• Title/Summary/Keyword: high-speed centrifugation

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Removing Lipemia in Serum/Plasma Samples: A Multicenter Study

  • Castro-Castro, Maria-Jose;Candas-Estebanez, Beatriz;Esteban-Salan, Margarita;Calmarza, Pilar;Arrobas-Velilla, Teresa;Romero-Roman, Carlos;Pocovi-Mieras, Miguel;Aguilar-Doreste, Jose-Angel;Commission on Lipoprotein and Vascular Diseases, Sociedad Espanola de Quimica Clinica
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.518-523
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    • 2018
  • Background: Lipemia, a significant source of analytical errors in clinical laboratory settings, should be removed prior to measuring biochemical parameters. We investigated whether lipemia in serum/plasma samples can be removed using a method that is easier and more practicable than ultracentrifugation, the current reference method. Methods: Seven hospital laboratories in Spain participated in this study. We first compared the effectiveness of ultracentrifugation ($108,200{\times}g$) and high-speed centrifugation ($10,000{\times}g$ for 15 minutes) in removing lipemia. Second, we compared high-speed centrifugation with two liquid-liquid extraction methods-LipoClear (StatSpin, Norwood, USA), and 1,1,2-trichlorotrifluoroethane (Merck, Darmstadt, Germany). We assessed 14 biochemical parameters: serum/plasma concentrations of sodium ion, potassium ion, chloride ion, glucose, total protein, albumin, creatinine, urea, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase, aspartate-aminotransferase, calcium, and bilirubin. We analyzed whether the differences between lipemia removal methods exceeded the limit for clinically significant interference (LCSI). Results: When ultracentrifugation and high-speed centrifugation were compared, no parameter had a difference that exceeded the LCSI. When high-speed centrifugation was compared with the two liquid-liquid extraction methods, we found differences exceeding the LCSI in protein, calcium, and aspartate aminotransferase in the comparison with 1,1,2-trichlorotrifluoroethane, and in protein, albumin, and calcium in the comparison with LipoClear. Differences in other parameters did not exceed the LCSI. Conclusions: High-speed centrifugation ($10,000{\times}g$ for 15 minutes) can be used instead of ultracentrifugation to remove lipemia in serum/plasma samples. LipoClear and 1,1,2-trichlorotrifluoroethane are unsuitable as they interfere with the measurement of certain parameters.

STUDY ON PLATELET RICH PLASMA CONCENTRATION ACCORDING TO PROCESSING METHOD (처리방법에 따른 혈소판 혈장의 농축도에 관한 연구)

  • Min, Seung-Ki;Kim, Hyung-Ju;Cha, Soo-Ryen
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.1
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    • pp.24-31
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    • 2005
  • Recently, Platelet rich plasma(PRP) is commonly used because it is now well known that platelets have many functions beyond that of simple hemostasis in aspect of containing autogenous source of several growth factors. It could be responsible for increasing cell mitosis, increasing collagen production, recruiting other cells to the site of injury, initiating vascular ingrowth, and inducing cell differentiation, enhancing bone formation capacity and easily handling to clinician. However, in spite of these clinical advantages, still the theory behind the use of PRP is compelling. This study was to determine preparation techniques used to increase the concentration of platelets and growth factors are all crucial steps in early wound healing of bone graft which may lead to a more rapid and denser bone regenerate. 200 volunteers were sampled and PRP were prepared according to each evaluation item in this study. Higher concentration of platelets have been gained in double centrifugation. 2000 and 2500 rpm showed proper concentration of platelets at first centrifugation and 5000 rpm in second. Timing for 2 minutes was showed good concentration of platelets in high and low centrifugation speed. It was better concentration of platelets in 20 or 30 ml volume during centrifugation. In histomorphologic findings, degrnulated and high concentraion of platelets were found in low centrifugation speed.

The Cell Survival and Differentiation after Transplantation, Which Harvest from Adult Rat Brain by High-speed Centrifugation Method

  • Kim, Jong-Tae;Yoo, Do-Sung;Woo, Ji-Hyun;Huh, Pil-Woo;Cho, Kyung-Sock;Kim, Dal-Soo
    • Journal of Korean Neurosurgical Society
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    • v.38 no.2
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    • pp.121-125
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    • 2005
  • Objective : Many recent reports have shown that the mature mammalian brain harbors multipotent stem cells, rendering the brain capable of generating new neurons and glia throughout life. Harvested stem cells from an adult rat are transplanted in order to evaluate the cell survival and differentiation. Methods : Using a percoll gradient with a high speed centrifugation method, we isolate neural stem/progenitor cells were isolated from the subventricular zone[SVZ] of a syngeneic adult Fisher 344 rats brain. For 14days expansion, the cultured cells comprised of a heterogeneous population with the majority of cells expressing nestin and/or GFAP. After expanding the SVZ cells in the presence of basic fibroblast growth factor-2, and transplanting then into the hippocampus of normal rats, the survival and differentiation of those cells were examined. For transplantation, the cultured cells were labeled with BrdU two days prior to use. In order to test their survival, the cells were transplanted into the dorsal hippocampus of normal adult Fisher 344 rats. Results : The preliminary data showed that at 7days after transplantation, BrdU+ transplanted cells were observed around the injection deposition sites. Immuno-fluorescent microscopy revealed that the cells co-expressed BrdU+ and neuronal marker ${\beta}$-tubulin III. Conclusion : The data demonstrate that the in vitro expanded SVZ cells can survive in a heterotypic environment and develop a neuronal phenotype in the neurogenic region. However more research will be needed to examine the longer survival time points and quantifying the differentiation in the transplanted cells in an injured brain environment.

DILUTION AND SHORT-TERM STORAGE OF COCK SPERMATOZOA BY INHIBITION OF MOTILITY WITH FRUCTOSE AT AMBIENT TEMPERATURE

  • Mohan, J.;Moudgal, R.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.705-709
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    • 1996
  • A simplified dilutor for cock spermatozoa at ambient temperature was achieved by adjusting the 5% concentration of fructose in isotonic saline. Motility of cock spermatozoa was arrested completely for maximum 6 hrs without affection the survivability of spermatozoa by employing this sugar. To study the effect of high concentration of fructose on fertility, sperm were inseminated with or without fructose at different hrs. Fructose from semen samples was removed by centrifugation. High fertility obtained in the hens inseminated with fructose free sperm (washed). In addition, washed sperm maintained the 85.00% fertility for 6 hrs in winter season ($17-21^{\circ}C$) and 82.67% fertility for 3 hrs in summer season ($31-35^{\circ}C$). Whereas control groups showed 47.33 and 25.33% fertility in winter and summer season respectively. No significant difference was found in percent motility and live counts between the control and washed experimental groups during winter season. However, these measures differed significantly in summer. Washing of cock spermatozoa more than once, high speed centrifugation and more duration for centrifugation proved harmful to fertility. It may be concluded that fructose (5%) can be used as a motility or metabolic inhibitor of spermatozoa for short-term storage of cock semen at ambient temperatures.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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The Production of Biopolymer by Zoogloea ramigera (Zoogloea ramigera에 의한 생물고분자 생산에 관한 연구)

  • 안대희;권해수정윤철
    • KSBB Journal
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    • v.7 no.3
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    • pp.166-171
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    • 1992
  • Zoogloea ramigera 115 was cultured for biopolymer production and its bioflocculant usages. Cultural conditions of the organism were examined with regard to high production of the microbial polysaccharide. The most suitable medium was found to contain glucose as a carbon source, $NaNO_3$ as a nitrogen source, and yeast extract as an organic nutrient. The initial pH of 6.0 proved to optimal. The biopolymer was extracted effectively using ultrasonication and high speed centrifugation, followed by propanol addition. Jar test results indicate that the polysaccharide produced by the organism is an effective flocculant.

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Clarification of Mixed Fruit and Vegetable Juices Using Various Clarification Methods (혼합과채주스의 청징 및 품질특성 비교)

  • Sohn, Kyung-Suck;Lee, Jun-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.5
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    • pp.629-634
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    • 2006
  • Clarified mixed fruit and vegetable (apple, carrot and orange) juices were prepared using different clarification methods including centrifugation, ultrafiltration (UF), and combined treatment with the blend ratio of 1:1:2, 1:2:1, and 2:1:1 (apple, orange and carrot). Effects of clarification methods on the clarification efficacy as well as other quality parameters were investigated. Clarification was improved with increase in centrifugation speed but was less affected by lowering temperature. Ultrafiltration process was very effective to produce clarified mixed fruit and vegetable juice. Combined treatment did not significantly improve the clarification efficacy since most clarification was already achieved during ultrafiltration process (p>0.05). L*- and b*-values increased while a*-value decreased significantly after clarification regardless of methods in all blend juices (p<0.05). Blend juices with high amount of orange or carrot were better in clarification efficacy and high amount of apple resulted in high turbidity in blend samples.

Evaluation and modification of alkaline lysis plasmid preparation method from Lactobacillus spp.

  • Lee, Deog-Yong;Seo, Yeon-Soo;Kang, Sang-Gyun;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.157-162
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    • 2007
  • Lactic acid bacteria (LAB) has been regarded as a useful microorganism and tried to manipulate plasmid DNA for increasing the usefulness. Although several methods have been developed to isolate plasmid DNA from Escherichia coli (E. coli), these methods were not sufficient to apply to LAB with exception of O'Sullivan's lysis method. So, we evaluated plasmid DNA extraction from LAB using general E. coli preparation methods and tried to improve the extraction yield and DNA purity by modifying O'Sullivan's alkaline lysis method. To improve the extraction yield, salt and carrier were added to precipitant and those were incubated at $-70{^{\circ}C}$. Only incubation at $-70{^{\circ}C}$ was the effective method of those modifications. Purity of plasmid DNA was improved by two times of each centrifugation and phenol/chloroform extraction. However, DNA was damaged by twice extraction with phenol/chloroform. Also, exclusion of ethidium bromide showed negative effect to purity. Additionally, it was recommended that improvement of the extraction yield may be due to centrifugation at high speed for more time and to dissolving complete DNA pellet before addition of 7.5 M ammonium acetate. Extraction using this modification produced higher quality of plasmid DNA.

The Effect of the Aging of Red Blood Cells on Rheological Properties and Hemolysis

  • Tomioka, Jun;Motokubo, Kazuhiro;Watanabe, Hisayoshi
    • Proceedings of the Korean Society of Tribologists and Lubrication Engineers Conference
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    • 2002.10b
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    • pp.371-372
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    • 2002
  • It is well known that red blood cells (RBCs) are suffered from chronic stresses in systemic circulation. The objective of this study is to clarify the effect of the aging of RBCs on rheological properties and hemolysis. Initially, RBCs age fractionation was performed by using a high-speed centrifugation (15[min] at 1500[G]), then young and aged RBCs were suspended in plasma to adjust the hematocrit level of 40[%]. After this pretreatment, the viscosity was measured by using a capillary type and a cone-plate type viscometers, respectively, and the hemolysis test was carried out by a seesaw type shaker. Results from these experiments showed that the viscosity of the aged RBCs measured by the capillary viscometer was increased by 10[%] as compared with that of the young RBCs. Under the condition of all shear zones, the viscosity of the aged RBCs was increased in case of using the cone-plate type viscometer. And the hemolytic level was increased twice as the aging. The data obtained in this study indicated that the ability of aggregation of RBCs was increased and the deformability of RBCs membrane got lower with the aging. Furthermore, it was exhibited that the fragility of RBCs ’ membrane was increased with the aging.

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STUDIES ON THE VIRUSES OF RADISH MOSAIC

  • KIM, Woon-Soo
    • Journal of Plant Biology
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    • v.6 no.2
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    • pp.9-21
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    • 1963
  • A mosaic diseased radish collected from the suburb of Seoul, in November, 1961 was used for studing the host range, physical properteis, purificaitiion, insect transmission, and electron microscopy. A Japanese strain of radish mosaic(RPV) was also used with Korean strain of radish mosaic (KRMV) for a comparative study. The two viruses, KRMA and RPV, were identified by the difference in host range, insect transmission and electron microscopy. The KRMA was severely infective on tobacco and Nicotiana glutinosa, while on Gomphrena globsa was immune to the virus. RPV produces necrotic local lesions on Gomphrena globosa but did not infect tobacco and N. glutinosa. Among varieties of radish, Seoul, Akamaroo, Akanagea, Koong-Joong showed more severe symptoms than Simoo, Minong, Paek-soo, which appeared to be fainly resistant. In a number of tests, it was found that the virus KRMA retained its infectivity until to a dilution of 1:2,000, heating at $58^{\circ}$ for 10 minutes, adn aging in vitro for 7 days at room temperature. The RPV was not inactivated until it was diluted to 1:2,000, heated to $56^{\circ}$, and aged for 6 days. The KRMV was readily transmitted by the aphid(Myzus persicae Sulz). The virus RPV was not transmitted by the aphid in a number of tests. Partialy purified viruses using ammonium acetate buffer, salting-out by ammonium sulfate and centrifugation of high and low speed were highly infective. Electron micrographs showed that the KRMV paticles are of spherical particles whereas the RPV particles are rod-shaped.

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