pp.190-196
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• 2020

Several factors, including genetic and environmental insults, impede protein folding and secretion in the endoplasmic reticulum (ER). Accumulation of unfolded or mis-folded protein in the ER manifests as ER stress. To cope with this morbid condition of the ER, recent data has suggested that the intracellular event of an unfolded protein response plays a critical role in managing the secretory load and maintaining proteostasis in the ER. Tauroursodeoxycholic acid (TUDCA) is a chemical chaperone and hydrophilic bile acid that is known to inhibit apoptosis by attenuating ER stress. Numerous studies have revealed that TUDCA affects hepatic diseases, obesity, and inflammatory illnesses. Recently, molecular regulation of ER stress in tooth development, especially during the secretory stage, has been studied. Therefore, in this study, we examined the developmental role of ER stress regulation in tooth morphogenesis using in vitro organ cultivation methods with a chemical chaperone treatment, TUDCA. Altered cellular events including proliferation, apoptosis, and dentinogenesis were examined using immunostaining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In addition, altered localization patterns of the formation of hard tissue matrices related to molecules, including amelogenin and nestin, were examined to assess their morphological changes. Based on our findings, modulating the role of the chemical chaperone TUDCA in tooth morphogenesis, especially through the modulation of cellular proliferation and apoptosis, could be applied as a supporting data for tooth regeneration for future studies.

• Journal of Yeungnam Medical Science
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• v.39 no.4
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• pp.322-331
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• 2022

Background: The combined use of biomaterials for regeneration may have great biological relevance. This study aimed to compare the regenerative potential of biphasic calcium phosphate (BCP) alone and with growth factor enamel matrix derivatives (EMDs) for the regeneration of intrabony defects at 1 year. Methods: This randomized controlled trial included 40 sites in 29 patients with stage II/III periodontitis and 2/3 wall intrabony defects that were treated with BCP alone (control group) or a combination of BCP and EMD (test group). BCP alloplastic bone grafts provide better bio-absorbability and accelerate bone formation. EMDs are commercially available amelogenins. Mean values and standard deviations were calculated for the following parameters: plaque index (PI), papillary bleeding index (PBI), vertical probing pocket depth (V-PPD), vertical clinical attachment level (V-CAL), and radiographic defect depth (RDD). Student paired and unpaired t-tests were used to compare the data from baseline to 12 months for each group and between the groups, respectively. The results were considered statistically significant at p<0.05. Results: At 12 months, the PI and PBI scores of the control and test groups were not significantly different (p>0.05). The mean V-PPD difference, V-CAL gain, and RDD difference were statistically significant in both groups at 12 months (p<0.001 for all parameters). Intergroup comparisons showed that the mean V-PPD reduction (2.13±1.35 mm), V-CAL gain (2.53±1.2 mm), and RDD fill (1.33±1.0 mm) were statistically significant between the groups at 12 months (p<0.001 for all parameters). Conclusion: BCP and EMDs combination is a promising modality for the regeneration of intrabony defects.

• Restorative Dentistry and Endodontics
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• v.31 no.6
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• pp.437-444
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• 2006

This study was aimed to elucidate the biological function of OD314 (Apin protein), which is related to ameloblast differentiation and amelogenesis. Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. We examined expression of the OD314 mRNA using in-situ hybridization during tooth development in mice. Expression of OD314 and several enamel matrix proteins were examined in the cultured ameloblast cell line up to 28 days by reverse transcription-polymerase chain reaction (RT-PCR) amplification. After inactivation and over-expression of the OD314 gene in ameloblast cell lines using U6 vectordriven RNA interference and CMV-OD314 construct, RT-PCR were performed to evaluate the effect of the OD314 during amelogenesis. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were more strongly expressed in ameloblast than odontoblast. 2. When ameloblast cells were cultured in the diffcrentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expressions of amelogenin and enamelin were gradually decreased according to the ameloblast differentiation. 3. Inactivation of OD314 by U6-OD314 siRNA construct down-regulated the expression of OD314, MMP-20, and tuftelin, whereas over-expression of OD314 by CMV-OD314 construct up-regulated the expression of OD314 and MMP-20 without change in tuftelin. These results suggest that OD314 is considered as an ameloblast-enriched gene and may play the important roles in ameloblast differentiation and mineralization.