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Overexpression of YbeD in Escherichia coli Enhances Thermotolerance

  • Kim, Sinyeon;Kim, Youngshin;Yoon, Sung Ho
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.401-409
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    • 2019
  • Heat-resistant microbial hosts are required for bioprocess development using high cell density cultivations at the industrial scale. We report that the thermotolerance of Escherichia coli can be enhanced by overexpressing ybeD, which was known to encode a hypothetical protein of unknown function. In the wild-type E. coli BL21(DE3), ybeD transcription level increased over five-fold when temperature was increased from $37^{\circ}C$ to either $42^{\circ}C$ or $46^{\circ}C$. To study the function of ybeD, a deletion strain and an overexpression strain were constructed. At $46^{\circ}C$, in comparison to the wild type, the ybeD-deletion reduced cell growth half-fold, and the ybeD-overexpression promoted cell growth over two-fold. The growth enhancement by ybeD-overexpression was much more pronounced at $46^{\circ}C$ than $37^{\circ}C$. The ybeD-overexpression was also effective in other E. coli strains of MG1655, W3110, DH10B, and BW25113. These findings reveal that ybeD gene plays an important role in enduring high-temperature stress, and that ybeD-overexpression can be a prospective strategy to develop thermotolerant microbial hosts.

Advancements of Common Gamma-Chain Family Cytokines in Cancer Immunotherapy

  • Alexandra A. Wolfarth;Swati Dhar;Jack B. Goon;Ugonna I. Ezeanya;Sara Ferrando-Martínez;Byung Ha Lee
    • IMMUNE NETWORK
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    • v.22 no.1
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    • pp.5.1-5.22
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    • 2022
  • The approval of immunotherapies such as checkpoint inhibitors (CPIs), adoptive cell therapies and cancer vaccines has revolutionized the way cancer treatment is approached. While immunotherapies have improved clinical outcome in a variety of tumor types, some cancers have proven harder to combat using single agents, underscoring the need for multi-targeted immunotherapy approaches. Efficacy of CPIs and cancer vaccines requires patients to have a competent immune system with adequate cell numbers while the efficacy of adoptive cellular therapy is limited by the expansion and persistence of cells after infusion. A promising strategy to overcome these challenges is combination treatment with common gamma-chain cytokines. Gamma-chain cytokines play a critical role in the survival, proliferation, differentiation and function of multiple immune cell types, including CD8 T-cells and NK cells, which are at the center of the anti-tumor response. While the short halflife of recombinant cytokines initially limited their application in the clinic, advancements in protein engineering have led to the development of several next-generation drug candidates with dramatically increased half-life and bioactivity. When combining these cytokines with other immunotherapies, strong evidence of synergy has been observed in preclinical and clinical cancer settings. This promising data has led to the initiation of 70 ongoing clinical trials including IL-2, IL-7, IL-15 and IL-21. This review summarizes the recent advancements of common gamma-chain cytokines and their potential as a cancer immunotherapy.

Efficient cell design and fabrication of concentration-gradient composite electrodes for high-power and high-energy-density all-solid-state batteries

  • Kim, Ju Young;Kim, Jumi;Kang, Seok Hun;Shin, Dong Ok;Lee, Myeong Ju;Oh, Jimin;Lee, Young-Gi;Kim, Kwang Man
    • ETRI Journal
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    • v.42 no.1
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    • pp.129-137
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    • 2020
  • All-solid-state batteries are promising energy storage devices in which high-energy-density and superior safety can be obtained by efficient cell design and the use of nonflammable solid electrolytes, respectively. This paper presents a systematic study of experimental factors that affect the electrochemical performance of all-solid-state batteries. The morphological changes in composite electrodes fabricated using different mixing speeds are carefully observed, and the corresponding electrochemical performances are evaluated in symmetric cell and half-cell configurations. We also investigate the effect of the composite electrode thickness at different charge/discharge rates for the realization of all-solid-state batteries with high-energy-density. The results of this investigation confirm a consistent relationship between the cell capacity and the ionic resistance within the composite electrodes. Finally, a concentration-gradient composite electrode design is presented for enhanced power density in thick composite electrodes; it provides a promising route to improving the cell performance simply by composite electrode design.

Effect of Ionizing Radiation and Mercury Chloride (II) on Cell Morphology in Yeast Cells Frequently and Temporarily Treated with Both Stressors (방사선과 염화수은의 일시 및 반복 복합 처리된 효모세포의 산화적 스트레스 적응과 형태 변화)

  • Kim, Su-Hyoun;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.28 no.2
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    • pp.101-107
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    • 2010
  • Metal ions are essential to life. However, some metals such as mercury are harmful, even when present at trace amounts. Toxicity of mercury arises mainly from its oxidizing properties. Ionizing radiation (IR) is an active tool for destruction of cancer cells and diagnosis of diseases, etc. IR induces DNA double strand breaks in the nucleus, In addition, it causes lipid peroxidation, ceramide generation, and protein oxidation in the membrane, cytoplasm and nucleus. Yeasts have been a commonly used material in biological research. In yeasts, the physiological response to changing environmental conditions is controlled by the cell types. Growth rate, mutation and environmental conditions affect cell size and shape distributions. In this work, the effect of IR and mercury chloride (II) on the morphology of yeast cells were investigated. Saccharomyces cerevisiae cells were treated with IR, mercury chloride (II) and IR combined with mercury chloride (II). Non-treated cells were used as a control group. Morphological changes were observed by a scanning electron microscope (SEM). The half-lethal condition from the previous experimental results was used to the IR combined with mercury. Yeast cells were exposed to 400 and 800 Gy at dose rates of 400Gy $hr^{-1}$ or 800 Gy $hr^{-1}$, respectively. Yeast cells were treated with 0.05 to 0.15 mM mercury chloride (II). Oxidative stress can damage cellular membranes through a lipidic peroxidation. This effect was detected in this work, after treatment of IR and mercury chloride (II). The cell morphology was modified more at high doses of IR and high concentrations of mercury chloride(II). IR and mercury chloride (II) were of the oxidative stress. Cell morphology was modified differently according to the way of oxidative stress treatment. Moreover, morphological changes in the cell membrane were more observable in the frequently stress treated cells than the temporarily stress treated cells.

Novel Properties for Endoglucanase Acquired by Cell-Surface Display Technique

  • Shi, Baosheng;Ke, Xiaojing;Yu, Hongwei;Xie, Jing;Jia, Yingmin;Guo, Runfang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1856-1862
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    • 2015
  • In order to improve the stability of endoglucanase under thermal and acidic conditions, the endoglucanase gene was fused to the N-terminus of the Saccharomyces cerevisiae pir gene, encoding the cell wall protein PIR. The fusion gene was transformed into Pichia pastoris GS115 for expression. A resulting strain with high expression and high activity was identified by examining resistance to Geneticin 418, Congo red staining, and quantitative analysis of enzyme activity. SDS-PAGE analysis revealed that the endoglucanase was successfully displayed on the yeast cell surface. The displayed endoglucanase (DEG) showed maximum activity towards sodium carboxyl methyl cellulose at approximately 275 IU/g cell dry weight. DEG exhibited greater than 60% residual activity in the pH range 2.5-8.5, higher than free endoglucanase (FEG), which had 40% residual activity at the same pH range. The highest tolerated temperature for DEG was 70℃, much higher than that of FEG, which was approximately 50℃. Moreover, DEG showed 91.1% activity at 65℃ for 120 min, while FEG only kept 77.8% residual activity over the same period. The half-life of DEG was 270 min at 65℃, compared with only 150 min for FEG. DEG could be used repeatedly at least three times. These results suggest that the DEG has broad applications as a yeast whole-cell biocatalyst, due to its novel properties of high catalytic efficiency, acid-thermal stabilities, and reusability.

Promoting Effects of Sanguinarine on Apoptotic Gene Expression in Human Neuroblastoma Cells

  • Cecen, Emre;Altun, Zekiye;Ercetin, Pinar;Aktas, Safiye;Olgun, Nur
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9445-9451
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    • 2014
  • Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

A Study on the Cytotoxicity of Lead in Cultured Fibroblasts (납(Pb)이 배양 섬유모세포에 미치는 세포독성에 관한 연구)

  • 정연태;최민규;김정중;문연자;김재민;백순기
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.122-130
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    • 1995
  • In order to evaluate the cytotoxicity of lead in cultures of Balb/c mouse 3T3 cell line, various cytotoxic assays were carried out after expose cells to various concentrations of lead nitrate. Cytotoxic assays using this study were included NR assay, MTT assay, measurement of LDH and protein, synthetic rate of DNA and UDS. Intrace!!ular Ca$^{2+}$ level was also measured. Light and electron microscopic studies were done for morphological changes of lead-treated cell cultures. The results were as follows; 1. The absorbances of NR and MTT were decreased dose-dependently, and NR, and MTT, values of lead nitrate were 3.4 mM and 1.5 mM, respectively. 2. Amount of LDH released into the medium was increased in dose-dependently and LDH activity at 5 mM concentration of lead nitrate was increased to 335 % of control. 3. Amount of total protein was decreased dose-dependently, and which was half of control at 2 mM concentration of lead nitrate. 4. The synthetic rate of DNA was decreased dose-dependently, and also which was remarkably decreased at 3 mM and 5 mM concentrations of lead nitrate. 5. The synthetic rate of UDS was increased at 1 mM concentration of lead nitrate, but which was remarkably decreased at 3 mM and 5 mM concentrations of lead nitrate. 6. Intrace!lular Ca$^{2+}$ level was remarkably increased at 1 mM concentration of lead nitrate, compared with control. 7. In light microscopy, number of cells and processes were decreased according to the increase of dosage of lead nitrate. Electron microscopic findings showed that many vacuoles and cisternal dilatation of rough endoplasmic reticulum were seen in the cytoplasm at 1 mM concentration of lead nittale. From the above results, high dosage treatment of lead nitrate (>3 mM) damaged genetic malerials and it also showed cytotoxicity in mouse 3T3 cell line cultures by injury of cell organelles and Ca$^{2+}$ channel.

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A Case of Intramedullary Spinal Cord Metastasis From Non-small Cell Lung Cancer (비소세포 폐암의 척수수질내 전이 1예)

  • Park, Sang-Woo;Wi, Hyung-Soo;Kim, Hoon-Soo;Cho, Jae-Hwa;Lee, Hong-Lyeol;Loh, John-Kyu;Ryu, Jeong-Seon
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.6
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    • pp.627-632
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    • 2002
  • Intramedullary spinal cord metastasis (ISCM) has rarely been reported in patients with carcinomas. In about half the ISCM reported the primary origins are lung cancer, with small cell lung cancer responsible for almost all reported cases. Thus, ISCM from small cell lung cancer is relatively well documented, but ISCM from non-small cell lung cancer is rarely diagnosed prior to the patients' demise, so very little data about such patients is available. Spine MRI is the most sensitive technique for diagnosing ISCM. ISCM are now being encountered with increasing frequency due to the increasing survival rates of lung cancer patients, and the development of new imaging technique. We reported a case of an ISCM from non-small cell lung cancer with a brief review of the literature.

Design of Denitrification Reactor by Using Permeabilized and Immobilized Paracoccus denitrificans (Permeabilized Paracoccus denitrificans를 이용한 고정화 균주의 탈질화 반응기 설계)

  • Yun, Mi-Sun;Song, Ju-Yeong;Park, Keun-Ho
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.100-105
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    • 2005
  • Removal of nitrogen compound from waste water is essential and often accomplished by biological process. Denitrification bacterium, Paracoccus denitrificans (KCTC 2350) is employed to estimate the denitrification ability and the characteristics. In the immobilized biological reactor system, the measurement of absolute amount of active strain in the reactor is comparatively difficult or impossible. In this. study, a reactor was designed with the unwoven texture wrapped peep holed plastic tube to calculate the absolute amount of active strain by comparing the activity of the permeabilized and or immobilized reactor and the free cell reactor The reactor system was continuous stirred tank reactor and the reaction rate of substrate consumption was assumed to satisfy the Michaelis-Menten equation. The effluent concentration of nitrate and nitrite was measured to estimate the apparent parameter of Michaelis-Menten equation. As a result, we found that the amount of immobilized active strain was figured out to be half of the total active strain in the reactor and the time required to be reached in the equilibrium state in the permeabilized and or immobilized reactor system was figured out to be shorter than that of the free cell reactor system.

Complexity Reduced CP Length Pre-decision Algorithm for SSS Detection at Initial Cell Searcher of 3GPP LTE Downlink System (3GPP LTE 하향링크 시스템의 초기 셀 탐색기 SSS 검출 시 복잡도 최소화를 위한 CP 길이 선 결정 알고리즘)

  • Kim, Young-Bum;Kim, Jong-Hun;Chang, Kyung-Hi
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.34 no.9A
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    • pp.656-663
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    • 2009
  • In 3GPP LTE system downlink, PSS (primary synchronization signal) and SSS (secondary synchronization signal) sequences are used for initial cell search and synchronization. UE (user equipment) detects slot timing, frequency offset, and cell ID by using PSS. After that it should detect frame timing, cell group ID, and CP length by using SSS. But in 3GPP LTE, there are two kinds of CP length, so we should operate FFT twice. In this paper, to minimize SSS detection complexity in cell searcher, we propose a CP length pre-decision algorithm that reduces the arithmetical complexity by half at most, with negligible performance degradation.