• Title/Summary/Keyword: 항원

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Separation of Egg White Using HPLC with Change of Mobile Phase and Temperature (HPLC에서 이동상 변화와 온도에 따른 난백의 분리)

  • Do, Jin-Sun;Song, Shin-Young;Cho, Ki-Jung;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.829-834
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    • 2011
  • Lysozyme in egg white functions as bacteriolysis agent and ovalbumin plays a role as antigen in immune system. Egg white analysis methods usually include electrophoresis, gel permeation chromatography and reversed-phase HPLC(RP-HPLC). Among them, RP-HPLC was selected for rapid analysis and C18 column(Agilent, USA) was used as HPLC column. Optimum conditions were searched by changing mobile phase and temperature. Capacity factor and resolution were calculated and compared for various elution conditions. In the isocratic elution, mobile phase volume ratio was changed from 30/70/0.1 to 60/40/0.1(Acetonitrile(ACN)/Distilled water(DW)/Trifluoroacetic acid(TFA)). ACN composition was increased by 10% and temperature was set as $20^{\circ}C$. In the gradient elution, ACN/DW ratio was changed from 10/90 to 60/40 during 20 minute and temperature was varied as 20, 30 and $40^{\circ}C$. In the isocratic elution, three peaks were separated at 50/50/0.1. Lysozyme and ovalbumin were confirmed as first and third peak in three peaks respectively. In the gradient elution, four peaks were separated at $30^{\circ}C$. Lysozyme and ovalbumin were confirmed as first peak and third peak in four peaks respectively.

The inhibition effects of Injinchunggantang on Hepatitis B and C Virus (인진청간탕(茵蔯淸肝湯)의 B형(型) 및 C형(型) 간염 바이러스에 대한 항(抗) 바이러스 효과(效果))

  • Kim, Su-Sung;Kim, Gi-Yeol;Yoon, CheoI-Ho;Seo, Woon-Gyo;Kim, Jong-Dae;Chung, Ji-Choen
    • The Journal of Dong Guk Oriental Medicine
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    • v.8 no.1
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    • pp.93-106
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    • 1999
  • For the purpose of investigate the inhibition effects of Injinchunggantang on Hepatitis and C Virus. The inhibition experiments were as follows : 1. The inhibition effects of Hepatitis B surface antigen production was remarkably increased in the pretreated group of Injinchunggantang compared with control group. 2. Amplified PCR products of HBV-DNA in culture media was considerably decreased in the pretreated group of Injinchunggantang compared with control group. 3. Results of ATP quantitation and ATPase inhibition percent was increased pretreated group of Injinchunggantang. Also ATPase activity of HCV helicase inhibition assay was increased with a dose dependent manner. These results may suggest that Injinchunggantang will have the inhibition effects of Hepatitis B and C Virus.

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Effects of Water-soaking and Mechanical and Chemical Scarifications on Seed germination of Hairy Vetch (Vicia villosa Roth) (헤어리베치 종자의 침지처리와 종피 파상 및 황산처리가 발아에 미치는 영향)

  • Kim, Min-Tae;Lee, Yong-Hwan;Jeon, Weon-Tai;Kim, Sook Jin;Yun, Da-Hae;Ku, Ja-Hwan;Song, Hana;Lee, Hyeon Bok;Seo, Myung Chul;Kang, Hang-Won
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.1
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    • pp.49-52
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    • 2013
  • Seed coat of hairy vetch is very thick and hard, and difficult to absorb water during germination. It requires much time that cotyledon comes out from seed coat. Therefore this experiment was carried out to increase the germination rate by enhancing water absorption through water-soaking of seed, chemical scarification by sulfuric acid, and mechanical scarification on seed coat. Water-soaking for 5 hours seemed to be highly effective in enhancing germination rate. Although not significant, water-soaking resulted in increase of germination rate by 9%. Effect of soaking temperature was not significant, but early germination rate was increased to be 5.0%, 31.7% and 48.3% at $10^{\circ}C$, $20^{\circ}C$ and $30^{\circ}C$, respectively. Mechanical scarification of seed coat led to a germination rate of 97% whereas intact seed showed that of 65%. Chemical scarification by sulfuric acid for 10min, 20min and 30min resulted in a germination rate of 76.7%, 74.7% and 96.0% respectively. It is clear that scarification increased germination rate.

Interrelationship of Matrix Metalloproteinase and $TNF-{\alpha}$ in Human Gingiva with Chronic Periodontitis associated to Type 2 Diabetes Mellitus (단순 만성 치주염 환자 및 2형 당뇨병을 가진 만성 치주염 환자의 치은조직에서 Matrix Metalloproteinase와 $TNF-{\alpha}$ 의 발현 양상 비교)

  • Kim, Doe-Heun;Park, Eei-Kyun;Shin, Hong-In;Cho, Je-Yeol;Suh, Jo-Young;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.409-425
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    • 2006
  • 치주질환의 병원균은 세포벽의 항원에 의하여 조직내 존재하는 mononuclear phagocytes가 활성화되어 cytokine들이 생성됨 으로써 치주 결체조직의 파괴를 진행시킨다. 이런 관련된 cytokine들은 순차적으로 상주하는 치은세포 및 대식세포가 Matrix metalloproteinase 합성을 하도록 유도하여 조직파괴를 시작한다. 이들 Matrix metalloproteinase중 MMP-2, MMP-9 (Gelatinase A,B)는 type IV collagen 및 변성된 interstitial collagen을 파괴하며 치주환자의 치은 열구액, 치은조직, 타액 네에서 높게 보고 되어왔다. 당뇨병은 치주질환의 위험요소중 하나로 달뇨 환자에서는 치주질환의 유병율이 일반인에 비해 높고 치주질환의 중증도도 더 심하여 진행도 빠르다고 알려져 있다. 그 병리 기전 중 하나로는 당뇨 환자에서는 치은 열구액 내 중성구 유래의 Matrix metalloproteinase의 활성 증가 및$TNF-{\alpha}$ 의 활성 증가가 추정되고 있다. 본 실험에서는 제2형 당뇨병 환자와 비당뇨 환자들에서 만성 치주염 부위의 치은 및 건강한 치은에서 염증매개체 중 하나인 MMP-2, MMP-9 및 $TNF-{\alpha}$ 의 발현에 대해 상호 비교 분석함으로서 염증, 혈당이 미치는 영향을 밝히고 제 2형 당뇨병 환자에서 심한 치주조직 파괴의 기전을 연구하고자 하였다. 경북대학교병원 치주과 내원환자 중 제2형 당뇨병 환자와 비당뇨 환자들 및 치주질환이 없는 건강인 대조군을 대상으로 여러 가지 환자요소, 임상 치주상태를 기록하고, 전신적으로 건강한 환자의 건강한 부위(n=8,Group 1), 전신적으로 건강한 환자으 만성 치주염 부위(n=8, Group 2), 제2형 당뇨병 환자의 만성 치주염 부위 (n=8,Group 3)에서 각각 변연치은을 채득하고 액화질소에 급속 동결하였다. Western blotting을 이용하여 각 조직 내 MMP-2, MMP-9 및 $TNF-{\alpha}$ 의 발현을 관찰, densitometer를 이용하여 상대적 발현을 정량, 각 조직의${\beta}-actin$을 이용하여 표준화하여 실험군과 대조군들의 평균치를 비교하였다. 비당뇨 환자들의 만성 치주염 부위 및 제2형 당뇨병 환자의 만성 치주염 부위에서 모두 건강 대조군에 비해 MMP-2와 MMP-9 의 발현이 증가되었다. 또한 MMP-2와 MMP-9는 2형 당뇨 환자의 만성 치주염 부위가 비당뇨 환자의 만성 치주염 부위보다 증가된 발현양상을 보였으며, $TNF-{\alpha}$ 발현 비교시 각 군간 유의성 있는 변화는 없었으나 2형 당뇨환자군에서 MMP-2 및 MMP-9의 증가와 함께 다소 증가 양상을 보였다. 결론적으로 본 실험에서 MMP-2 및 MMP-9의 증가가 만성 치주염 및 2형 당뇨 환자에서의 만성치주염에서 비당뇨환자 보다 MMP-2, MMP-9의 증가양상을 보여 주었으며 $TNF-{\alpha}$ 가 2형 당뇨환자의 만성치주염 진행과정에 기여인자로써 역할을 하는 것으로 생각된다.

Endometrial Cell Culture: Isolation, Characterization, and Immortalization (자궁내막 세포의 분리 및 배양 그리고 불멸화된 세포주의 확립)

  • Hong, In-Sun;Kim, Seok-Hyun;Koong, Mi-Kyoung;Jun, Jin-Hyun;Lee, Yong-Soon;Kang, Kyung-Sun
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.4
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    • pp.317-324
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    • 2003
  • 목 적: 본 실험의 목적은 자궁내막세포를 분리 및 배양법 확립과 함께 불멸화 시키는 것이다. 방 법: 자궁내막에서 상피세포(epithelial cells)와 기질세포(stromal cells)의 분리는 Satyawaroop 등(1979)의 방법에 기초를 두었다. 자궁내막에서 상피세포와 기질세포의 순수 분리도를 확인하고, 불멸화된 기질세포에서 SV40 large T antigen을 확인하기 위하여 면역형광 염색(immunocytochemistry)과 Western blot 기법을 이용하였다. 정상 기질세포의 경우 subconfluence (60%) 상태에서 transfection을 진행하였다. 순수 분리된 plasmid DNA와 Qiagen 사의 superfect를 이용하여 transfection을 실시하였다. 결 과: 본 연구에서 우리는 두 가지 형태의 자궁내막 세포의 분리 및 배양에 성공하였다. 상피세포는 다면체의 형태를 띠며, 선(grandular)조직의 조각으로부터 나선형으로 자란다.기질 세포는 길쭉한 형태를 띠며, 상피세포에 비해 오래 살고, 빠르게 증식하여 나란한 형태로 배열된 세포 다발(cell bundle)을 형성한다. 이렇게 분리된 세포들은 95%의 균질성을 보였으며, 면역형광염색과 western blot을 통해 확인 하였다. 한편 SV40(Simian Virus 40) large T 항원을 암호화 하고 있는 염기 서열을 포함한 플라스미드 벡터로 안정적인 트랜스펙션을 시킴으로써 불멸화 된 자궁내막의 기질 세포주를 확립하였다. 불멸화 된 세포는 그 세포가 유래한 정상의 세포와 동일한 표현형을 가지고 있었다. 결 론: 본 연구에서, 우리는 자궁내막에서 상피세포(epithelial cells)과 기질세포(stromal cells)를 분리하여 배양법을 확립하였다. 동시에 SV40 large T antigen을 이용하여 불멸화된 세포주를 확립하였다. 이렇게 확립된 세포주는 자궁의 생리작용 연구 및 자궁내막증(Endometriosis)과 자궁암(Endometrial cancer) 등과 같은 여러 자궁관련 질병 연구에 많은 도움이 될 것으로 사료된다.

The Effects of Progesterone on Cell Mediated Immunity to Trophoblast in Woman with Recurrent Pregnancy Loss (반복 유산환자의 말초혈액 단핵구와 태반항원을 체외 공동 배양시 세포 매개 면역반응에 프로게스테론이 미치는 영향)

  • Choi, Bum-Chae;Hill, Joseph A.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.399-405
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    • 1997
  • Progesterone is necessary for successful pregnancy and had immunosuppressive properties. Peripheral blood mononuclear cells (PBMC) from many women with unexplained recurrent spontaneous abortion responded to trophoblast extract in vitro by prolifertion and releasing soluble, heat-labile factors that are toxic to mouse embryos (embryotoxic factors). Accumulating evidence suggests that T Helper (Th)-1 type immunity to trophoblast is correlated with embryotoxic factor production and is associated with pregnancy loss, while Th2-type immunity is associated with successful gestation. The objective of this study was to determine whether progesterone can inhibit Th1-type cytokine secretion (IFN-${\gamma}$, TNF-${\alpha}$) by trophoblast-activated peripheral blood mononuclear cells from 23 nonpregnant women (age 25-35) with unexplained recurrent abortion (median 5, range 3 to 15)who otherwise produce embryotoxic factors in response to trophoblast. We also determined whether progesterone affected Th2-type cytokines (IL-4, IL-10) in this system in vitro and if IL-10 (1,500 pg/mL) could inhibit Th1-type immunity to trophoblast. IFN-${\gamma}$ was detected in 17 of 23 (74%) trophoblast stimulated PBMC culture supernatants ($77.94{\pm}23.79$ pg/mL) containing embryotoxic activity. TNF-${\alpha}$ was detected in 19 (83%) of these same supernatants ($703.15{\pm}131.36$ pg/mL). In contrast, none of the supernatants contained detectable levels of IL-4 or IL-10. Progesterone ($10^{-5}$, $10^{-7}$, $10^{-9}$M) inhibited Th1-type immunity in a dose dependent manner, but had no effect on Th2-type cytokine secretion. The inhibitory effects of progesterone were abrogated with RU486, but did not affect Th2-type cytokine secretion in trophoblast-activated cell cultures. IL-10, like progesterone also inhibited Th1-type cytokine secretion but had no effect on Th2-type cytokines. These data suggest that therapies designed to suppress Th1-type cytokine secretion in women with recurrent abortion who have evidence of Th1-type immunity to trophoblast may be efficacious in preventing pregnancy loss and should be tested in appropriately designed clinical trials.

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Development of Direct Competitive Enzyme-Linked Immunosorbent Assay using Monoclonal Antibody (MAb) against Sulfamthazine (SMZ) and Establishment of Application Condition for Milk Sample (설파메타진에 단클론성 항체를 이용한 직접경쟁효소면역분석법의 개발과 우유 시료 적용 조건 확립)

  • Shim, Won-Bo;Mun, Chun-Sun;Kim, Jung-Sook;Choe, Ju-Mi;Kim, Ji-Hun;Park, Seon-Ja;Kang, Sung-Jo;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.176-182
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    • 2006
  • Sensitive and specific monoclonal antibody (MAb) was produced from hybridoma (1H11-5) obtained by fusion of myeloma cell (V653) and spleen cell isolated from mouse immunized sulfamthazine (SMZ)-HG-KLH. Direct competitive ELISA was developed for rapid detection of SMZ in milk samples using MAb against SMZ with optimized conditions between MAb and SMZ-HG-HRP conjugate, and applicable conditions for analysis of milk samples were established. Detection range of immunoassay was 0.1 to 100 ppb. Recoveries from spiked raw milk and processed milk samples averaged 82.1-120.7 and 82.1-97.1%, respectively.

Induction of Rice Allergen-Specific IgE Synthesis by KU8l2 Cells (KU812세포에 의한 쌀 알레르겐 특이적 IgE항체 합성의 유도)

  • Shim, Sun-Yup;Katakura, Yoshinori;Shirahata, Sanetaka
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1492-1496
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    • 2007
  • In vitro IgE class switching could be induced through co-culture of CD40L-expressing KU812 cells and CD40-expressing B cells in the presence of IL-4 or IL-13. It has been generated several B cell lines, which produce rice allergen (RA)-specific IgM antibody by in witγo immunization (IVI) using peripheral blood lymphocyte (PBL). In this study, induction of RA-specific IgE antibody by KU812 cells was attempted. Before co-culture, we determined the CD40 expression in RA-specific B cell lines, RA9G11 and the CD40 ligand (CD40L) expression in activated KU812 cells by treatments with phorbol myristate acetate (PMA) and ionomycin for 6 hrs. Flow cytometric analysis shown that RA9G11 and activated KU812 cells expressed high level of CD40 and CD40L, respectively. RA9G11 cells were cultured with activated KU812 cells for 12 days in the presence of IL-4 for IgE class switching. Mature $C{\varepsilon}$ mRNA level and RA-specific IgE spot forming cells (SFC) were observed in all culture condition, and especially, high level of RA-specific IgE synthesis was determined the same ratio of RA9G11 and activated KU812 cells in the presence of 50U IL-4. Therefore, induction of RA-specific IgE synthesis by activated KU812 cells can be contributed in the application for allergic therapy and prevention.

The observation about occurrence rate of HBs Antigen to waitress entertaing at restaurant business (일부 접객업소 여성 종사자들에 있어서의 간염 B 항원 발현빈도에 관한 관찰)

  • 윤기은;김태전;원종만;나동진
    • Journal of Environmental Health Sciences
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    • v.6 no.1
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    • pp.47-52
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    • 1979
  • Up to the present there were the blood transfusion and the use of apparatus not to sterilize at surgical operation and medical treatments in well known infectious ways of hepatitis B virus. But all its ways were still not surely known. As the other ways of hepatitis B virus infection, it was suggested that contagion such as kissing, sexual contact, homosexuality, and varieties discharged out human body urine, stool, tear, salivary, menstrual blood, Vaginal discharge etc, and low economic status, unsanitary environmental life, alcohol and smoking related to hepatitis B virus infection. From 800 waitress who frequently contact with a lot of drinker with low economic status and unclear environmental life, this study were investigated HBs Ag (Hepatitis B surface antigen) known marker for hepatitis B virus infection in their serum, and a few conditions of their private life in order to known dangerous rate b~ing exposed to the source of hepatitis B virus infection as comparison with control group. The results were summarlized as following 1. The positive rate of HBs Ag (4.3%) was significantly higher in waitress than in control group(1.1%). 2. In waitress old, 20-24 ages group was the most as 59.5%, and positive rate of HBs Ag was trend of higher in twenties than thirties. 3. Among the waitress, one that it was less than a year were the most (62.4%) in the period to work at restaurant business, and positive rate of HBs Ag was trend of higher in propotion to period to work at there. 4. Among the waitress, one who entertained to beer hall was the most (46%), and they were trend of higher in positive rate of HBs Ag (6.3%) than other waitress. 5. Among the waitress, one to drink with smoking, and only to drink, and only to smoke, and not to drink and smoke 66.1%, 21.5%, 4.6% and 7.8%. Espically in one to drink with smoking, ther was trend of indicating the high positve rate of HBs Ag, and it was made suspicion of the relationship of alcohol smoking, contact with a lot of drinker, unclear environment to hepatitis B virus infection. In the above results, it was found that dangerous rate being exposed in hepatitis B virus infection was high in the waitress. Therefore there are required for active preventions against hepatitis B virus infection them. Also as it is possible to be infectious source in public health that waitress infected to viral hepatitis, it is thought that appropriate rules about them.

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Nutrients Removal Efficiency by Vegetation Density on Constructed Wetland from Small Watershed (소수계 유역 인공습지에서 식생 밀도 차이에 다른 영양염류 제거효율)

  • Ko, Jee-Yeon;Kang, Hang-Won;Lee, Jae-Sang;Kim, Chun-Song;Sakadevan, K.;Bavor, H.J.
    • Korean Journal of Environmental Agriculture
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    • v.22 no.4
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    • pp.266-272
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    • 2003
  • This study was conducted to evaluate effects of 2 constructed wetlands with different vegetation states (plumpton park wetland and Woodcroft park wetland) for reducing non-point source pollution from small watershed consisted of residential and agricultural area in suburban district of Sydney, Australia. The total nitrogen and phosphate removal efficiency of Plumpton park constructed wetland, composed of stable and dense vegetation, were 38.3% and 26.2% and Woodcroft park constructed wetland having still poor vegetation due to the short time to settle down transplanted plants after construction, showed relatively low removal efficiency of 20.2% and 14.0%. The removal efficiency of inorganic nutrients such as $NH_4-N$, $NO_3-N$, $PO_4^{-3}$ were higher than total nitrogen and phosphate because plants and microorganisms in rhizosphere of constructed wetlands took up inorganic nutrients shortly. According to the type of wetland inflow, the nutrients removal efficiency of storm water flow was lower than base flow.