• 미생물학회지
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• 제30권6호
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• pp.524-527
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• 1992

경기도 의왕시에 위치한 백운 저수지에서 분리된 cyanobacteria 인 Synechococcus sp. 로부터 한 종의 cyanophage 를 분리하여 ultrafiltration, differential centrifugation 과 sucrose density gradient centrifugation 을 이용하여 정제하였다. 분리된 cynophage 는 transmission electron microscope (TEM) 를 이용하여 100,000 배로 관찰한 결과 직경이 89 nm인 isometric head 와 tail 의 전체 길이가 111 nm 의 contractile tail 을 가진 phage 로서 Myoviridae 에 속함을 알 수 있었다. 생화학적 특징으로는 20~$40^{\circ}C$ 와 pH 5-10에서 50% 이상의 안정성을 유지하며 $30^{\circ}C$ 와 pH 9 에서 최고의 infectivity 를 나타내었다.

• Parasites, Hosts and Diseases
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• 제32권4호
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• pp.243-248
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• 1994

질편모충 항원 분석의 일환으로 막항원의 분석을 시도하였다 수확 세척된 질편모충 HY-l의 homogenate를 sucrose step-gradient를 이용하여 differential centrifugation하였으며 25%/45%의 sucrose 경계면으로녁터 막분회을 얻었다. 분리된 막분회은 transmission electron microscopy를 통하여 순수 분리되었는지 확인하였고 효소면역 전기영동 이적법(EITB)을 이용하여 항원성을 관찰하였으며 그 결과는 다음과 같았다. 분리된 막분획은 투과전자 현미경상에서 extended sheet나 concentric vesicle의 형태가 거의 균질하게 분포하고 있었으며 막 분획에서 특징적으로 나타나는 trilaminal appearance를 보여 질편모충의 막분획이 순수 분리된 것으로 간주할 수 있었다. 분리된 막분획은 EITB상에 토끼의 항혈청과 반응하였을 때 46, 60, 110, 120, 130 및 150 kfDa에서 항원성이 있는 반응대가 관찰되었으며 N-hydroxysuccinimido-biotic으로 표지하여 분리된 표면항원의 녁획과 비교하였을 때 60 KDa의 항원 분획이 서로 일치하는 것으로 나타났다. 따라서 60 kfDa 의 항원 분획은 료면 항원임을 확신할 수 있었다.

• Tuberculosis and Respiratory Diseases
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• 제45권1호
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• pp.176-183
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• 1998

연구배경: 기관지폐포세척술로 얻은 기관지폐포세척액의 총세포 숫자와 감별숫자는 특발성폐섬유증을 포함한 간질성 폐질환의 진단과 예후 및 치료방침의 결정에 유용한 검사방법이다. 그러나 기관지폐포세척액 처리 방법에 대한 표준 방법 (standard method)이 아직 정립되어 있지 않고, 병원마다 처리 방법이 상이한 경우가 많아 검사 결과의 해석에 어려움이 많은 실정이다. 이에 본 연구에서는 기관지폐포세척액의 처리방법 중에서 면거즈 (cotton gauze)로 걸르는 방법, 세척액의 원심 분리 후 재부유하는 방법, 기관지폐포세척액의 보관 시간등이 총세포숫자와 감별숫자에 미치는 영향을 관찰하였다. 방 법: 활동성 폐질환이 없는 6명의 사람에 대해서 기관지내시경을 하여 얻은 기관지폐포세척액을 모아서 siliconized glass tube에 분주한 후, 0, 1, 2, 4겹의 면거즈 (pore size: 1mm)로 걸른후 일부를 trypan blue로 염색한 후 hemocytometer로 총세포숫자를 계수하고 일부는 cytocentrifuge한 후 Wright-Giemsa stain을 하여 감별숫지를 하였다. 그리고 일부의 세척액을 두 겹의 거즈로 걸른 후에 원심분리(400g, 30min) 하고 RPMI1640으로 재부유액을 만들어 총세포숫자와 감별숫자를 하였다. 그리고 나머지는 $4^{\circ}C$ 냉장고에 보관하였다가 2시간, 24시간, 48시간 후에 각각 같은 방법으로 총세포숫자와 감별숫자를 하였다. 결 과: 기관지폐포세척액을 면거즈로 거르지 않은 경우와 여러겹의 면거즈로 걸른 경우 총세포숫자와 감별숫자에 차이가 없었으나 기관지폐포세척액을 면거즈로 거르지 않았을 경우 두 예(33%)에서 점액사 때문에 세포가 뭉치고 겹쳐져서 총세포숫자와 감별숫자가 어려웠다. 기관지폐포세척액을 원심분리하였을 경우 총세포숫자는 감소하였으나 ($24{\pm}18%$), 감별숫자에는 차이가 없었다. 보관시간에 따른 총세포숫자에 있어서 2시간 보관했을 경우에는 총세포숫자의 감소가 없었으나 24시간 이상 보관시 유의하게 감소하였으며 (24시간: $28{\pm}21%$, 48시간: $41{\pm}24%$), 감별숫자에 있어서는 보관시간에 따른 차이는 없었으나 48시간 보관시에는 세포변성으로 감별숫자가 어려웠다. 결 론: 기관지폐포세척액의 총세포숫자와 감별숫자를 위한 기관지폐포세척액의 처리과정에서 면거즈로 걸러서 두시간 아내에 처리하는 것이 좋으며 원심분리는 하지 않는 것이 좋을 것으로 생각된다.

• 한국식품과학회지
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• 제19권5호
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• pp.441-445
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• 1987

발아중인 대두콩에서의 lipoxygenase의 세포내 위치를 차별 원심 분리법과 밀도 구배법을 이용하여 규명하였다. 차별 원심 분리법을 사용하였을때, lipoxygenase활성은 거의 supernatant fraction에서 나타났으며, 1.5%의 활성만이 particulate fractions에서 나타났다. 밀도 구배법에서는 lipoxygenase와 acid phosphatase의 활성이 1.19, 1.23, $1.25g/cm^3$의 높은 밀도들에서 일치하였으며, 이것은 발아중 가수분해되는 단백질체내에 이 효소가 국재되어지는 것으로 사료되어진다. 다른 세포내 소기관인미토콘드리아, ER, 그리고 glyoxysomes에 lipoxygenase가 존재한다는 증거는 보이지 않았다.

• 한국식품영양학회지
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• 제8권4호
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• pp.307-317
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• 1995

The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

• 농촌의학ㆍ지역보건
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• 제17권1호
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• pp.46-55
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• 1992

A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from P.westermani antigen. Differential centrifugation of the homogenate yield three particulate and one soluble fractions ; the $480{\times}G$ pellet(Pw1), the $7650{\times}G$ pellet(Pw2) treated with n-butanol(Pw3), and $100000{\times}G$ supernatnat(Pw4). Comparison of antigenicity of these antigens, based upon differential centrifugation, to that of saline extract of P. westermani worm(SEP) was performed by SDS-PAGE and immunoblot techniques. The results obtained were as follows : 1) The ratio of absorbance value of ELISA against paragonimiasis positive pool sera to that of negative sera was highest when using Pw3 as antigen and that was lowest using Pwl. 2) Silver stained and SDS-PAGE of each antigen showed 34 and 13Kd band as common antigen band, but Pw2 didn't show clear band. 3) By immunoblot 55 and 34Kd bands using SEP and Pw4 showed strong positive reaction without cross reaction with sera from other helmenthic infections. Using Pw3, 10Kd band was observed as specific band. In conclusion, Pw3($100000{\times}G$ pellet urea soluble, treated with n-butanol) and Pw4($100000{\times}G$ supernatant) were usable for ELISA and immunoblot technique.

• Parasites, Hosts and Diseases
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• 제21권1호
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• pp.41-48
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• 1983

The activity and distribution of aspartate aminotransferase (EC 2.6. 1. 1) and alanine aminotransferase (EC 2.6.1.2) in adult Fascicle hepatica have been studied. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of GOT and GPT was measured by the method of Reitman and Frankel. Isozyme patterns of those enzyme were also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity of aspartate and alanine aminotransferase was about 0.55 unit and 0.92 unit per 1g of Fascicle hepatica, respectively. 2. The activity of those enzymes was relatively low compared with those in mammalian tissues. 3. The distribution of aspartate aminotransferase in the subcellular organelles showed that 71% of the activity was in cytosolic, 24% in mitochondrial and 5% was in nuclear fraction. 4. About 22% of the total alanine aminotransferase activity was found in the mitochondrial fratstion, about 66% in the cytosolic fraction. 5. Aspartate aminotransferase from cytosolic fraction was separated into two types of isozymes, whereas alanine aminotransferase from cytosolic fraction gave only one active peak on DEAE-cellulose column chromatography.

• 약학회지
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• 제38권1호
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• pp.12-19
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• 1994

Capsaicin(8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component of Capsicum fruits. This work is directed to the capsaicin-hydrolyzing enzyme playing a key role in the rate limiting and critical step of capsaicin metabolism. In order to get precise information on the enzyme's subcellular location, rat liver homogenate was divided into six subcellular fractions by differential centrifugation technique: crude nuclear pellet, PNS(post nuclear supernatant) fraction, lysosomal pellet, cytosol, Tris wash fraction, micrisomes. Capsaicin-hydrolysing enzyme activity was analysed by high performance liquid chromatography(HPLC). This enzyme was found at the highest specific activity in the microsomal fraction and co-distributed with marker enzymes of the endoplasmic reticulum, NADPH-cytochrome c reductase and nucleoside diphosphatase. This is compatible with the result of ninhydrin color reaction of vanillylamine, primary metabolite of capsaicin hydrolysis, on thin layer chromatography(TLC). This enzyme is most active at pH $8.0{\sim}9.0$. Definite subcellular location of this enzyme will make it easy to proceed with further study.

• Asian-Australasian Journal of Animal Sciences
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• 제22권6호
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• pp.885-892
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• 2009

To investigate host defense mechanisms for protecting the mammary gland from mastitis infection, the membrane fraction of mammary tissues from Holstein cows was purified by differential velocity centrifugation, and then the sodium dodecyl sulfate-polyacrylamid gel electrophoresis (SDS-PAGE) separated proteins were identified by ion trap mass spectrometer equipped with a Surveyor high performance liquid chromatography (HPLC) system. A total of 183 proteins were identified. Bioinformatics software was applied to analyse physicochemical characteristics of the identified proteins and to predict biochemical function. These data may provide valuable information to investigate the mechanisms of mammary gland milk secretion and infectious disease, and enable a clear identification of proteins and potential protein targets for therapies.

• Applied Biological Chemistry
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• 제20권2호
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• pp.175-181
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• 1977

돼지 백혈구(白血球) Iysoromal enzyme의 latency를 서로 다른 농도의 sucrose용액(0.0125-0.25M)으로서 조사하고 각(各) sedimentation fraction에 분포되어 있는 효소들의 specific activity, pH optima 및 activation energy를 측정하였다.