• Title/Summary/Keyword: induction technique

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Quantitative Analysis of DNA Single-strand Breaks in EL 4 cells and Mouse Spleen Lymphocytes after Irradiation (방사선에 의한 EL 4 백서 백혈병 세포 및 정상 백서 비장 임파구 DNA Single-Strand Breaks의 정량적 분석과 측정)

  • Yoo Seong Yul;Cho Chul Koo;Koh Kyung Hwan;Park Woo Yoon;Park Young Hwan;Kim Sung Ho;Kim Tae Hwan;Chung In Yong
    • Radiation Oncology Journal
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    • v.8 no.2
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    • pp.137-144
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    • 1990
  • The filter elution technique was used to assay Co-60 $\gamma$ ray-induced DNA single-strand breaks(SSB) in EL 4 mouse leukemia cell and mouse spleen lymphocyte. The lymphocytes were stimulated with lipopolysaccharide (LPS, 20 $\mug/ml$) to label [${^3}H$] thymidine. EL 4 cells and lymphocytes in suspension were exposed at $0^{\circ}C$ to 0 Gy, 1 Gy, 5 Gy,10 Gy of Co-60 radiation and elution procedure was performed at PH 12.1. The number of DNA single-strand breaks increased with increasing doses of $\gamma$ rays. The strand scission factor (SSF) was estimated in each experiment (eluted volume 21 ml). The slope for EL 4 cells was $0.01301\pm0.00096\;Gy^{-1}(n=5)$ and the slope for lymphocytes was $0.01097\pm0.00091\;Gy^{-1}(n=5)$. The slopes were significantly different (P<0.005). Thus EL 4 cells were more sensitive to induction of DNA SSB by ionizing radiation than lymphocytes.

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Effect of electrode coating and applied voltage of pulsed electric fields (PEF) on sludge solubilization (펄스전기장 (Pulsed Electric Fields)의 전극 코팅과 인가 전압에 따른 슬러지의 가용화 효과)

  • Um, Se-Eun;Chang, In-Soung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.10
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    • pp.16-23
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    • 2018
  • Alternative technologies for sludge treatment and disposal need to be developed urgently because the amount of produced production has increased continuously. In this study, Pulsed Electric Fields (PEF) technique was applied for sludge solubilization and the performance was evaluated. The PEF equipped with electrodes coated by epoxy resin and teflon was inducted to the activated sludge suspension, and the effect of the coating materials on the solubilization was determined. In addition, the effect of the applied voltage on the solubilization yield was investigated as the applied voltage was increased from 6 to 12 and 15 kV. Sludge solubilization was not observed when the epoxy-coated electrode was used for PEF induction regardless of the applied voltage. However, sludge solubilization occurred when 12 and 15 kV were applied to the teflon-coated electrodes. The MLSS decreased to 9%, and the soluble-COD increased to 496% when the applied voltage was 15 kV. But sludge solubilization did not happen under 6 kV condition. The corona discharge was observed at applied voltages of 12 and 15 kV, (Ed- sorry but I cannot understand the following highlight) but if 6 kV, strongly indicating that the corona discharge make the sludge solubilized, which suggests that the critical voltage for sludge solubilization lies between 6 and 12 kV. Consequently, proper selection of electrode-coating materials and the applied voltage of PEF could lead to sludge solubilization by corona discharge.

Semi-Quantitative Analyses of Hippocampal Heat Shock Protein-70 Expression Based on the Duration of Ischemia and the Volume of Cerebral Infarction in Mice

  • Choi, Jong-Il;Kim, Sang-Dae;Kim, Se-Hoon;Lim, Dong-Jun;Ha, Sung-Kon
    • Journal of Korean Neurosurgical Society
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    • v.55 no.6
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    • pp.307-312
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    • 2014
  • Objective : We investigated the expression of hippocampal heat shock protein 70 (HSP-70) infarction volume after different durations of experimental ischemic stroke in mice. Methods : Focal cerebral ischemia was induced in mice by occluding the middle cerebral artery with the modified intraluminal filament technique. Twenty-four hours after ischemia induction, both hippocampi were extracted for HSP-70 protein analyses. Slices from each hemisphere were stained with 2,3,5-triphenyltetrazolium chloride (2%), and infarction volumes were calculated. HSP-70 levels were evaluated using western blot and enzyme-linked immunosorbent assay (ELISA). HSP-70 subtype (hsp70.1, hspa1a, hspa1b) mRNA levels in the hippocampus were measured using reverse transcription-polymerase chain reaction (RT-PCR). Results : Cerebral infarctions were found ipsilateral to the occlusion in 10 mice exposed to transient ischemia (5 each in the 30-min and 60-min occlusion groups), whereas no focal infarctions were noted in any of the sham mice. The average infarct volumes of the 2 ischemic groups were $22.28{\pm}7.31mm^3$ [30-min group${\times}$standard deviation (SD)] and $38.06{\pm}9.53mm^3$ (60-min group${\times}$SD). Western blot analyses and ELISA showed that HSP-70 in hippocampal tissues increased in the infarction groups than in the sham group. However, differences in HSP-70 levels between the 2 infarction groups were statistically insignificant. Moreover, RT-PCR results demonstrated no relationship between the mRNA expression of HSP-70 subtypes and occlusion time or infarction volume. Conclusion : Our results indicated no significant difference in HSP-70 expression between the 30- and 60-min occlusion groups despite the statistical difference in infarction volumes. Furthermore, HSP-70 subtype mRNA expression was independent of both occlusion duration and cerebral infarction volume.

Cytotoxicities of Tumor-specific T Lymphocytes Primed by Glioma Apoptotic Body - or Glioma Cell Lysate-pulsed Dendritic Cells

  • Kim, Jong-Tae;Chung, Dong-Sup;Kwak, Seung-Won;Han, Young-Min;Park, Young-Sup;Kim, Moon-Chan
    • Journal of Korean Neurosurgical Society
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    • v.38 no.2
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    • pp.126-131
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    • 2005
  • Objective : The choice of tumor antigen for dendritic cell[DC]-loading has still been an unresolved problem in the DC-based vaccine strategies against malignant gliomas that has not been found well-characterized tumor specific antigens. In this study, we compare tumor-specific T cell response induced by glioma apoptotic body[GAB]-pulsed DCs to response induced by glioma cell lysate-pulsed ones quantitatively. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor and interleukin[IL]-4 from peripheral blood mononuclear cells[PBMCs] of HLA-A2 positive healthy donors were cultured. Each GABs and glioma cell lysate generated from HLA-A2 positive T98G glioblastoma cells were co-incubated with DCs. $CD8^+$ T lymphocytes isolated from PBMCs of same donors were cultured in media containing IL-2 and either stimulated by GAB- or lysate-pulsed DCs three times at a weekly interval. The interferon[IFN]-${\gamma}$ concentrations of each cell culture supernate were measured by enzyme immunoassay technique. Cytolytic activity of the generated cytotoxic $CD8^+$ T cells either stimulated with GAB- or lysate-pulsed DCs was determined by a standard 4-h $^{51}Cr$-release assay. Results : IFN-${\gamma}$ production and cytolytic activity of effector T cells stimulated by GAB-pulsed DCs were significantly higher than those of T cells stimulated by lysate-pulsed ones. Conclusion : These results indicate the choice of antigen is a critical determinant in the induction of antitumor immunity against malignant glioma. Antigen preparations from GABs represent a promising alternative to glioma cell lysate in DC-based glioma vaccine strategies.

Biochemical Responses in Olive Flounder, Paralichthys olivaceus Fed Diet Supplemented with Fermented Aquaculture Sewage (양식장 배출물 발효물의 사료첨가에 따른 넙치, Paralichthys olivaceus의 생화학적 반응)

  • Jee, Jung-Hoon;Moon, Sang-Wook;Kim, Se-Jae;Lee, Young-Don;Keum, Yoo-Hwa;Kang, Ju-Chan
    • Journal of fish pathology
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    • v.18 no.2
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    • pp.187-195
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    • 2005
  • Effluent of aquaculture industry has caused a growing concern regarding its environmental impact. We assessed the use of flounder farming sewage as supplement of diet, to minimize the impact of aquaculture on the environment or also establish the technique for the recycling of effluent sediment derived from land-based seawater fish farm. In order to investigate the effects of a fermented aquaculture waste on biochemical responses of olive flounder (Paralichthys olivaceus), fermented products of aquaculture wastes were used as test compounds that cause hepatic and renal stress through the induction of oxidative stress in liver and kidney. Hepatosomatic index (HSI), glutathione content and glutathione dependent enzyme were not significantly different and no correlation was found within the different types of fermentation condition or supplement concentration, except for significant increases in 50% fermentation group and 50% concentration group in case of glutathione peroxidase activity and HSI value, respectively. These results showed addition of fermented aquaculture sewage may be an economic artificial sources of diet for fish aquaculture practices without affecting the function and safety in view of biochemical examination.

On the Use of Q Methodology in Research on the Subjective Perceptions of Election Campaign Advertising (Q방법론을 활용한 공직선거 광고의 주관적 수용인식 유형)

  • Yang, Chang-Hoon;Lee, Jei-Young
    • The Journal of the Korea Contents Association
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    • v.13 no.1
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    • pp.115-126
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    • 2013
  • The purpose of this study is to understand the utility of election campaign advertising by applying Q methodology that draw upon schematic model in subjectivity study. A survey was carried out among college students to classify the 16 selected Q-statements into a normal distribution using a 7 point scale. The collected data was analyzed using QUANL program, and principal component analysis using varimax rotation was used to identify the types of perceived utility of election campaign advertising. Type I can be categorized by a strong concern for the advertising technique improvement, Type II can be categorized by advocating for interest induction and Type III can be categorized by the truth inducement. The use of Q methodology provides insights into audience perceptions on the utility of election campaign advertising that would not be available through traditional methodologies and offers a foundation for audience involvement to address and overcome concerns about the utility of advertising for election campaign.

Development of a cylindrical ultrasound applicator for Intracavitary Hyperthermia (강내온열 치료를 위한 원통형 초음파 치료기 개발)

  • Lee, Rena J.;Hyunsuk Suh
    • Progress in Medical Physics
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    • v.13 no.2
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    • pp.79-84
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    • 2002
  • In this study, a cylindrical ultrasound applicator is developed for the treatment of vagina and rectum in combination with high dose rate brachytherapy. A cylindrical transducer (PZT-8, 1=1.5 cm, thickness=1.5mm OD=2.5 cm) was used as an energy source for induction of hyperthermia. Three single-element applicators were constructed to examine the performance of the PZT material. Vector impedance was measured to determine driving frequency. The efficiencies of the elements were determined using a radiation force technique to evaluate the feasibility of using the applicator as a hyperthermia source. A multi-element ultrasound applicator was designed using the PZT-8 material for the treatment of vagina. Results from the vector impedance measurements showed maximum magnitude at 1.78, 1.77, and 1.77 MHz for applicator 1,2, and 3, respectively. The radiation force measurements showed that the acoustic power of 40 watts was obtained in all three elements. The average efficiencies of the elements were 61.4, 65.2, and 54.0% for element 1, 2, and 3, respectively. The designed ultrasound hyperthermia applicator could be used in combination with high dose rate brachytherapy for the treatment of vagina and rectum. The use of this applicator with intracavitary brachytherapy could offer improved tumor control by increasing radiosensitiyity of the tumor.

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Efficient Gene Delivery into Hematopoietic Stem Cells by Intra-Bone Marrow Injection of Retrovirus (IBM 이식을 통한 골수 조혈 줄기 세포에의 효과적인 유전자 도입)

  • Lee, Byun-Joo;Lee, Yong-Soo;Kim, Hye-Sun;Kim, Yu-Kyung;Kim, Jae-Hwan;Park, Jin-Ki;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.9-14
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    • 2008
  • Efficient gene transfer into hematopoietic stem cells is a great tool for gene therapy of hematopoietic disease. Retrovirus have been extensively used for gene delivery and gene therapy. However, current in vitro gene transfer has some obstacles suck as induction of differentiation loss of self-renewal capacity, and down-regulation of homing efficiency for in vitro hematopoietic stem cells transplantation. To overcome these problems, we developed efficient in vitro retroviral transfer technique by direct intra-bone marrow injection (IBM). We identified effective retrovirus gene transfer in bone marrow hematopoietic cells in vitro. Two weeks after retrovirus transfer via IBM injection, we observed stable EGFP gene expression in bone marrow, lymph node, spleen, and liver cells. In addition, $6.4{\pm}2.7%$ of hematopoietic stem/progenitor cells were expressed EGFP transgene from flow cytometry analysis. Our results demonstrate that in vitro retrovirus gene transfer via IBM injection can provide a viable alternative to current or moo gene transfer approach.

Production of secondary metabolites by tissue culture of Artemisia annua L. (Artemisia annua L.의 조직배양을 이용한 이차대사 산물의 생산)

  • Kim, Nam-Cheol;Kim, Jeong-Gu;Lim, Hyung-Joon;Hahn, Tae-Ryong;Kim, Soo-Un
    • Applied Biological Chemistry
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    • v.35 no.2
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    • pp.99-105
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    • 1992
  • Artemisia annua contains the antimalarial principle, artemisinin. The possibility of the production of this compound through tissue culture technique was studied. The optimum combinations of hormones for the induction of callus were p-chlorophenoxyacetic acid(pcPA) and 6-benzylaminopurine(BAP) or pcPA and N-isopentenylaminopurine(2iP) in 0.05 mg/l each. For the growth of callus, the same combination of pcPA and BAP was optimum in concentrations of $1.0\;{\mu}M\;and\;0.5\;{\mu}M$, respectively, and the optimal concentration of sucrose was also found to be 2%(w/v). Tissue culture from the crown gall grew faster than normal callus. In the suspension culture broth and the cells of normal callus or Agrobacterium-transformed tumors, arteannuic acid and 11,12-dihydroarteannuic acid were found together with common phytosterols, whereas artemisinin was not found.

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Bioactivity-guided isolation of ginsenosides from Korean Red Ginseng with cytotoxic activity against human lung adenocarcinoma cells

  • Yu, Jae Sik;Roh, Hyun-Soo;Baek, Kwan-Hyuck;Lee, Seul;Kim, Sil;So, Hae Min;Moon, Eunjung;Pang, Changhyun;Jang, Tae Su;Kim, Ki Hyun
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.562-570
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    • 2018
  • Background: Lung cancer is the leading cause of cancer-related death worldwide. In this study, we used a bioactivity-guided isolation technique to identify constituents of Korean Red Ginseng (KRG) with antiproliferative activity against human lung adenocarcinoma cells. Methods: Bioactivity-guided fractionation and preparative/semipreparative HPLC purification were used with LC/MS analysis to separate the bioactive constituents. Cell viability and apoptosis in human lung cancer cell lines (A549, H1264, H1299, and Calu-6) after treatment with KRG extract fractions and constituents thereof were assessed using the water-soluble tetrazolium salt (WST-1) assay and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, respectively. Caspase activation was assessed by detecting its surrogate marker, cleaved poly adenosine diphosphate (ADP-ribose) polymerase, using an immunoblot assay. The expression and subcellular localization of apoptosis-inducing factor were assessed using immunoblotting and immunofluorescence, respectively. Results and conclusion: Bioactivity-guided fractionation of the KRG extract revealed that its ethyl acetate-soluble fraction exerts significant cytotoxic activity against all human lung cancer cell lines tested by inducing apoptosis. Chemical investigation of the ethyl acetatesoluble fraction led to the isolation of six ginsenosides, including ginsenoside Rb1 (1), ginsenoside Rb2 (2), ginsenoside Rc (3), ginsenoside Rd (4), ginsenoside Rg1 (5), and ginsenoside Rg3 (6). Among the isolated ginsenosides, ginsenoside Rg3 exhibited the most cytotoxic activity against all human lung cancer cell lines examined, with $IC_{50}$ values ranging from $161.1{\mu}M$ to $264.6{\mu}M$. The cytotoxicity of ginsenoside Rg3 was found to be mediated by induction of apoptosis in a caspase-independent manner. These findings provide experimental evidence for a novel biological activity of ginsenoside Rg3 against human lung cancer cells.