• 제목/요약/키워드: Sucrose gradient

검색결과 100건 처리시간 0.025초

질편모충의 표면항원 분석 (Identification of surface antigen of Trichomonas vaginalis)

  • 민득영;임미혜
    • 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.

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개 말초혈액(末稍血液)에서 monocytes 분리(分離) (Separation of monocytes from canine peripheral blood)

  • 김정배;이방환
    • 대한수의학회지
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    • 제29권2호
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    • pp.33-39
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    • 1989
  • Pure separation of various leukocytes is required for the assessment of their roles in immunological and phisiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at $37^{\circ}C$ for 2 hours. 1. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. 2. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability-rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particulary showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes.

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면역결핍 바이러스 입자의 비특이적 성질 (Unusual Features of Human Immunodeficiency Virus Type-1 Virion)

  • 신차균
    • 대한바이러스학회지
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    • 제26권1호
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    • pp.107-114
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    • 1996
  • 본 연구는 인간면역결핍바이러스의 입자를 비이온성 계면활성제로 처리할 때 바이러스 입자구조에서 분리되어 방출되는 바이러스 구조단백질들의 분포를 sucrose gradient로 분석하여, 바이러스 입자를 구성하는 바이러스 구조단백질과 바이러스입자의 생물리학적 특성을 연구하였다. 바이러스입자들을 0.16% NP40 (Nonidet P-40)으로 처리할 때, 바이러스 capsid 단백질과 바이러스 막 단백질 (membrance protein)들은 다른 바이러스 구성성분들과 잘 분리되었다. 계면활성제처리에서 방출되지 않은 구성 성분들은 matrix 단백질, nucleocapsid 단백질, reverse transcriptase, integrase 및 바이러스 RNA genome로써, 이들은 subviral 구조를 형성한다. 이러한 결과는 상대적으로 다른 바이러스들의 capsid 단백질과 면역 결핍 바이러스의 capsid 단백질 (p24)를 비교할 때, 면역결핍바이러스의 capsid 단백질은 바이러스핵을 형성할 때, capsid 단백질 사이의 결합력이 매우 약한 것으로 추정된다. 또한 바이러스 조절단백질의 하나인 vpr 단백질을 함유하는 바이러스입자를 NP40 처리하여 분석하였을 때, vpr 단백질은 subviral 구조에 존재하는 것으로 나타났다.

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유행성 폐출혈열 환자에서 분리된 병원체에서 관찰된 다양한 형태들간의 연관성 (Relatedness Between Different Morphologies of L. interrogans Isolated from a Patient with Epidemic Pulmonary Hemorrhagic Fever)

  • 이태윤;김경원;김계성
    • 대한미생물학회지
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    • 제22권3호
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    • pp.219-224
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    • 1987
  • A strain of Leptospira interrogans(UM-19) was isolated in 1984 from a patient with epidemic pulmonary hemorrhagic fever. The bacteria has been characterized morphologically by a polymorphism showing spiral forms, short and long rods in culture. In order to study the biological relatedness among these forms of the bacteria, the short rods were separated from others by means of continuous sucrose density gradient centrifugation, and cultured at various temperatures such as $10^{\circ}C$, $15^{\circ}C$, $30^{\circ}C$ and $37^{\circ}C$. It was revealed that cultivation or subcultivation of the short rods has resulted in morphological changes observed by darkfield examination, silver stain, and scanning electron microscope showing spiral forms as well as short and long rods.

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Phycobilisome composition in Chondrus crispus (Gigartinales, Rhodophyta) from a wild type strain and its vegetatively derived green mutant

  • Cornish, M. Lynn;O' Leary, Stephen J.B.;Garbary, David J.
    • ALGAE
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    • 제28권1호
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    • pp.121-129
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    • 2013
  • Intact phycobilisomes from a wild-type red Chondrus crispus and its vegetatively derived green mutant were isolated by centrifugation through a discontinuous sucrose density gradient. Pigment composition was subsequently characterized by spectrophotometry. Vegetative thalli of the two strains grown together for six months in the laboratory resulted in different pigment profiles. Two pigmented phycobilisome bands appeared in the sucrose gradient of the wild-type alga, a purple coloured one, and a pink one, whereas only a single blue band appeared in the gradient of the green mutant. Spectrophotometric and fluorescence analyses identified the phycobiliprotein composition of the purple band as the typical phycoerythrin-phycocyanin-allophycocyanin complement in the wild-type, but there was no detectable phycoerythrin present in the blue band of the green mutant. Sodium dodecyl sulphate, preparative polyacrylamide gel electrophoresis analysis confirmed the presence of allophycocyanin subunits in all extracts, but firm evidence of an R-phycoerythrin linker polypeptide in the blue band was missing. These results highlight the ability of C. crispus to adapt to a phycoerythrin deficiency by adjusting light harvesting pigment ratios.

Separation of soil Organic Debris using Sucrose-ZnCl2 Density Gradient Centrifugation

  • Jung, Seok-Ho;Chung, Doug-Young;Han, Gwang-Hyun
    • 한국토양비료학회지
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    • 제45권1호
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    • pp.30-36
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    • 2012
  • The active fraction of soil organic matter, which includes organic debris and light organic fraction, plays a major role in nutrient cycling. In addition, particulate organic matter is a valuable index of labile soil organic matter and can reflect differences in various soil behaviors. Since soil organic matter bound to soil mineral particles has its density lower than soil minerals, we partitioned soil organic matter into debris ($<1.5g\;cm^{-3}$), light fraction ($1.5-2.0g\;cm^{-3}$), and heavy fraction ($>2.0g\;cm^{-3}$), based on high density $ZnCl_{2-}$ sucrose solutions. Generally, partitioned organic bands were clearly separated, demonstrating that the $ZnCl_{2-}$ sucrose solutions are useful for such a density gradient centrifugation. The available gradient ranges from 1.2 to $2.0g\;cm^{-3}$. Although there was not a statistically meaningful difference in organic debris and organomineral fractions among the examined soils, there was a general trend that a higher content of organic debris resulted in a higher proportion of light organomineral fraction. In addition, high clay content was associated with increased fraction of light organomineals. Partitioning of soil organic carbon revealed that carbon content is reduced in the heavy fraction than in the light fraction, reflecting that the light fraction contains more fresh and abundant carbon than the passive resistant fraction. It was also found that carbon contents in the overall organic matter, debris, light fraction, and heavy fractions may differ considerably in response to different farming practices.

자당 및 탄산음료 섭취 후 생성되는 구강 내 치아우식 유발성 유기산의 농도 차이 (The concentration differences of dental caries induced organic acids which are produced after intake of sucrose and carbonated drinks)

  • 박정은;장종화
    • 한국치위생학회지
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    • 제17권3호
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    • pp.381-394
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    • 2017
  • Objectives: This study aims to evaluate carbonated drinks induced dental caries with qualitative analysis and to compare with oral organic acids including lactate, acetate, propionate, formate, butyrate, pyruvate and valerate which cause caries when taking either 10% sucrose drinks or carbonated drinks. Methods: Saliva was collected from six study subjects before and after (start, 5, 10, 30 minutes) taking water intake upon (A) 10% sucrose intake, (B) 10% sucrose intake, and (C) carbonated drink intake, then they were centrifuged at 1,200 rpm followed by removing bacteria and enzymes with syringe filtering, performing a qualitative analysis with HPLC conductivity detection (GP50 gradient pump, ED 50 detector) after saliva pre-treatment under isocratic 100 mM NaOH mobile phase. Results: Higher risk of dental caries was evaluated in order of C>B>A, with the results of total oral organic acids' concentration, lactates of organic acids and organic acids produced after 5 minutes from the 3 types of drinks intake. Conclusions: Carbonated beverages were estimated to develop higher dental caries induction than beverages containing 10% sucrose because of the high organic acid concentration in the mouth after its intake.

Density Gradient를 이용한 식품소재를 커들란의 분리공정개발 (Development of Curdlan Separation Process with Density Gradient Centrfugation)

  • 김봉영;이중헌
    • KSBB Journal
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    • 제16권5호
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    • pp.523-525
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    • 2001
  • 커들란은 용액의 pH 7.0에서 salt 상태로 존재하기 때문에 커들란만이 액상에 녹아 있는 경우에는 분리공정은 원심분리를 함으로써 분리가 가능하다. 그러나, 세포외 다당류로 생산된 커들란읜 세포, 염 의 무기물이 섞여 있기 때문에 이러한 물질을 제거하는 것이 커들란 분리의 중요한 변수가 되었다. 커들란의 생산을 위한 분리에 관한 보고는 원심분리 방법을 이용한 것으로 알려져 있음 세포 및 불순물 등을 제거하여 순수한 커들란을 분리하기 위해 pH swing separation (pH가 높은 경우 커들란이 용액에 녹는 성질을 이용)을 이용하였으나 원심분리를 수 차례 반복해야 되기 때문에 소요되는 동력이 많이 필요하여 에너지 절감형 및 오염절감형의 새로운 공정의 개발이 필요하게 되었다. 온 연구에서는 pH swing separation 에서 동력비가 많은 드는 단점을 감소시키기위하여 커들란의 물리 화학적인 특성을 고려한 분리공정을 개발하였다. 향후 공정 개선을 위한 지속적인 연구가 필요하며 완성시 경제적이 공정을 개발하는 것이 가능할 것이다.

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해양에서 분리한 Vibrio parahaemolyticus Phage의 특성 (Characterization of a Vibrio parahaemolyticus Phage Isolated from Marine)

  • 윤선옥;주성아;허문수;정초록;주진우
    • 대한미생물학회지
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    • 제34권5호
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    • pp.423-433
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    • 1999
  • A novel bacteriophage, designated as VPP97, that infects the strains of Vibiro parahaemolyticus (hallophilic, Gram-negative bacterium) isolated most commonly from marine environments, has been discovered, and several of its properties have been determined. The plaques were clear and sized $0.6{\sim}1.0\;mm$ in diameter. The virion forms a single band on 70% sucrose gradient and ${\rho}1.50$ CsCl gradient by sucrose gradient centrifugation and CsCl gradient centrifugation respectively. It has a hexagonal head and a relatively long tail, as shown by electron microscopy. Vibrio alginolyticus, Vibrio fluvialis and Vibrio furnissii were also sensitive to this phage. It was almost totally inactivated at $70^{\circ}C$ and at pH below 5 or over 10. The nucleic acid of VPP97 is composed of DNA. The VPP97 had 9 specific structural proteins sized between 21.5 kDa and 97.4 kDa on SDS-PAGE. When V. parahaemolyticus cultures were treated with either phage VPP97 or one of the several antibiotics for 2 hours, the viable number of V. parahaemolyticus treated with the phage VPP97 is lower than that treated with chloramphenicol, erythromycin or penicillin, but not lower than that treated with tetracycline. Mice that have responded to the phage treatment revealed the lower numbers of V. parahaemolyticus in small intestine and less damage on small intestine compared to the untreated mice. Therefore, we suggest that the phage treatment appears effective to the infection by V. parahaemolyticus.

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