• 제목/요약/키워드: Cell Characterization

검색결과 1,776건 처리시간 0.032초

Novel High-Throughput DNA Part Characterization Technique for Synthetic Biology

  • Bak, Seong-Kun;Seong, Wonjae;Rha, Eugene;Lee, Hyewon;Kim, Seong Keun;Kwon, Kil Koang;Kim, Haseong;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • 제32권8호
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    • pp.1026-1033
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    • 2022
  • This study presents a novel DNA part characterization technique that increases throughput by combinatorial DNA part assembly, solid plate-based quantitative fluorescence assay for phenotyping, and barcode tagging-based long-read sequencing for genotyping. We confirmed that the fluorescence intensities of colonies on plates were comparable to fluorescence at the single-cell level from a high-end, flow-cytometry device and developed a high-throughput image analysis pipeline. The barcode tagging-based long-read sequencing technique enabled rapid identification of all DNA parts and their combinations with a single sequencing experiment. Using our techniques, forty-four DNA parts (21 promoters and 23 RBSs) were successfully characterized in 72 h without any automated equipment. We anticipate that this high-throughput and easy-to-use part characterization technique will contribute to increasing part diversity and be useful for building genetic circuits and metabolic pathways in synthetic biology.

Developing an interface strength technique using the laser shock method

  • James A. Smith;Bradley C. Benefiel;Clark L. Scott
    • Nuclear Engineering and Technology
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    • 제55권2호
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    • pp.432-442
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    • 2023
  • Characterizing the behavior of nuclear reactor plate fuels is vital to the progression of advanced fuel systems. The states of pre- and post-irradiation plates need to be determined effectively and efficiently prior to and following irradiation. Due to the hostile post-irradiation environment, characterization must be completed remotely. Laser-based characterization techniques enable the ability to make robust measurements inside a hot-cell environment. The Laser Shock (LS) technique generates high energy shockwaves that propagate through the plate and mechanically characterizes cladding-cladding interfaces. During an irradiation campaign, two Idaho National Laboratory (INL) fabricated MP-1 plates had a fuel breach in the cladding-cladding interface and trace amounts of fission products were released. The objective of this report is to characterize the cladding-cladding interface strengths in three plates fabricated using different fabrication processes. The goal is to assess the risk in irradiating future developmental and production fuel plates. Prior LS testing has shown weaker and more variability in bond strengths within INL MP-1 reference plates than in commercially produced vendor plates. Three fuel plates fabricated with different fabrication processes will be used to bound the bond strength threshold for plate irradiation insertion and assess the confidence of this threshold value.

구리에 의해 유도된 VBNC 대장균의 특성 (Characterization of Viable But Nonculturable Condition of Escherichia coli Induced with Copper)

  • 구형근;박상열;김숙경
    • 한국미생물·생명공학회지
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    • 제36권3호
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    • pp.209-214
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    • 2008
  • VBNC(Viable but nonculturable)란 생존에 불리한 환경하에서 살아 있으나 일반 영양배지에서 자라지 못하는 미생물의 상태를 나타낸다. 본 연구는 구리를 이용해 Escherichia coli에서 VBNC를 유도하고 이의 특성을 살펴보았다. 구리를 처리한 후 전통적인 평판 배양법에 의한 집락 형성계수(colony forming unit, CFU)를 측정한 결과 배양되지는 않으나, Live/Dead BacLight bacterial viability kit 염색 후 유세포계수기로 측정한 결과 살아있는 미생물로 계수되어 VBNC 상태가 확인하였다. VBNC 유도된 미생물로부터 genomic DNA와 RNA를 분리하고 이들의 안정성을 관찰하였는데 DNA에 비해 RNA의 붕괴가 많이 진행되었음을 확인할 수 있었고 RNA의 붕괴는 특정크기로 붕괴되는 것으로 관찰되었다. 또한 생물전용 투과전자현미경(Bio-Transmission Electron Microscope, Bio-TEM)을 통해 VBNC 세포의 형태를 관찰하였는데 VBNC 상태에서는 정상상태에 비해 periplasmic space가 온전하지 못하고 세포내막과 세포 외막이 분리되었으며 세포질의 양이 현저히 감소됨이 관찰되었다.

Obtainment and Characterization of Brain Tumor Cell Using Vasopressin-SV40 T Ag Transgenic Mouse

  • Kim, Sung-Hyun;Lee, Eun-Ju;Kim, Myoung-Li;Park, Jun-Hong;Cho, Kyoungin;Jung, Boo-Kyung;Kim, Hee-Chul;Hwnag, Sol-Ha;Lee, Hoon-Taek;Ryoo, Zae-Young
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.105-105
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    • 2003
  • In previous reports, pVPSV.IGR2.1 transgenic mouse were described that brain tumor and lymphoma by reason of Vasopressin-SV40 T antigen. In this study, we produced pVPSV.IGR3.6 transgenic mouse that used pVPSV.IGR3.6 vector. Expression of transgene was vary different in transgenic mouse. We obtained 6 transgenic mouse line, moreover they had died at the age of 2-6 weeks without transmitting the transgene to their offspring, and had tumorigenesis on same location with pVPSV.IGR2.1 transgenic mouse. Only a founder mouse was investigated for expression of fusion gene. Here we extended this transgenic approach to the study of tumor progression. From the mouse, we confirmed brain tumor cell, after then cultured for investigate characterization. In this report, we demonstrate that reduction of survival rate in transgenic mouse fused vasopressin gene length, acquisition of brain tumor cell, composition with astrocyte cells and neuronal cells. Finally, cells had no change with increase of passage.

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Characterization of Brain Tumor Cell using Vasopressin-SV40 T Ag Transgenic Mouse

  • Kim, Sung-Hyun;Lee, Eun-Ju;Kim, Myoung-Ok;Park, Jun-Hong;Kyoungin-Cho;Jung, Boo-Kyung;Kim, Hee-Chul;Hwang, Sol-Ha;Lee, Hoon-Taek
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.44-44
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    • 2003
  • In previous reports, pVPSV.IGR2.1 transgenic mouse were described that brain tumor and lymphoma by reason of Vasopressin-SV40 T antigen. In this study, we produced pVPSV.IGR3.6 transgenic mouse that used pVPSV.IGR3.6 vector. Expression of transgene was vary different in transgenic mouse. We obtained 6 transgenic mouse line, moreover they had died at the age of 2~6 weeks without transmitting the transgene to their offspring, and had tumorigenesis on same location with pVPSV.IGR2.1 transgenic mouse. Only a founder mouse was investigated for expression of fusion gene. Here we extended this transgenic approach to the study of tumor progression. From the mouse, we confirmed brain tumor cell, after then cultured for investigate characterization. In this report, we demonstrate that reduction of survival rate in transgenic mouse fused vasopressin gene length, acquisition of brain tumor cell, composition with astrocyte cells and neuronal cells. Finally, cells had no change with increase of passage.

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Fabrication and Characterization of BCP Nano Particle Loaded PCL Fiber and Their Biocompatibility

  • Nguyen, Thi-Phuong;Lee, Byong-Taek
    • 한국재료학회지
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    • 제20권7호
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    • pp.392-400
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    • 2010
  • The electrospinning process was established as a promising method to fabricate nano and micro-textured scaffolds for tissue engineering applications. A BCP-loaded PCL micro-textured scaffold thus can be a viable option. The biocompatibility as well as the mechanical properties of such scaffold materials should be optimized for this purpose. In this study, a composite scaffold of poly ($\varepsilon$-caprolactone) (PCL)-biphase calcium phosphate (BCP) was successfully fabricated by electrospinning. EDS and XRD data show successful loading of BCP nano particles in the PCL fibers. Morphological characterization of fibers shows that with a higher loaded BCP content the fiber surface was rougher and the diameter was approximately 1 to 7 ${\mu}m$. Tensile modulus and ultimate tensile stress reached their highest values in the PCL- 10 wt% BCP composite. When content of nano ceramic particles was low, they were dispersed in the fibers as reinforcements for the polymer matrix. However, at a high content of ceramic particles, the particles tend to agglomerate and lead to decreasing tensile modulus and ultimate stress of the PCL-BCP composite mats. Therefore, the use of nano BCP content for distribution in fiber polymer using BCP for reinforcement is limited. Tensile strain decreased with increasing content of BCP loading. From in vitro study using MG-63 osteoblast cells and L-929 fibroblast like cells, it was confirmed that electrospun PCL-BCP composite mats were biocompatible and that spreading behavior was good. As BCP content increased, the area of cell spreading on the surface of the mats also increased. Cells showed the best adherence on the surface of composite mats at 50 wt% BCP for both L-929 fibroblast-like cells and MG-63 osteoblast cell. PCL- BCP composites are a promising material for application in bone scaffolds.

실리카 화합물을 함유한 PVA/PSSA-MA 전해질 막의 제조 및 특성과 연료전지로의 응용 (Preparation and Characterization of PVA/PSSA-MA Electrolyte Membranes Containing Silica Compounds for Fuel Cell Application)

  • 변홍식;김대훈;이병성;이보성;윤석원;임지원
    • 멤브레인
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    • 제18권4호
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    • pp.336-344
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    • 2008
  • 본 연구에서는 Poly(vinyl alcohol) (PVA)을 주쇄부(base polymer)로 하여 화학적 가교를 실시하기 위하여 Poly(styrene sulfonic acid-co-maleic acid) (PSSA-MA)를 첨가하고, 실리카와 술폰산기를 함유한 3-(trihydroxysilyl)-1-propanesulfonic acid (THS-PSA)의 함량변화와 열 가교온도 변화를 통하여 직접 메탄을 연료전지(DMFC)에 적용하기 위한 고분자 전해질 막 소재에 관한 연구를 실시하였다. 제조된 막을 Nafion 115와 함께 비교하기 위하여 동일한 조건에서 함수율, 열 중량 분석(TGA), 이온교환용량, 이온전도도 및 메탄올 투과도를 실시하였다. 실험을 통하여 THS-PSA의 함량과 열 가교 온도변화에 따라서 메탄올 투과도가 $10^{-6}$에서 $10^{-8}\;cm^2/s$로 감소된 결과를 얻었으며, 이온전도도는 $10^{-3}$에서 $10^{-2}\;S{\cdot}cm^{-1}$으로 향상되어 Nafion 115보다 우수한 결과를 나타내었다.

Characterization of intracellular Ca2+ mobilization in gefitinib-resistant oral squamous carcinoma cells HSC-3 and -4

  • Kim, Mi Seong;Kim, Min Seuk
    • International Journal of Oral Biology
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    • 제46권4호
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    • pp.176-183
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    • 2021
  • Oral squamous cell carcinoma (OSCC) metastasis is characterized by distant metastasis and local recurrence. Combined chemotherapy with cisplatin and 5-fluorouracil is routinely used to treat patients with OSCC, and the combined use of gefitinib with cytotoxic drugs has been reported to enhance the sensitivity of cancer cells in vitro. However, the development of drug resistance because of prolonged chemotherapy is inevitable, leading to a poor prognosis. Therefore, understanding alterations in signaling pathways and gene expression is crucial for overcoming the development of drug resistance. However, the altered characterization of Ca2+ signaling in drug-resistant OSCC cells remains unclear. In this study, we investigated alterations in intracellular Ca2+ ([Ca2+]i) mobilization upon the development of gefitinib resistance in human tongue squamous carcinoma cell line (HSC)-3 and HSC-4 using ratiometric analysis. This study demonstrated the presence of altered epidermal growth factor- and purinergic agonist-mediated [Ca2+]i mobilization in gefitinib-resistant OSCC cells. Moreover, Ca2+ content in the endoplasmic reticulum, store-operated calcium entry, and lysosomal Ca2+ release through the transient receptor potential mucolipin 1, were confirmed to be significantly reduced upon the development of apoptosis resistance. Consistent with [Ca2+]i mobilization, we identified modified expression levels of Ca2+ signaling-related genes in gefitinib-resistant cells. Taken together, we propose that the regulation of [Ca2+]i mobilization and related gene expression can be a new strategy to overcome drug resistance in patients with cancer.

Screening of Lactobacilli Derived from Fermented Foods and Partial Characterization of Lactobacillus casei OSY-LB6A for Its Antibacterial Activity against Foodborne Pathogens

  • Chung, Hyun-Jung;Yousef, Ahmed E.
    • Preventive Nutrition and Food Science
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    • 제14권2호
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    • pp.162-167
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    • 2009
  • Various fermented foods were screened in search of food-grade bacteria that produce bacteriocins active against Gram-negative pathogens. An isolate from a mold-ripened cheese presented antibacterial activity against Gram-positive and Gram-negative bacteria. The most active isolate was identified as Lactobacillus casei by a biochemical method, ribotyping, and membrane lipid analysis, and was designated as OSY-LB6A. The cell extracts of the isolate showed inhibition against Escherichia coli p220, E. coli O157, Salmonella enerica serovar Enteritidis, Salmonella Typhimurium, and Listeria monocytogenes. The antibacterial nature of the cell extract from the isolate was confirmed by eliminating the inhibitory effects of acid, hydrogen peroxide, and lytic bacteriophages. The culture supernatant and cell extract retained antibacterial activity after heating at $60{\sim}100^{\circ}C$ for $10{\sim}20$ min. The activity of the cell extract from Lb. casei was eliminated by pronase and lipase. Finally, the cell extract showed a bactericidal mode of action against E. coli in phosphate buffer solution, but it was bacteriostatic in broth medium and food extracts.

Selenization of CIG Precursors Using RTP Method with Se Cracker Cell

  • Kang, Young-Jin;Song, Hye-Jin;Cho, You-Suk;Yoon, Jong-Man;Jung, Yong-Deuk;Cho, Dea-Hyung;Kim, Ju-Hee;Park, Su-Jung
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제43회 하계 정기 학술대회 초록집
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    • pp.426-426
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    • 2012
  • The CIGS absorber has outstanding advantages in the absorption coefficient and conversation efficiency. The CIGS thin film solar cells have been researched for commercialization and increasing the conversion efficiency. CIG precursors were deposited on the Mo coated glass substrate by magnetron sputtering with multilayer structure, which is CuIn/CuGa/CuIn/CuGa. Then, the metallic precursors were selenized under high Se pressure by RTP method which included. Se vapor was supplied using Se cracker cell instead of toxic hydrogen selenide gas. Se beam flux was controlled by variable reservoir zone (R-zone) temperature during selenization process. Cracked Se source reacted with CIG precursors in a small quantity of Se because of small size molecules with high activation energy. The CIGS thin films were studied by FESEM, EDX, and XRD. The CIGS solar cell was also developed by layering of CdS and ZnO layers. And the conversion efficiency of the CIGS solar cell was characterization. It was reached at 6.99% without AR layer.

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