• Annals of Laboratory Medicine
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• 제38권6호
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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.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권1호
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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.

• Journal of Korean Neurosurgical Society
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• 제38권2호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제9권6호
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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.

• 핵의학기술
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• 제18권1호
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• pp.153-157
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• 2014

Anti double-stranded DNA (Anti-ds DNA)항체의 검출은 SLE의 진단에 중요하며 American College of Rheumatologists의 SLE 진단기준에 포함되어 있다. 또한 SLE 질병의 활성도와 Anti-ds DNA 항체 수준의 상관성이 보고되어 있으며 Anti-ds DNA 항체 정량검사는 SLE의 치료 전, 후 추적에 매우 유용하다. Anti-ds DNA 항체의 검출을 위한 검사 방법으로 방사면역측정법(radioimmunoassay, RIA)을 이용한 Farr assay, Crithidia luciliae를 이용한 면역형광 측정법(immunofluorescence Test, CLIFT), 효소면역측정법(enzyme-linked immunosorbent assay, ELISA), 화학발광면역 측정법(chemiluminescence immunoassay, CLIA)이 있다. 본원에서 방사면역측정법(radioimmunoassay, RIA)으로 Anti-ds DNA 항체 검사 과정에서 lipemic한 검체의 경우 침전물의 형성이 원활하지 않거나 침전물도 같이 흡입되는 상황이 발생 하였고, 이러한 문제점을 해결하기 위해 lipemic 검체가 분석 결과에 미치는 영향 정도를 평가하였다. 2012년 9월부터 2013년 2월까지 Anti-ds DNA 항체 검사가 의뢰된 검체 중 lipemic한 검체(n=81)를 선택하여 마이크로 원심분리기로 전처리(고속 원심분리: 14,000 rpm 5 mins)한 후 동시에 Anti-ds DNA 항체 검사(Anti-ds DNA kit, Trinity Biotech, Ireland)를 시행하였다. 실험군 1 (lipemic 검체의 Anti-ds DNA 항체 농도 ${\leq}7IU/mL$)에서 y=0.3636x+4.7322, $R^2=0.0238$, Pearson 상관계수는 0.154, paired t-test (P=0.003), Difference (%) mean 65.7의 결과를 얻었으며 통계적으로 유의한 차이를 보였다. 그러나 실험군 2 (lipemic 검체의 Anti-ds DNA 항체 농도 ${\geq}8IU/mL$)에서 y=0.9837x+0.2982, $R^2=0.994$, Pearson 상관계수는 0.997, paired t-test (P=0.181), Difference (%) mean -5.53을 보였고 통계적으로 유의한 차이가 없음을 확인할 수 있었다. 임상에서 SLE (systemic lupus erythematosus)의 진단에 중요한 역할을 하는 Anti-ds DNA 항체 검사는 lipemic 검체의 영향을 배제하기 위해서 반드시 전처리(고속 원심분리: 14,000 rpm 5 mins) 과정을 통해 혈청 내 지질 성분을 제거한 후 검사를 시행하여야 한다.

• ALGAE
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• 제39권2호
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• pp.109-127
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• 2024

High production and efficient harvesting of microalgae containing high omega-3 levels are critical concerns for industrial use. Aeration can elevate production of some microalgae by providing CO₂ and O₂. However, it may lower the production of others by generating shear stress, causing severe cell damage. The mixotrophic dinoflagellate Gymnodinium smaydae is a new, promising microalga for omega-3 fatty acid production owing to its high docosahexaenoic acid content, and determining optimal conditions and methods for high omega-3 fatty acid production and efficient harvest using G. smaydae is crucial for its commercial utilization. Therefore, to determine whether continuous aeration is required, we measured densities of G. smaydae and the dinoflagellate prey Heterocapsa rotundata in a 100-L semi-continuous cultivation system under no aeration and continuous aeration conditions daily for 9 days. Furthermore, to determine the optimal conditions for harvesting through centrifugation, different rotational speeds of the continuous centrifuge and different flow rates of the pump injecting G. smaydae + H. rotundata cells into the centrifuge were tested. Under continuous aeration, G. smaydae production gradually decreased; however, without aeration, the production remained stable. Harvesting efficiency and the dry weights of omega-3 fatty acids of G. smaydae + H. rotundata cells at a rotational speed of 16,000 rpm were significantly higher than those at 2,000-8,000 rpm. However, these parameters did not significantly differ at injection pump flow rates of 1.0-4.0 L min^-1. The results of the present study provide a basis for optimized production and harvest conditions for G. smaydae and other microalgae.

• KSBB Journal
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• 제7권3호
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• pp.166-171
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• 1992

생물응집체 개발을 목적으로 Zoogloea ramigera115를 이용하여 균주배양 및 생물고분다 생성실험을 시시하였다. 다당류 최고 생성률을 얻을 수 있는 최적기질로 탄소원으로는 glucose, 질소원으로는 $NaNO_3$ 유기영양원으로는 yeast extract가 선택되었다. 초기 pH가 6일 때 가장 높은 세포성장과 점성을 나타내었다. 생성돈 생물고분자는 ultrasonication과 고속 원심분리를 거친 후 propanol 첨가에 의해 효율적으로 추출되었다. 생물고분자는 jae test에 의한 응집실험에서 높은 침강성을 보여 응집제 사용가능성을 확인하였다.

• 한국식품영양과학회지
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• 제35권5호
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• pp.629-634
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• 2006

혼합과채 청징주스 제조를 위한 실험적 기초자료를 제시하기 위하여 원심분리, 한외여과처리, 혼합처리를 이용하여 주스를 제조하고 청징방법에 따른 청징효과 및 품질특성을 비교하였다. 원심분리를 이용하여 혼합과채주스를 청징하였을 경우 회전속도가 5,000에서 10,000 rpm으로 증가함에 따라 청징효과가 30% 이상 개선되었으나 온도에 대한 영향은 단일주스와는 달리 미미하였다. 한외여과를 이용한 경우 모든 시료에서 탁도값의 범위가 $0.001{\sim}0.003$으로 나타나 공정조건에 관계없이 청징효과가 매우 우수한 것으로 나타났다. 한외여과의 경우 탁도 이외에 투과플럭스를 고려하여 볼 때 최적 공정조건은 150 kPa, $45^{\circ}C$로 나타났다. 혼합처리할 경우 한외여과 경우와 유사하게 모든 혼합시료에서 99% 이상의 청징 개선효과를 얻을 수 있었는데, 이는 한외여과공정을 통하여 이미 대부분의 청징이 이루어졌음을 의미한다. 청징방법에 따른 색도 변화에서 L*값은 모든 시료에서 대조구보다 현저하게 증가했으며 a*값 및 ${\Delta}E$값은 한외여과 시료에서 높았으며, b*값은 원심분리 시료에서 가장 높게 나타났다. 가용성 고형분은 원심분리된 시료에서 가장 많이 보존되었으나 모든 시료에서 청징 후 감소하는 경향을 보였으며 청징공정에 따른 pH 변화는 적은 것으로 나타났다.

• 대한수의학회지
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• 제47권2호
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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.

• 한국윤활학회:학술대회논문집
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• 한국윤활학회 2002년도 proceedings of the second asia international conference on tribology
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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.