• Title/Summary/Keyword: core reconstitution

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Preparation and Reconstitution of Core-shell Type Nanoparticles of Poly(ε -caprolactone)/Poly(ethyleneglycol)/Poly(ε -caprolactone) Triblock Copolymers

  • Jeong, Young-Il;Ryu, Jae-Gon;Kim, Young-Hoon;Kim, Sung-Ho
    • Bulletin of the Korean Chemical Society
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    • v.23 no.6
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    • pp.872-879
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    • 2002
  • One of the improtant characteristics of core-shell type nanoparticles is the long-term storage and reuse as an aqueous injection solution when required. For this reason, reconstruction of lyophilized core-shell type nanoparticles is considered to be essential . BAB type triblock copolymers differ from AB type diblock copolymers, which contain the A block as a hydrophilic part and the B block as a hydrophobic part. by not being easily redistributed into phosphate-buffered saline (PBS, pH 7.4, 0.1 M). Therefore, lyophilized core-shell type nanoparticles of CEC triblock copolymer were reconstituted using a somication process with a bar-type sonicator in combination with a freezing-thawing process. Soncation for 30s only resuspended CEC nanoparticles in PBS; their particle size distribution showed a monomodal pattern with narrow size distribution. The bimodal size distribution pattern and the aggregates were reduced by further sonication for 120 s but these nanoparticles showed a wide size distribution. The initial burst of drug release was increased by reconstitution process. The reconstitution of CEC core-shell type nanoparticles by freezing-thawing resulted in trimodal distribution pattern and formed aggregates, although freezing-thawing process was easier than sonication . Drug release form CEC nanoparticles prepared by freezing-thawing was slower than from the original dialysis solution. Although core-shell typenanoparticles of CEC triblock copolymers were not easily performed. Cytotoxicity testing of core-shell type nanoparticles of CEC-2 triblock copolymers containing clonazepam (CNZ) was performed using L929 cells. Cytotoxicity of CNZ was decreased by incorporation into nanoparticles.

In Vitro Formation of Protein Nanoparticle Using Recombinant Human Ferritin H and L Chains Produced from E. coli

  • RO HYEON SU;PARK HYUN KYU;KIM MIN GON;CHUNG BONG HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.254-258
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    • 2005
  • We have conducted in vitro reconstitution study of ferritin from its subunits FerH and FerL. For the reconstitution, FerH was produced from an expression vector construct in Escherichia coli and was purified from a heat treated cell extract by using one-step column chromatography. FerL was expressed as inclusion bodies. The denatured form of FerL was obtained by a simple washing step of the inclusion bodies with 3 M urea. The reconstitution experiment was conducted with various molar ratios of urea-denatured FerH and FerL to make the ferritin nanoparticle with a controlled composition of FerH and FerL. SDS-PAGE analysis of the reconstituted ferritins revealed that the reconstitution required the presence of more than 40 molar$\%$ of FerH in the reconstitution mixture. The assembly of the subunits into the ferritin nanoparticle was confmned by the presence of spherical particles with diameter of 10 nm by the atomic force microscopic image. Further analysis of the particles by using a transmission electron microscope revealed that the reconstituted particles exhibited different percentages of population with dense iron core. The reconstituted ferritin nanoparticles made with molar ratios of [FerH]/[FerL]=l00/0 and 60/40 showed that 80 to $90\%$ of the particles were apoferritin, devoid of iron core. On the contrary, all the particles formed with [FerH]/[FerL]=85/ 15 were found to contain the iron core. This suggests that although FerH can uptake iron, a minor portion of FerL, not exceeding $40\%$ at most, is required to deposit iron inside the particle.

Recombinant Human L-ferritin from Saccharomyces cerevisiae: Molecular Characterization and Synthesis of Iron Oxide Nanoparticles (효모에서 생산한 재조합 human L-ferritin의 생화학적 특성 및 나노입자의 철산화물 합성)

  • Kim, Kyung-Suk
    • KSBB Journal
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    • v.26 no.2
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    • pp.119-125
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    • 2011
  • In the synthesis of nanoparticles, much attention has been paid to regulating the particle size. There has been a possible evident that using the central cavity (core) of the protein ferritin has a greatly significant influence on it because the core can generate the nanometer-sized mineral particles of variable metal ions. In this report, recombinant human L-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. The cDNA for human ferritin L chain was also expressed in another host such as Escherichia coli, and the properties of recombinant L-ferritins were compared. From isoelectric focusing experiment, the L-ferritin from the recombinant yeast showed no indication of N-glycosylation. Some post-translational modifications other than N-glycosylation were speculated in the L-ferritins from yeast. A difference was made in the L-ferritins in their iron uptake rates and the initial rate of the L-ferritin from yeast was slightly increased. The reconstitution yield and size distribution of the core minerals were analyzed in the L-ferritins by transmission electron microscopy. The L-ferritin from yeast with higher reconstitution yield (54.5%) showed slightly larger sizes (mean 6.92 nm) with narrower size distribution than the L-ferritin from E. coli. It is, in conclusion, speculated that L-ferritin from yeast is relatively superior to the other, in view of the size of nanoparticle and its relative homogeneity.

Reconstitution of Iron Cores in Horse Spleen and Yeast-derived Recombinant Human H- and L-chain Ferritins

  • Kim, Sung-Won;Jo, Min-Young;Yokota, Yasuhiro;Chung, Yun-Jo;Park, Chung-Ung;Kim, Kyung-Suk
    • Bulletin of the Korean Chemical Society
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    • v.25 no.2
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    • pp.237-242
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    • 2004
  • Recombinant human ferritin homopolymers (rHF and rLF) were successfully produced in the Saccharomyces cerevisiae Y2805, which was transformed with human ferritin H or L-chain genes, respectively. In order to characterize the molecular properties of the recombinant ferritins in relation to mineralization, the proteins were isolated and apoferritins were prepared. The apoferritins were reconstituted with 2000 Fe atoms per protein molecule under various experimental conditions (the concentration of the protein, the buffer concentration of the MOPS buffer, the total volume of the reaction and the reconstitution method). The structure and composition of the iron cores formed in the ferritins were examined using transmission electron microscopy. The recombinant ferritins behaved in a similar manner to other mammalian ferritins in accumulating iron in the core. Proteins of rHF and rLF showed varying reconstitution yields of 37-72% depending on the reaction conditions. In general, the rHF showed higher reconstitution yield than the rLF at the protein concentrations and the reaction volumes we examined. Iron cores with a similar mean particle size were obtained in the rHF, rLF and horse spleen ferritin reconstituted at a protein concentration of 1.0 mg/mL. Electron diffraction of all the three ferritins showed 2-3 diffuse lines, with d-spacings corresponding to those of the mineral ferrihydrite with a limited crystallinity.

The Methodology in Historical Demography at the Cambridge Group (역사 인구학 방법론의 현황)

  • 이흥탁
    • Korea journal of population studies
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    • v.12 no.2
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    • pp.56-68
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    • 1989
  • The methodology in historical demography comprises the three core areas the family reconstitution method at the Institut National d' Ftudes Demographiques(I.N.E.D), the back projection at the Cambridge Group for the History of PopuJation and Social Struc-ture(HPSS). and the household-pattern analysis at the Cambridge Group and at the California Institute of Technology. The paper presents an outline of the family reconstitu-tion method and discusses the problems, both theoretical and methodological, arising from the problematic back projection vis-a-vis the usual inverse projection developed by Ronald D. Lee at Berkeley. Recent developments in the tield of the generalized inverse projection method designed 10 supplement the defects in the back projection and the inverse projection are presented. and for ease of explanation of the parish register data for the family reconstitution form (FRE). pre-modern Korean household register data are presented along with the parish register data of England and Wales that constitute the backbone of historical demography in pre-modern Europe. Possibilities of exploring the household pattern analysis method based on the Laslett-Hammel classification scheme for the mid-eighteenth-century Korean household register data are suggested.

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Crystal Structure of Ferrihydrite Nanoparticles Synthesized in Ferritin

  • Kim, Sung-Won;Seo, Hyang-Yim;Lee, Young-Boo;Park, Young-Seog;Kim, Kyung-Suk
    • Bulletin of the Korean Chemical Society
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    • v.29 no.10
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    • pp.1969-1972
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    • 2008
  • In this study, horse spleen apoferritins were induced to form biominerals using up to 3000 Fe atoms per protein molecule. The morphology and crystallinity of the nanometer-sized biominerals formed in the ferritins were then analyzed using field emission-energy filtering-transmission electron microscopy (FE-TEM). The ferritins were found to have reconstitution yields of 60-70% in the experiments. The mean core size of the ferritins varied somewhat with protein concentrations, indicating that crystal growth in ferritins could be controlled via protein concentrations. The core mineral size increased with the amount of Fe used. Lattice fringes of the core, associated with good crystallinity, were found in all samples. The lattice fringe images of a single domain ferrihydrite mineral appeared frequently in the (011) planes (d-spacing of 0.246 nm) under [100] zone axis in all samples of this study. In addition, the lattice image occasionally revealed fringes corresponding to the (100) planes (d = 0.254 nm) from the [001] zone axis, indicating the characteristic pattern of hexagonal crystal lattice. Diffraction patterns in the minerals identified as ferrihydrite were fitted well into the space group of $P3_{1c}$.

Partial Purification of Factors for Differential Transcription of the rrnD Promoters for Ribosomal RNA Synthesis in Streptomyces coelicolor

  • Hahn, Mi-Young;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.534-540
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    • 2007
  • The Streptomyces coelicolor A3(2) genome contains six operons (rrnA to F) for ribosomal RNA synthesis. Transcription from rrnD occurs from four promoters (p1 to p4). We found that transcripts from the p1 and p3 promoters were most abundant in vivo in the early exponential phase. However, at later phases of exponential and stationary growth, transcripts from the p1 promoter decreased drastically, with the p3 and p4 transcripts constituting the major forms. Partially purified RNA polymerase supported transcription from the p3 and p4 promoters, whereas pure reconstituted RNA polymerase with core enzyme (E) and the major vegetative sigma factor ${\sigma}^{HrdB}$ ($E{\cdot}{\sigma}^{HrdB}$) did not. In order to assess any potential requirement for additional factor(s) that allow transcription from the p3 and p4 promoters, we fractionated a partially purified RNA polymerase preparation by denaturing gel filtration chromatography. We found that transcription from the p3 and p4 promoters required factor(s) of about 30-35 kDa in addition to RNAP holoenzyme ($E{\cdot}{\sigma}^{HrdB}$). Therefore, transcription from the p3 and p4 promoters, which contain a consensus -10 region but no -35 for ${\sigma}^{HrdB}$ recognition, are likely to be regulated by transcription factor(s) that modulate RNA polymerase holoenzyme activity in S. coelicolor.

EXTENSION OF OPERATIONAL LIFE-TIME OF WWER-440/213 TYPE UNITS AT PAKS NUCLEAR POWER PLANT

  • Katona, Tamas Janos;Ratkai, Sandor
    • Nuclear Engineering and Technology
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    • v.40 no.4
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    • pp.269-276
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    • 2008
  • Operational license of WWER-440/213 units at Paks NPP, Hungary is limited to the design lifetime of 30 years. Prolongation by additional 20 years of the operational lifetime is feasible. Moreover, enhancement of the reactor thermal power by 8% will increase both the net power output and the competitiveness of the plant. Paks NPP is a pioneer considering the power up-rate and preparation of long-term operation of WWER-440/213 design. Systematic preparatory work for long-term operation of Paks NPP has been started in 2000. A regulatory framework and a comprehensive engineering practice have been developed. According to the authors view, creation of a gapless engineering system via consequent application of best practices, and feed-back of experiences together with proper consideration of WWER-440/V213 features are the decisive elements of ensuring the safety of long-term operation. That systematic engineering approach is in the focus of recent paper. Key elements of justification and measures for ensuring the safety of long-term operation of Paks NPP WWER-440/213 units are identified and discussed. These are the assessment of plant condition and review of adequacy of ageing management programmes, also the review, validation and reconstitution of time limited ageing analyses as core tasks of licence renewal.

A Study on the Reconstitution of CPTED Theoretical Model - Focused on the Analysis of Sustainable Development - (CPTED 이론 모델의 재구성에 관한 연구 - 지속가능한 발전 분석을 중심으로 -)

  • Liu, He;Hong, Kwan-Seon
    • The Journal of the Korea Contents Association
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    • v.20 no.7
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    • pp.302-315
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    • 2020
  • The issue concerning how to integrate with the needs of sustainable development according to Crime Prevention Through Environmental Design(CPTED) theory still continues in the development process of the third-generation theory at the present stage. In addition, the issue is under worldwide controversy and discussion. What is more noteworthy is that the viewpoints under debate and discussion have an influence on the mutual effect and relationship between the theoretical model and the principles in the theoretical model, and the influence can never be underestimated. After a mastery of the overall context of theoretical development, it is believed in this study that the identity of CPTED theory development can provide a diverse understanding dimension and a communication method between the environment and environmental users. On the basis of identity the development of CPTED theory, the crime prevention design mode of CPTED needs to connect different dimensions of sustainability, with the design goal of livability as the starting point, the Top-down model to adjust the safety of the environment. By no means can the environmental condition be improved by solely relying on the previous physical design method in virtue of Bottom-up model. Therefore, in this study, the identity of CPTED will be set as the core combined with Top-down crime prevention design model to propose a visual proposal for the reconstitution of the theoretical model, rather than supplementing the application of the contents or principles at a certain stage. Hence, it is expected to provide reference and enlightenment for the sustainable development of CPTED theory.