• 제목/요약/키워드: Percoll

검색결과 96건 처리시간 0.029초

적조 와편모조류, Cochlodinium polykrikoides의 순수분리 및 성장 (Axenic Culture Production and Growth of a Dinoflagellate, Cochlodinium polykrikoides)

  • 서필수;이상준;김윤;이정호;김학균;이재동
    • 한국수산과학회지
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    • 제31권1호
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    • pp.71-76
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    • 1998
  • C. polykrikoides의 항생제에 대한 특이성을 알아보기 위해 C. polykrikoides의 항생제에 대한 $50\%$ 생존시간을 조사하였다. C. polykrikoides는 tetracycline과 thlorarnphe-nicol에 대해서는 아주 민감하였고, polymixin-B, ampicil-lin, penicillin-G, dihydrostreptomycin, neomycin에 대 해 서 는 저항성이 큰 선택적 특이성을 가졌다. C. polykrikoides에 민감한 항생제인 tetracycline과 chloramphenicol의 경우는 사멸곡선을 분석하여 사멸에 영향을 주지 않는 농도를 확인했으며, 또한, 농도에 따른 C. polykrikoidess에 대한 항생제의 작용특성을 분석하였다. 박테리아에 오염된 C. polykrikoides 배양액을 percoll 원심분리로 박테리아를 제거하기 전에 C. polykrikoides에 저항성이 큰 항생제에 대한 박테리아 군집의 감수성 시험을 행하여 박테리아 제거를 위한 적당한 항생제와 농도를 결정 한 뒤, percoll 원심분리로 박테리아와 곰팡이를 제거하고, 조체를 회수한 후, 남아있는 소량의 박테리아를 감수성이 큰 항생제를 연속적으로 처리 (antibiotic cascade)하여 C. polykrikoides의 순수배양(axonic culture)이 가능하게 되었다. 순수배양한 C. polykrikoides은 밀도가 약 740cells/ml 이상에서 대수증식기에 접어들었고 최대 5,800cells/ml까지 증식하였다. C. polykrikoides의 하루당 분열율 k 값은 저밀도 세포 상태에서 아주 정확한 증식지표를 제시하였고, 대수증식기 전에 반드시 최대의 k 값 변화를 가진 세포의 밀도가 존재하였으며, 우리는 이 최대 k 값 변화를 가진 밀도를 정확한 적조 예보와 초기에 적조제어를 위한 모델로서 사용될 수 있음을 제시한다. 결론적으로 본 연구에서 확립된 C. polykrikoides의 순수분리에 의한 배양방법은 C. polykrikoides 뿐만 아니라 다른 적조 생물의 독성학적, 면역학적, 생화학적, 생태학적, 분자생물학적 연구를 위해 편리하게 이용할 수 있을 것이다.

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Efficiency of Phototransduction Cascade in Carp Cones

  • Tachibanaki, Shuji;Tsushima, Sawae;Kawamura, Satoru
    • Journal of Photoscience
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    • 제9권2호
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    • pp.44-46
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    • 2002
  • In the vertebrate retina, rods mediate twilight vision and cones daylight vision. Rods have been purified easily from the retina, and thus the phototransduction mechanism in rods is now well documented. However, it has not been possible to purify cones in large quantities, and therefore, the knowledge on the mechanism in cones is limited. Here we report purification of carp (Cyprinus carpio) cones with a stepwise Percoll gradient. Using purified cells, we compared the phototransduction mechanism between rods and cones. The results showed that both transducin activation and phosphodiesterase activation are less effective, and visual pigment phosphorylation is faster in cones. These differences explain lower light-sensitivity and briefer photoresponse time course in cones.

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Theiler's virus 에 감염된 마우스의 척수 신경교세포배양과 면역세포학적 관찰 (Culture of glial cells isolated from the spinal cord of demyelinating mice infected with Theiler's virus:An immunocytochemical study)

  • 신태균
    • 대한수의학회지
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    • 제31권2호
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    • pp.155-161
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    • 1991
  • The mechanisms of demyelination in Theiler's murine encephalomyelitis virus (TMEV)-induced chronic central nervous system(CNS) disease are still unclear and are probably multifactoral. This study was intended to culture spinal cord cells isolated from TMEV-induced demyelinating mice. By Percoll density centrifugation of enzymatically dissociated tissue, the cells were collected and then cultured on poly-L-lysine-coated plastic coverslips for 2 weeks. Oligodendrocytes, astrocytes and macrophages were identified using cell-type specific markers. Viral antigens were not present in oligodendrocytes and in astrocytes by double immunofluorescence. Affected mouse oligodendrocytes had less capacities of sheet formation and galactocerebroside immunoreactivity than those of control cell 3. These findings support the hypothesis that immune mediated mechanisms play an important role in the process of demyelination in this animal model.

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Sprague-dawley(SD) 랫드에서 natural killer cell의 분리 ·동정 및 형태적 특징 (Isolation and morphological characterization of natural killer cell in the sprague-dawley(SD) rats)

  • 강경선;이영순
    • 대한수의학회지
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    • 제32권2호
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    • pp.245-250
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    • 1992
  • This study was performed to demonstrate the presence of large granular lymphocyte(LGL) in Sprague-Dawley(SD) rats and morphologically observe NK cell and also establish the method of isolation of natural killer cell in SD rats. By percoll discontinuous density gradients centrifugation, highly enriched LGL population were shown to fraction 2(border line between 44.2% and 50.8%). LGL were shown to bind selectively to YAC1 mouse lymphoma cell. This fraction expressed very high NK cell cytolysis. Therefore, we thought that LGL have NK activity in SD rats. The Morphology of rat LGL is very similar to that of human LGL. These cells have an eccentric kidney-shaped nucleus. Their most distinctive feature was their cytoplasmic azurophilic granules. Another distinguishing feature of rat LGL was their high cytoplasmic : nuclear ratio. It was concluded that LGL played a role part in mediating natural killer activity in this species.

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Inhibitory Action of Phenylpropanoids on Histamine Release from Rat Peritoneal Mast Cells

  • Lee, Jin-Hee;Lee, Ji-Yun;Kim, Youn-Joung;Kim, Tae-Doo;Yoon, Mi-Yun;Sim, Sang-Soo;Kim, Chang-Jong
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.194.1-194.1
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    • 2003
  • Phenylpropanoids originating from vegetable kingdom have some biological activity. In this experiments, effect of phenylpropanoids on the histamine release from mast cells were studied in vitro. Rat peritoneal mast cells were isolated by the discontineous gradients of Percoll and their histamine release by stimulation of compound 48/80 and A23187 at a concentration of 6.0 $\mu\textrm{g}$/$m\ell$ were determined. (omitted)

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The Use of Bull Round Spermatids for Producing Reconstructed Embryos

  • S.A. Ock;D.O. Kwack;Park, G.J.;S.Y. Choe
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.133-133
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    • 2003
  • Recently, sperm has been used as a vector to carry exogenous genes for the production of transgenic animals. However, the success in cattle is low, due to deficiencies in oocyte activation and sperm decondensation caused by high disulphide bond (S=S) content in mature sperm. This study was carried out to develop an effective method for producing transgenic animals with round spermatids (RS). Two methods of embryo production - electric fusion (EC) or intracyto-plasmic injection (IC) and three activation treatments were compared. RS were isolated from bull testes by Percoll density gradients (20, 35, 40, 45 and 90%). Fusion between ooplast and RS was performed with a single DC electric pulse (1.0 KV/cm, 45 sec) in 0.28 M mannitol solution supplemented with 100 M CaCl2 and 100 M MgCl$_2$. (중략)

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신장 피질 기저막 소포에서 p-Aminohippurate 이동의 특성에 대한 연구 -음이온의 효과- (Characteristics of p-Aminohippurate (PAH) Transport in Basolateral Membrane Vesicles of Rabbit Kidney Cortex -Effect of Anion-)

  • 이상호;정진섭;김용근
    • The Korean Journal of Physiology
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    • 제20권2호
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    • pp.225-235
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    • 1986
  • 가토 근위세뇨관에서 Percoll gradient에 의해 분리한 기저막소포(basolateral membrane vesicle)에서 PAH 이동에 미치는 여러 음이온들의 영향을 rapid filtration technique을 이용하여 관찰하였다. NA-dependent PAH 축적은 용액내 $SO_4$$SSO_3$에 의하여 유의하게 억제되었으나 $Cl,\;SCN,\;HPO_4,\;acetate$ 및 oxalate에 의해서는 영향을 받지 않았다. 용액내의 $SO_4$는 상경적으로 PHA 이동을 억제하였다. 소포내 음이온을 부하시킨 후 PAH 이동을 측정했을 때 $acetate,\;SO_4$ 및 PAH의 부하에 의해 PAH 이동은 유의하게 증가되었으며, 소포내 부하되는 $SO_4$농도가 증가함에 따라 PAH 축적은 증가되었다. $SO_4$에 의한 PAH의 trans-stimulation은 Na 농도 경사 존재시 더욱 증가되었으며 이들은 2 mM probenecid 및 1mM SITS에 의해 억제되었다. 이들 결과들은 신피질세뇨관 세포의 기저막에 PAH/음이온 교환에 의해 PAH가 이동되는 기전이 존재한다는 것을 가르킨다. 그러나 PAH 이동에 영향이 없는 음이온들은 PAH/음이온 교환에 기질로써 작용하지 않기 때문인지 아니면 이들 음이온들의 높은 투과성에 인해 나타난 결과인지는 더욱 추구해 보아야 확인될 것으로 생각된다.

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Inhibititory Activity of Flavonoid Acetates on the Histamine Release from IgE-Sensitized Mast Cells

  • Shin, Wha-Woo;Kim, Chun-Re;Choi, Se-Woong;Kim, Sung-Jin;Cho, Seung-Kil;Kim, Chang-Jong
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1996년도 춘계학술대회
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    • pp.255-255
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    • 1996
  • Flavonoids are the large class of plant-derived polyphenolic compounds with a broad spectrum of biological actions including anti-inflammatory, anti-allergic and anti-asthmatic activities. It has been reported that quercetin and some other flavonoids inhibit the histamine release from mast cells. Disodium cromoglycate, a kind of biflavonoid, have been used as a anti-asthmatic agent. Therefore, the following study was undertaken to characterize tile structure-activity relationships of flavonoids in the inhibition of histamine release, from IgE-sensitized mast cells. Flavonoid acetate were synthesized by addition of acetic anhydride to the flavonoids dissolved in pyridine. According In the Mota method, IgE-rich serum was made in rats sensitized with Pertussis vaccine and ovalbumin. Rat abdominal mast cells were harvested and purified over tile discontineous Percoll gradients, Mast cells were preincubated in RPMI with IgE-rich serum and further incubated with flavonoids. Histamine contents released from mast cells were determined fluorospectrophotometrically after stimulation with ovalbumin. (omitted)

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Change in Cell Size and Buoyant Density of Pseudomonas diminuta in Response to Osmotic Shocks

  • Lee, So-Hee;Cho, Yu-Ree;Choi, Yong-Jin;Kim, Chan-Wha
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.326-328
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    • 2001
  • Pseudomonas diminuta (ATCC 19146) has been typically used in the bacterial challenge test for validation of the sterilizing filtration process. Cell size is critical for determining the retention characteristics of membrane filters with pore-size of $0.2{\mu}m$. The changes of cell sizes after osmotic shocks at 150, 260, 500, and 700 mosM were measured by a particle size analyzer and the changes of their buoyant densities were analyzed with a Percoll gradient. The results indicated that there were no significant differences when cells were cultured in 260 mosM medium and osmotically shocked at 500 and 700 mosM. However, the osmotically shocked cells at 150 mosM showed a 38% increase of the cell size compared to the cells at 260 mosM. From these study, we concluded that the worst case condition for validation of a sterilizing filter would be 500 mosM, not because of changes in the cell size, but due to decrease in cell viability under those conditions.

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체외생산 소 초기배 할구세포의 Embedding Matrix에서의 발생능력 (Development Ability of Bovine Early Embryo Blastomere In Vitro in Embedding Matrix)

  • 이홍준;서승운;이상호;송해범
    • 한국수정란이식학회지
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    • 제10권2호
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    • pp.171-175
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    • 1995
  • This study was performed to establish the condition and the methods for the techniques of insertion the isolated blastomere cells into cytoplasm, in order to research the develop-mental ability of bovine embryo blastomere cells in vitro produced. After 24h in vitro ovary maturation with the ovaries from a slaughter house, in vitro fertilization was performed to the vital sperms which their mobility were decided by percoll gradient method, with 2~8 cell stage embryos, the blastomeres were isolated in $Ca^2$+. $Mg^2$+-free PBS, and following that embedded into agar and alginate solution, respectively. The rates of in vitro develop-ment are as follows ; in agar embedded 11 among 120(9.2%) 1 /2~1 /3 blastomers cleaved and 6 among 93(6.5%) 1 /4~1 /8 blastomeres cleaved. In sodium alginate-embedded 14 among 84(16.7%) 1 /2~1 /3 blastomeres cleaved and 6 among 85(7.1%) 1 /4~1 /8 blastomeres cleaved. In case of Na-alginate, the rate of the cells were better than those of agar. The results suggest that the techniques for embeeding the isolated blastomeres into gel may help cloning of bovine early embryo without nuclear transplantation.

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