• Title/Summary/Keyword: 계통추출법

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Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

버섯의 항암효과 -영자 균사체 다당류의 항종양 활성-

  • 이준우
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.13-14
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    • 1997
  • 영지의 약리효과는 triterpenoid계의 저분자와 polysaccharide인 고분자로 대별할 수 있다. 저분자 물질은 항염증작용, 항알러지작용, 간암세포 성장 억제작용, 혈압 강하작용, 혈소판 응집저해작용 등이 있으며, 고분자 물질은 혈압강하작용, 정혈작용, 고지혈증 개선작용, 혈당 강하작용, 면역 증강작용 및 항종양작용 등이 있는 것으로 알려졌다. 현재까지의 영지에 관한 연구는 주로 자실체에 대한 연구가 주류를 이루고 있으며 이들의 산지, 환경조건, 채집시기 및 실험자들에 따라 약리 효과 등에 많은 차이가 있다. 이들에 대한 효과가 영지의 대표적 약리효과라고 언급하기엔 부족한 점이 많으므로 약리효과가 우수하고 일관성이 있는 영지 유래 다당류를 얻기 위한 방법으로서는 균사체의 대량배양법 및 분리방법이 확립 되어야만 한다. 본 연구에서는 새로운 항암활성 다당류의 개발을 위해 국내 자생 영지를 채집하여 검색 및 계통적 분류를 행하였으며 분리된 균주의 최적 액체배양 조건, 다당류 분리 조건, 다당류의 물리화학적 특성 및 약리활성 등에 관한 연구를 실시하였다. 그 중 약리활성이 우수한 G- lucidum IY009 균주를 선발 하였다. 구조적 특징과 종양 면역 활성과의 관계 규명을 위해, G- lucidum IY009 배양균사체로부터 다당류를 T-AS, U-AS, M-AS, S 및 H로 분획 하였다. 추출조건에 따른 다당류 GLG의 수율은 열수 추출 분획이 2.98g/l로 가장 높았으며, U-AS및 T-AS에서 각각 2.32g/l, 2.07g/l이었다. GLG의 특성을 조사하기 위하여 수용성과 비수용성으로 분리한 결과 수용성 분획은 비수용성 분획보다 높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정 혈청단백을 증가시켰으며, 항체 생성능의 증가가 관찰되어 effect T 세포의 활성화가 나타나고 있음을 알 수 있었다. T-AS는 생체내 투여시에 대식세포의 탐식능이 증진되었으며, 대식세포 기능 저해제에 의한 대식세포의 기능 저해 현상이 회복되었다. 이와 같은 결과들로부터, T-AS의 항암 활성은 활성화된 보체 성분 및 당 수용체들이 존재하는 대식세포의 개입을 시사한다.

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Neuroprotective effects of phenolic compounds isolated from Spiraea prunifolia var. simpliciflora (조팝나무(Spiraea prunifolia var. simpliciflora)로부터 분리한 페놀 화합물의 신경세포 보호효과)

  • Oh, Seon Min;Choi, Doo Jin;Kim, Hyoung-Geun;Lee, Jae Won;Lee, Young-Seob;Lee, Jeong-Hoon;Lee, Seung-Eun;Kim, Geum-Soog;Baek, Nam-In;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.397-403
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    • 2018
  • The leaves of Spiraea prunifolia were extracted with 80% aqueous MeOH and the concentrates were partitioned into EtOAc, n-BuOH, and $H_2O$ fractions. The repeated $SiO_2$ or ODS column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of two phenolic glucosides. The chemical structures of these compounds were determined as isosalicin (1) and crenatin (2) based on spectroscopic analyses including Nuclear magnetic resonance and MS. Extracts were analyzed using UPLC-MS/MS providing a short analysis time within 5 min using MRM technique. The concentration of crenatin was higher as 9.53 mg/g and isosalicin was lower as 0.65 mg/g. Neuroprotective effects of these compounds against hydrogen peroxide ($H_2O_2$)-induced neurotoxicity were evaluated. The results showed that exposure to $H_2O_2$ induced morphological changes, cell death and neurotoxicity in SK-N-MC cells. However, pretreatment with crenatin resulted in inhibition of morphological change, reduction of loss of cell viability and attenuation of neuronal damage. These results suggested that neuroprotective effect of crenatin isolated from S. prunifolia can be a good candidate for the development of health beneficial foods which can ameliorate the degenerative neuronal disease caused by oxidative stress.

Application of Natural Dyes for Developing Colored Wood Furniture (I) - Color Variation by Extraction Methods of Natural Dyes - (색채 목가구재 개발을 위한 천연염료의 이용에 관한 연구 (제1보) - 천연염료의 추출 방법에 따른 색채 변화 연구 -)

  • Moon, Sun-Ok;Kim, Chul-Hwan;Kim, Jae-Ok;Kim, Jong-Gab
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.5
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    • pp.75-85
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    • 2004
  • The natural dyes from Gardenia jasminoides, Carthamus tinctorius L., Rhus javanica, Lithospermum erythrorhizon, Caesalpinia sappan L., and Castanea crenata were extracted under different pH in distilled water, As the pH in distilled water went from acid to alkali, the much deeper colors in the same color tone were generated from the individual plant species. Before dyeing, wood species were treated by different mordants including AI, Cu, Cr and Fe for color-fixing between wood and the natural dyes. Each mordant could develop independent color on the surface of the woods. The wood species dyed by the natural dyes created deep-tone colors according to higher pH and temperature of the dyeing solution, leading to deeper penetration of the dyes into the wood tissues. Finally through the computer modelling of natural-dyed wood furniture, it was confirmed that the colored furniture can adequately be compatible with the current interior spaces of diverse colors.

Collection, Identification and Hepatic Effect of Native Cordyceps militaris (새로운 번데기 동충하초의 수집, 동정 및 간기능에 미치는 효과)

  • Lee, Ki-Won;Nam, Byung-Hyouk;Jo, Wool-Soon;Oh, Su-Jung;Kang, Eun-Young;Cui, Yong;Lee, Jae-Yun;Cheon, Sang-Cheol;Jeong, Min-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Entomopathogenic fungus Cordyceps militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. In this study, we investigated the effect of the extract of C. militaris (MPUN8501), which was identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA, on the function of liver. C. militaris powder was extracted using hot water extracts method as time, volume and temperature and using method as differential polarity of organic solvent. Each fraction was tested for the improvement of hepatic enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity. The BuOH extracts (CME) had highest activity which was used for the test of toxicity and efficacy of C. militaris. The enhancing effect of CME on the activity of ADH and ALDH was much more than medicine, drink, natural tea etc. Thus CME promoted the resolution of alcohol and acetaldehyde in rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters (levels of sGOT and sGPT) and histological observation. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintaining normal hepatic activity as a functional health food.

Influence of Health-Related Quality of Life (EQ-5D) on Oral Health Problems in the Elderly in the National Health and Nutrition Survey (국민건강영양조사에서 나타난 노인의 건강관련 삶의 질(EQ-5D)이 구강건강 문제에 미치는 영향)

  • Ju, On-Ju;Kim, In-Ja;Son, Ju-Lee
    • Journal of the Korea Convergence Society
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    • v.11 no.1
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    • pp.309-315
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    • 2020
  • The purpose of this study was to examine the influence of the health-related quality of life(EQ-5D) on oral health problems in an effort to provide some information on health promotion programs geared toward the elderly and on the right direction of related policy setting. Raw data from the 7th National Nutrition Survey were used, and systematic sampling was conducted on elderly subjects over 65 years. EQ-5D and chewing problems increased in the group without exercise ability, in the group with pain discomfort, and in the group with anxiety depression (p<0.05). EQ-5D and speaking problems increased in terms of odds ratio(OR) in the group without exercise ability, in the group with pain discomfort, and more in terms of articulating difficulties in the anxiety depression group (p<0.05). The results of this study show a direct correlation between health-related quality of life (EQ-5D) and the oral health of the elderly. As it's mandatory to promote the oral health of elderly people to improve their EQ-5D To promote rural societies should provide oral health education and institutions for continuous oral health care.

Evaluation of Various Oligotrophic Media for Cultivation of Previously Uncultured Soil Bacteria (난배양성 토양세균의 배양법 평가 및 신 분류군의 순수분리)

  • Kim, Do-Hyoung;Lee, Sang-Hoon;Cho, Jae-Chang
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.352-357
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    • 2008
  • We evaluated cultivation methods to obtain pure cultures of previously uncultivated bacteria from soil. Soil bacteria (suspensions) were inoculated onto various oligotrophic media with one of the following additives: 1) soil extract; 2) anthraquinone disulfonate (humic acid analogue); 3) acyl homoserine lactones (quorum-signaling compounds); 4) catalase (for the protection of bacteria from exogenous peroxides). After the relatively long period (60 days) of incubation with elevated concentrations of $CO_2$ (5%, v/v), the media containing catalase showed the highest colony count. We purified 147 randomly selected colonies from the media and the isolates were subjected to the phylogenetic analyses of their 16S rRNA gene sequences. Phylogenetic analysis revealed that approximately 30% of the isolates might belong to novel species or novel family, suggesting that the media and incubation conditions used could be useful for the cultivation of as-yet-uncultured bacteria. Especially, bacteria belonging to the phylum Acidobacteria, ubiquitous bacterial taxon known as an uncultured bacterial group (at least difficult to culture from environmental samples), were successfully cultured in this study.

The Correlation Analysis between Occurrence of Seedlings of Korean fir (Abies koreana E.H. Wilson) and Understory on the Eastern Slope of Mt. Halla (한라산국립공원 동사면의 구상나무숲 내 주요 하층식물이 구상나무 유묘 발생에 미치는 상관성 분석)

  • Lee, Jong-Won;Lee, Su-Hong;Im, WonTaek;Kang, Shin-Ho
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.491-501
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    • 2021
  • This study was carried out to investigate the occurrence of seedlings of Korean fir (Abies koreana) in Korean fir forest located on the eastern slope of Mt. Halla National Park. Between June and August in 2016, the systematic sampling method was applied to 125 plots of 5 m × 5 m quadrats and field survey was came out to measure the characteristics of sites and understory vegetation. Correlation analysis was applied to the variables quantified from the data collected. Total number of flora found at the study site is 31 in families, 44 in genera, and 56 in species. The main lower layer vegetation was investigated such as Jeju dwarf bamboo, Toothed club moss (Lycopodium serratum), Pacific fir moss (Lycopodium chinense), Climbing hydrangea (Hydrangea petiolaris) etc. Among these plant species, the vegetation which had a high positive correlation with the frequency and the coverage of seedlings of Korean fir were Jeju dwarf bamboo, Toothed club moss and Pacific fir moss.

Antioxidant and Nitrite Scavenging Effect of Soybean, Meju and Doenjang (대두, 메주 및 된장의 항산화 효과와 아질산염 소거 효과)

  • 최계선;임선영;최재수
    • Journal of Life Science
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    • v.8 no.5
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    • pp.473-478
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    • 1998
  • The radical scavenging activity of the methanol extracts was in the order of doenjang〉 meju〉 soybean. The metha-nol extracts were further fractionated with dichloromethane, ethyl acetate, butanol and water in sequence. Among the solvent fractions of the methanol extracts, the ethyl acetate fraction demonstrated high levels of radical scave-nging activity. On the other hands, the nitrite scavenging activity of the methanol extrats was in the order of doen-jang〉soybean〉meju. All fractions from doenjang were exhibited the strong nitrite scavenging effect. Especially, $H_2O$, EtOAc and BuOH fractions with 6gmg/10$m\ell$ concentration exhibited much stronger nitrite scavenging effect than that of L-ascorbic acid. The ethyl acetate fraction showed remarkable antioxidant and nitrite scavenging effects compared with the other fractions obtained from soybean, meju and doenjang. The ethyl acetate fraction of doenjang was fur-ther purified by repeated chromoatograpic methods and isolated three compounds. There are identified as genistin, dai-dzein and genistein on the basis of spectral data.

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Error analysis and performance test of the volumetric interferometer for three dimensional coordinate measurements (삼차원 좌표 측정을 위한 부피 간섭계의 오차분석 및 성능평가)

  • 이혁교;주지영;김승우
    • Korean Journal of Optics and Photonics
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    • v.13 no.6
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    • pp.521-529
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    • 2002
  • We have recently proposed the new concept of a phase-measuring volumetric interferometer that enables us to accurately measure the xyz-coordinates of the probe without metrology frames. The interferometer is composed of a movable target and a fixed photo-detector array. The target is made of point diffraction sources to emit two spherical wavefronts, whose interference is monitored by an array of photo-detectors. Phase shifting is applied to obtain the precise phase values of the photo-detectors. Then the measured phases are fitted to a geometric model of multilateration so as to determine the xyz-location of the target by minimizing least square errors. The proposed interferometer has been designed and built with a volumetric uncertainty of less than 1.0 $\mu\textrm{m}$ within a cubic working volume of side 120 mm. Here, in this paper, we also present error sources, an evaluated uncertainty, and test results from the prototype system. The self-calibration of two-dimensional precision metrology stages is applied to test the performance of the interferometer.