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A STUDY OF APIN-PROTEIN INTERACTIONS USING PROTEIN MICROARRAY (Protein microarray를 이용한 APin-단백질의 상호작용에 관한 연구)

  • Park, Joo-Cheol;Park, Sun-Hwa;Kim, Heung-Joong;Park, Jong-Tae;Youn, Seong-Ho;Kim, Ji-Woong;Lee, Tae-Yeon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.32 no.5
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    • pp.459-468
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    • 2007
  • Protein microarray or protein chips is potentially powerful tools for analysis of protein-protein interactions. APin cDNA was previously identified and cloned from a rat odontoblast cDNA library. The purpose of this study was to investigate the APin-protein interactions during ameloblast differentiation. Protein microarray was carried with recombinant APin protein and MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein were selected among 74 interacting proteins. Immortalized ameloblast cells (ALCs) were transfected with pCMV-APin construct and U6-APin siRNA construct. After transfection, the expression of the mRNAs for four proteins selected by protein micoarrays were assessed by RT-PCR. The results were as follows: 1. APin expression was increased and decreased markedly after its over-expression and inactivation, respectively. 2. Over-expression of the APin in the ALCs markedly down-regulated the expression of MEF2 and Aurora kinase A, whereas their expression remained unchanged by its inactivation. 3. Expression of BMPR-IB and EF-hand calcium binding protein were markedly increased by the over-expression of the APin in the ALCs, whereas expression of BMPR-IB remained unchanged and expression of EF-hand calcium binding protein was markedly decreased by its inactivation. These results suggest that APin plays an important role in ameloblast differentiation and mineralization by regulating the expression of MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein.

Experiment Report about the Effect on the Anti-cancer of Herbal-acupuncture with Evodiae Fructus Infusion Solution (오수유(吳茱萸)약침의 항암효과에 대한 실험적 연구)

  • Cha, Kwan-Bae;Kim, Yoon-Sik;Yoo, Ho-Ryong;Jo, Hyun-Kyung;Oh, Young-Seon;Seol, In-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.5
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    • pp.1261-1270
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    • 2006
  • This study was done to investigate the effect of anti-cancer, anti-metastasis and immune response improvement of herbal-acupuncture with Evodiae Fructus herbal infusion solution(EF-HAS). To study the effects of anti-cancer, anti-metastasis and immune response improvement of EF-HAS, we injected EF-HAS into Chung-wan(CV12) of C57BL/6 mice implanted intravenously with Bl6-Fl0 melanoma. Then, we have examined the effect on the increasing of spleen cells, the effect on the number of CD25$^+$/CD4$^+$, CD8$^+$/CD3e$^+$, CD69$^+$/B220$^+$, NK1.1$^+$/CD3e$^+$ cells in mice's PBMCs, the effect on the pulmonary colony number, and the effect on median survival time(MST) and increase of life span(ILS) of C57BL/6 mice implanted intravenously with Bl6-Fl0 melanoma. The following results were obtained; in the experiment groups treated with EF-HAS, the spleen cell proliferation in C57BL/6 mice, the percentage of CD25$^+$/CD4$^+$, CD8$^+$/CD3e$^+$, CD69$^+$/B220$^+$, NK1.1$^+$/CD3e$^+$ cells in C57BL/6 mice's PBMCs, and MST and ILS of C57BL/6 mice implanted intravenously with Bl6-Fl0 melanoma were significantly increased compared with that of the control group. And in the experiment groups treated with EF-HAS, the pulmonary colony number of C57BL/6 mice implanted intravenously with Bl6-Fl0 melanoma was decreased significantly compared with that of the control group. These results support a role for EF-HAS might De usefully applied in treatment of cancer.

Effect of Evodiae fructus Methanol Extract on Virulence-Related Genes' Expression of Helicobacter pylori (오수유 메탄올 추출물이 Helicobacter pylori 병원성 관련 유전자 발현에 미치는 영향)

  • Yang, Ji Yeong;Lee, Pyeongjae;Kim, Jong-Bae
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.316-322
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    • 2019
  • Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

Distribution of Organic Matter and Trace Metals in Surface Sediments and Ecological Risk Assessment in the Tongyeong Coast (통영연안 표층퇴적물에서의 유기물과 미량금속 분포 특성 및 생태위해성 평가)

  • YANG, WON HO;LEE, HYO JIN;KIM, GI BEUM
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.21 no.4
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    • pp.125-133
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    • 2016
  • We investigated the distribution of organic matter and trace metals in surface sediment from Tonyeong coast. To determine the status of trace metal pollution, we also conducted an ecological risk assessment. Relatively high concentration of TN (total nitrogen), TOC (total organic carbon), and AVS (acid volatile sulfide) was found in surface sediment located in the narrow channel (site 35-38). Spatial distribution of Cd, Cr, Ni, Co, Hg, and Zn in surface sediment was similar and high Cu concentrations were found in narrow channel. The assessment of heavy metal pollution was derived using the Enrichment factors (EF). The enrichment factor indicated that Cd was no enrichment (EF<1), Pb, Cr, Ni, Co, Zn, and Hg were minor enrichment (1

Genetic Monitoring of Plant Growth Promoting Rhizobacterium (PGPR), Bacillus subtilis AH18 using Multiplex PCR in Field Soil (Multiplex PCR을 이용한 생물방제균 Bacillus subtilis AH18의 토양내 Genetic Monitoring)

  • Woo, Sang-Min;Lim, Jong-Hui;Jeong, Hee-Young;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.1-9
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    • 2009
  • The genetic monitoring method was developed for the rapid detection of the PGPR and biocontrol agent, B. subtilis AH18 in red-pepper field soil by multiplex PCR using sid, aec and cel gene primers. The monitoring of B. subtilis AH18 in the soil was carried by amplified a 2,3-dihydro-2,3-dihydroxy benzoate dehydrogenase [EC: 1. 3. 1. 28]gene (sid - 794 bp : EF408238) which is a key enzyme of siderophore synthesis, an auxin efflux carrier gene (aec - 1,052 bp : EF408239) and a cellulase gene (cel - 1,582 bp : EF070194). The natural un sterilized soil was inoculated with B. subtilis AH18 to determine the sensitivity ($1.8\times10^5$ cfu/g) of multiplex PCR for the rapid dectection and then the strain was monitored successfully in rhizosphere or non-rhizosphere soil of red-pepper cultural soil. At 3 weeks after the treatment, density of the strain was monitored more abundantly in rhizosphere soil.

Evaluation of indirect N2O Emission from Nitrogen Leaching in the Ground-water in Korea (우리나라 농경지에서 질소의 수계유출에 의한 아산화질소 간접배출량 평가)

  • Kim, Gun-Yeob;Jeong, Hyun-Cheol;Kim, Min-Kyeong;Roh, Kee-An;Lee, Deog-Bae;Kang, Kee-Kyung
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1232-1238
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    • 2011
  • This experiment was conducted to measure concentration of dissolved $N_2O$ in ground-water of 59 wells and to make emission factor for assessment of indirect $N_2O$ emission at agricultural sector in agricultural areas of Gyeongnam province from 2007 to 2010. Concentrations of dissolved $N_2O$ in ground-water of 59 wells were ranged trace to $196.6{\mu}g-N\;L^{-1}$. $N_2O$ concentrations were positively related with $NO_3$-N suggesting that denitrification was the principal reason of $N_2O$ production and $NO_3$-N concentration was the best predictor of indirect $N_2O$ emission. The ratio of dissolved $N_2O$-N to $NO_3$-N in ground-water was very important to make emission factor for assessment of indirect $N_2O$ emission at agricultural sector. The mean ratio of $N_2O$-N to $NO_3$-N was 0.0035. It was greatly lower than 0.015, the default value of currently using in the Intergovernmental Panel on Climate Change (IPCC) methodology for assessing indirect $N_2O$ emission in agro-ecosystems (IPCC, 1996). It means that the IPCC's present nitrogen indirect emission factor ($EF_{5-g}$, 0.015) and indirect $N_2O$ emission estimated with IPCC's emission factor are too high to use adopt in Korea. So we recommend 0.0034 as national specific emission factor ($EF_{5-g}$) for assessment of indirect $N_2O$ emission at agricultural sector. Using the estimated value of 0.0034 as the emission factor ($EF_{5-g}$) revised the indirect $N_2O$ emission from agricultural sector in Korea decreased from 1,801,576 ton ($CO_2$-eq) to 964,645 ton ($CO_2$-eq) in 2008. The results of this study suggest that the indirect Emission of nitrous oxide from upland recommend 0.0034 as national specific emission factor ($EF_{5-g}$) for assessment of indirect $N_2O$ emission at agricultural sector.

Screening of Antioxidative Effect and Suppressive Effect of LDL Oxidation of Euryale ferox Salisbury (Euryale ferox Salisbury의 항산화효과 및 LDL 산화 억제효과 탐색)

  • Kim, Young-Hwan;Lee, Min-Ja;Lee, Hye-Sook;Kim, Jung-Guk;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.92-99
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    • 2011
  • Topical natural antioxidants are a useful strategy for the prevention of oxidative stress mediated cardiovascular disease including atherosclerosis. From the viewpoint of this underlying principle, the screening of natural plant extracts with scavenging activity for pro-oxidant reactive species is a primary requirement for the development of new topical antioxidant formulations. Euryale ferox Salisbury (EF) is botanical name and it's pharmaceutical name is EURYALES SEMEN (ES). The stems and branchs of EF have been used as a traditional herbal medicine for the treatment of dysentery, diarrhea, leucorrhoea, incontinence and paralysis of joint. In this study, the antioxidant activity of extract from EF was studied in vitro methods by measuring the antioxidant activity and free radical scavenging activity by TEAC and DPPH, measuring the scavenging effects on reactive oxygen species (ROS) [superoxide anion, hydroxyl radical] and on reactive nitrogen species (RNS) [nitric oxide and peroxynitrite] as well as measuring the inhibitory effect on $Cu^{2+}$-induced human LDL oxidation. The EF extracts were found to have a potent scavenging activity, as well as an inhibitory effect on LDL oxidation. In conclusion, the EF extracts have antioxidative effects in vitro system, which can be used for developing pharmaceutical drug against oxidative stress and chronic degenerative disease such as atherosclerosis.

Important Features for Protein Foldings in Two Acyl Carrier Proteins from Enterococcus faecalis

  • Seoyeong Yoo;Jiwon Yeon;Eunhee Kim;Yangmee Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.1
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    • pp.10-16
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    • 2024
  • The emergence of multi-drug resistant Enterococcus faecalis raises a serious threat to global public health. E. faecalis is a gram-positive intestinal commensal bacterium found in humans. E. faecalis can endure extreme environments such as high temperature, pressure, and high salt, which facilitates them to cause infection in hospitals. E. faecalis has two acyl carrier proteins, AcpA (EfAcpA) in de novo fatty acid synthesis (FAS) and AcpB (EfAcpB) which utilizes exogenous fatty acids. Previously, we determined the tertiary structures of these two ACPs and investigated their structure-function relationships. Solution structures revealed that overall folding of these two ACPs is similar to those of other bacterial ACPs. However, circular dichroism (CD) experiments showed that the melting temperature of EfAcpA is 76.3℃ and that of EfAcpB is 79.2℃, which are much higher than those of other bacterial ACPs. In this study, to understand the origin of their structural stabilities, we verified the important residues for stable folding of these two ACPs by monitoring thermal and chemical denaturation. Hydrogen/deuterium exchange and chemical denaturation experiments on wild-type and mutant proteins revealed that Ile10 of EfAcpA and Ile14 of EfAcpB mediate compact intramolecular packing and promote high thermostability and stable folding. E. faecalis may maximize efficiency of FAS and increase adaptability to the environmental stress by having two thermostable ACPs. This study may provide insight into bacterial adaptability and development of antibiotics against multi-drug-resistant E. faecalis.

Expression of Human KCNE1 Gene in Zebrafish (Zebrafish에서 인간 KCNE1 유전자 발현에 관한 연구)

  • Park, Hyeon Jeong;Yoo, Min
    • Journal of Life Science
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    • v.27 no.5
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    • pp.524-529
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    • 2017
  • This study was aimed to produce a transgenic zebrafish expressing the human KCNE1 gene. Initially, the entire CDS of the human KCNE1 gene was amplified from a human genomic DNA sample by polymerase chain reaction using a primer set engineered with restriction enzyme sites (EcoRI, BamHI) at the 5' end of each primer. The resultant 402 bp KCNE1 amplicon flanked by EcoR1 and BamH1 was obtained and subsequently cloned into a plasmid vector pPB-CMVp-EF1-GreenPuro. The integrity of the cloned CDS sequence was confirmed by DNA sequencing analysis. Next, the recombinant vector containing the human KCNE1 (pPB-CMVp-hKCNE1-EF1-GreenPuro) was introduced into fertilized eggs of zebrafish by microinjection. Successful expression of the recombinant vector in the eggs was confirmed by the expression of the fluorescence protein encoded in the vector. Finally, in order to assure that the stable expression of the human KCNE1 gene occurred in the transgenic animal, RNAs were extracted from the animal and the presence of KCNE1 transcripts was confirmed by RT-PCT as well as DNA sequencing analysis. The study provides a methodology to construct a useful transgenic animal model applicable to the development of diagnostic technologies for gene therapy of LQTS (Long QT Syndrome) as well as tools for cloning of useful genes in fish.

A Study on Shear Capacity and Behavior of Large Sized Concrete Anchorage System (대형 콘크리트 앵커시스템의 전단성능 및 거동특성에 관한 연구)

  • Kim, Kang Sik;Shin, Sung Woo;Lee, Kwang Soo
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.15 no.5
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    • pp.82-91
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    • 2011
  • In this study, 24 prototype specimens were tested to find out the shear behavior and strength of large anchorage system exceeding 50mm(2") in anchor bolt diameter($d_0$) and 635mm(25") in effective embedment depth($h_{ef}$) not addressed by ACI349-06 Appendix B. Test variables are anchor bolt diameter($d_0$ = 63.5, 76.2, 88.9mm), effective embedment depth($h_{ef}$=635, 762mm), and edge distance($c_1$=381, 508, 762mm). Concrete compressive strength is constant($f_{ck}$=38MPa). Test results ($V_{test}$) were overestimated by $V_{aci06}$(shear strength by ACI 349-06) and $V_{ccd}$(shear strength by CCD method). In large anchorage system exceeding 50mm(2") of anchor bolt diameter($d_0$) and 635mm(25") of anchor bolt effective embedment depth($h_{ef}$), the bolt diameter variation and effective embedment depth($h_{ef}$) has no influence on the shear strenth, But, according to the analysis results of the feature ratio on edge distance($c_1$) and anchor bolt diameter, the feature ratio become smaller, which means anchor bolt diameter is bigger, predicted ratio of test results and predicted equation is larger. It was found that anchor bolt diameter is immediate cause of deterioration in the shear capacity of large anchorage system. To improve and extend the validity of current design recommendations further theoretical and numerical work is needed.