• Title/Summary/Keyword: Phase identification

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Production and identification of antisera against mu-opioid receptor using synthetic peptide epitope (Synthetic peptide를 이용한 mu-opioid receptor에 대한 항혈청의 생산과 검정)

  • Lee, Jang-hern;Kwon, Young-bae;Han, Ho-jae
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.45-54
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    • 1999
  • In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398 of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide synthesizer. The specificity and identification of the antisera were tested by analysis of transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR. MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as endoplasmic reticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity was abolished when anti-MOR sera were preincubated with the peptide against which they were raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which was determined to be within the last amino acid, 391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was observed in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain areas showed the mirrored pattern observed in auto radiographic studies of mu-opioid binding as well as a pattern similar to that seen by is situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future.

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Isolation and Identification of GABA-producing Microorganism from Chungkookjang (GABA 함량이 높은 청국장을 발효하는 균주의 분리 및 동정)

  • Mann, So-Yon;Kim, Eun-Ah;Lee, Ga-Young;Kim, Ro-Ui;Hwang, Dae-Youn;Son, Hong-Joo;Kim, Dong-Seob
    • Journal of Life Science
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    • v.23 no.1
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    • pp.102-109
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    • 2013
  • To isolate GABA-producing microorganisms, 1,500 strains were isolated from different Chungkookjang samples and screened. From these strains, 20 were selected for further analyses based on a protease and slime-producing activity test. The MC 31 strain showed the highest GABA concentration in Chungkookjang and was used in this study. MC 31 was identified as Bacillus subtilis by an API 50CHB kit and 16S rDNA sequences analysis and named as B. subtilis MC 31. B. subtilis MC 31 showed exponential growth up to 12 hours at $37^{\circ}C$ in LB broth, and it reached a stationary phase after 24 to 36 hours of incubation. B. subtilis MC 31 showed maximum GABA content at 72 hours after incubation at $40^{\circ}C$.

Rapid separation and identification of 31 major saponins in Shizhu ginseng by ultra-high performance liquid chromatography-electron spray ionization-MS/MS

  • Sun, Ting-Ting;Liang, Xin-Lei;Zhu, He-Yun;Peng, Xu-Ling;Guo, Xing-Jie;Zhao, Long-Shan
    • Journal of Ginseng Research
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    • v.40 no.3
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    • pp.220-228
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    • 2016
  • Background: Among the various ginseng strains, Shizhu ginseng is endemic to China, mainly distributed in Kuandian Manchu Autonomous County (Liaoning Province, China); however, not much is known about the compounds (especially saponins) in Shizhu ginseng. Methods: A rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with MS/MS (UHPLC-MS/MS) method was developed to separate and identify saponins in Shizhu ginseng. Results: The separation was carried out on a Waters ACQUITY UPLC BEH $C_{18}$ column ($100mm{\times}2.1mm$, $1.7{\mu}m$) with acetonitrile and 0.1% formic acid aqueous solution as the mobile phase under a gradient elution at $40^{\circ}C$. The detection was performed on a Micromass Quattro Micro API mass spectrometer equipped with electrospray ionization source in both positive and negative modes. Under the optimized conditions, a total of 31 saponins were identified or tentatively characterized by comparing retention time and MS data with related literatures and reference substances. Conclusion: The developed UHPLC-MS/MS method was suitable for identifying and characterizing the chemical constituents in Shizhu ginseng, which provided a helpful chemical basis for further research on Shizhu ginseng.

Quality Monitoring of Specification of Crataegi Fructus in the Korean Pharmacopoeia and Studies HPLC Standard Chromatogram (산사(山楂)의 규격 기준 모니터링 및 HPLC 표준크로마토그램 연구)

  • Kim, Kyoung Hee;Kim, Sun Mi;Lee, Young Jong;Baek, Wan Sook
    • The Korea Journal of Herbology
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    • v.32 no.1
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    • pp.55-62
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    • 2017
  • Objectives : Crataegi Fructus has been used as traditional medicines for more digestion action, amenorrhea due to blood stasis, and hyperlipemia. The aim of this study was to compare of Crataegi Fructus in South Korea collected during three years according to the standards in monographs of the Korean Pharmacopoeia Eleventh edition (KP 11). Methods : Crataegi Fructus was carried out identification test (Qualitative reaction, Thin layer chromatography), heavy metal test, and total ash registered at KP. Add to we tested loss on dry, contents of ethanol-soluble extracts, and HPLC profiling. Results : Identification test (TLC) was on comparing with ursolic acid standard solution in $R_f$ value, all samples showed red purple spot ($R_f$ value 0.9). Ursolic acid spot in $R_f$ value 0.35 showed by changing mobile phase condition. Heavy metals showed contents for Pb, Cd, As, and Hg range of 0.0 ~ 0.5 ppm, 0.0 ~ 0.2 ppm, 0.0 ~ 0.3 ppm, and 0.0 ~ 0.1 ppm. Loss on drying was ranged from 5.5 to 11.9 %, total ash was between the range 2.7 ~ 4.0 %. Contents of ethanol-soluble extracts was ranged from 17.8 to 44.9 %. The content of chlorogenic acid was ranged from 0.0 to 0.1 % based on the chlorogenic acid standard curve. Conclusion : We have verified the current specification standard of Crataegi Fructus and standard that is not set. We hope that it will help the standardization of Crataegi Fructus.

The Development of a Computer-Assisted HACCP Program for the Microbiological Quality Assurance in Hospital Foodservice Operations (병원급식의 미생물적 품질보증을 위한 HACCP 전산프로그램의 개발)

  • Kwak, Tong-Kyung;Ryu, Kyung;Choi, Seong-Kyung
    • Journal of the Korean Society of Food Culture
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    • v.11 no.1
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    • pp.107-121
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    • 1996
  • This study was carried out to develop the computer-assisted Hazard Analysis and Critical Control Point (HACCP) program for a systematic approach to the identification, assessment and control of hazards for foodservice manager to assure the microbiological quality of food in hospital foodservice operations. Sanitation practices were surveyed and analyzed in the dietetic department of 4 hospitals. Among them, one 762-bed general hospital was selected as standard model to develop computer-assisted HACCP program. All data base files and processing programs were created by using Foxpro package for easy access of HACCP concept. HACCP program was developed based on the methods suggested by NACMCF, IAMFES and Bryan. This program consisted of two parts: the pre-stage for HACCP study and the implementation stage of the HACCP system. 1. Pre-stage for HACCP study includes the selection of menu item, the development of the HACCP recipe, the construction of a product flow diagram, and printing the HACCP recipe and a product flow diagram. A menu item for HACCP study can be selected from the menu item lists classified by cooking methods. HACCP recipe includes ingredients, their amount and cooking procedure. A flow diagram is constructed based on the HACCP recipe. The HACCP recipe and a product flow diagram are printed out. 2. Implementation of HACCP study includes the identification of microbiological hazards, the determination of critical control points, the establishment of control methods of each hazard, and the complementation of data base file. Potentially hazardous ingredients are determined and microbiological hazards are identified in each phase of the product flow. Critical control points (CCPs) are identified by applying CCP decision trees for ingredients and each process stage. After hazards and CCPs are identified, criteria, monitoring system, corrective action plan, record-keeping system and verification methods are established. When the HACCP study is complemented, HACCP study result forms are printed out. HACCP data base file can be either added, corrected or deleted.

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Distance Sensing of Moving Target with Frequency Control of 2.4 GHz Doppler Radar (2.4 GHz 도플러 레이다의 주파수 조정을 통한 이동체 거리 센싱)

  • Baik, Kyung-Jin;Jang, Byung-Jun
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.30 no.2
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    • pp.152-159
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    • 2019
  • In general, a Doppler radar can measure only the velocity of a moving target. To measure the distance of a moving target, it is necessary to use a frequency-modulated continuous wave or pulse radar. However, the latter are very complex in terms of both hardware as well as signal processing. Moreover, the requirement of wide bandwidth necessitates the use of millimeter-wave frequency bands of 24 GHz and 77 GHz. Recently, a new kind of Doppler radar using multitone frequency has been studied to sense the distance of moving targets in addition to their speed. In this study, we show that distance sensing of moving targets is possible by adjusting only the frequency of a 2.4 GHz Doppler radar with low cost phase lock loop. In particular, we show that distance can be sensed using only alternating current information without direct current offset information. The proposed technology satisfies the Korean local standard for low power radio equipment for moving target identification in the 2.4 GHz frequency band, and enables multiple long-range sensing and radio-frequency identification applications.

Identification of Void Diameters for Cast-Resin Transformers (몰드변압기의 보이드 결함 크기 판별)

  • Jeong, Gi-woo;Kim, Wook-sung
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2022.10a
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    • pp.570-573
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    • 2022
  • This paper presents the identification of void diameters for a cast-resin transformer using an artificial neural network (ANN) model. A PD signal was measured by the Rogowski coil sensor which has the planar and thin structures fabricated on a printed circuit board (PCB), and the PD electrode system was fabricated to simulate a PD defect by a void. In addition, void samples with different diameters were fabricated by injecting air in a cylindrical aluminum frame using a syringe during the epoxy curing process. To identify the diameter of void defects, PD characteristics such as the discharge magnitude, pulse count, and phase angle were extracted and back propagation algorithm (BPA) was designed using virtual instrument (VI) based on the Labview program. From the experimental results, the BPA algorithm proposed in this paper has over 90% accurate rate to identify the diameter of void defects and is expected to use reference data of maintenance and replacement of insulation for cast-resin transformers in the on-site PD measurement.

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Two-stage crack identification in an Euler-Bernoulli rotating beam using modal parameters and Genetic Algorithm

  • Belen Munoz-Abella;Lourdes Rubio;Patricia Rubio
    • Smart Structures and Systems
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    • v.33 no.2
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    • pp.165-175
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    • 2024
  • Rotating beams play a crucial role in representing complex mechanical components that are prevalent in vital sectors like energy and transportation industries. These components are susceptible to the initiation and propagation of cracks, posing a substantial risk to their structural integrity. This study presents a two-stage methodology for detecting the location and estimating the size of an open-edge transverse crack in a rotating Euler-Bernoulli beam with a uniform cross-section. Understanding the dynamic behavior of beams is vital for the effective design and evaluation of their operational performance. In this regard, modal parameters such as natural frequencies and eigenmodes are frequently employed to detect and identify damages in mechanical components. In this instance, the Frobenius method has been employed to determine the first two natural frequencies and corresponding eigenmodes associated with flapwise bending vibration. These calculations have been performed by solving the governing differential equation that describes the motion of the beam. Various parameters have been considered, such as rotational speed, beam slenderness, hub radius, and crack size and location. The effect of the crack has been replaced by a rotational spring whose stiffness represents the increase in local flexibility as a result of the damage presence. In the initial phase of the proposed methodology, a damage index utilizing the slope of the beam's eigenmode has been employed to estimate the location of the crack. After detecting the presence of damage, the size of the crack is determined using a Genetic Algorithm optimization technique. The ultimate goal of the proposed methodology is to enable the development of more suitable and reliable maintenance plans.

Transcriptional Regulation of a DNA Repair Gene in Saccharomyces cerevisiae

  • Jang, Yeon-Kyu;Sancar, Gwen-B.;Park, Sang-Dai
    • Proceedings of the Zoological Society Korea Conference
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    • 1998.10b
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    • pp.113-113
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    • 1998
  • In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

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Isolation and Identification of Antimicrobial Compounds against Helicobacter pylori from Rosemary (Rosmarinus officinalis L.) Extracts (로즈마리(Rosmarinus officinalis L.) 추출물로부터 Helicobacter pylori에 대한 항균물질 분리 및 동정)

  • Yoon, So-Jung;Kim, Jin-Sung;Jo, Bun-Sung;Kim, Jeung-Hoan;Lee, Sun-Ho;Ahn, Bong-Jeun;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.159-165
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    • 2011
  • Rosmarinus officinalis extracts had a significant antimicrobial activity against Helicobacter pyori. Total phenolic contents and inhibition zone of rosemary extracts were estimated to be 25.7 mg/g and 14 mm at $200{\mu}g/mL$ of phenolic contents, respectively. The Sephadex LH-20 and MCI-gel CHP-20 column chromatographic separations for the phenolic extracts from R. officinalis leaves led to isolation of five acids, whose structures were determined as protocatechuic acid (A), coumaric acid (B), caffeic acid (C), chlorogenic acid (D), and rosmarinic acid (E), from interpretation of spectroscopic data including nagative fast atom bombardment (FAB)-mass, $^1H$-NMR, $^{13}C$-NMR, and IR. All isolated compounds were tested for antimicrobial activity against H. pyori. The purified single compound showed less antimicrobial activity against H. pylori than the mixed purified compounds, which generate A+B, A+E, C+D, C+E (each $200{\mu}g/disc$) excellent as large clear zone by synergy effect. These results indicate rosemary extracts are preventive agents against H. pyori.