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Control of Trophoblast Gene Expression and Cell Differentiation

  • Cheon, Jong-Yun
    • 대한생식의학회:학술대회논문집
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    • 2001.03a
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    • pp.195-205
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    • 2001
  • 태반 영양배엽 (trophoblast)은 포유동물의 발생과정 중 가장 먼저 분화되는 세포로서, 자궁환경내에서 배아가 착상, 발생, 및 분화하기 위해서 반드시 필요한 태반을 형성하는 색심적인 세포이다. 영양배엽 세포의 분화과정중의 결함은 배아의 사산이나 임신질환 등의 치명적 결과를 초래한다. 하지만, 영양배엽 세포의 분화를 조절하는 분자생물학적인 메카니즘은 아직 규명되지 않고 있다. 영양배엽 세포의 분화를 조절하는 경로를 규경하기 위한 선결과제는 분화된 영양배엽 세포에서만 발현하는 많은 유전자들이 밝혀져야만 한다. 본 연구팀은 최근에 분화된 영양배엽 세포에서만 발현하는 두 종류의 새로운 유전자들을 찾았다. 한 종류는 homeobox를 보유하고 있는 조절 유전자 Psx이고, 다른 한 종류는 임신호르몬인 태반 프로락틴 라이크 단백질 유전자 PLP-C${\beta}$이다. 본 연구과제의 목표는 이들 유전자의 기능과 조절 메카니즘을 규명함으로써, 영양배엽 세포의 분화를 조절하는 조절경로를 밝히는 것이다. 이를 위하여 다음과 같은 일련의 연구를 수행할 것이다. 1) Psx 유전자가 분화된 영양배엽 세포에서만 발현케 하는 조절 메카니즘을 규명하기 위해 functional assays, in vitro footprinting, gel mobility shift assays, 생쥐형질전화, UV crosslinking, Southwestern blot 등의 방법을 통해 Psx 유전자의 cis-acting 요인과 trans-acting factor를 밝혀 분석한다. 2) 영양배엽 세포의 분화조절 경로를 규명하기 위해 random oligonuclotide library screening, DD-PCR, subtractive screening 등의 방법을 이용하여 Psx 유전자에 의해 조절되는 하부유전자를 밝힌다. 3) Psx 유전자를 knock-out시켜 영양배엽 세포가 발달 및 분화하는데 미치는 역할을 밝힌다. 4) Yeast two-hybrid screening방법을 이용하여 태반 프로락틴 유전자의 수용체를 찾아 이들의 신호전달 기전을 밝힌다. 제1차년 연구결과로서, mouse와 rat으로부터 각각 Psx 유전자의 genomic DNA를 클로닝하여, 유전자 구조를 비교한 결과, mouse Psx (mPsx2)는 4개의 exons으로 이루어져 있는 반면에, rat Psx (Psx3)는 3개의 exons으로 구성되어 있었다. 즉, rPsx3는 mPsx2의 exon1이 없었다. Notrhern blot과 in situ hybridization 분석에 의해 mouse와 rat에서 Psx 유전자가 다르게 발현 조절되는 현상을 밝혔다. 실제로 mPsx2와 rPsx3의 5'-flanking지역을 클로닝하여 염기서열 분석 결과 전혀 homology를 찾을 수 없었다. 또한, 이들 각각 promoter의 activity를 luciferase reporter를 이용하여 조사한 결과 Rcho-1 trophoblast cells에서 각기 다른 activity를 보여 주는 것을 발견하였다. Psx 유전자의 transcription start sites는 Primer extension에 의해 밝혔다. 또한 Psx2 유전자를 knock-out 시키기 위해 targeting vector를 Osdupde1에 제작하였다. 본 과제를 시작할 때 새로운 프로락틴 유전자 하나를 클로닝하여 이 유전자를 PLP-I라고 이름을 붙였다. 이 후 이 유전자 (PLP-I)는 PLP-C${\beta}$라고 이름을 붙이게 되었다. Mouse PLP-C${\beta}$ 유전자의 counterpart를 rat에서 찾아 염기서열을 비교한 결과 mouse와 rat에서 PLP-C${\beta}$유전자의 homology는 약 79% (amino acid level)였다. 본 연구과정을 통해 또 하나의 새로운 PLP-C subfamily member를 mouse로부터 클로닝 하였고, 이 유전자를 PLP-C${\gamma}$라 하였다. PLP-C${\beta}$와 PLP-C${\gamma}$의 발현 유형은 Northern blot과 in 냐셔 hybridization 분석에 의해 태반의 제한된 spongitrophoblast와 trophoblast giant cells에서만 발현하는 것을 밝혔다. 놀랍게도 이들 두 새로운 유전자는 alternative splicing에 의해 두 종류의 isoform이 있음을 밝혔다. PLP family member 유전자로서 splicing에 의한 isoforms을 보여 주는 유전자로는 PLP-C${\beta}$와 PLP-C${\gamma}$가 최초이다. 이들 isoform mRNAs의 발현 유형은 RT-PCR 방법을 이용하여 규명하였다. 또 하나의 새로운 발견은 PLP-C${\beta}$와 PLP-C${\gamma}$가 독특한 유전자 구조를 갖고 있었다. 즉, PLP-C${\beta}$는 exon3의 alternative splicing에 의해 5개 혹은 6개의 exons을 갖는 two isoforms이 생긴다. 반면에 PLP-C${\gamma}$는 exon2가 alternative splcing이 되면서 7개의 exons을 갖거나 6개의 exons을 갖는 isoforms을 만든다. 그리고, PLP-C${\gamma}$의 promoter activity를 trophoblast Rcho-l${\gamma}$ 세포주를 이용하여 PLP-C${\gamma}$ 의 1.5 kb 5'-flanking 지역이 trophoblast-specific promoter activity를 갖고 있음을 밝혔다. PLP-C${\gamma}$ 유전자의 transcription start site는 Primer extension에 의해 밝혔다. 제 1차 년도의 연구결과를 토대로, 2차년에서는 다음단계의 연구를 수행하고자 한다. 즉, 1) mPsx2와 rPsx3의 promoter를 비교분석 함으로서 mouse와 rat에서 Psx 유전자가 다르게 조절되는 메카니즘 규명, 2) Psx와 PLP-C 유전자의 promoter에 있는 cis-acting elements 탐색, 3) Psx2와 Psx3의 단백질을 이용하여 이들이 binding하는 target sequence 규명, 4) 제작한 Psx2 targeting vector를 이용하여 ES cells에서 Psx2 유전자 knock-out, 5) Psx 유전자를 과발현시키는 세포주를 만들고 Psx에 의해 조절되는 유전자 탐색, 6) 새로 밝히 PLP-C members 유전자들의 조절기전을 Rcho-1 세포주를 이용하여 여러 거지 성장인자와 다른 호르몬에 대한 반응을 탐색, 7) Psx와 PLP-C${\gamma}$ 유전자의 chromosomal mapping 등을 밝힐 것이다.

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GnRH Antagonist Versus Agonist Flare-up Protocol in Ovarian Stimulation of Poor Responder Patients (저 반응군의 체외수정시술을 위한 과배란유도에 있어 GnRH Antagonist 요법과 GnRH Agonist Flare Up 요법의 효용성에 관한 연구)

  • Ahn, Young-Sun;Yeun, Myung-Jin;Cho, Yun-Jin;Kim, Min-Ji;Kang, Inn-Soo;Koong, Mi-Kyoung;Kim, Jin-Yeong;Yang, Kwang-Moon;Park, Chan-Woo;Kim, Hye-Ok;Cha, Sun-Hwa;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.125-131
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    • 2007
  • Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.

Antioxidant Activity of Rhododendron brachycarpum D. Don Extracts and Its Skin Hydration Effect Measure (만병초 추출물의 항산화활성과 보습효과 측정)

  • Park, Jung-Ok;Lim, Gyu-Nam;Park, Su-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.157-165
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    • 2010
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase and elastase of Rhododendron brachycarpum D. Don extracts were investigated. And the moisturizing effect of cream containing R. brachycarpum D. Don extract were investigated by clinical trial. The ethyl acetate fraction of R. brachycarpum D. Don extract (1.83 ${\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of R. brachycarpum D. Don extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The 50 % extract fraction (0.064 ${\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of R. brachycarpum D. Don on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The R. brachycarpum D. Don extracts suppressed photohemolysis in a concentration dependent manner (1 ~ 10 ${\mu}g/mL$). The inhibitory effects ($IC_{50}$) of R. brachycarpum D. Don extracts on tyrosinase were determined with ethyl acetate fraction of R. brachycarpum D. Don extract (70.5 ${\mu}g/mL$) and aglycone fraction of extract (122.40 ${\mu}g/mL$). The inhibitory effects ($IC_{50}$) on elastase were determined with ethyl acetate of R. brachycarpum D. Don extract (43.50 ${\mu}g/mL$) and aglycone fraction of extract (20.73 ${\mu}g/mL$). The cream containing the ethyl acetate fraction of R. brachycarpum D. Don extracts was formulated for skin hydration effect and transepidermal water loss (TEWL). The cream containing R. brachycarpum D. Don extract was applied to the right lower arm. After 180 min, the water contents in skin were increased by 1 ~ 4 % than the placebo cream. And TEWL of parts was decreased as 7.7 $g/m^2h$ (experimental cream) and 8.9 $g/m^2h$ (placebo cream) respectively. These results indicate that extract/fractions of R. brachycarpum D. Don can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And inhibitory activity on tyrosinase of the aglycone fraction could be applicable to new functional cosmetics for whitening and anti-wrinkle products. Also the increase of skin hydration of the cream containing extract could be applicable to new functional cosmetics for antiaging.

A Clinical Study of Hoarseness (애성을 주소로 한 후두질환의 임상 통계적 고찰)

  • 윤완규;조규모;송기준;김홍수
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1983.05a
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    • pp.6.4-7
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    • 1983
  • Hoarseness is the most common and early symptom in laryngeal diseases. A clinico - statistical analysis was performed on 228 cases with chief complaint of hoarseness at the department of otolaryngology, Jeonbug National University during the past 3 years from January 1980 to December 1980. 1) The number of the patient with hoarseness were 228 cases (2.3 %) among total outpatient of 10110 cases. 2) Among the 228 cases with hoarseness, male were 115 cases and female were 113 cases, so sex ratio was nearly same. 3) The underlying diseases causing hoarseness in order of frequency were acute laryngitis 43 cases (18.9 %), chronic laryngitis 36 cases (15.8 %), vocal nodule 30 cases(13.2 %), vocal polyp 30 cases (13.2 %), vocal cord paralysis 26 cases (11.4 %), laryngeal carcinoma 18 cases (7.9 %) and laryngeal tuberculosis 15 cases (6.6 %). There were other diseases of larynx in 30 cases (13.2 %). 4) The incidence of age distribution in order of frequency were 4th decade (26.8 % ), 3rd decade (18.9%), 5th decade (17.1 %), 2nd decade (15.8 %) and 6th decade (9.6 %). 5) The duration from onset to consultation in order of frequency were 11 days-1 month (22.8 %), 1 month-3 months(19.7%), 3 months -6 months (18.9 %), within 10 days (13.6 %), 6 months-l year (13.2 % ), 1 year -3 years(7. 9 %) and 3 years over. 6) The duration from onset to consultation for underlying diseases were as follows ; acute laryngitis was within 10 days, chronic laryngitis was 3 months - 6 months, vocal nodule was 1 month-3 months, vocal polyp was 3 months -1 year, vocal cord paralysis was 11 days -1 month. 7) Associated symptoms with hoarseness in order of frequency were sore throat (25 %), sputum (8.8 %), swallowing difficulty (7.9 %) and dyspnea (6.1 %). But 84 cases (36.8 %), there were no other symptoms except hoarseness.

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Detection with a SWNT Gas Sensor and Diffusion of SF6 Decomposition Products by Corona Discharges (탄소나노튜브 가스센서의 SF6 분해생성물 검출 및 확산현상에 관한 연구)

  • Lee, J.C.;Jung, S.H.;Baik, S.H.
    • Journal of the Korean Vacuum Society
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    • v.18 no.1
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    • pp.66-72
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    • 2009
  • The detection methods are required to monitor and diagnose the abnormality on the insulation condition inside a gas-insulated switchgear (GIS). Due to a good sensitivity to the products decomposed by partial discharges (PDs) in $SF_6$ gas, the development of a SWNT gas sensor is actively in progress. However, a few numerical studies on the diffusion mechanism of the $SF_6$ decomposition products by PD have been reported. In this study, we modeled $SF_6$ decomposition process in a chamber by calculating temperature, pressure and concentration of the decomposition products by using a commercial CFD program in conjunction with experimental data. It was assumed that the mass production rate and the generation temperature of the decomposition products were $5.04{\times}10^{-10}$ [g/s] and over 773 K respectively. To calculate the concentration equation, the Schmidt number was specified to get the diffusion coefficient functioned by viscosity and density of $SF_6$ gas instead rather than setting it directly. The results showed that the drive potential is governed mainly by the gradient of the decomposition concentration. A lower concentration of the decomposition products was observed as the sensors were placed more away from the discharge region. Also, the concentration increased by increasing the discharge time. By installing multiple sensors the location of PD is expected to be identified by monitoring the response time of the sensors, and the information should be very useful for the diagnosis and maintenance of GIS.

Quality Properties of Capsule Type Meju Prepared with Aspergillus oryzae (Aspergillus oryzae를 이용한 캡슐형 메주의 품질특성)

  • 최재훈;권선화;이상원;남상해;최상도;박석규
    • Food Science and Preservation
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    • v.10 no.3
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    • pp.339-346
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    • 2003
  • In order to improve some problems such as contamination of undesirable mold, mycotoxin production and excessive drying on the surface of traditional meju, we developed a capsule type-meju(CM) coated with steamed black bean and Aspergillus oryzae - rice koji(0.3%, w/w) mixture to surface of traditional meju and fermented at 25$^{\circ}C$ for 14 days under 80% relative humidity. Contamination of undesirable mold on the surface of CM was not detected within 2 weeks and some genus Penicillium molds on control meju without koji were found naturally after 12 days of fermentation. The moisture content of meju was showed to be in the range of 34.7 - 29.4% being 32.7%(w/w) of mean value. Titratable acidities in CMs prepared with black bean(BCM) and soybean(SCM) were much higher than that in control meju, and BCM was similar to SCM. Free sugar content in BCM(123.98 mg%) was 10 times and 2.1 times higher than that in control meju(15.02 mg%) and SCM(59.85 mg%), respectively. Amino type nitrogen content in control meju was 147.00 mg% and its content in BCM(255.50 mg%) was 1.37 times higher than that in SCM(187.25 mg%). Total organic acid content in BCM(95.98 mg%) and SCM(1l9.98 mg%) were much higher than that in control meju(26.44 mg%), and then lactic and malic acid contents were markedly changed according to capsulation of meju. Lightness value(L) of Hunter color index was much higher in BCM than in SCM. Fatty acid composition of CM was not different as compared to control meju. Total free amino acid content in BCM(1039.70 mg%) was 4.4 times and 2.4 times higher than that in control meju(236.45 mg%) and SCM(556.07 mg%), respectively.

Quality characteristics of commercial liquid type yogurt in Korea (국내 시판 액상요구르트의 품질특성)

  • Lee, Ji Hyun;Park, Hye Young;Won, Ju In;Park, Hye In;Choi, In Duck;Lee, Seuk Ki;Park, Ji Young;Joe, Dong Hwa;Jeon, Yong Hee;Oh, Sea Kwan;Han, Sang Ik;Choi, Hye Sun
    • Food Science and Preservation
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    • v.24 no.6
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    • pp.865-870
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    • 2017
  • Yogurt is produced by fermentation of milk using bacteria known as "yogurt cultures". Most of these bacteria are probiotics such as Lactobacillus delbrueckii subsp. bulgaricus, Lb. rhamnosus, Streptococcus thermophilus, and Bifidobacterium. The domestic fermented milk market is increasing, and about 30 companies are producing yogurt. The purpose of this study was to analyze the quality characteristics of domestic commercial liquid yogurt. Total 30types of commercial yogurts were sampled and their physicochemicial properties, including pH, sugar content, acidity, viscosity, and microbial characteristics of lactic acid bacteria counts, were measured. Commercial liquid type yogurt showed a pH of 4.5, sugar content of 7.4-21.2%, total acid content of 0.4-0.9%, and viscosity of 0.1-250 cP. In terms of microbial populations, lactic acid bacteria counts were 7.2-11.3 log CFU/mL and anaerobic lactic acid bacteria counts were 8.0-11.5 log CFU/mL. The quality characteristics differed depending on the constituents of the sample and the microorganisms used. These results are related to the quality characteristics of yogurts and are useful for identifying new trends in the domestic fermented milk industry.

Effect of Temperature and Various Pre-treatments on Germination of Hippophae rhamnoides Seeds (갈매보리수나무 종자의 온도 및 여러 가지 전처리에 따른 발아반응)

  • Choi, Chung-Ho
    • Korean Journal of Plant Resources
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    • v.25 no.1
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    • pp.132-141
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    • 2012
  • This study was carried out to test seed germination responses to temperatures and pre-treatments in Hippophae rhamnoides, which has many abilities in antioxidant activity, soil improvement and erosion control. H. rhamnoides seeds were placed at 10, 15, 20, 25, 30 and $35^{\circ}C$ under light condition. As the results, germination percentage (GP) was the highest at 15 and $20^{\circ}C$, and mean germination time (MGT), germination rate (GR) and germination value (GV) were the highest at $25^{\circ}C$. Quadratic and linear regression model were used to determine the cardinal temperatures such as base ($T_b$), maximum ($T_m$) and optimum ($T_o$) temperature for germination. In quadratic regression model using PG, $T_b$, $T_m$ and $T_o$ was estimated as 0.6, 36.4 and $18.5^{\circ}C$, respectively, and temperature range for germination was $35.8^{\circ}C$. In linear regression model using GR, $T_b$, $T_m$ and $T_o$ was estimated as 8.3, 35.4 and $25.3^{\circ}C$, respectively, and temperature range for germination was $27.2^{\circ}C$. Germination properties were investigated after H. rhamnoides seeds were treated by prechilling (1, 2, 4, 6 and 8 weeks), stratification (2, 4, 6 and 8 weeks), solid matrix priming (seed : carrier : water = 5 : 1 : 7, 8, 9 and 10), osmo-priming (-0.25, -0.5, -1.0 and -1.5 MPa) and calcium chloride ($CaCl_2$) -priming (100, 200, 300 and 400 mM). The highest GP was observed in $CaCl_2$ 300 and 400 mM treatments, and MGT was the shortest in stratification 6 and 8 weeks treatments. GR and GV were the highest and GP was the second highest when seeds were prechilled for 1 and 2 weeks. Consequently, prechilling 1 or 2 weeks treatment was considered as the appropriate method when we contemplate qualitative and quantitative effects in seedling production.

Total Polyphenol Contents and Antioxidant Activities of Methanol Extracts from Vegetables produced in Ullung Island (울릉도산 산채류 추출물의 총 폴리페놀 함량 및 항산화 활성)

  • Lee, Syng-Ook;Lee, Hyo-Jung;Yu, Mi-Hee;Im, Hyo-Gwon;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.233-240
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    • 2005
  • To discover new functional materials using edible plants, antioxidant activities of methanol extracts from various parts of seven wild vegetables were investigated in vitro. Total polyphenol contents, determined by Folin-Denis method, varied from 16.74 to $130.22{\mu}g/mg$. Radical-scavenging activities of methanol extracts were examined using ${\alpha},\;{\alpha}-diphenyl-{\beta}-pirrylhydrazyl$ (DPPH) radicals and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay. Inhibition effects on peroxidation of linoleic acid determined by ferric thiocyanate (FTC) method and on oxidative degradation of 2-deoxy-D-ribose in Fenton-type reaction system were dose-dependent. Athyrium acutipinulum Kodama (leaf and rood), Achyranthes japonica (Miq.) Nakai (seed), and Solidago virga-aurea var. gigantea Nakai (root) showed relatively high antioxidant activities in various systems.

A Study on Improving Perception of Subject-Special Librarian and Subject-Special Services: Focusing on Survey Targeting Students Who Take 'Medical Terms and MeSH' and 'Health & Medical Information Sources' (주제전문사서와 주제전문서비스에 대한 인식제고를 위한 연구 - '의학용어와 MeSH', '보건의학정보원' 수강생 대상 설문조사를 중심으로 -)

  • Oh, Eui-Kyung
    • Journal of the Korean Society for Library and Information Science
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    • v.50 no.1
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    • pp.437-456
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    • 2016
  • This study, based on the need for department of LIS education program that targets training of subject-special librarian, focuses on proving that such education can improve perception of subject-special librarian and strengthen career decision-making. This study is composed of two parts. First, this study investigates case studies about subject-special librarian cultivation. Second, this study includes a survey about subject-special librarian, improving understanding about subject-special librarian, and improving career decision-making which was carried out targeting students who take 'Medical Terms and MeSH', and 'Health & Medical Information Sources' courses. Results are as follows. First, most of the respondents felt the need for medical librarian and medical information services (88.7%), understood the role of medical librarian (92.5%), got to know more about the types and functions of medical information services (100%), and came to be more aware of the self-esteem a medical librarian feels as a professional. Second, relatively, the percentage of respondents who demonstrated the willingness to work at a medical library (45.3%) or work in the field of medical information services (26.4%) was noticeably low. Third, there were no statistical difference in whether the students took related courses or not, number of courses they took and difference in their school year, and in regard of the level of understanding of medical librarian and medical information services, those who took courses demonstrated higher understanding than did those who did not take those courses. Also, those who took the courses showed stronger willingness to work as a medical librarian. Fourthly, the curriculum carried out by the department of library and information service was effective in understanding medical librarian and medical information services, but makes students more cautious about deciding whether or not to start their career at a medical library. Lastly, it was recommended that there is a need to have the curriculum updated in order to help students overcome the difficulty and unfamiliarity they feel about medical subject.