• 제목/요약/키워드: continuous centrifugation

검색결과 7건 처리시간 0.025초

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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농도분배에 따라 분리한 사람 적혈구 막에 관한 연구 (Studies of Density-Fractionated Human Ervthrocvte Membranes)

  • 정종문
    • 한국동물학회지
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    • 제37권4호
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    • pp.597-604
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    • 1994
  • Membranes obtained from the normal human RBC population were separated by continuous sucrose density gradient centrifugation and the density-fractionated membranes were then examined for changes in molecular markers. This study focuses on changes of (i) the membrane protein profile, (ii) differences in membrane-associsted enzyme activities, and (iii) the amount of autologous IgG bound. The following observations were made: (i) ratios for band 4. la over the sum of bands (4. la + 4.Ib) ranged from 0.58 to 0.79 for membranes of lowest density; (ii) significant changes in bound glyceraldehyde-3-phosphate dehydrogenase and acetvlcholinesterase activities were found; (iii) the amounts of autolosous IgG's attached to the red blood cells was highest in the membrane fraction of lowest density.

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해바라기 유(油)의 탈납 (Dewaxing of Sunflower Seed Oil)

  • 이준식
    • 한국식품과학회지
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    • 제11권2호
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    • pp.112-117
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    • 1979
  • 해바라기유의 재래식 탈납공정 경제성을 높이기 위해서 여러가지 유화제(0.2% sodium bexametaphosphate, 0.05% sodium lauryl sulfate, 0.001% dioctyl sodium sulfosuccinate)를 이용한 새로운 탈납공정을 개발하였다. 해바라기 원유를 회분식 (batch process)과 연속식(continuous process)으로 유화제 수용액과 유화시킨후 원심분리기로 분리시켰을 때 재래식 탈납공정에 비견할 만한 탈납을 할 수있음을 알았다. 탈납 공정에서 위에 설명한 탈납정도(degree of dewaxing)외에 또 하나의 중요인자는 수율(dewaxirg loss)인바 회분식 공정의 경우에는 재래식 탈납공정의 수율과 같았으나 연속식 공정의 경우에는 개량할 여지가 있음을 알았다. 실험결과에 의하면 회분식 공정의 경우 탈납전 원유에 함유된 납(wax)의 다과에 관계없이 수율은 $0.62{\sim}0.82%$로 재래식 공정과 같았으나 연속식 공정인 경우에는 탈납전 원유에 함유된 납의 양이 적은 경우와 많은 경우 각각 2.28%와 5.68%로 현격한 차이를 보였다. 또한 연속식 공정의 경우 원유와 유화제를 유화시키는 방법에 따라 수율에 많은 영향이 미치는 것으로 밝혀졌으며 contactor와 static mixer를 썼을때 각각 5.2%와 2.8%이었다.

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Autocrine Regulation of Gonadotropin-releasing Hormone (GnRH) Operates at Multiple Control levels of GnRH Gene Expression in GT1-1 Neuronal Cells

  • Jin Han;Sehyung Cho;Woong Sun;Kyungjin Kim
    • Animal cells and systems
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    • 제2권4호
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    • pp.483-488
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    • 1998
  • We previously found that a potent gonadotropin-releasing hormone (GnRH) agonist, buserelin, decreases GnRH promoter activity together with GnRH mRNA level, providing evidence for an autoregulatory mechanism operating at the level of GnRH gene transcription in immortalized GT1-1 neuronal cells. To examine whether agonist-induced decrease in GnRH mRNA level requires the continuous presence of buserelin, we performed a pulse-chase experiment of buserelin treatment. Short-term exposure (15 min) of GT1-1 neuronal cells to buserelin ($10{\mu}M$) was able to decrease GnRH mRNA levels when determined 24 h later. When GT1-1 cells were treated with buserelin ( $10{\mu}M$) for 30 min and then incubated for 1, 3, 6, 12, 24, and 48 h after buserelin removal, a significant decrease in GnRH mRNA levels was observed after the 12 h incubation period. These data indicate that inhibitory signaling upon buserelin treatment may occur rapidly, but requires a long time (at least 12 h) to significantly decrease the GnRH mRNA level. To examine the possible involvement of de novo synthesis and/or mRNA stability in buserelin-induced decrease in GnRH gene expression, actinomycin D ($5{\mu}m/ml$), a potent RNA synthesis blocker, was co-treated with buserelin. Actinomycin D alone failed to alter basal GnRH mRNA Revel, but blocked the buserelin-induced decrease in GnRH mRNA level at 12 h of post-treatment. These data suggest that buserelin may exert its inhibitory action by altering the stability of GnRH mRNA. Moreover, a polvsomal RNA separation by sucrose gradient centrifugation demonstrated that buserelin decreased the translational efficiency of the transcribed GnRH mRNA. Taken together, these results clearly indicate that GnRH agonist buserelin acts as an inhibitory signal at multiple levels such as transcription mRNA stability, and translation.

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Full validation of high-throughput bioanalytical method for the new drug in plasma by LC-MS/MS and its applicability to toxicokinetic analysis

  • Han, Sang-Beom
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2006년도 추계학술대회
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    • pp.65-74
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    • 2006
  • Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

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Ginsenoside Re Enriched Fraction (GS-F3K1) from Ginseng Berries Ameliorates Ethanol-Induced Erectile Dysfunction via Nitric Oxide-cGMP Pathway

  • Pyo, Mi Kyung;Park, Kwang-Hyun;Oh, Myeong Hwan;Lee, Hwan;Park, Young Sik;Kim, Na Young;Park, So Hee;Song, Ji Hye;Park, Jong Dae;Jung, Se-Hee;Lee, Bong-Gun;Won, Beom Young;Shin, Ki Young;Lee, Hyung Gun
    • Natural Product Sciences
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    • 제22권1호
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    • pp.46-52
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    • 2016
  • Erectile dysfunction (ED) is a highly prevalent disorder that affects millions of men and considered to be an early symptom of atherosclerosis and a precursor of various systemic vascular disorders. The aim of the present study was to prepare ginsenoside Re enriched fraction (GS-F3K1, ginsenoside Re 10%, w/w) from ginseng berries flesh and to investigate the enhanced activities of GS-F3K1 on alcohol-induced ED. GS-F3K1 was prepared by the continuous liquid and solid separating centrifugation and circulatory ultrafiltration from ginseng berries flesh. GS-F3K1 was administered for 5 weeks in ethanol-induced ED rat by oral administration of 20% ethanol. To investigate the effects of GS-F3K1 on ED model, the levels of nitrite expression, cyclic guanosine monophosphate (cGMP) and erectile response of the penile corpus cavernosum of rat were measured. The erectile response of the corpus cavernosum was restored after GS-F3K1 administration, to a level similar to the normal group. The level of nitrite and cGMP expression in the corpus cavernosum of GS-F3K1-administered male rats was increased significantly compared to positive control group. GS-F3K1 from ginseng berries should effectively restore ethanol-induced ED in male rats and could be developed as a new functional food for the elderly men.