• Title/Summary/Keyword: Signal Folding

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Gene Expression Analyses in Hypothalami of Immobilization-stressed and BoshimgeonbiTang-treated Mice Using cDNA Microarray (구속 스트레스 (immobilization stress)를 가한 rat의 hypothalamus에서의 유전자 발현 및 포심건비탕의 항스트레스 효과에 관한 cDNA microarray 분석)

  • Lee Han Chang;Yeam Mi Jung;Kim Gun Ho;Choi Kang Duk;Lee Seoung Hee;Shim Insop;Lee Hye Jung;Hahm Dae Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.6
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    • pp.1393-1403
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    • 2003
  • The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-Tang on the stress were studied with cDNA microarray analyses on hypothalamus using an immobilization-stress mouse as stress model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hours once a day, and this challenge was repeated for seven consecutive days. The body weights of the immobilization-stress mice were diminished about 25 percent degree as compared to normal ones. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with CyDye/sup TM/ fluorescence dyes (Amersham Bioscience Co., NJ), and then hybridized to cDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix 4000 series scanner and GenePix Pro/sup TM/ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis- and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosynthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 3.5 fold. The 20 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Hspe1 (protein folding), Aatk (apoptosis), Dffa (apoptosis), Itgb1 (cell adhesion), Vcam1 (cell adhesion), Fkbp5 (protein folding), BDNF (neuron survival) were restored to the normal one by the treatment of Boshimgeonbi-Tang.

Virtual Screening and Testing of GSK-3 Inhibitors Using Human SH-SY5Y Cells Expressing Tau Folding Reporter and Mouse Hippocampal Primary Culture under Tau Cytotoxicity

  • Chih-Hsin Lin;Yu-Shao Hsieh;Ying-Chieh Sun;Wun-Han Huang;Shu-Ling Chen;Zheng-Kui Weng;Te-Hsien Lin;Yih-Ru Wu;Kuo-Hsuan Chang;Hei-Jen Huang;Guan-Chiun Lee;Hsiu Mei Hsieh-Li;Guey-Jen Lee-Chen
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.127-138
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    • 2023
  • Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (TauRD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC50 of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 TauRD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

Development of a Finite Element Model for Studying the Occupant Behavior of a Mid-Size Truck with a Driver Side Airbag (운전석 에어백을 장착한 중형 트럭의 승객거동해석을 위한 유한요소 모델의 개발)

  • 홍창섭;오재윤;이대창
    • Journal of the Korean Society for Precision Engineering
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    • v.17 no.4
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    • pp.220-225
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    • 2000
  • This paper develops a finite element model for studying occupant behavior of a mid-size truck equipped with a driver side airbag. The developed model simulates an occupant behavior using PAM-CRASH/PAM-SAFE in super computer SP2. The model is developed based on a sled test. A 50% hybrid dummy III is used for measuring head and chest accelerations and femur loads, and major injury coefficients such as HIC, CA and femur load. Inferior components such as foot rest, seat, kneebolster, crash pad, etc. are roughly modeled and defined by a rigid material model. And contact type II is used for detecting a contact with dummy. Contact type II definition uses force-deflection relationship of each body Such components as steering column which directly affect on the occupant injuy are modeled in detail and defined by an elastic-plastic material model. Airbag cushion is modeled using rivet elements. Airbag cover groove is modeled using rivet elements. Airbag tether is modeled as nonlinear bar elements. Airbag model has two vent holes to ventilating the exploded gas. Airbag is folded close to the real airbag folding procedure, and folded cautiously in order not to have initial penetration. A vehicle pulse acquired from 31mph frontal barrier test is used as input signal for the simulation. The simulation conditions are tuned to the sled test ones. The measured dummy accelerations and major injury coefficients, and filmed dummy behavior and airbag inflation process using high speed camera are compared to the simulation results to verify the developed finite element model.

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Effects of Boshimgeonbi-tang on Gene Expression in Hypothalamus of Immobilization-stressed Mouse (보심건비탕(補心健脾湯) 투여가 Stress 유발 Mouse의 Hypothalamus 유전자 발현에 미치는 영향)

  • Lee Seoung-Hee;Chang Gyu-Tae;Kim Jang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1585-1593
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    • 2005
  • The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-tang on the stress were studied with cDNA microarray analyses, RT-PCR on hypothalamus using an immobilization-stress mice as an animal model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hrs once a day, and this challenge was repeated for seven· consecutive days. In the change of body weight it showed that the Boshimgeonbi-tang is effected recovery on weight loss caused by the immobilization-stress. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with $CyDye^{TM}$ fluorescence dyes and then hybridized to CDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix4000 series scanner and GenePix $Pro^{TM}$ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis-, stress protein, transcriptional factor, and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosysthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 5.5 fold. The 11 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Ogg1 (DNA repair), Aatk (apoptosis), Dffa (apoptosis), Fkbp5 (protein folding) were restored to the normal one by the treatment of Boshimgeonbi-tang.

Proteomic Analysis of the Aging-related Proteins in Human Normal Colon Epithelial Tissue

  • Li, Ming;Xiao, Zhi-Qiang;Chen, Zhu-Chu;Li, Jian-Ling;Li, Cui;Zhang, Peng-Fei;Li, Mao-Yu
    • BMB Reports
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    • v.40 no.1
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    • pp.72-81
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    • 2007
  • In order to screen the aging related proteins in human normal colon epithelia, the comparative proteomics analysis was applied to get the two-dimensional electrophoresis (2-DE) profiles with high resolution and reproducibility from normal colon epithelial tissues of young and aged people. Differential proteins between the colon epithelia of two age groups were found with PDQuest software. The thirty five differential protein-spots were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and database searching. Among them there are sixteen proteins which are significantly up-regulated in the colonic mucosal epithelia of young people group, which include ATP synthase beta chain, electron transfer flavoprotein alpha-subunit, catalase, glutathione peroxidase 1, annexin A2 and heat shock cognate 71 kDa protein, etc.; There are nineteen proteins which are significantly up-regulated in the colonic mucosal epithelia of aged people group, which include far upstream element-binding protein 1, nucleoside diphosphate kinase B, protein disulfide-isomerase precursor and VDAC-2, etc.. The identified differential proteins appear to be involved in metabolism, energy generation, chaperone, antioxidation, signal transduction, protein folding and apoptosis. The data will help to understand the molecular mechanisms of human colon epithelial aging.

Cloning and Characterization of Bombyx mori Cyclophilin A

  • Kim, Sung-Wan;Yun, Eun-Young;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Kwon, O-Yu;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.2
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    • pp.223-229
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    • 2011
  • Cyclophilins are originally identified as cytosolic binding protein of the immunosuppressive drug cyclosporine A. They have an activity of peptidyl prolyl cis/trans-isomerases (PPIase), which may play important roles in protein folding, trafficking, assembly and cell signaling. In this study, we report the cloning and characterization of a Bombyx mori cyclophilin A (bCypA) cDNA. The full-length cDNA of bCypA consist of 947 nucleotides with a polyadenylation signal sequence AATAAA and contain an open reading frame of 498 nucleotides encoding a polypeptide of 166 amino acids. The deduced amino acid sequence of bCypA shares a central peptidyl prolyl cis/trans-isomerase and a cyclosporin-A-binding domain with other cyclophilin sequences. Relative quantification real-time (RT) PCR analysis shows that mRNA transcripts of bCypA are detected in all the investigated tissues and highest expression level in the skin of 3-day-old 5 instar larva. Also, bCypA had PPIase activity on the proline-containing peptides. Accordingly, we suggest that bCypA is a new member of the cyclophilin A (CyPA) family and will be useful for quality control of bioactivity recombinant proteins with proline-containing peptides.

Raw Spectrum Analysis of operated UHF-Wind Profiler Radar in South Korea (국내 운용 UHF-윈드프로파일러 레이더의 원시 스펙트럼 분석)

  • Lee, Kyung-Hun;Kwon, Byung-Hyuk;Kim, Yu-Jin;Lee, Geon-Myeong
    • The Journal of the Korea institute of electronic communication sciences
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    • v.17 no.5
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    • pp.767-774
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    • 2022
  • In this paper raw spectrum data were analyzed to suggest the moving forward of performance evaluation and quality control of wind profilers of four manufacturers operating in South Korea. For the analysis, the profile of the spectrum averaged by season and the profile of four statistical values (minimum, average, median, and maximum) calculated by Power Spectrum Density (PSD) were used. The quality of spectrum data was the best for LAP-3000, followed by YKJ3, PCL-1300, and CLC-11-H. In Cheorwon and Chupungnyeong, where PCL-1300 was installed, the variability of the spectrum due to ground clutter and non-meteorological signals was large, so ground clutter removal and signal processing such as moving average and multi-peak were required. In Gunsan and Paju, where CLC-11-H was installed, DC (Direct Current) bias and propagation folding were found, so it is necessary to remove the DC bias and limit the effective altitude for observation.

Differential Proteomic Analysis of Chinese fir Clone Leaf Response to Salicylic Acid

  • Yang, Mei;Lin, Sizu;Cao, Guangqiu
    • Journal of Forest and Environmental Science
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    • v.26 no.2
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    • pp.83-94
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    • 2010
  • Chinese fir (Latin name: Cunninghaimia lanceolata) is one of the major commercial coniferous trees. Most of Chinese fir forests are managed in successive rotation sites, which lead productivity to decline. Autotoxicity is the important reason for soil degradation of Chinese fir plantation, especially, phenolic acids are considered as the major allelopathic toxins which induce autotoxicity in Chinese fir rotation stands. We performed here proteomic approach to investigate the response of proteins in Chinese fir leaves to salicylic acid. The tube plantlets of Chinese fir clone were treated with 120 mg/L salicylic acid for 1, 3 and 5th day. 2-DE, coupled with MALDI-TOF-TOF/MS, was used to separate and identify the responsive proteins. We found 12, 7, and 12 candidate protein spots that were up- or down-regulated by at least 2.5 fold after 1, 3, and 5th day of the stress, respectively. Of these protein spots, 16 spots were identified successfully. According to the putative physiological functions, these proteins were categorized into five classes (1) the proteins involved in protein stability and folding, including 26S proteome, Grp78, Hsp70, Hsp90 and PPIase; (2) the protein involved in photosynthesis and respiration, including OEC 33 kDa subunit, GAPDH; (3) the protein related to cell endurance to acid, F-ATPase; (4) the protein related to cytoskeleton, tubulin; (5) the protein related to protein translation: prolyl-tRNA synthetase. These results give new insights into autotoxic substance stress response in Chinese fir leaves and provide preliminary footprints for further studies on the molecular signal mechanisms induced by the stress.

Disturbance of $\alpha$-Amylase Secretion from Bacillus amyloliquefaciens Cells by the Treatment of Puromycin and Magnesium (Bacillus amyloliquefaciens에서 Puromycin 과 Magnesium에 의한 $\alpha$-Amylase 의 분비저해)

  • 안순자;김순옥;이동희;송방호
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.412-420
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    • 1989
  • To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

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Broadening the Understanding of Sixteenth-century Real Scenery Landscape Painting: Gyeongpodae Pavilion and Chongseokjeong Pavilion (16세기(十六世紀) 실경산수화(實景山水畫) 이해의 확장 : <경포대도(鏡浦臺圖)>, <총석정도(叢石亭圖)>를 중심으로)

  • Lee, Soomi
    • MISULJARYO - National Museum of Korea Art Journal
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    • v.96
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    • pp.18-53
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    • 2019
  • The paintings Gyeongpodae Pavilion and Chongseokjeong Pavilion were recently donated to the National Museum of Korea and unveiled to the public for the first time at the 2019 special exhibition "Through the Eyes of Joseon Painters: Real Scenery Landscapes of Korea." These two paintings carry significant implications for understanding Joseon art history. Because the fact that they were components of a folding screen produced after a sightseeing tour of the Gwandong regions in 1557 has led to a broadening of our understanding of sixteenth-century landscape painting. This paper explores the art historical meanings of Gyeongpodae Pavilion and Chongseokjeong Pavilion by examining the contents in the two paintings, dating them, analyzing their stylistic characteristics, and comparing them with other works. The production background of Gyeongpodae Pavilion and Chongseokjeong Pavilion can be found in the colophon of Chongseokjeong Pavilion. According to this writing, Sangsanilro, who is presumed to be Park Chung-gan (?-1601) in this paper, and Hong Yeon(?~?) went sightseeing around Geumgangsan Mountain (or Pungaksan Mountain) and the Gwandong region in the spring of 1557, wrote a travelogue, and after some time produced a folding screen depicting several famous scenic spots that they visited. Hong Yeon, whose courtesy name was Deokwon, passed the special civil examination in 1551 and has a record of being active until 1584. Park Chung-gan, whose pen name was Namae, reported the treason of Jeong Yeo-rip in 1589. In recognition of this meritorious deed, he was promoted to the position of Deputy Minister of the Ministry of Punishments, rewarded with the title of first-grade pyeongnan gongsin(meritorious subject who resolved difficulties), and raised to Lord of Sangsan. Based on the colophon to Chongseokjeong Pavilion, I suggest that the two paintings Gyeongpodae Pavilion and Chongseokjeong Pavilion were painted in the late sixteenth century, more specifically after 1557 when Park Chung-gan and Hong Yeon went on their sightseeing trip and after 1571 when Park, who wrote the colophon, was in his 50s or over. The painting style used in depicting the landscapes corresponds to that of the late sixteenth century. The colophon further states that Gyeongpodae Pavilion and Chongseokjeong Pavilion were two paintings of a folding screen. Chongseokjeong Pavilion with its colophon is thought to have been the final panel of this screen. The composition of Gyeongpodae Pavilion recalls the onesided three-layered composition often used in early Joseon landscape paintings in the style of An Gyeon. However, unlike such landscape paintings in the An Gyeon style, Gyeongpodae Pavilion positions and depicts the scenery in a realistic manner. Moreover, diverse perspectives, including a diagonal bird's-eye perspective and frontal perspective, are employed in Gyeongpodae Pavilion to effectively depict the relations among several natural features and the characteristics of the real scenery around Gyeongpodae Pavilion. The shapes of the mountains and the use of moss dots can be also found in Welcoming an Imperial Edict from China and Chinese Envoys at Uisungwan Lodge painted in 1557 and currently housed in the Kyujanggak Institute for Korean Studies at Seoul National University. Furthermore, the application of "cloud-head" texture strokes as well as the texture strokes with short lines and dots used in paintings in the An Gyeon style are transformed into a sense of realism. Compared to the composition of Gyeongpodae Pavilion, which recalls that of traditional Joseon early landscape painting, the composition of Chongseokjeong Pavilion is remarkably unconventional. Stone pillars lined up in layers with the tallest in the center form a triangle. A sense of space is created by dividing the painting into three planes(foreground, middle-ground, and background) and placing the stone pillars in the foreground, Saseonbong Peaks in the middle-ground, and Saseonjeong Pavilion on the cliff in the background. The Saseonbong Peaks in the center occupy an overwhelming proportion of the picture plane. However, the vertical stone pillars fail to form an organic relation and are segmented and flat. The painter of Chongseokjeong Pavilion had not yet developed a three-dimensional or natural spatial perception. The white lower and dark upper portions of the stone pillars emphasize their loftiness. The textures and cracks of the dense stone pillars were rendered by first applying light ink to the surfaces and then adding fine lines in dark ink. Here, the tip of the brush is pressed at an oblique angle and pulled down vertically, which shows an early stage of the development of axe-cut texture strokes. The contrast of black and white and use of vertical texture strokes signal the forthcoming trend toward the Zhe School painting style. Each and every contour and crack on the stone pillars is unique, which indicates an effort to accentuate their actual characteristics. The birds sitting above the stone pillars, waves, and the foam of breaking waves are all vividly described, not simply in repeated brushstrokes. The configuration of natural features shown in the above-mentioned Gyeongpodae Pavilion and Chongseokjeong Pavilion changes in other later paintings of the two scenic spots. In the Gyeongpodae Pavilion, Jukdo Island is depicted in the foreground, Gyeongpoho Lake in the middle-ground, and Gyeongpodae Pavilion and Odaesan Mountain in the background. This composition differs from the typical configuration of other Gyeongpodae Pavilion paintings from the eighteenth century that place Gyeongpodae Pavilion in the foreground and the sea in the upper section. In Chongseokjeong Pavilion, stone pillars are illustrated using a perspective viewing them from the sea, while other paintings depict them while facing upward toward the sea. These changes resulted from the established patterns of compositions used in Jeong Seon(1676~1759) and Kim Hong-do(1745~ after 1806)'s paintings of Gwandong regions. However, the configuration of the sixteenth-century Gyeongpodae Pavilion, which seemed to have no longer been used, was employed again in late Joseon folk paintings such as Gyeongpodae Pavilion in Gangneung. Famous scenic spots in the Gwandong region were painted from early on. According to historical records, they were created by several painters, including Kim Saeng(711~?) from the Goryeo Dynasty and An Gyeon(act. 15th C.) from the early Joseon period, either on a single scroll or over several panels of a folding screen or several leaves of an album. Although many records mention the production of paintings depicting sites around the Gwandong region, there are no other extant examples from this era beyond the paintings of Gyeongpodae Pavilion and Chongseokjeong Pavilion discussed in this paper. These two paintings are thought to be the earliest works depicting the Gwandong regions thus far. Moreover, they hold art historical significance in that they present information on the tradition of producing folding screens on the Gwandong region. In particular, based on the contents of the colophon written for Chongseokjeong Pavilion, the original folding screen is presumed to have consisted of eight panels. This proves that the convention of painting eight views of Gwangdong had been established by the late sixteenth century. All of the existing works mentioned as examples of sixteenth-century real scenery landscape painting show only partial elements of real scenery landscape painting since they were created as depictions of notable social gatherings or as a documentary painting for practical and/or official purposes. However, a primary objective of the paintings of Gyeongpodae Pavilion and Chongseokjeong Pavilion was to portray the ever-changing and striking nature of this real scenery. Moreover, Park Chung-gan wrote a colophon and added a poem on his admiration of the scenery he witnessed during his trip and ruminated over the true character of nature. Thus, unlike other previously known real-scenery landscape paintings, these two are of great significance as examples of real-scenery landscape paintings produced for the simple appreciation of nature. Gyeongpodae Pavilion and Chongseokjeong Pavilion are noteworthy in that they are the earliest remaining examples of the historical tradition of reflecting a sightseeing trip in painting accompanied by poetry. Furthermore, and most importantly, they broaden the understanding of Korean real-scenery landscape painting by presenting varied forms, compositions, and perspectives from sixteenth-century real-scenery landscape paintings that had formerly been unfound.