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Synthesis and Evaluation of F-18 Labeled 2'-Deoxy-2'-fluoro-5-methyl-1-β-L-arabinofuranosyluracil (L-[18F]FMAU)

  • Jo, Nam-Hyun;Moon, Byung-Seok;Hong, Su-Hee;An, Gwang-Il;Choi, Tae-Hyun;Cheon, Gi-Jeong;Cho, Jung-Hyuck;Yoo, Kyung-Ho;Lee, Kyo-Chul;Oh, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.28 no.12
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    • pp.2449-2453
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    • 2007
  • L-[18F]FMAU ([18F]1b) was prepared from the precursor 2-O-[(trifluoromethyl)-sulfonyl]-1,3,5-tri-Obenzoyl- α-L-ribofuranose, by coupling the radioactive fluoro-sugar with the corresponding silylated thymine in 4 steps. The final products, including the α and β anomers, were purified using reverse phase HPLC with an appropriate solvent (5% CH3CN/H2O) at a flow rate of 3.0 mL/min. The total elapsed time of synthesis was about 180-200 min from EOB. The α/β anomeric ratio of the compounds was about 1:9, and the radiochemical purity of the product (β-form) was >98% with decay-corrected yields of 25-35%. All radioactive samples were confirmed using co-injection with pure non-radioactive analogues in every step. In the cellular uptake in vitro test of herpes simplex virus-thymidine kinase (HSV1-TK) gene expressed cells, the percent uptake of injected dose (%ID) of L- and D-FMAU was 37.28 and 65.86, respectively after 240 min incubation. However, the relative uptake (MCA-TK/MCA cellular uptake ratio) of L-FMAU was higher than that of D-FMAU (%ID of L-FMAU, 0.36 and D-FMAU, 0.93 after 240 min incubation in MCA cells). This means that L-FMAU will show better specific HSV1-TK gene expressed cell uptake for selective HSV1-TK gene imaging.

REMINERALIZATION EFFECTS BY APF GEL AND CPP-ACP PASTE ON INCIPIENT ARTIFICIAL CARIES OF ENAMEL (법랑질 인공우식병소에서 APF gel과 CPP-ACP 제제의 재광화 효과)

  • Kang, Ji-Sun;Lee, Nan-Young;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.4
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    • pp.684-691
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    • 2008
  • Many operations have been carried out using the fluoride ion to prevent and reverse dental caries. It certainly encourages remineralization and also prevents dental caries. However, the remineralization developed by these means is superficial only leaving lower levels of demineralized lesion with a degree of porosity and incomplete mineralization. We must consider its toxic effects when it is ingested for overdose. The CPP-ACP paste is able to remineralize the incipient lesion and has no harmful effects when it is ingested, because it was made form casein which is from the protein of milk. The purpose of this article is to compare the remineralization effects between APF gel and the CPP-ACP paste. we applicated the APF gel and CPP-ACP paste on the artificial enamel carious lesion. After 14 days, we measured the surface microhardness and observed the remineralized lesion under polarized light microscope. The results were as follows : 1. The surface microhardness of group III was the highest, followed by group II, and I(p<0.05). 2. The surface microhardness of group III was significantly higher than those of group I, and II(p<0.05). 3. We could observe thin and irregular remineralization layer of group II, and regular and moderate remineralization layer of group III under polirized light microscope.

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Expression of Nucleocapsid Protein Gene of Maaji Virus and Use of the Protein as an Immunodiagnostic Antigen of Hemorrhagic Fever with Renal Syndrome (마지바이러스 Nucleocapsid Protein 유전자의 발현과 신증후 출혈열 진단용 항원으로의 이용)

  • Lee, Pyung-Woo;Kim, Yun-Cheol;Paik, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.77-90
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    • 1996
  • Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

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INFLUENCE OF DIETARY PROTEIN ON THE FRACTIONATION OF SELENIUM IN THE RUMEN OF SHEEP

  • Serra, A.B.;Serra, S.D.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.5
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    • pp.557-562
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    • 1996
  • The effects of diets differing in protein content through soybean meal supplementation on ruminal fractionation of selenium (Se) were studied. A $3{\times}3$ Latin square design was used with three Japanese Corriedale wethers (45 kg average body weight), three periods, and three dietary treatment. The three dietary treatments were : Diet 1, without soybean meal supplementation (14% crude protein, CP); Diet 2, with 10% soybean meal supplementation (16.5% CP); and Diet 3, with 20% soybean meal supplementation (19% CP). All the diets had a Se supplementation in the form of sodium selenite at 0.2 mg Se/kg dietary DM. The Se supplement and the concentrate mixture were fed only in the morning before the hay was given. Daily feeding schedule for gay was set at 09:00 and 17:00 h. On the final day of collection period, ruminal fluid samples were obtained at 0.5, 2, 6, 12 and 24 h post-feeding starting at 09:00 h. Total ruminal fluid Se was markedly higher (p<0.05) in Diet 3 than those in Diets 1 and 2 at almost all sampling time except at 24 h. The proportion of Se in soluble protein to the total ruminal Se was higher (p< 0.05) in Diet 3 (40%) followed by Diet 2 (28%) and Diet 1 (21%). The proportion of free inorganic Se to the total ruminal Se was the reverse, especially after two hours where Diet 1 (p<0.05) was higher than the other diets. Bacterial Se was lower (p < 0.05) in Diet 1 than those in Diets 2 and 3 at any sampling time. The highest was observed at 2 h postprandially in all diets with a value of 421, 556, $655{\mu}g/kg$ bacterial DM for Diet 1, 2 and 3, respectively. No differences (p>0.05) were observed on ruminal pH, ammonia and total nolatile fatty acids although increasing protein supplementation tended to decline the ruminal pH and increase ruminal ammonia. This study concludes that increasing dietary protein content by soybean meal supplementation can affect the ruminal Se metabolism.

Manikin Model Study on Reproducibility and Accuracy of Maxillofacial Measurements Determined by Stereocamera: Comparative Study of Direct Anthropometry, Digitizer and Stereophotogrammetery (스테레오 카메라를 이용한 안면부 측정의 재현성과 정확도에 대한 마네킨을 이용한 연구: 직접 인체계측, Digitizer, Stereophotogrammetry의 비교 연구)

  • Jeoung, Youn-Wook;Yang, Ji-Woong;Chung, Kwang;Kook, Min-Suk;Oh, Hee-Kyun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.34 no.1
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    • pp.17-25
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    • 2012
  • Purpose: Recently, a three dimensional approach to hard and soft tissues of the maxillofacial area has been widely used. This study was to evaluate the reproducibility and accuracy of a stereocamera compared to actual measurement methods using a digital caliper and digitizer. Methods: The stereoscopies of 7 head dummies with different sizes and shapes were obtained using a Di3D system (Dimensional Imaging, Glasgow, UK) after marking reference points on facial areas. From the obtained stereoscopy, 10 measurements representing the width, height and depth of each of the facial sections of the dummy were measured twice using a three dimensional reverse engineering software program (RapidForm$^{TM}$ 2006, Inus, Seoul, Korea). The x, y, and z coordinates of each of the three dimensional measurements were obtained and distances between two points were calculated. All procedures were repeated twice. The actual measurement method was performed twice, directly on dummies, using a digital caliper and values were compared with the previously determined values. Results: The results were as follows. In the ANOVA analysis, there were no significant statistical differences among the three measurement methods. In the Bonferroni analysis, with adjustments applied for multiple comparisons, there was no difference between actual measurement methods using a digitizer and a digital caliper. However, there was some difference between using a stereocamera and actual measurement methods using a digitizer and a digital caliper in values of $Ex_{Rt}-Ex_{Lt}$, $En_{Rt}-En_{Lt}$, $Ala_{Rt}-Ala_{Lt}$, $Ch_{Rt}-Ch_{Lt}$, G-Pg', $Ala_{Rt}$-Prn, $Ala_{Rt}$-Prn. The mean value for technical error in measurement (TEM) in Di3D (0.98 mm) was slightly higher than for a digital caliper (0.17 mm) and a digitizer (0.30 mm). In an intraclass correlation coefficient (ICC) there were no significant differences among the three measurement methods, but the Di3D system with the stereocamera showed relatively lower reproducibility compared to actual measurement methods using a digitizer and a digital caliper. Conclusion: These results indicate that some complementary measures may be needed to improve accuracy and reproducibility in the Di3D system with stereocamera.

A Study on the Growth intention of Korean and Chinese Co-Founding Startup Companies (한국인과 중국인의 공동창업기업 성장의도에 대한 연구)

  • Kim, A-Hyun;Moon, Jun Hwan;Lee, Jae Bum
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.15 no.3
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    • pp.145-158
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    • 2020
  • Because individuals come together to form a society, society has characteristics that are different or similar. Furthermore, as globalization and language acquisition in other countries have been activated, the frequency of studying abroad has increased. While Korea also studies abroad, the number of students coming to Korea from other countries continues to increase, increasing. In particular, when there was interest in start-up businesses, the factors were discovered through exploratory research in order to identify factors affecting the level of growth in start-ups when they had different nationalities. In order to conduct exploratory research, the government wanted to check more in-depth information through semi-structured interviews with co-founder companies composed of Koreans and Chinese. The main keywords were repeated or emphasized continuously during the interview, and other keywords were obtained through additional questions. As a result, it has been confirmed that self-acceptance, cultural distance, entrepreneurship, knowledge quality and growth are very large keywords in the co-founding start-up of different countries. The proposition was established as having a relationship of justice with self-acceptance, cultural distance and entrepreneurship as independent variables and with the degree of growth as dependent variables. In particular, in the case of co-founding with different nationalities, the most important knowledge quality was represented the effect of reverse U in each relationship (the relationship between self-acceptance, cultural distance, entrepreneurship and growth).

Induction of Apoptosis by Samgibopae-tang in Human Non-small-cell Lung Cancer Cells (인체폐암세포 NCI-H460 및 A549의 apoptosis 유발에 미치는 삼기보배탕의 영향)

  • Heo, Man-Kyu;Heo, Tae-Yool;Kim, Ki-Tak;Byun, Mi-Kwon;Kim, Jin-Young;Sim, Sung-Heum;Kim, Koang-Lock;Kam, Cheol-Woo;Park, Dong-Il
    • The Journal of Internal Korean Medicine
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    • v.28 no.3
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    • pp.473-491
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    • 2007
  • Objectives : This study was designed to investigate the antiproliferative activity of the water extract of Samgibopae-tang (SGBPT) in NCI-H460 and A549 non-small-cell lung cancer cell lines Methods : In this study, we measured the subsistence, form of NCI-H460 and A549 non-small-cell lung cancer cell by hemocytometer and DAPI staining. In each cell, we analyzed DNA fragmentation. reverse transcription-polymerase chain reaction and measured activity of caspase-3, caspase-8 and caspase-9. Results and Conclusions : We found that exposure of A549 cells to SGBPT resulted in growth inhibition in a dose-dependent manner. butSGBPT did not affect the growth of NCI-H460 cells. The antiproliferative effect by SGBPT treatment in A549 cells was associated with morphological changes. SGBPT treatment partially induced the expression of DR5 cells and the expression of Faswas markedly increased in both transcriptional and translational levels in A549 cells. SGBPT treatment partially induced the expression of Bcl-2, Bcl-XL and the expression of Bid was markedly decreased in translational levels in A549 cells. However, SGBPT treatment did not affect the expression of IAP family in A549 orNCI-H460 cells. SGBPT treatment partially induced the expression of caspase-3, caspase-8, caspase-9 activity which markedly increased in a dose-dependent manners in A549 cells. The fragmental development of PARP and ${\beta}$-catenin protein was observed in A549 cells by SGBPT treatment. SGBPT treatment induced the expression of PLC-${\gamma}1$ protein which decreased in A549 cells. SGBPT treatment partially induced the expression of DFF45/ICAD which markedly increased in a dose-dependent manner in A549 cells. Taken together. these findings suggested that SGBPT-induced inhibition of human lung carcinoma did not affect NCI-H460 cell growth. However, SGBPT-induced inhibition of human lung carcinoma A549 cell growth was associated with the induction of death receptor and mitochondrial pathway. The results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of SGBPT.

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DEVELOPMENT OF MICROPOROUS CALCIUM PHOSPHATE COATED NERVE CONDUIT FOR PERIPHERAL NERVE REPAIR (말초신경 재건을 위한 인회석 박막 코팅 미세공성 신경재생관(nerve conduit)의 개발)

  • Lee, Jong-Ho;Hwang, Soon-Jeong;Choi, Won-Jae;Kim, Soung-Min;Kim, Nam-Yeol;Lee, Eun-Jin;Ahn, Kang-Min;Myung, Hoon;Seo, Byoung-Moo;Choi, Jin-Young;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Hyun-Man
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.3
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    • pp.151-156
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    • 2003
  • This study was performed to develop a useful nerve conduit which provides favorable environment for Schwann cell viability and proliferation. Milipore membrane of $0.45{\mu}m$ pore size was selected because it permits nutritional inflow from the outside of the conduit and prevents from invading the fibrotic tissue into the conduit. The membrane was rolled and sealed to form a conduit of 2mm diameter and 20mm length. To improve the axonal regeneration and to render better environment for endogenous and exogenous Schwann cell behaviour, the microgeometry and surface of conduit was modified by coating with thin film of calcium phosphate. Cellular viability within the conduit and attachment to its wall were assessed with MTT assay and SEM study. Milipore filter conduit showed significantly higher rate of Schwann cell attachment and viability than the culture dish. However, the reverse was true in case of fibroblast. Coating with thin film of low crystalline calcium phosphate made more favorable environment for both cells with minimal change of pore size. These findings means the porous calcium phosphate coated milipore nerve conduit can provide much favorable environment for endogenous Schwann cell proliferation and exogenous ones, which are filled within the conduit for the more advanced strategy of peripheral nerve regeneration, with potential of reducing fibrotic tissue production.

Validated HPLC Method for the Pharmacokinetic Study of Atenolol and Chlorthalidone Combination Therapy in Korean Subjects

  • Kang, Hyun-Ah;Kim, Hwan-Ho;Kim, Se-Mi;Yoon, Hwa;Cho, Hea-Young;Oh, Seaung-Youl;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.5
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    • pp.331-338
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    • 2006
  • A rapid, selective and sensitive reverse-phase HPLC methods for the determination of atenolol and chlorthalidone in human serum and whole blood were validated, and applied to the pharmacokinetic study of atenolol and chlorthalidone combination therapy. Atenolol and an internal standard, pindolol, were extracted from human serum by liquid-liquid extraction, and analyzed on a $\mu$-Bondapak C18 $10-{\mu}$ column in a mobile phase of methanol-0.01 M potassium dihydrogenphosphate(30:70, v/v, adjusted to pH 3.5) and fluorescence detection(emission: 300 nm, excitation: 224 nm). Chlorthalidone and an internal standard, probenecid, were extracted form human whole blood by liquid-liquid extraction, and analyzed on a Luna C18 $5-{\mu}$ column in a mobile phase of acetonitrile containing 77% 0.01 M sodium acetate and UV detection at 214 nm. These analysis were performed at three different laboratories using the same quality control(QC) samples. The chromatograms showed good resolution, sensitivity, and no interference by human serum and whole blood, respectively. The methods showed linear responses over a concentration range of 10-1,000 ng/mL for atenolol and 0.05-20 ${\mu}g/mL$ for chlorthalidone, with correlation coefficients of greater than 0.999 at all the three laboratories. Intra- and inter-day assay precision and accuracy fulfilled international requirements. Stability studies(freeze-thaw, short-, long-term, extracted sample and stock solution) showed that atenolol and chlorthalidone were stable. The lower limit of quantitation of atenolol and chlorthalidone were 10 ng/mL and 0.05 ${\mu}g/mL$, respectively, which was sensitive enough for pharmacokinetic studies. These methods were applied to the pharmacokinetic study of atenolol and chlorthalidone in human volunteers following a single oral administration of Hyundai $Tenoretic^{\circledR}$ tablet(atenolol 50 mg and chlorthalidone 12.5 mg) at three different laboratories.

Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells

  • Kim, Seo Hwa;Baek, Moon Seong;Yoon, Dong Sik;Park, Jong Seol;Yoon, Byoung Wook;Oh, Byoung Su;Park, Jinkyeong;Kim, Hui Jung
    • Tuberculosis and Respiratory Diseases
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    • v.77 no.2
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    • pp.73-80
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    • 2014
  • Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.