• Title/Summary/Keyword: electroporation

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Irreversible Electroporation in Patients with Pancreatic Cancer :Angel and Devil (비가역적 전기 천공술을 시행한 국소 진행형 췌장암 환자 2례)

  • Keum, Jiyoung;Lee, Hee Seung;Kang, Huapyong;Jo, Jung Hyun;Chung, Moon Jae;Park, Jeong Youp;Park, Seung Woo;Song, Si Young;Bang, Seungmin
    • Journal of Digestive Cancer Reports
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    • v.7 no.1
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    • pp.26-30
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    • 2019
  • Pancreatic cancer has a very poor prognosis. Complete surgical resection remains the only current curative treatment. Locally advanced pancreatic cancer (LAPC) is considered as unresectable because of involvement of celiac and/or mesenteric vessels. The treatment of LAPC is a challenge. Current guidelines suggest systemic therapy. However, the majority of patients will never experience conversion to surgical resection. Thus, in these patients, ablation is an alternative therapy for local control, which causes local destruction while ideally avoiding injury to surrounding healthy tissue. Irreversible electroporation (IRE) is an energy delivery system, effective in ablating tumors by inducing irreversible membrane destruction of cells. IRE demonstrated to be safe in previous studies. However, it is not free from complications, even serious. Here, we reported two cases of the IRE in LAPC patients.

Quantification of DNA Delivery Efficiency Labeled with Fluorescent Dye in Digital Electroporation System (디지털 전기천공시스템에서 형광 염료로 표지 된 DNA 전달 효율의 정량화)

  • Bae, Seo Jun;Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.58 no.3
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    • pp.450-457
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    • 2020
  • In the previous study, there was a big difference between the tendency of the delivery efficiency of Yo-Pro-1 and the expression efficiency of the CFP gene, but there was a problem that could not provide a clue to this problem. Therefore, this study aimed to present a clue to this problem by quantifying and comparing the delivery efficiency after labeling DNA using a fluorescent dye, which was one of the methods for quantifying biomolecules. As a fluorescent dye for labeling, Yo-Pro-1 was used, and the delivery efficiency of the fluorescent dye Yo-Pro-1 and the labeled DNA was compared. The delivery efficiency of Yo-Pro-1 and labeled DNA according to the voltage condition of the digital electroporation system was maximized at 96 V, and the delivery efficiency tended to decrease as the voltage increased further. In the comparison of the delivery efficiency of Yo-Pro-1 and labeled DNA according to the number of voltage application conditions, the delivery efficiency was maximized at the number of 8 voltage application times for both delivery materials, and the delivery efficiency tended to decrease as the number of voltage application increases further. Through the two results, it was confirmed that the delivery efficiency using Yo-Pro-1 in the digital electroporation system represents the delivery efficiency of the system well. In addition, through the results of this study, the difference between the tendency of the delivery efficiency of Yo-Pro-1 and the expression efficiency of the CFP gene shown in the preceding study was not the result of the difference in the delivery efficiency of the delivery material, but it can be predicted to be due to a problem with the expression process of the genetic material that had been delivered.

Comparison of Surface Characteristics According to 3D Printing Methods and Materials for the Fabrication of Microfluidic Systems (미세유체시스템 제작을 위한 3D 프린팅 방식 및 소재 별 표면특성 비교)

  • Bae, Seo Jun;Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.57 no.5
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    • pp.706-713
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    • 2019
  • In this study, basic research was conducted to provide guidelines for selecting printers and materials suitable for each application case by analyzing 3D printing method and surface characteristics of materials suitable for microfluidic system. We have studied the surface characteristics according to the materials for the two typical printing methods: The most commonly used method of Fused Deposition Modeling (FDM) printing and the relatively high resolution method of Stereolithography (SLA) printing. The FDM prints exhibited hydrophilic properties before post - treatment, regardless of the material, but showed hydrophobic properties after post - treatment with acetone vapor. It was confirmed by the observation of surface roughness using SEM that the change of the contact angle was due to the removal of the surface structure by post-treatment. SLA prints exhibited hydrophilic properties compared to FDM prints, but they were experimentally confirmed to be capable of surface modification using hydrophobic coatings. It was confirmed that it is impossible to make a transparent specimen in the FDM method. However, sufficient transparency is secured in the case of the SLA method. It is also confirmed that the electroporation chip of the digital electroporation system based on the droplet contact charging phenomenon was fabricated by the SLA method and the direct application to the microfluidic system by demonstrating the electroporation successfully.

Immortalization of Rat Kidney Glomerular Mesangial Cell and Its Coculture with Glomerular Epithelial Cell

  • Toshinobu Kida;Sachi Fujishima;Masatoshi Matsumra;Wang, Pi-Chao
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.92-98
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    • 2000
  • Mesangial cell has several key roles in thee control of glomerular function: it partocipates in the regulation of glomerular filtration rate, macromolecular clearance, and as both a source and target of numerous hormones and autocrines. Many of these insights into mesangial cell function have been obtained by studying mesangial cells in culture. However, no suitble cell lines have established yet. We here reported the immortalization of rat kidney glomeruar mesangial cell by transfection of E6 and E7 genes of human papillomavirus type 16 (HPV-16) via electroporation and lipofection. The reslts showed that only electroporation could transfect the genes to mesangial cells and the transfected cells maintained the viability for longer than 6 months. Fluorescence microscopic observation showed that cellular contractility and phagocytosis, which are the two main phenotypes of mesangial cells with rat glomerular epithelial cells showed that the growth of mesangial dells was suppressed by epithelial cell, but the growth of epithelisl cells was enhanced by mesangial cells. Moreover, Such results may imply that the glomerular cell-cell interaction plays an important role in the regulation of cell proliferation and differentiation.

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Expression of ${\alpha}$-Galactosidase Gene from Leuconostoc mesenteroides SY1 in Lactobacillus brevis 2.14

  • Lee, Kang-Wook;Park, Ji-Yeong;Park, Jae-Yong;Chun, Ji-Yeon;Kim, Jeong-Hwan
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1115-1118
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    • 2008
  • ${\alpha}$-Galactosidase gene (aga) from Leuconostoc mesenteroides SY1 was expressed in a heterologous host, Lactobacillus brevis 2.14 using an Escherichia coli-Leuconostoc shuttle vector, pSJE. pSJEaga (pSJE carrying aga) was introduced into Lactobacillus brevis 2.14 by electroporation and transformation efficiency was $1.1{\times}10^3$ per ${\mu}g$ DNA. L. brevis transformants (TFs) showed higher ${\alpha}$-galactosidase (${\alpha}$-Gal) activities than cells containing pSJE. Transcription levels of aga in L. brevis 2.14 grown on different carbon sources (1%, w/v) were examined by slot blot analysis. Aga transcript levels and ${\alpha}$-Gal activities were higher in cells grown on melibiose, raffinose, and galactose than cells on glucose, sucrose, and fructose. Western blot result showed that L. brevis 2.14 harboring pSJEaga produced much more ${\alpha}$-Gal when grown on melibiose than on glucose.

Leakage of Cellular Materials from Saccharomyces cerevisiae by Ohmic Heating

  • Yoon, Sung-Won;Lee, Chung-Young-J.;Kim, Ki-Myung;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.183-188
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    • 2002
  • The ohmic heating of foods for sterilization provides a shorter come-up time compared to conventional thermal processes. The electric fields as well as the heat generated by ohmic heating facilitate germicidal effects. In the present study, the effect of ohmic heating on the structure and permeability of the cell membrane of yeast cells, Saccharomyces cerevisae, isolated from Takju (a traditional Korean rice-beer), was investigated. The ohmic heating was found to translocate intracellular protein materials out of the cell wall, and the amount of exuded protein increased significantly as the electric field increased from 10 to 20 V/cm. As higher frequencies were applied, more materials were exuded. Compared to conventional heating, more amounts of proteins and nucleic acids were exuded when these cells were treated with ohmic heating. The molecular weights of the major exuded proteins ranged from 14 kDa to 18 kDa, as analyzed by Tricine-SDS PAGE. A TEM study also confirmed the leakage of cellular materials, thus indicating irreversible damage to the cell wall by ohmic heating. It was, therefore, concluded that the electric fields generated by ohmic heating induced electroporation, causing irreversible damage to the yeast cell wall and promoting the translocation of intracellular materials.

Comparison of Ectopic Gene Expression Methods in Rat Neural Stem Cells

  • Kim, Woosuk;Kim, Ji Hyeon;Kong, Sun-Young;Park, Min-Hye;Sohn, Uy Dong;Kim, Hyun-Jung
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.1
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    • pp.23-30
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    • 2013
  • Neural stem cells (NSCs) have the ability to proliferate and differentiate into various types of cells that compose the nervous system. To study functions of genes in stem cell biology, genes or siRNAs need to be transfected. However, it is difficult to transfect ectopic genes into NSCs. Thus to identify the suitable method to achieve high transfection efficiency, we compared lipid transfection, electroporation, nucleofection and retroviral transduction. Among the methods that we tested, we found that nucleofection and retroviral transduction showed significantly increased transfection efficiency. In addition, with retroviral transduction of Ngn2 that is known to induce neurogenesis in various types of cells, we observed facilitated final cell division in rat NSCs. These data suggest that nucleofection and retroviral transduction provide high efficiency of gene delivery system to study functions of genes in rat NSCs.

Controllable Movement of the Azobenzene Linked Crown Ether Conjugated Liposome

  • Seo, Eun-Seok;Kim, Soo-Hyun;Kim, Jin-Seok;Kim, Byung-Kyu
    • 제어로봇시스템학회:학술대회논문집
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    • 2005.06a
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    • pp.1158-1162
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    • 2005
  • Drug delivery systems have been developed to reduce the side toxicity of drugs by localizing them in the site of action. But it depends on the circulation of the blood and it doesn't have the function of locomotive mechanism of itself for searching for the region of disease. However, this problem could be solved by nanobot which have the locomotive function. So, we mimic the movement of cell that can move in a human body. In this paper, to polymerize the encapsulated actin within the liposome, electroporation technique is employed. In order to optimize polymerization and depolymerization of the liposome, we compare the time of polymerization and depolymerization by concentration of crown ether. we synthesis the liposome which contain azobenzene Linked crown Ether conjugated Actin protein. Azobenze linked crown ether holds the K+ ion by exposure of UV light and this disturbs the actin polymerization. In result, UV light could control the liposome growth. Finally, we could develop the liposome robot and control the growth and degeneration of the liposome by external stimuli such s UV light. The merit of the controlling by UV light doesn't need to inject proteins which induce polymerization and depolymerization of actin protein.

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Protein Engineering of Deoxynucleoside Kinase from Lactobacillus acidophilus: Effect of Site-Directed Mutagenesis on Microbial Growth

  • Park, Inshik;Kim, Eun-Ae;Bang, Keuk-Seung;Kim, Seok-Hwan;Kim, Gi-Nahm;Lee, Min-Kyung;Kil, Ji-Oeun
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.79-81
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    • 2001
  • Deoxynucleoside kinases exist as heterodimeric pairs specific for deoxyadenosine/deoxyguanosine kinase (dAK/dGK) and deoxyadenosine/deoxycytidine kinase (dAK/dCK). The aspartic acid-84 in dGK was mutated to alanine, asparagine and glutamic acid by site-directed mutagenesis. The mutation resulted in a drastic decease in dGK activity compared to the unmodified cloned enzyme while it increased production of dAK activity. The mutated dak/dgk genes, which synthesize tandem deoxyadenosine/deoxyguanosine kinase, were inserted back to the Lactobacillus acidophilus and Lactococcus lactis by electroporation to determine the effect of site-directed mutation of he enzymes on the microbial growth. However, no significant change was observed in cell growth and lactic acid production between wild type and mutant lactic acid bacteria.

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