• Title/Summary/Keyword: Block formation

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Recombinant Glargine Insulin Production Process Using Escherichia coli

  • Hwang, Hae-Gwang;Kim, Kwang-Jin;Lee, Se-Hoon;Kim, Chang-Kyu;Min, Cheol-Ki;Yun, Jung-Mi;Lee, Su Ui;Son, Young-Jin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1781-1789
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    • 2016
  • Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrix-assisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.

The Comparison of the Spontaneous Zona Pellucida Hardening and PMA-induced Zona Pellucida Hardening during in Vitro Culture of the Mouse Oocytes (생쥐 난자 배양시의 자발적 투명대 경화 현상과 PMA에 의한 투명대 경화 현상 비교)

  • Kang, Hye-Na;Bae, In-Ha;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.1
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    • pp.99-110
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    • 1994
  • One consequence of fertilization in mammals is an increased resistance of the zona pellucida (ZP) to proteases and various chemical reagents. This phenomenon has been called 'zona pellucida hardening' (ZPH), and it is generally accepted that it is caused by the secretory products of cortical granules released by the egg at fertilization. ZP of mouse oocytes maturing in vitro in a chemically defined medium becomes progressively more resistant to solubilization by chymotrypsin ("Spontaneous" ZP hardening). In the present study, it was aimed to find the specificity of spontaneous ZPH in relation to its possible relevance to the cortical reaction and the physiological block to polyspermy. When a maturation inhibitors, cAMP analog(dbcAMP) and phosphodiesterase inhibitor (IBMX) was added to culture medium, it prevent spontaneous ZPH of mouse oocyte during in vitro culture. Thus spontaneous ZPH requires GVBD, since it is prevented by those agents, which inhibit GVBD in vitro. However, culture for 3 hours in the presence of PMA(lOng/ml), a protein kinase C activator, resulted in ZPH without GVBD, thus suggesting that ZPH may be regulated independently apart from the event of GVBD. Pretreatment of mouse oocyte with FBS result in partially inhibitory effect on subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhibitory effect on the spontaneous ZPH, but subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhinbitory effect on the spontaneous ZPII, but had no inhibitory effect on PMA-induced ZPH. Treatment with a microfilament formation blocker(cytochalasin-B) at 1${\mu}g$/ml concentration, resulted in the excellent inhibitory effect on spontaneous ZPH. However cytochalasin-B did not inhibit PMA-induced ZPH. Thus this suggesting that spontaneuse ZPH had a different mechanism from PMA-induced ZPH.

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Starting Transients in Dual-Mode Scramjet Engine (이중 모드 스트램제트 엔진의 시동 천이 과정)

  • Choi, Jeong-Yeol;Noh, Jin-Hyun;Byun, Jong-Ryul;Lim, Jin-Shik
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2011.11a
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    • pp.981-984
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    • 2011
  • A high-resolution numerical study is carried out to investigate the transient process of the combustion and the shock-train developments in an ethylene-fueled direct-connect dual-mode scramjet combustor. Following the fuel injection, air-throttling is applied at the expansion part of the combustor to provide mass addition to block the flow to subsonic speed. The ignition occurs several ms later when the fuel and air are mixed sufficiently. The pressure build up by the combustion leads to the shock train formation in the isolator section that advances to the exit of the intake nozzle. Then, the air-throttling is deactivated and the exhaust process begins and the situation before the air-throttling is restored. Present simulation shows the detailed processes in the dual-mode scramjet combustor for better understanding of the operation regimes and characteristics.

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Synthesis, Characterization and in vitro Anti-Tumoral Evaluation of Erlotinib-PCEC Nanoparticles

  • Barghi, Leila;Asgari, Davoud;Barar, Jaleh;Nakhlband, Aylar;Valizadeh, Hadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10281-10287
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    • 2015
  • Background: Development of a nanosized polymeric delivery system for erlotinib was the main objective of this research. Materials and Methods: Poly caprolactone-polyethylene glycol-polycaprolactone (PCEC) copolymers with different compositions were synthesized via ring opening polymerization. Formation of triblock copolymers was confirmed by HNMR as well as FT-IR. Erlotinib loaded nanoparticles were prepared by means of synthesized copolymers with solvent displacement method. Results: Physicochemical properties of obtained polymeric nanoparticles were dependent on composition of used copolymers. Size of particles was decreased with decreasing the PCL/PEG molar ratio in used copolymers. Encapsulation efficiency of prepared formulations was declined by decreasing their particle size. Drug release behavior from the prepared nanoparticles exhibited a sustained pattern without a burst release. From the release profiles, it can be found that erlotinib release rate from polymeric nanoparticles is decreased by increase of CL/PEG molar ratio of prepared block copolymers. Based on MTT assay results, cell growth inhibition of erlotinib has a dose and time dependent pattern. After 72 hours of exposure, the 50% inhibitory concentration (IC50) of erlotinib hydrochloride was appeared to be $14.8{\mu}M$. Conclusions: From the obtained results, it can be concluded that the prepared PCEC nanoparticles in this study might have the potential to be considered as delivery system for erlotinib.

Handwritten Image Segmentation by the Modified Area-based Region Selection Technique (변형된 면적기반영역선별 기법에 의한 문자영상분할)

  • Hwang Jae-Ho
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.43 no.5 s.311
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    • pp.30-36
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    • 2006
  • In this paper, a new type of written image segmentation based on relative comparison of region areas is proposed. The original image is composed of two distinctive regions; information and background. Compared with this binary original image, the observed one is the gray scale which is represented with complex regions with speckles and noise due to degradation or contamination. For applying threshold or statistical approach, there occurs the region-deformation problem in the process of binarization. At first step, the efficient iterated conditional mode (ICM) which takes the lozenge type block is used for regions formation into the binary image. Secondly the information region is estimated through selecting action and restored its primary state. Not only decision of the attachment to a region but also the calculation of the magnitude of its area are carried on at each current pixel iteratively. All region areas are sorted into a set and selected through the decision parameter which is obtained statistically. Our experiments show that these approaches are effective on ink-rubbed copy image (拓本 'Takbon') and efficient at shape restoration. Experiments on gray scale image show promising shape extraction results, comparing with the threshold-segmentation and conventional ICM method.

An Assessment of Image Analysis of Longitudinal Bone Changes (시간경과에 따른 골변화의 영상 분석에 관한 연구)

  • Kim Young-Jin;Kim Jae-Duk
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.73-86
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    • 1997
  • This study was performed to assess the analyzing methods developed to detect clinically and quantitatively longitudinal bone changes. Through preliminary experiment, accuracy of Cu-Eq value conversion to the mass of HA was examined. For main experiment, 15 intraoral radiograms taken at soon, 1st, 2nd, 4th, and 6th week after implantation of mixture in extracted sites of 3 cases were used. We took the radiograms with copper step wedge as test object and HA phantom X -ray taking was standardized by using Rinn XCP device customized directly to the individual dentition with resin bite block. The images inputted by Quick scanner into computer were digitized and analyzed by NIH image program. The stability of the copper equivalent transformation and the usefulness of two analyzing methods by ROI and Reslice were examined. Obtained results as follows: 1) On the Cu equivalent images, the coefficient of variation in the measurement of Cu-Eq. value of ROI ranged from 0.05 to 0.24 and showed high reproducibility. 2) All results obtained by resliced contiguous image were coincident with those obtained from the assessment by ROI and formation of plot profile. 3) On the stacked and resliced image at the line of interest, we could analyze directly and quantitatively the longitudinal changes at several portions by plot profile and qualitatively by surface plot. 4) Implant area showed marked resorption till 2 weeks after implantation and showed significant increase in Cu-Eq. value at 6th week(p<0.01) and periapical area showed increase in Cu-Eq. value at 6th week compared to after-operation's.

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Dissolution behavior and early bone apposition of calcium phosphate-coated machined implants

  • Hwang, Ji-Wan;Lee, Eun-Ung;Lee, Jung-Seok;Jung, Ui-Won;Lee, In-Seop;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.43 no.6
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    • pp.291-300
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    • 2013
  • Purpose: Calcium phosphate (CaP)-coated implants promote osseointegration and survival rate. The aim of this study was to (1) analyze the dissolution behavior of the residual CaP particles of removed implants and (2) evaluate bone apposition of CaP-coated machined surface implants at the early healing phase. Methods: Mandibular premolars were extracted from five dogs. After eight weeks, the implants were placed according to drilling protocols: a nonmobile implant (NI) group and rotational implant (RI) group. For CaP dissolution behavior analysis, 8 implants were removed after 0, 1, 2, and 4 weeks. The surface morphology and deposition of the coatings were observed. For bone apposition analysis, block sections were obtained after 1-, 2-, and 4-week healing periods and the specimens were analyzed. Results: Calcium and phosphorus were detected in the implants that were removed immediately after insertion, and the other implants were composed mainly of titanium. There were no notable differences between the NI and RI groups in terms of the healing process. The bone-to-implant contact and bone density in the RI group showed a remarkable increase after 2 weeks of healing. Conclusions: It can be speculated that the CaP coating dissolves early in the healing phase and chemically induces early bone formation regardless of the primary stability.

Salmonella Promotes ASC Oligomerization-dependent Caspase-1 Activation

  • Hwang, Inhwa;Park, Sangjun;Hong, Sujeong;Kim, Eun-Hee;Yu, Je-Wook
    • IMMUNE NETWORK
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    • v.12 no.6
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    • pp.284-290
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    • 2012
  • Innate immune cells sense and respond to the cytoplasmic infection of bacterial pathogens through NLRP3, NLRC4 or AIM2 inflammasome depending on the unique molecular pattern of invading pathogens. The infection of flagellin- or type III secretion system (T3SS)-containing Gram-negative bacteria such as Salmonella enterica serovar Typhimurium (S. typhimurium) or Pseudomonas aeruginosa (P. aeruginosa) triggers NLRC4-dependent caspase-1 activation leading to the secretion of proinflammatory cytokines such as interleukin-1-beta (IL-$1{\beta}$) and IL-18. Previous studies have shown that apoptosis-associated speck-like protein containing a CARD (ASC) is also required for Salmonella-induced caspase-1 activation, but it is still unclear how ASC contributes to the activation of NLRC4 inflammasome in response to S. typhimurium infection. In this study, we demonstrate that S. typhimurium triggers the formation of ASC oligomer in a potassium depletion-independent manner as determined by in vitro crosslinking and in situ fluorescence imaging. Remarkably, inhibition of potassium efflux failed to block Salmonella-promoted caspase-1 activation and macrophage cell death. These results collectively suggest that ASC is substantially oligomerized to facilitate the activation of caspase-1 in response to S. typhimurium infection. Contrary to NLRP3 inflammasome, intracellular potassium depletion is not critical for NLRC4 inflammasome signaling by S. typhimurium.

Influence of the Novel Urease Inhibitor Hydroquinone on Growing Lamb Nitrogen Utilization

  • Zhang, Y.G.;Shan, A.S.;Bao, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.992-997
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    • 2002
  • Two in vivo experiments were conducted to evaluate the effect of novel urease inhibitor hydroquinone (HQ) on ammonia release rate from urea hydrolysis, nitrogen balance, nutrient digestibility and efficiency of microbial protein synthesis. In Exp. 1, twelve crossbred cannulated lambs were randomly assigned within initial body weight block to one of four HQ treatments, which included 0 (control), 30, 60 or 80 mg HQ/kg DM intake. Ammonia concentration and pH of ruminal fluid were immediately measured at 0, 2, 4, 6 and 8 h after feeding. Increasing the dose of HQ tended (p<0.15) to linearly decrease NH3 formation. The ammonia peak concentration (2 h post-feeding) in animals receiving HQ was approximately one-half of that in animals not receiving HQ (p<0.01), and a relatively sustained ammonia release could be obtained at the dose of 30 or 60 mg HQ/kg DM. In Exp. 2, sixteen intact crossbred lambs (weight $40{\pm}0.8kg$) were used in a $2{\times}2$ factorial design experiment. The four rations consisting of soybean meal-based (SBM) or urea-based (Urea) nitrogen source with or without HQ (S1, S0, U1 and U0) were fed in digestion and N balance trials. Apparent digestibility of major nutrients except that of ADF was not affected by either nitrogen source or addition of HQ. Regardless of nitrogen source, supplementation of HQ significantly improved ADF digestibility (p<0.05). The various ration had no effects on N metabolism in the presence of HQ. There was significant difference between total purine derivatives (PD), estimated efficiency of microbial N synthesis (p<0.05) and urea-N excretion (p<0.01) in the urine for the SBM ration and for the Urea ration. However, HQ had little influence on efficiency of microbial N synthesis as proportion of daily intake of total tract digestible OM (p>0.05). No interactions between main nitrogen source and HQ were measured throughout the trial. Results of this study suggest that addition of HQ to ration may improve ADF digestion with having no negative effect on N metabolism and microbial protein production.

A Statistical Modeling for the Economic Interpretation of Centrality in the International Arms Export (세계 무기 수출 중심성에 관한 통계적 분석과 경제적 의미)

  • Park, Joonsoo;Kim, Sung-Chul
    • The Journal of Society for e-Business Studies
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    • v.25 no.1
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    • pp.177-202
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    • 2020
  • We propose the statistical modeling and empirical results that can be utilized to identify and interpret the structural factors of international arms exports in recent years. The building blocks of research comprise the following questions; which would be the explanatory variables for the changing trend of international arms exports, whether the statistical significance can be verified on those variables and how those are interpreted for the future policy making purpose. We use the dataset of top 40 countries from SIPRI's Arms Transfers Database and analyze several regression models which consist of explanatory variables derived from research hypotheses. The most noticeable result is that the national fiscal reserve is shown to have consistent influence on the arms exports changes. UN security council members' group also has dominant power to make a formation of arms exports market block. Furthermore, gross domestic product and net exports volume in the national economy would seem to be related to changes of international arms exports in post-2000 period as well.