• Title/Summary/Keyword: pE194

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Potential Control of Foodborne Pathogenic Bacteria by Pediococcus pentosaceus and Lactobacillus graminis Isolated from Fresh Vegetables

  • Gonzalez-Perez, C.J.;Vargas-Arispuro, I.;Aispuro-Hernandez, E.;Aguilar-Gil, C.L.;Aguirre-Guzman, Y.E.;Castillo, A.;Hernandez-Mendoza, A.;Ayala-Zavala, J.F.;Martinez-Tellez, M.A.
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.183-194
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    • 2019
  • The consumption of fresh vegetables has been related to recurrent outbreaks of foodborne diseases (FBD) worldwide. Therefore, the development of effective alternative technologies is necessary to improve the safety of these products. This study aimed to isolate and identify epiphytic lactic acid bacteria (LAB) from fresh fruits and leafy vegetables and characterize their antagonistic capacity due to their ability to produce bacteriocins or antibacterial compounds. For this, 92 LAB isolates from fruits and leafy vegetables were screened for antagonistic activity. Two strains with the highest and broadest antagonistic activities were selected for further characterization; one from cantaloupe melon (strain CM175) and one from cilantro leaves (strain C15). The cell-free supernatants (CFS) of CM175 and C15 were found to exhibit antagonistic activity against FBD-causing pathogens. The CM175 and C15 strains were identified as Pediococcus pentosaceus and Lactobacillus graminis, respectively. Notably, the P. pentosaceus CM175 CFS stopped the growth of Salmonella Typhimurium, Salmonella Saintpaul, Staphylococcus aureus, and Listeria monocytogenes, and delayed Escherichia coli O157:H7 growth. Moreover, L. graminis C15 CFS delayed the growth of all indicator pathogens, but did not completely stop it. Organic acids and bacteriocin-like molecules were determined to be possibly exerting the observed antagonistic activity of the identified LAB strains. Thus, application of the antagonistic compounds produced by Pediococcus pentosaceus and Lactobacillus graminis could be a novel and ecological strategy in developing antimicrobial biopreservatives for the food industry and mitigating FBD by reducing the biological contamination in fruit and vegetable orchards, mainly via their potential in controlling both gram-negative and gram-positive pathogenic bacteria.

Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • Korean Journal of Veterinary Service
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    • v.44 no.4
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    • pp.185-194
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    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

Analysis of nucleotide sequence of a novel plasmid, pILR091, from Lactobacillus reuteri L09 isolated from pig

  • Lee, Deog-Yong;Kang, Sang-Gyun;Rayamajhi, Nabin;Kang, Milan;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.441-449
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    • 2008
  • The genus Lactobacillus is the largest of the genera included in lactic acid bacteria and is associated with mucosal membranes of human and animal. Only a few Lactobacillus plasmid-encoded functions have been discovered and used. In this study, a novel plasmid (pILR091) was isolated from a wild L. reuteri isolated from pig and described the characteristics of its replicons, genetic organization, and relationship with other plasmids. After digestion of the plasmid, pILR091, with SalI, plasmid DNA was cloned into the pQE-30Xa vector and sequenced. The complete sequence was confirmed by the sequencing of PCR products and analyzed with the Genbank database. The isolate copy number and stability were determined by quantitative-PCR. The complete sequence of L. reuteri contained 7,185 nucleotides with 39% G-C content and one cut site by two enzymes, SalI and HindIII. The similar ori sequence of the pC194- rolling circle replication family (TTTATATTGAT) was located 63 bp upstream of the protein replication sequence, ORF 1. Total of five ORFs was identified and the coding sequence represented 4,966 nucleotides (70.4%). ORF1 of pILR091 had a low similarity with the sequence of pTE44. Other ORFs also showed low homology and E-values. The average G-C content of pILR091 was 39%, similar with that of genomic DNA. The copy number of pILR091 was determined at approximately 24 to 25 molecules per genomic DNA. These results suggested that pILR091 might be a good candidate to construct a new vector, which could be used for cloning and expression of foreign genes in lactobacilli.

Aerobic Capacity and Ventilatory Response During Incremental Exercise in Elite High School Cyclist (점진부하 운동에서 중고교 엘리트 사이클 선수들의 유산소능력과 폐환기 반응)

  • Lee, Dae-Taek;Bae, Yoon-Jung
    • Journal of Life Science
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    • v.20 no.3
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    • pp.437-443
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    • 2010
  • This study was designed to examine the aerobic capacity and ventilatory response during an incremental exercise in elite high school cyclists. Twelve boys ($17{\pm}1\;yr$, $175{\pm}5\;cm$, $70{\pm}9\;kg$) participated in anthropometric measurements, incremental exercise testing, and pulmonary function tests. During incremental exercise testing using a cycle ergometer, their maximal oxygen uptake ($VO_2max$), maximal power output, ventilation, ventilatory equivalents for oxygen ($V_E/VO_2$) and carbon dioxide ($V_E/VCO_2$), respiratory rate, and tidal volume were measured. Time variables such as inspiratory time (Ti), expiratory time (Te), breathing time (Tb), and inspiratory duty cycle (Ti/Tb), as well as inspiratory flow rate ($V_T$/Ti) were assessed. Pulmonary function of vital capacity (FVC), forced expiratory volume in one second ($FEV_1$), $FEV_1$/FVC, and peak expiratory flow were evaluated. Their $VO_2max$, maximal heart rate, and Wmax were $57.5{\pm}3.9\;ml{\cdot}kg^{-1}{\cdot}min^{-1}$, $194.1{\pm}8.6\;beat{\cdot}min^{-1}$, and 452 W, respectively. $VO_2max$ was not related to any anthropometric parameters. Most ventilatory variables progressively increased with exercise intensity. As intensity increased, Ti, Tb, Tb decreased while Ti/Tb was maintained. Below an intensity of 250 W, height, weight, body mass index, and body surface were highly correlated with $V_T$/Ti and Ti/Tb (p<0.05). Collectively, $VO_2max$ appeared to be lower than adult cyclists, suggesting a different pattern of ventilatory control as age advances. Morphological characteristics were not related to $VO_2max$ in the population. Time variables of ventilatory response seemed to be related only at an exercise intensity level of less than 250 W. $V_T$/Ti may be related to exercise endurance capacity, but Ti/Tb was similar to adult cyclists.

The Rapid Detection of Pathogens in Organically Grown Vegetables Using PCR-DGGE (PCR-DGGE를 이용한 유기농 채소의 유해 미생물 신속 검지)

  • Kwon, Oh Yeoun;Son, Seok Min
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.370-375
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    • 2011
  • In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

FREE-FLOATING PLANETS, THE EINSTEIN DESERT, AND 'OUMUAMUA

  • Gould, Andrew;Jung, Youn Kil;Hwang, Kyu-Ha;Dong, Subo;Albrow, Michael D.;Chung, Sun-Ju;Han, Cheongho;Ryu, Yoon-Hyun;Shin, In-Gu;Shvartzvald, Yossi;Yang, Hongjing;Yee, Jennifer C.;Zang, Weicheng;Cha, Sang-Mok;Kim, Dong-Jin;Kim, Seung-Lee;Lee, Chung-Uk;Lee, Dong-Joo;Lee, Yongseok;Park, Byeong-Gon;Pogge, Richard W.
    • Journal of The Korean Astronomical Society
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    • v.55 no.5
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    • pp.173-194
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    • 2022
  • We complete the survey for finite-source/point-lens (FSPL) giant-source events in 2016-2019 KMTNet microlensing data. The 30 FSPL events show a clear gap in Einstein radius, 9 𝜇as < 𝜃E < 26 𝜇as, which is consistent with the gap in Einstein timescales near tE ~ 0.5 days found by Mróz et al. (2017) in an independent sample of point-source/point-lens (PSPL) events. We demonstrate that the two surveys are consistent. We estimate that the 4 events below this gap are due to a power-law distribution of free-floating planet candidates (FFPs) dNFFP/d log M = (0.4 ± 0.2) (M/38 M)-p/star, with 0.9 ≲ p ≲ 1.2. There are substantially more FFPs than known bound planets, implying that the bound planet power-law index 𝛾 = 0.6 is likely shaped by the ejection process at least as much as by formation. The mass density per decade of FFPs in the Solar neighborhood is of the same order as that of 'Oumuamua-like objects. In particular, if we assume that 'Oumuamua is part of the same process that ejected the FFPs to very wide or unbound orbits, the power-law index is p = 0.89 ± 0.06. If the Solar System's endowment of Neptune-mass objects in Neptune-like orbits is typical, which is consistent with the results of Poleski et al. (2021), then these could account for a substantial fraction of the FFPs in the Neptune-mass range.

Efficacy of 1:1000 Diluted Heparin versus 0.9% Normal Saline for Maintenance of Intermittent Intravenous Locks (희석된 헤파린과 생리식염수가 간헐적 말초정맥장치의 폐색, 유지기간 및 정맥염 발생에 미치는 영향)

  • Park Mi-Mi;Kim Chang-Hee;Cho Eun-Suk;Lee Mi-Jung;Kim Hae-Suk
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.7 no.2
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    • pp.208-221
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    • 2000
  • The objective of this research was to compare the flushing effects using 1:1,000 diluted heparin or 0.9% normal saline in relation to needle gauge and frequency of IV medications. The comparative categories were clotting, duration of patency, and incidence of phlebitis. The design of research was a Nonequivalent Control group, Post test, Nonsynchrorized Design. The independent variable was 0.9% normal saline flushing the IV locks and the dependent variables were clotting, duration of patency, and incidence of phlebitis. Subjects were medical-surgical inpatients over 15 years old and with peripherally placed IVs who were hospitalized in a university medical center. Exclusion criteria included foreigners and those who were rejected for this research. The final sample for data analysis included 295 IV sites in 194 patients; 154 were in the saline group and 141 were in the heparin group. Subjects were assigned to have IV locks is flushed with 0.9% normal saline in the experimental group and to have IV lock flushed with 1:1,000 diluted heparin(100 units) in the control group. In order to increase reliability, the nurses who were assigned to the units involved in the study received an explanation on the standard method for locking an IV, recording method for observational data and criteria for the detection of phlebitis. Data were collected for a period of 2 weeks, from March 16, 2000 to March 29, 2000. Total duration of IV was defined the time of IV insertion to the time of discontinuation. Phlebitis was defined as the presence of the following : pain, swelling. erythema at the insertion site. Chi-square was used to determine the association between the degree of clotting, duration of patency, and incidence of phlebitis for the diluted heparin or the normal saline and needle gauge and frequency of IV medications. The results are summarized as follows : (a) There was difference in the degree of clotting between two groups($X^2=5.882$, p=.015). (b) There was no difference in the degree of duration of patency between two groups($X^2=2.439$, p=.295). (c) There was no difference in the incidence of phlebitis between two groups($X^2=0.190$, p=.663). (d) There was difference in the degree of clotting($X^2=6.209$, p=.013) and in the degree of duration of patency($X^2=6.978$, p=.031) according the needle guage between the two groups. (e) There was difference in the incidence of phlebitis($X^2=5.008$, p=.025) according to the frequencies of IV injection between two groups.

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Anti-Proliferative Activity of OD78 Is Mediated through Cell Cycle Progression by Upregulation p27kip1 in Rat Aortic Vascular Smooth Muscle Cells

  • Tudev, Munkhtsetseg;Lim, Yong;Park, Eun-Seok;Kim, Won-Sik;Lim, Il-Ho;Kwak, Jae-Hwan;Jung, Jae-Kyung;Hong, Jin-Tae;Yoo, Hwan-Soo;Lee, Mi-Yea;Pyo, Myoung-Yun;Yun, Yeo-Pyo
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.187-194
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    • 2011
  • Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.

Hydrolysis of Arabinoxylo-oligosaccharides by α-ʟ-Arabinofuranosidases and β-ᴅ-Xylosidase from Bifidobacterium dentium

  • Lee, Min-Jae;Kang, Yewon;Son, Byung Sam;Kim, Min-Jeong;Park, Tae Hyeon;Park, Damee;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.32 no.2
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    • pp.187-194
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    • 2022
  • Two α-ʟ-arabinofuranosidases (BfdABF1 and BfdABF3) and a β-ᴅ-xylosidase (BfdXYL2) genes were cloned from Bifidobacterium dentium ATCC 27679, and functionally expressed in E. coli BL21(DE3). BfdABF1 showed the highest activity in 50 mM sodium acetate buffer at pH 5.0 and 25℃. This exo-enzyme could hydrolyze p-nitrophenyl arabinofuranoside, arabino-oligosaccharides (AOS), arabinoxylo-oligosaccharides (AXOS) such as 32-α-ʟ-arabinofuranosyl-xylobiose (A3X), and 23-α-ʟ-arabinofuranosyl-xylotriose (A2XX), whereas hardly hydrolyzed polymeric substrates such as debranched arabinan and arabinoxylans. BfdABF1 is a typical exo-ABF with the higher specific activity on the oligomeric substrates than the polymers. It prefers to α-(1,2)-ʟ-arabinofuranosidic linkages compared to α-(1,3)-linkages. Especially, BfdABF1 could slowly hydrolyze 23,33-di-α-ʟ-arabinofuranosyl-xylotriose (A2+3XX). Meanwhile, BfdABF3 showed the highest activity in sodium acetate at pH 6.0 and 50℃, and it has the exclusively high activities on AXOS such as A3X and A2XX. BfdABF3 mainly catalyzes the removal of ʟ-arabinose side chains from various AXOS. BfdXYL2 exhibited the highest activity in sodium citrate at pH 5.0 and 55℃, and it specifically hydrolyzed p-nitrophenyl xylopyranoside and xylo-oligosaccharides (XOS). Also, BfdXYL2 could slowly hydrolyze AOS and AXOS such as A3X. Based on the detailed hydrolytic modes of action of three exo-hydrolases (BfdABF1, BfdABF3, and BfdXYL2) from Bf. dentium, their probable roles in the hemiceullose-utilization system of Bf. dentium are proposed in the present study. These intracellular exo-hydrolases can synergistically produce ʟ-arabinose and ᴅ-xylose from various AOS, XOS, and AXOS.

K-Ar and $^{40}$ Ar/$^{39}$ Ar Ages from Metasediments in the Okcheon Metamorphic Belt and their Tectonic Implication (옥천 변성대 변성퇴적암의 K-Ar및 $^{40}$ Ar/$^{39}$ Ar 연대와 그 의의)

  • 김성원;오창환;이덕수;이정후
    • The Journal of the Petrological Society of Korea
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    • v.12 no.2
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    • pp.79-99
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    • 2003
  • Muscovite and biotite from 52 metasediments and 5 granites in the Hwasan area, the southwest of the Okcheon metamorphic belt and the Miwon-Jeungpyeong area, central Okcheon metamorphic belt were dated by the K-Ar and $^{40}$ Ar/$^{39}$ Ar methods. Muscovite and biotite ages from metapelitic and psammitic rocks (metasediments) of the Boeun and Pibanryeong units in the Hwasan area are concentrated in the mid-Jurassic (149-180 Ma). K-Ar and $^{40}$ Ar/$^{39}$ Ar ages for metapelitic and psammitic rocks of the Boeun and Pibanryeong units in the Miwon-Jeungpyeong area show complicated age distribution. Muscovite and biotite ages are classified by three groups, 142-194 Ma, 216-234 Ma, and 241-277 Ma. Younger (Cretaceous) ages occur only in metasediments close to Cretaceous granitic rocks in the southeastern region and the older ages of 216-277 Ma are restricted to the middle Part of the Jeungpyeong area. Most ages in the other area of the central Okcheon metamorphic belt fall between 142-194 Ma (Jurassic). K-Ar and $^{40}$ Ar/$^{39}$ Ar ages for granite from the northern part in the both the southwest and central Okcheon metamorphic belt also gave middle Jurassic ages (156-168 Ma). The similar ages from both metasediments and granites in the study areas indicate simultaneous cooling of both rocks to 300-350$^{\circ}C$ during the middle Jurassic. The state of graphitization of carbonaceous material of all metasediments in the study areas Indicates fully ordered graphite falling within a small range, from 3.353 to 3.359 ${\AA}$, which indicate amphibolite facies regional metamorphism. In the southern sector of the Boeun unit from the Hwasan area, metamorphic grade indicated by mineral paragenesis during regional intermediate-P/T metamorphism is greenschist facies. Whereas, the $d_{002}$ values for carbonaceous materials in the same sector show fully ordered graphite (ca. 500$^{\circ}C$) indicating amphibolite facies. This result with the concentration of mica ages of metasediments into the middle Jurassic, the presence of low-P/T thermal metamorphic zone (>500$^{\circ}C$) in the metasediments close to the Jurassic granite and the regional intrusion of Jurassic granites and their middle Jurassic intrusion and cooling ages may indicate the low-P/T regional thermal event during the early(\ulcorner)-middle Jurassic after main intermediate-P/T metamorphism which formed main mineral assemblage regionally in the study area. The regional thermal event failed, however, to reset the mineral assemblage of regional intermediate-P/T metamorphism except for narrow aureole (1-2 km) around Jurassic granite because e duration of thermal effect was relatively short by repid cooling of the Jurassic granite. In the middle part of the Jeungpyeong area, central Ogcheon metamorphic belt, muscovite and biotite K-Ar ages from 5 samples are 263-277 Ma and 241-249 Ma, respectively. An intermediate-P/T metamorphism is currently accepted to have occurred between 280 and 300 Ma. Therefore, the muscovite and biotite ages can be interpreted as cooling ages after Ml metamorphism indicating rapid cooling to ca 350$^{\circ}C$ between 280-300 Ma and 263-271 Ma, and biotite ages indicate slower cooling to ca. 300$^{\circ}C$ between 263-277 Ma and 241-249 Ma. However, more detail study is needed to confirm why the Permian to Triassic ages occur only in the middle Part of the Jeungpyeong area.a.