• Title/Summary/Keyword: sonication

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Isolation of Putative in vivo Hoxc8 Downstream Target Genes Using ChIP-Cloning Method

  • Chung, Hyun-Joo;Kang, Myeng-Mo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.14 no.1
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    • pp.47-53
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    • 2008
  • Hox genes are known to be transcription factors controlling vertebrate pattern formation along the anteroposterior body axis by regulating many target gene expressions during vertebrate embryogenesis. In order to isolate in vivo Hox responsive target genes, ChIP-cloning technique has been applied using Hoxc8 antibody. Here murine embryo of day 11.5 post coitum (E11.5) highly expressing Hoxc8 gene was used after removing head and tail portions where Hoxc8 is rarely expressing. After fixation with formaldehyde, the chromatin DNAs harboring bound proteins were isolated. After sonication, about 0.5- to 1 Kb chromatin DNAs were immunoprecipitated with anti Hoxc8 antibody. After removing the bound proteins with proteinase K, DNAs were isolated, cloned into the pBluescsript II SK vector, and then sequenced. Total 33 random clones sequenced were anlalyzed to be located at 12 different genomic regions. Among these, 8 turned out to be introns and 4 were intergenic regions localized in random chromosomes. The base composition of total cloned genomic sequences (6608 bp) were AT-rich, i.e., 40% GC. When the Hoxc8 core binding sites, such as TAAT, ATTA, TTAT, and ATAA were analyzed total number of 55, 45, 54, and 55 were found, respectively, which are than twice as many as expected number of 26. Although this in silico analysis does not mean that the ChIP-cloned sequence is real Hoxc8 regulatory element in vivo, these results strongly imply that the DNA fragments cloned through chromatin immunoprecipitation could be very much likely the putative Hoxc8 downstream target genes.

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Sonochemical Grafting of Poly(vinyl alcohol) onto Multiwall Carbon Nanotubes in Water (초음파를 이용한 PVA에 의한 다중벽 탄소나노튜브의 수상 그래프팅)

  • Kim, Yeongseon;Baeck, Sung Hyeon;Shim, Sang Eun
    • Polymer(Korea)
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    • v.38 no.3
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    • pp.378-385
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    • 2014
  • Multiwall carbon nanotubes (MWCNTs) were modified with a water soluble polymer, poly(vinyl alcohol), PVA, using a simple ultrasonic wave in water. Under the irradiation of ultrasound, PVA chains were severed as macroradicals and instantly grafted onto the surface of MWCNTs due to the radical scavenging effect of MWCNTs. To control the grafting PVA onto MWCNTs, the ultrasonication power and irradiation time were changed from 300 to 500 W and from 10 to 50 min, respectively. The grafted PVA onto MWCNTs was confirmed by FTIR, TGA, SEM, and TEM. Dispersion stability of the modified MWCNTs was monitored by Turbiscan. The amount of grafted PVA on MWCNTs increased with the increase in the sonication power and irradiation time. The grafted PVA on MWCNTs induced the improved dispersion stability of the modified MWCNTs in water. These findings exhibit that ultrasound can be readily used for the grafting polymer chains on MWCNTs.

Relation between the Concentration of Hexagonal Boron Nitride Nano-Sheets Dispersed in Pure Water and Their Width and Height (초순수 용매 내 육방정 질화붕소 나노시트의 농도와 크기의 관계)

  • Cho, Dae-Hyun;Park, Miyoung;Ha, Seonghun
    • Tribology and Lubricants
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    • v.35 no.6
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    • pp.343-349
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    • 2019
  • According to a report in 2011, hexagonal boron nitride demonstrated good solubility in pure water, even without surfactants or organic functionalization. Hexagonal boron nitride nanosheets are an effective lubricant additive, and their solubility in pure water has motivated lubrication engineers to utilize aqueous solutions containing these nanosheets as water-based lubricants. In this study, we measure the width and height of the hexagonal boron nitride nanosheets dispersed in pure water by using the Zetasizer and atomic force microscopy. Without surfactants or functionalization, aqueous solutions containing 0.10, 0.07, 0.05, and 0.01 wt% of hexagonal boron nitride nanosheets are synthesized via sonication-assisted hydrolysis. The Zetasizer provides only a one-dimensional size of approximately 410 nm, regardless of the concentration of the solution. Thus, it does not allow the estimation of the shape of the nanosheet. To acquire the three-dimensional size of the nanosheets, atomic force microscopy is employed. The aqueous solutions containing 0.10, 0.07, 0.05, and 0.01 wt% of the hexagonal boron nitride nanosheets show average values of 740, 450, 700, and 610 nm in width, and 37, 26, 33, and 32 nm in thickness, respectively. No significant trend is observed between the concentration of the solution and size of the nanosheets. Therefore, when preparing a water-based lubricant, it may be appropriate to adjust conditions such as ultrasonication time rather than the concentration.

Field Emission Characteristics of Double-walled Carbon Nanotubes Related with Hydrochloric Acid Treatment (이중벽 탄소나노튜브의 염산처리 시간에 따른 전계방출 특성 평가)

  • Jung, Da-Mi;Sok, Jung-Hyun
    • Journal of the Korean Vacuum Society
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    • v.20 no.1
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    • pp.70-76
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    • 2011
  • High-quality double-walled carbon nanotubes (DWCNTs) were synthesized by catalytic decomposition method at $800^{\circ}C$ using Tetrahydrofuran. The as-synthesized DWCNTs typically have catalytic impurities and amorphous carbon, which were removed by two-step purification process, consisting of thermal oxidation and H2O2, HNO3, HCl treatment. The DWCNT suspension was prepared by dispersing the purified DWCNTs in an aqueous sodium dodecylbenzenesulfonate solution with horn-type sonication. This was then sprayed on ITO glass to fabricate CNT field emitters. The quality of purified DWCNTs was estimated with X-ray diffraction and Thermal Gravity Analysis. The field emission properties were improved by increasing the process time of HCl treatment.

Evaluation of Environmental Mutagens-Complex Mixture in Diesel Exhaust Respirable Particulate Matter

  • Kim, Soung-Ho;Ryu, Byung-Tak;Jang, Hyoung-Seok;Kim, Yun-Hee;Lee, Do-Han;Han, Kyu-Tae;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.194-194
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    • 2003
  • The International Agency for Research on Cancer (IARC, 1989) has classified whole diesel exhaust as probably carcinogenic to humans. Diesel exhaust particulate matter (DPM) adsorbs different chemical substances including PAHs and nitroarenes. DPM is emphasized because it is a major component of diesel exhaust, it is suspected of contributing to a health hazard. Diesel exhaust is a complex mixture of carbon particles and associated organics and inorganics, and it is not known what fraction or combination of fractions cause the health effects [cancer effects, noncancer effects (respiratory tract irritation/inflammation and changes in lung function)] that have been observed with exposure to diesel exhaust. In order to identify which chemical classes are responsible for the majority of the observed biological activities, we performed a particular biological/chemical analysis. Respirable particulate matter (PM2.5: <2.5mm) was collected from diesel engine exhaust using a high-volume sampler equipped with a cascade impactor. Particulate oganic matter was extracted by the dichloromethane/sonication method and the crude extract was fractionated according to EPA recommended procedure into seven fractions by acid-base partitioning and silica gel column chromatography. We examined genotoxic potentials of diesel exhaust particulate matter using novel genotoxicity tests, which are rapid, simple and sensitive methods for assessing DNA-damage at the DNA and chromosomal level (comet assay, in vitro MN test and Ames test). Higher genotoxic potency was observed in non polar fractions and several PAHs were detected by GC-MS, such as 1,2,5,6 dibenzanthracene, chrysene, 1,2-benzanthracene, phenanthrene and fluoranthene.

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Preparation and analysis of nickel-coated alumina by sonochemistry (음향화학법으로 니켈을 코팅한 알루미나의 제조 및 분석)

  • Kim, Jin-Woo;Choi, Sung-Woo;Lee, Chang-Seop
    • Analytical Science and Technology
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    • v.24 no.2
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    • pp.61-68
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    • 2011
  • Ni-coated alumina was prepared by sonochemical method. To increase an efficiency of Ni coating on alumina, amorphous alumina was prepared by sol-gel method and Ni was coated to fine particles of alumina. Ni-coated alumina was prepared from various calcination temperatures ($500^{\circ}C$, $1,000^{\circ}C$), concentrations of Ni solution (0.01 M~0.2 M) and sonochemical reaction times (30 min, 2h). The prepared fine particles were characterized by X-Ray Diffractometer (XRD), Scanning Electron Microscope (SEM), Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES), and Particle Size Analyzer (PSA). The coating amount of Ni increased, as Ni concentration and ultrasonication time increased. The maximum amount of Ni was coated to fine particles of alumina, when Ni-coated alumina was prepared with 0.1 M concentration of Ni solution for 2 h of sonication time at $1000^{\circ}C$ of calcination temperature. The average particle size was in the range of 835.9 to 986.7 nm.

Synthesis and Crystallization of Hydrazinium Nitroformate(HNF) as Eco-friendly Oxidizer (친환경 산화제 HNF 합성 및 결정화 연구)

  • Kim, Jina;Kim, Min Jun;Min, Byoung Sun
    • Journal of the Korean Society of Propulsion Engineers
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    • v.19 no.4
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    • pp.69-76
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    • 2015
  • Recently, environmental sustainability of the transitional explosives and propellants is an issue of growing importance in energetic materials. For examples, ammonium perchlorate(AP) as an solid propellants oxidizer could create a poisonous gas and atmospheric pollutions, such as HCl. Among the several oxidizers, hydrazinium nitroformate(HNF) is an effective candidate substance for eco-friendly oxidizer, which has high density, pressure index, and less smog generating property during combustion for the Divert and Attitude Control System(DACS). This study was confirmed a synthesis through various conditions, was performed for the essential data of solubility the crystallization process. Also, crystallization process such as cooling, drowning-out and sonication were performed.

Appropriate In Vitro Methods for Genotoxicity Testing of Silver Nanoparticles

  • Kim, Ha Ryong;Park, Yong Joo;Shin, Da Young;Oh, Seung Min;Chung, Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.28
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    • pp.3.1-3.8
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    • 2013
  • Objectives We investigated the genotoxic effects of 40-59 nm silver nanoparticles (Ag-NPs) by bacterial reverse mutation assay (Ames test), in vitro comet assay and micronucleus (MN) assay. In particular, we directly compared the effect of cytochalasin B (cytoB) and rat liver homogenate (S9 mix) in the formation of MN by Ag-NPs. Methods Before testing, we confirmed that Ag-NPs were completely dispersed in the experimental medium by sonication (three times in 1 minute) and filtration ($0.2{\mu}m$ pore size filter), and then we measured their size in a zeta potential analyzer. After that the genotoxicity were measured and especially, S9 mix and with and without cytoB were compared one another in MN assay. Results Ames test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains revealed that Ag-NPs with or without S9 mix did not display a mutagenic effect. The genotoxicity of Ag-NPs was also evaluated in a mammalian cell system using Chinese hamster ovary cells. The results revealed that Ag-NPs stimulated DNA breakage and MN formation with or without S9 mix in a dose-dependent manner (from $0.01{\mu}g/mL$ to $10{\mu}g/mL$). In particular, MN induction was affected by cytoB. Conclusions All of our findings, with the exception of the Ames test results, indicate that Ag-NPs show genotoxic effects in mammalian cell system. In addition, present study suggests the potential error due to use of cytoB in genotoxic test of nanoparticles.

Soluble Expression and Purification of Receptor Activator of Nuclear Factor-Kappa B Ligand Using Escherichia coli

  • Park, Sol-Ji;Lee, Se-Hoon;Kim, Kwang-Jin;Kim, Sung-Gun;Kim, Hangun;Choe, Han;Lee, Sang Yeol;Yun, Jung-Mi;Cho, Jae Youl;Chun, Jiyeon;Choi, Kap Seong;Son, Young-Jin
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.274-279
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    • 2015
  • Receptor activator of nuclear factor-kappa B ligand (RANKL) is a critical factor in osteoclastogenesis. It makes osteoclasts differentiate and multinucleate in bone remodeling. In the present study, RANKL was expressed as a soluble maltose binding protein (MBP)-fusion protein using the Escherichia coli maltose binding domain tag system (pMAL) expression vector system. The host cell E. coli DH5α was cultured and induced by isopropyl β-D-1-thiogalactopyranoside for rRANKL expression. Cells were disrupted by sonication to collect soluble MBP-fused rRANKL. The MBP-fusion rRANKL was purified with MBP Trap affinity chromatography and treated with Tobacco Etch Virus nuclear inclusion endopeptidase (TEV protease) to remove the MBP fusion protein. Dialysis was then carried out to remove binding maltose from the cleaved rRANKL solution. The cleaved rRANKL was purified with a second MBP Trap affinity chromatography to separate unsevered MBP-fusion rRANKL and cleaved MBP fusion protein. The purified rRANKL was shown to have biological activity by performing in vitro cell tests. In conclusion, biologically active rRANKL was successfully purified by a simple two-step chromatography purification process with one column.

Evaluation of Methods for Cyanobacterial Cell Lysis and Toxin (Microcystin-LR) Extraction Using Chromatographic and Mass Spectrometric Analyses

  • Kim, In S.;Nguyen, Giang-Huong;Kim, Sung-Youn;Lee, Jin-Wook;Yu, Hye-Weon
    • Environmental Engineering Research
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    • v.14 no.4
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    • pp.250-254
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    • 2009
  • Contamination of microcystins, a family of heptapeptide hepatotoxins, in eutrophic water bodies is a worldwide problem. Due to their poisoning effects on animals and humans, there is a requirement to characterize and quantify all microcystins present in a sample. As microcystins are, for most part, intracellular toxins produced by some genera of cyanobacteria, lysing cyanobacterial cells to release all microcystins is considered an important step. To date, although many cell lysis methods have been used, little work has been conducted comparing the results of those different methods. In this study, various methods for cell lysis and toxin extraction from the cell lysates were investigated, including sonication, bead beating, freeze/thaw, lyophilization and lysing with TritonX-100 surfactant. It was found that lyophilization, followed by extraction with 75% methanol, was the most effective for extracting toxins from Microcystis aeruginosa cells. Another important step prior to the analysis is removing impurities and concentrating the target analyte. For these purposes, a C18 Sep-Pak solid phase extraction cartridge was used, with the percentage of the eluent methanol also evaluated. As a result, methanol percentages higher than 75% appeared to be the best eluting solvent in terms of microcystin-leucine-arginine (MC-LR) recovery efficiency for the further chromatographic and mass spectrometric analyses.