• Journal of Periodontal and Implant Science
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• 제34권4호
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• pp.759-769
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• 2004

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the expression of extracellular matrix proteins in primary rat calvarial cells in Vitro. In the control group, cells was cultured with BGjb media. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Then each group was characterized by examining alkaline phosphatase activity at 3 and 7 days, and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. Synthesis of type I collagen (COL-I), osteocalcin (OCN), bone sialoprotein (BSP) was evaluated by RT-PCR at 14 days. The results were as follows: 1. Alkaline phosphatase activity was significantly higher in the concentration of chitosan 0.01mg/ml, 0.1mg/ml and 1.0mg/ml compared to control (p<0.05). 2. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1mg/ml and 1.0mg/ml than the control. 3. At 14 day culture, the expression of OCN was increased by chitosan in concentration of 1.0 mg/ml and 2.0 mg/ml. These results suggested that chitosan in concentration of 0.1 and 1,0 mg/ml stimulate the extracellular matrix of primary rat calvarial cells and may facilitate the formation of bone.

• 식물병연구
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• 제16권3호
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• pp.312-315
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• 2010

본 연구는 애기장대에서 무름병에 대한 저항성 유전자를 탐색하고자 2만여개의 T-DNA 삽입 돌연변이군을 이용하여 Pcc에 대한 스크리닝을 수행하고 이 방법을 소개한 연구다. 1차 선발을 통하여 15개의 저항성 line과 20개의 감수성 line을 선발하였으며, 이로부터 2차 선발하여 3개의 저항성 line과 4개의 감수성 line을 선발하였고, 최종적으로 3차 선발을 통하여 1개의 감수성 line (Atstp1)을 선발할 수 있었다. 현재 Atstp1을 이용해 flanking sequencing 하여 유전자를 탐색하고 있으며, 앞으로 클로닝을 통하여 다양한 무름병 저항성 식물 개발에 유용하게 이용될 것으로 기대한다.

• Biomolecules & Therapeutics
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• 제11권2호
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• pp.139-144
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• 2003

Ganoderma lucidum is commonly known as medically potent mushroom, which has been widely used in China and other oriental countries for the treatment of various diseases, including cancer. In this report, we investigated the anti-oxidant and protective effect of Ganodema lucidum extract (GLE) against the DNA damage induced by free radical and U.V. In the assay of cell growth inhibition, the inhibitory cell growth rate induced by hydroxyl radical was dose-dependently decreased by GLE. This results support that GLE has a detoxifying activity against cytotoxicity of hydroxyl radical in E. coli cell. GLE also protected ColE1 plasmid DNA damage in the concentration of 200$\mu\textrm{g}$ per reaction on the DNA fragmentation assay. The nuclear tailing by hydrogen peroxide in single cell gel electrophoresis(SCGE) was decreased by GLE in the concentration of 50$\mu\textrm{g}$/ml. These data indicate that Ganoderma lucidum has an anti-oxidative activity to hydrogen peroxide. The mutation rate after irradiation of U.V. was reduced by 50$\mu\textrm{g}$/ml GLE and total number of Rif (Rifampicin) resistant mutants was decreased in a concentration dependent manner when added the GLE exogenously in a culture media. According to the results, it is likely that GLE has not only an anti-oxidative activity to hydroxyl radical but also an anti-mutagenic activity to U.V. mutagenesis.

• 생명과학회지
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• 제33권7호
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• pp.555-564
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• 2023

본 연구에서는 이너뷰티용 소재로서의 활용가능성을 확인하기 위하여 생물전환 가시오가피-진피 혼합추출물(B-MEAC)의 항산화 활성, 엘라스타제 억제활성 및 피부보습 효과 등을 평가하였다. B-MEAC의 항산화 활성을 측정한 결과 DPPH radical을 농도의존적으로 소거하여 SC₅₀이 156.1±0.82 ㎍/ml로 나타났다. 또한, 엘라스타제 억제활성을 측정한 결과 B-MEAC 처리 농도 의존적으로 엘라스타제 활성을 억제하는 것으로 확인되었다(p<0.001). 경구투여한 B-MEAC가 마우스 피부 보습에 미치는 영향을 확인하기 위하여 피부 수분량과 경피수분손실량을 측정한 결과 건조유도 된 대조군에 비하여 피부 수분량은 증가하고(p<0.001), 경피수분손실량은 감소하는 것으로 나타났다(p<0.01). 또한, 건조유도 된 마우스 피부의 교원섬유 변화에 미치는 영향을 Masson's trichrome 염색을 통해 관찰한 결과 B-MEAC를 투여한 군에서 콜라겐의 양이 대조군에 비해 상대적으로 증가하여 푸른색으로 보다 진하게 염색되었다. 또한, 건조유도 된 마우스 피부의 보습 기능에 미치는 영향을 확인하기 위하여 보습 관련 인자들의 발현을 Western blot 기법으로 확인한 결과 B-MEAC를 투여한 군에서 대조군에 비하여 MMP-1 단백질의 발현량은 감소하였고, COL1A1, HAS2, filaggrin, AQP3 단백질 발현량은 회복되는 것으로 나타났다(p<0.001). 따라서 B-MEAC의 보습효과는 엘라스타제 활성 억제, MMP-1 발현 감소, 콜라겐 생성 촉진, 피부 보습 관련 단백질인 HAS2 및 AQP3 발현 증가, 피부 장벽기능의 중요인자인 filaggrin 발현 증가를 통한 수분량 증가와 TEWL 감소에 의해 나타나는 것으로 이너뷰티 소재로서의 개발 가능성을 제시한다.

• Clinics in Shoulder and Elbow
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• 제25권4호
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• pp.296-303
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• 2022

Background: A previous study reported that hyperlipidemia increases the incidence of tears in the rotator cuff tendon and affects healing after repair. The aim of our study was to compare the gene and protein expression of torn rotator cuff tendons in patients both with and without hypercholesterolemia. Methods: Thirty patients who provided rotator cuff tendon samples were classified into either a non-hypercholesterolemia group (n=19, serum total cholesterol [TC] <200 mg/dL) and hypercholesterolemia group (n=11, serum TC ≥240 mg/dL) based on their concentrations of serum TC. The expression of various genes of interest, including COL1A1, IGF1, IL-6, MMP2, MMP3, MMP9, MMP13, TNMD, and TP53, was analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, Western blot analysis was performed on the proteins encoded by interleukin (IL)-6 and TP53 that showed significantly different expression levels in real-time qRT-PCR. Results: Except for IGF1, the gene expression levels of IL-6, MMP2, MMP9, and TP53 were significantly higher in the hypercholesterolemic group than in the non-hypercholesterolemia group. Western blot analysis confirmed significantly higher protein levels of IL-6 and TP53 in the hypercholesterolemic group (p<0.05). Conclusions: We observed an increase in inflammatory cytokine and matrix metalloproteinase (MMP) levels in hypercholesterolemic patients with rotator cuff tears. Increased levels of IL-6 and TP53 were observed at both the mRNA and protein levels. We suggest that the overexpression of IL-6 and TP53 may be a specific feature in rotator cuff disease patients with hypercholesterolemia.

• International Journal of Oral Biology
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• 제39권3호
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• pp.145-151
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• 2014

During bone remodeling, there is requirement of differentiation of osteoblastic cells. Previously, we identified proteins differentially expressed in soft tissue during bone healing. Of these proteins, we focused the effect of LTF on differentiation of osteoblast. In order to analyze the osteogenic ability of LTF, we treated conditioned media collected from human LTF-stably transfected HEK293T cells into osteoblastic MC3T3-E1. The results showed that the activity and expression of alkaline phosphatase were increased in MC3T3-E1 cells treated with conditioned media containing LTF in dose- and time-dependent manner. At the same time, we observed the significant increase of the expression of osteoblastic genes, such as ALP, BSP, COL1A1, and OCN, and along with matrix mineralization genes, such as DMP1 and DMP2, in LTF conditioned media-treated groups. Moreover, the result of treating recombinant human LTF directly into osteoblastic MC3T3-E1 showed the same pattern of treating conditioned media containing LTF. Our study demonstrated that LTF constitutively enhances osteoblastic differentiation via induction of osteoblastic genes and activation of matrix mineralization in MC3T3-E1 cells.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.357-366
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• 2018

Periodontitis, an infective disease caused by oral pathogens and the intrinsic aging process, results in the destruction of periodontal tissues and the loss of alveolar bone. This study investigated whether Boesenbergia pandurata extract (BPE) standardized with panduratin A exerted anti-periodontitis effects, using an aging model representative of naturally occurring periodontitis. In aged rats, the oral administration of BPE ($200mg{\cdot}kg^{-1}{\cdot}day^{-1}$) for 8 weeks significantly reduced the mRNA and protein expression of $interleukin-1{\beta}$, nuclear factor-kappa B, matrix metalloproteinase (MMP)-2, and MMP-8 in gingival tissues (p < 0.01). In alveolar bone, histological analysis with staining and micro-computed tomography revealed the attenuation of alveolar bone resorption in the BPE-treated aged group, which led to a significant reduction in the mRNA and protein expression of nuclear factor of activated T-cells c1 (NFATc1), c-Fos, tartrate-resistant acid phosphatase, and cathepsin K (p < 0.01). BPE not only increased the expression of osteoblast differentiation markers, such as alkaline phosphate, and collagen type I (COL1A1), but also increased the ratio of osteoprotegerin to RANKL. Collectively, the results strongly suggested that BPE is a natural resource for the prevention or treatment of periodontal diseases.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.222-228
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• 2001

Vibrio metschnikovii strain RH530 is a non-pathogenic, industrially-important alkaline protease producer which has been isolated from wastewater. In this paper, we report on the transformation of this strain by using the method of electroporation. A field strength of $7.5\;kVcm^{-1}$ and $25\;{\mu}F$, and using a 0.2-cm cuvette, appeared to be the optimal conditions for electroporation of the cells with the recombinant pSBCm plasmid carrying the vapK alkaline protease gene and the ColE1 replicon. Cells were subjected to osmotic shock in order to remove extracelluar DNase, and adding 200 mM of sucrose to electroporation buffer cells showed an increased transformation efficiency. Maximum efficiency of transformation was obtained at an early exponential growth phase. Using all of the conditions mentioned above, we routinely obtained a transformation efficiency of more than $10^4{({\mu}g\;plasmid\;DNA)}^{-1}$. The stability of the plasmid pSBCm in V. metschnikovii RH530 was 25% after 18h of growth (27 generations) in the medium without antibiotic selection. The insertion of the par locus to the pSBCm increased the stability of the plasmid up to 42% without selective pressure. The increase in plasmid stability was accompanied by the increase in the productivity of alkaline protease in the recombinant V. metschnikovii strain RH530. Determining optimal conditions for the transformation of the industrially-important, nonpathogenic Vibrio strain, and the improvement of plasmid stability by introducing the par locus into the high copy number plasmid vector, will allow the development of procedures involved in the genetic manipulation of this strain, particularly for its use in the production of industrial enzymes such as alkaline protease.

• Journal of Korean Dental Science
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• 제9권1호
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• pp.9-18
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• 2016

Purpose: Wnt signaling plays an essential role in the dental epithelium and mesenchyme during tooth morphogenesis. Deletion of the Wntless (Wls) gene in odontoblasts appears to reduce canonical Wnt activity, leading to inhibition of odontoblast maturation. However, it remains unclear if autonomous Wnt ligands are necessary for differentiation of dental pulp cells into odontoblast-like cells to induce reparative dentinogenesis, one of well-known feature of pulp repair to form tertiary dentin. Materials and Methods: To analyze the autonomous role of Wls for differentiation of dental pulp cells into odontoblast-like cells, we used primary dental pulp cells from unerupted molars of Wls-floxed allele mouse after infection with adenovirus for Cre recombinase expression to knockout the floxed Wls gene or control GFP expression. The differentiation of dental pulp cells into odontoblast-like cells was analyzed by quantitative real-time polymerase chain reaction. Result: Proliferation rate was significantly decreased in dental pulp cells with Cre expression for Wls knockout. The expression levels of Osterix (Osx), runt-related transcription factor 2 (Runx2), and nuclear factor I-C (Nfic) were all significantly decreased by 0.3-fold, 0.2-fold, and 0.3-fold respectively in dental pulp cells with Wls knockout. In addition, the expression levels of Bsp, Col1a1, Opn, and Alpl were significantly decreased by 0.7-fold, 0.3-fold, 0.8-fold, and 0.6-fold respectively in dental pulp cells with Wls knockout. Conclusion: Wnt ligands produced autonomously are necessary for proper proliferation and odontoblastic differentiation of mouse dental pulp cells toward further tertiary dentinogenesis.

• 한국미생물·생명공학회지
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• 제26권5호
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• pp.393-399
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• 1998

고추역병균(Phytophthora capsici)의 성장을 in vitro에서 저해하는 길항균 Bl4를 한국토양으로부터 분리 동정하여 Enterobacter속임을 밝혔고 Pl::Tn5 lac에 의해 transposon돌연변이를 유기하여 길항력 강화주와 약화주들의 염색체내에 Tn5 lac이 각기 상이한 위치에 무작위로 삽입되었음을 southern hybridization에 의해 확인하였다.