• Title/Summary/Keyword: 3D Reconstitution

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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.

Utilization of Reconstituted High-tannin Sorghum in the Diets of Broiler Chickens

  • Kumar, Vinod;Elangovan, A.V.;Mandal, A.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.538-544
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    • 2005
  • The present experiment was conducted to assess the effect of reconstitution (R) on utilization of red sorghum (S) in diets of broiler chickens. Day-old broiler chicks (n=360) were randomly divided into 36 groups of 10 chicks each, and 9 dietary treatments were allotted to 4 groups (replicates) in a completely randomized design. Out of the 9 treatments, one was corn-soy based control (D1). The rest of the treatments were diets consisting of four levels (25, 50, 75 and 100% part of corn) of raw red sorghum (S25-S100) or four levels of reconstituted red sorghum (RS25-RS100). The tannin content reduced from 2.3% to 1.6% after reconstitution of red sorghum. Body weight gain reduced significantly (p<0.01) in diets containing unprocessed red sorghum beyond 33% in diet or reconstituted red sorghum at any level. However, during finishing growth phase the birds receiving either processed or unprocessed sorghum (barring S75) had statistically similar gains in body weight. During over all growth phase (0-6 wk), live weight gains in all the dietary treatments did not differ statistically. Feed intake during 0-6 wk was significantly higher (p<0.05) in diets containing sole red sorghum than corn-soy based control diets Feed conversion ratio during 0-3 wk period in control and unprocessed red-sorghum diets were similar but statistically poorer (p<0.01) FCR emanated from reconstituted groups, while during 3-6 wk of age FCR was poorer (p<0.05) in diets containing 75% red sorghum, either processed or unprocessed. FCR, in overall growth phases, in control diet was statistically similar to the groups fed diets containing up to 33% unprocessed or 16% reconstituted group. The carcass traits and yield of organs did not differ (p>0.05) due to the various levels of red-sorghum. It was concluded that though the tannin content was reduced by 30% by the reconstitution process, but this did not give any additional advantage in broiler performance. More over, red-sorghum can be used effectively up to 33% in diet replacing 50% of corn after proper adjustment of proteins, energy and amino acids.

Improved human hematopoietic reconstitution in HepaRG co-transplanted humanized NSG mice

  • Kim, Jin;Ryu, Bokyeong;Kim, Ukjin;Kim, Chang-Hwan;Hur, Gyeung-Haeng;Kim, C-Yoon;Park, Jae-Hak
    • BMB Reports
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    • v.53 no.9
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    • pp.466-471
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    • 2020
  • Several humanized mouse models are being used to study humanspecific immune responses and diseases. However, the pivotal needs of fetal tissues for the humanized mice model have been huddled because of the demand for ethical and medical approval. Thus, we have verified the hematopoietic and immunomodulatory function of HepaRG and developed a new and easy humanized mouse model to replace the use of fetal liver tissue. HepaRG co-transplanted Hu-NSG mice significantly increased CD45+ lymphocytes and CD19+ B cells and CD3+ T cells than normal Hu-NSG, suggesting enhanced reconstitution of the human immune system. These results have improved the applicability of humanized mice by developing new models easily accessible.

Optimization of 1D 1H Quantitative NMR (Nuclear Magnetic Resonance) Conditions for Polar Metabolites in Meat

  • Kim, Hyun Cheol;Ko, Yoon-Joo;Kim, Minsu;Choe, Juhui;Yong, Hae In;Jo, Cheorun
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.1-12
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    • 2019
  • The objective of this study was to establish an optimized 1D $^1H$ quantitative nuclear magnetic resonance (qNMR) analytical method for analyzing polar metabolites in meat. Three extraction solutions [0.6 M perchloric acid, 10 mM phosphate buffer, water/methanol (1:1)], three reconstitution buffers [20 mM 3-morpholinopropane-1-sulfonic acid, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, phosphate buffer], and two pulse programs (zg30, noesypr1d) were evaluated. Extraction with 0.6 M perchloric acid and 20 mM phosphate resulted in a stable baseline and no additional overlap for quantifying polar metabolites in chicken breast. In qNMR analysis, zg30 pulse program (without water-suppression) showed smaller relative standard deviation (RSD) and faster running time than noesypr1d (water-suppression). High-performance liquid chromatography was compared with qNMR analyses to validate accuracy. The zg30 pulse program showed good accuracy and lower RSD. The optimized qNMR method was able to apply for beef and pork samples. Thus, an optimized 1D $^1H$ qNMR method for meat metabolomics was established.

Three-dimensional geostatistical modeling of subsurface stratification and SPT-N Value at dam site in South Korea

  • Mingi Kim;Choong-Ki Chung;Joung-Woo Han;Han-Saem Kim
    • Geomechanics and Engineering
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    • v.34 no.1
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    • pp.29-41
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    • 2023
  • The 3D geospatial modeling of geotechnical information can aid in understanding the geotechnical characteristic values of the continuous subsurface at construction sites. In this study, a geostatistical optimization model for the three-dimensional (3D) mapping of subsurface stratification and the SPT-N value based on a trial-and-error rule was developed and applied to a dam emergency spillway site in South Korea. Geospatial database development for a geotechnical investigation, reconstitution of the target grid volume, and detection of outliers in the borehole dataset were implemented prior to the 3D modeling. For the site-specific subsurface stratification of the engineering geo-layer, we developed an integration method for the borehole and geophysical survey datasets based on the geostatistical optimization procedure of ordinary kriging and sequential Gaussian simulation (SGS) by comparing their cross-validation-based prediction residuals. We also developed an optimization technique based on SGS for estimating the 3D geometry of the SPT-N value. This method involves quantitatively testing the reliability of SGS and selecting the realizations with a high estimation accuracy. Boring tests were performed for validation, and the proposed method yielded more accurate prediction results and reproduced the spatial distribution of geotechnical information more effectively than the conventional geostatistical approach.

Grim Stimulates Diap1 Poly-Ubiquitination by Binding to UbcD1

  • Yoo, Soon Ji
    • Molecules and Cells
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    • v.20 no.3
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    • pp.446-451
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    • 2005
  • Diap1 is an essential Drosophila cell death regulator that binds to caspases and inhibits their activity. Reaper, Grim and Hid each antagonize Diap1 by binding to its BIR domain, activating the caspases and eventually causing cell death. Reaper and Hid induce cell death in a Ring-dependent manner by stimulating Diap1 auto-ubiquitination and degradation. It was not clear that how Grim causes the ubiquitination and degradation of Diap1 in Grim-dependent cell death. We found that Grim stimulates poly-ubiquitination of Diap1 in the presence of UbcD1 and that it binds to UbcD1 in a GST pull-down assay, so presumably promoting Diap1 degradation. The possibility that dBruce is another E2 interacting with Diap1 was examined. The UBC domain of dBruce slightly stimulated poly-ubiquitination of Diap1 in Drosophila extracts but not in the reconstitution assay. However Grim did not stimulate Diap1 poly-ubiquitination in the presence of the UBC domain of dBruce. Taken together, these results suggest that Grim stimulates the poly-ubiquitination and presumably degradation of Diap1 in a novel way by binding to UbcD1 but not to the UBC domain of dBruce as an E2.

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.

Extraction of Subject Size in Still Image Using Floor Pattern (바닥 패턴을 이용한 단일영상 내의 피사체 크기 추출)

  • Hwang, Min-Gu;Kim, Dong-Min;Har, Dong-Hwan
    • The Journal of the Korea Contents Association
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    • v.11 no.4
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    • pp.11-17
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    • 2011
  • This paper aims to realize the information of a subject existing in a still image with objective values. To attain the goal, this research takes the vanishing point that a 2D still image has as the basis and recomposes the still image into a 3D image using a 3D program. Also, in order to set up the axis of the camera necessary to recompose a 3D image, this paper used the lens angle of view that the image has and floor patterns as well. The 3D image completed in this way can measure the size and distance of all subjects in the floor patterns if the size value of a particular reference subject is known, and through this, it can be possible to acquire basic information of a subject that can be either a criminal or a clue in the images of CCTVs or some criminal scene.

The Utility Evaluation of Reconstructed 3-D Images by Maximum Intensity Projection in Magnetic Resonance Mammography and Cholangiopancreatography

  • Cho, Jae-Hwan;Lee, Hae-Kag;Park, Cheol-Soo;Kim, Ham-Gyum;Baek, Jong-Geun;Kim, Eng-Chan
    • Journal of Magnetics
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    • v.19 no.4
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    • pp.365-371
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    • 2014
  • The aim of this study was to evaluate the utility of 3-D images by comparing and analyzing reconstructed 3-D images from fast spin echo images of MRI cholangiopancreatography (MRCP) images using maximum intensity projection (MIP) with the subtraction images derived from dynamic tests of magnetic resonance mammography. The study targeted 20 patients histologically diagnosed with pancreaticobiliary duct disease and 20 patients showing pancreaticobiliary duct diseases, where dynamic breast MR (magnetic resonance) images, fast spin echo imaged of pancreaticobiliary duct, and 3-D reconstitution images using a 1.5T MR scanner and 3.0T MR scanner were taken. As a result of the study, the signal-to-noise ratio in the subtracted breast image before and after administering the contrast agent and in the reconstructed 3-D breast image showed a high ratio in the reconstructed image of lesional tissue, relevant tissue, and fat tissue. However, no statistically meaningful differences were found in the contrast-to-noise ratio of the two images. In the case of the MRCP image, no differences were found in the ratios of the fast spin echo image and reconstructed 3-D image.

Studies on Hemicellulase System in Aspergillus niger ( I ) - Purification and reconstitution of D-xylanase - (Aspergillus niger의 Hemicellulase계 효소에 관한 연구-D-xylanase계 효소의 정제와 재조합)

  • Park, Yang-Do;Lee, Hee-Jong;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.23-32
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    • 1983
  • In the present study, two kinds of D-xylanases (1, 4-$\beta$-D-xylan xylanohydrolase (EC 3.2.1.8) were purified and characterized from crude extract of Aspergillus niger KG79. Xylanase I was most active at pH 5.0, whereas xylanse II at pH 4.0 Both enzymes demonstrated their maximum activity at 45$^{\circ}C$. They were relatively stable between pH 4.0 and 6.0 at 3$0^{\circ}C$ for 6 hours. Molecular weight of xylanse I and II were 12, 500 and 11, 500, respectively. Michaelis-Menten constants of xylanse I and II were 0.28% and 0.26% of xylan, respectively. Both enzymes could degrade commercial D-xylan to xylose, xylobiose, and xylotriose to the degree of about 10% of total reducing power. Xylanse I could, however, liberate arabinose from barley straw xylan in addition to xylose and xylooligasaccharides more rapidly than xylanase II. The degree of hydrolysis was about 25%. The reconstituted D-xylanase system with purified xylanases and $\beta$-xylosidase degraded commercial xylan and barley straw xylan to the degree of 28% and 54% respectively. The limit of hydrolysis by the enzymes was suggested to be resulted from the physical structure of the substrate.

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