• Title/Summary/Keyword: cell lysis rate

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A Continuous Electrical Cell Lysis Chip using a DC Bias Voltage for Cell Disruption and Electroosmotic Flow (한 쌍의 전극으로 전기 삼투 유동과 세포 분쇄 기능을 동시에 구현한 연속적인 세포 분쇄기)

  • Lee, Dong-Woo;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.32 no.10
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    • pp.831-835
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    • 2008
  • We present a continuous electrical cell lysis chip, using a DC bias voltage to generate the focused high electric field for cell lysis as well as the electroosmotic flow for cell transport. The previous cell lysis chips apply an AC voltage between micro-gap electrodes for cell lysis and use pumps or valves for cell transport. The present DC chip generates high electrical field by reducing the width of the channel between a DC electrode pair, while the previous AC chips reducing the gap between an AC electrode pair. The present chip performs continuous cell pumping without using additional flow source, while the previous chips need additional pumps or valves for the discontinuous cell loading and unloading in the lysis chambers. The experimental study features an orifice whose width and length is 20 times narrower and 175 times shorter than the width and length of a microchannel. With an operational voltage of 50 V, the present chip generates high electric field strength of 1.2 kV/cm at the orifice to disrupt cells with 100% lysis rate of Red Blood Cells and low electric field strength of 60 V/cm at the microchannel to generate an electroosmotic flow of $30{\mu}m/s{\pm}9{\mu}m/s$. In conclusion, the present chip is capable of continuous self-pumping cell lysis at a low voltage; thus, it is suitable for a sample pretreatment component of a micro total analysis system or lab-on-a-chip.

Bio-inspired Cell Deformability Monitoring Chips Based on Strain Dependent Digital Lysis Rates (미소유로의 길이에 따른 통과세포의 파괴율을 바탕으로 한 생체모사 세포 변형성 검사칩에 관한 연구)

  • Youn, Se-Chan;Lee, Dong-Woo;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.32 no.10
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    • pp.844-849
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    • 2008
  • We present a novel cell deformability monitoring chip based on the digitally measured cell lysis rate which is dependent on the areal strain of the cell membrane. This method offers simple cell deformability monitoring by automated high-throughput testing system. We suggest the filter design considering the areal strain imposed on the cell membrane passing through the filter array having gradually increased orifice length. In the experiment using erythrocytes, we characterized the cell deformability in terms of average fracture areal strain which was $0.24{\pm}0.014\;and\;0.21{\pm}0.002$ for normal and chemically treated erythrocytes, respectively. We also verified that the areal strain of 0.15 effectively discriminates the deformability difference of normal and chemically treated erythrocytes, which can be applied to the clinical situation. We compared the lysis rates and their difference for the samples from different donors and found that the present chips can be commonly used without any calibration process. The experimental results demonstrate the simple structure and high performance of the present cell deformability monitoring chips, applicable to simple and cost-effective cell aging process monitoring.

A New Method for Determination of Enzyme Reaction and Activity of Lysozyme with UV-Spectrophotometer (UV-분광광도계를 이용한 새로운 Lysozyme의 효소반응 및 활성측정법 연구)

  • Kim, Woon-Soo;Kim, Yong-Wook;Kim, Woo-Sik
    • Applied Chemistry for Engineering
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    • v.9 no.6
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    • pp.857-863
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    • 1998
  • A simple and new experimental method for determination of lysozyme-M. lysodeikticus cell lysis reaction and lysozyme activity was suggested using Beer's law. The UV transmittance of the solution changed with the concentration of M. lysodeikticus and the relationship between the UV transmittance and M. lysodeikticus cell concentration followed Beer's Law. In addition, it was experimentally proven that the UV transmittance of the solution was not influenced by the lysozyme concentration and product of the lysis reaction. During the lysozyme-M. lysodeikticus cell lysis reaction, thus, M. lysodeikticus cell concentration in the solution could be measured in-situ by UV-spectrophotometer. By using these experimental data, kinetic Parameters of the Michaelis-Menten equation for the lysozyme-M. lysodeikticus cell 1ysis reaction was simply determined The maximum reaction rate constant ($k_3$) and Michaelis-Menten constants were $0.1734sec^{-1}$ and $9.83{\times}10^{-6}M$ respectively. The activity of the lysozyme could also be obtained with this experiment because the lysis reaction rate of the 1ysozyme depended on its activity.

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Lysis Action of Lauric Acid on Bacillus subtilis var. niger (Bacillus subtilis var. niger에 대한 Lauric Acid의 용균작용)

  • 조석금;조효현
    • The Korean Journal of Food And Nutrition
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    • v.3 no.1
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    • pp.23-28
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    • 1990
  • Effect of lauric acid and whir derivatives on the growth of Bacillus subtilis var niger was studied. Lauric acid showed the strongest inhibition among the fatty acids tested, Lysis rate of lauric acid proved to be greatly sensitibility against logarithmic phase cells but was not so influenced by cell concentration. On the other hand, lauric acid was inhibited lysis activity when the pH shift from 7.0 to 5.5

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A Mathematical Model for the Whole Ripening Process of Cheddar Cheese (체다치즈의 숙성 전과정에 대한 수학식)

  • 김중균
    • KSBB Journal
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    • v.9 no.1
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    • pp.72-84
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    • 1994
  • A model to explain the observed kinetics in a whole process of Cheddar-cheese ripening has been developed. It includes growth and lysis of cells in the cheese matrix, cell-wall bound protelnases and intracellular dipeptidases that are released into cheese upon cell lysis, and the production of dipeptides and amino acids from casein in cheese. Model simulations have been conducted to figure out the crucial factors in the process of the cheese ripening. The influential factors have been found to be the cell numbers and the dipeptidase activity at the beginning of the cheese ripening, and the cell-lysis rate of cheese starters. The simulation results have also suggested the use of a mixed culture as well as the experimental screening for a more suitable organism as a cheese starter hence, the model shows how to accelerate the cheese ripening.

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Fatal Tumor Lysis Syndrome During Chemotherapy in Small Cell Lung Cancer (소세포폐암에서 항암화학요법 중 발생한 치명적 종양용해증후군 1예)

  • Kook, Eun Hee;Kim, Min Soo;Ahn, Se Han;Jeon, Se Young;Yoon, Jung Ho;Han, Min Sung;Kim, Cheol Hyeon;Lee, Jae Cheol
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.3
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    • pp.215-218
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    • 2008
  • Tumor lysis syndrome is a life-threatening complication of anti-cancer therapy that typically occurs in patients with large, rapidly growing and treatment-sensitive tumors such as high-grade lymphomas and acute leukemias. However, its incidence in solid tumors has been known to be very low. Tumor lysis syndrome in solid tumors has a high mortality rate owing to the lack of prophylactic therapy to prevent this complication. We report a case of fatal tumor lysis syndrome developed during chemotherapy in extensive-stage small cell lung cancer, along with a brief review of the relevant literature considering the rarity of this manifestation in solid tumor.

Studies on Sexing of Bisected Mouse Embryos by Rat H-Y Antibody (Rat H-Y 항체에 의한 생쥐 분할란의 성 조절에 관한 연구)

  • 정장용;박희성;박충생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.179-187
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    • 1991
  • This experiment was carried out to develop a new technique of identifying XX of XY-bearing bisected embryos prior to implantation by immunological method. H-Y antiserum prepared in inbred Wastar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the concentraton or affected H-Y antibody was also investigated by culturing embryos under the concentration or affected H-Y antibody and culture rate. However, production of live young or sex rates of male and female from embryos transferred with psudopregnant. The biological test with the morula stage embryos showed that H-Y antibody was formed in all female rats immunized with spleen cell, but it was formed only in 80% female rats immunized with the antigen. When the bisected mouse embryos were cultured in vitro for 5~6 hours in morula stage, of 457 bisected embryos 81.4% of then were developed to the blastocyst stage. When the concentration rate of complement to H-Y antiserum varied from 1.0~5.0${mu}ell$, the lysis-rate of embryo was 19.5 to 67.3%. The concentration rate of complement did not influence the lysis-rate of embryos(P<0.05). The morphology embryos of bisected, zona-free and intact embryos showed the embryos lysis rate of 58.6, 42.7 and 48.5% respectively(P<0.05). Pregnancy rate were 50.0, 45.5 and 57.1% in psudopregnant recipient transferred with bisected, zona-free and intact blastocyst embryos. However, production of live youngs, sexual rate of male or female was 24(50.0:50.0), 22(45.5:55.5) and 36(58.3:41.7)mice, but affected and non affected half embryos with H-Y antiserum treatment was 23.1 and 26.7%. Also production of live youngs and sexual rate was 14(92.9:7.1) and 17(17.6:82.4)mice in affected and non affected half embryos in H-Y antiserum treatment(P<0.05).

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Effect of Electric fusion Methods on Cell Fusion Rate and Embryo Development by Somatic Cell Nuclear Transfer in Korean Native Cattle(KNC) (한우 체세포를 이용한 핵이식에서 전기융합 방법이 융합율 및 배발달율에 미치는 영향)

  • 김은국;김정욱
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.171-178
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    • 2003
  • This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM+0.05% FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3% BSA or 10% FBS at 37$^{\circ}C$, 90% $N_2$, and 5% $CO_2$in incubator for 6 days. The fusion rates were 51.6%(chamber) and 68.9%(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7%, 11.5%), respectively. The cleavage rates were significantly different between the chamber method(73.2%) and needle method(80.3%), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1%, 61.1% and 18.4%, 26.3% respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7%) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

Effects of Stireed Tank Bioreactor Scale-up on Cell Growth and Alkaloids Production in Cell Cultures of Eschscholtzia californica (탱크 교반형 생물반응기의 scale-up이 Eschscholtzia californica 세포생장 및 알칼로이드 생성에 미치는 영향)

  • 유병삼;변상요
    • KSBB Journal
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    • v.13 no.6
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    • pp.700-705
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    • 1998
  • Studies were made to investigate effects of the scale-up of stirred tank bioreactors on cell growth and alkaloids production for suspension cultures of Eschscholtzia californica. In the 1.5 L STR, cell lysis was observed at 110 rpm or higher agitation speed. The agitation speed of 30 L STR was 43.7 rpm to maintain the same shear stress developed in 1.5 L STR of 100 rpm. As a result of scale-up from 1.5 L to 30 L STR, the specific growth rate was decreased from 0.12 to 0.07 day-1. The alkaloids productivity was also decreased from 0.24 to 0.14 mg/L-day. Changes of mixing performance and oxygen transfer were studied to explain the decrease of cell growth and alkaloids production. Decreased oxygen transfer rate coefficient(KLa) and increased mixing time by the scale-up was observed at various aeration rates.

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Inhibition of Autolysis by Lipase LipA in Streptococcus pneumoniae Sepsis

  • Kim, Gyu-Lee;Luong, Truc Thanh;Park, Sang-Sang;Lee, Seungyeop;Ha, Jung Ah;Nguyen, Cuong Thach;Ahn, Ji Hye;Park, Ki-Tae;Paik, Man-Jeong;Pyo, Suhkneung;Briles, David E.;Rhee, Dong-Kwon
    • Molecules and Cells
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    • v.40 no.12
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    • pp.935-944
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    • 2017
  • More than 50% of sepsis cases are associated with pneumonia. Sepsis is caused by infiltration of bacteria into the blood via inflammation, which is triggered by the release of cell wall components following lysis. However, the regulatory mechanism of lysis during infection is not well defined. Mice were infected with Streptococcus pneumoniae D39 wild-type (WT) and lipase mutant (${\Delta}lipA$) intranasally (pneumonia model) or intraperitoneally (sepsis model), and survival rate and pneumococcal colonization were determined. LipA and autolysin (LytA) levels were determined by qPCR and western blotting. S. pneumoniae Spd_1447 in the D39 (type 2) strain was identified as a lipase (LipA). In the sepsis model, but not in the pneumonia model, mice infected with the ${\Delta}lipA$ displayed higher mortality rates than did the D39 WT-infected mice. Treatment of pneumococci with serum induced LipA expression at both the mRNA and protein levels. In the presence of serum, the ${\Delta}lipA$ displayed faster lysis rates and higher LytA expression than the WT, both in vitro and in vivo. These results indicate that a pneumococcal lipase (LipA) represses autolysis via inhibition of LytA in a sepsis model.