• Title/Summary/Keyword: T7 expression system

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Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo (자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과)

  • Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.559-566
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    • 2017
  • This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.

Software development for the visualization of brain fiber tract by using 24-bit color coding in diffusion tensor image

  • Oh, Jung-Su;Song, In-Chan;Ik hwan Cho;Kim, Jong-Hyo;Chang, Kee-Hyun;Park, Kwang-Suk
    • Proceedings of the KSMRM Conference
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    • 2002.11a
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    • pp.133-133
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    • 2002
  • Purpose: The purpose of paper is to implement software to visualize brain fiber tract using a 24-bit color coding scheme and to test its feasibility. Materials and Methods: MR imaging was performed on GE 1.5 T Signa scanner. For diffusion tensor image, we used a single shot spin-echo EPI sequence with 7 non-colinear pulsed-field gradient directions: (x, y, z):(1,1,0),(-1,1,0),(1,0,1),(-1,0,1),(0,1,1),(0,1,-1) and without diffusion gradient. B-factor was 500 sec/$\textrm{mm}^2$. Acquisition parameters are as follows: TUTE=10000ms/99ms, FOV=240mm, matrix=128${\times}$128, slice thickness/gap=6mm/0mm, total slice number=30. Subjects consisted of 10 normal young volunteers (age:21∼26 yrs, 5 men, 5 women). All DTI images were smoothed with Gaussian kernel with the FWHM of 2 pixels. Color coding schemes for visualization of directional information was as follows. HSV(Hue, Saturation, Value) color system is appropriate for assigning RGB(Red, Green, and Blue) value for every different directions because of its volumetric directional expression. Each of HSV are assigned due to (r,$\theta$,${\Phi}$) in spherical coordinate. HSV calculated by this way can be transformed into RGB color system by general HSV to RGB conversion formula. Symmetry schemes: It is natural to code the antipodal direction to be same color(antipodal symmetry). So even with no symmetry scheme, the antipodal symmetry must be included. With no symmetry scheme, we can assign every different colors for every different orientation.(H =${\Phi}$, S=2$\theta$/$\pi$, V=λw, where λw is anisotropy). But that may assign very discontinuous color even between adjacent yokels. On the other hand, Full symmetry or absolute value scheme includes symmetry for 180$^{\circ}$ rotation about xy-plane of color coordinate (rotational symmetry) and for both hemisphere (mirror symmetry). In absolute value scheme, each of RGB value can be expressed as follows. R=λw|Vx|, G=λw|Vy|, B=λw|Vz|, where (Vx, Vy, Vz) is eigenvector corresponding to the largest eigenvalue of diffusion tensor. With applying full symmetry or absolute value scheme, we can get more continuous color coding at the expense of coding same color for symmetric direction. For better visualization of fiber tract directions, Gamma and brightness correction had done. All of these implementations were done on the IDL 5.4 platform.

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Cloning and Transcription Analysis of Sporulation Gene (spo5) in Schizosaccharomyces pombe (Schizosaccharomyces bombe 포자형성 유전자(spo5)의 Cloning 및 전사조절)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.112-118
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    • 2002
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.

Mitotic-Specific Methylation in the HeLa Cell through Loss of DNMTs and DMAP1 from Chromatin

  • Kim, Kee-Pyo;Kim, Gun-Do;Kang, Yong-Kook;Lee, Dong-Seok;Koo, Deog-Bon;Lee, Hoon-Taek;Chung, Kil-Saeng;Lee, Kyung-Kwang;Han, Yong-Mahn
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.27-27
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    • 2003
  • A diversified and concentrative approach of methylation player can be one of the most powerful studies in the understanding of global epigenetic modifications. Previous studies have suggested that DNA methylation contributes to transcriptional silencing through the several DNA methylation-mediated repression systems by hypermethylation, including methyltransferases (DNMTs), DNA methyltransferase association protein 1 (DMAPl), methyl-CpG binding domain (MBD), and histone deacetylases (HDACs). Assembly of these regulatory protein complexes act sequentially, reciprocally, and interdependently on the newly composed DNA strand through S phase. Therefore, these protein complexes have a role in coupling DNA replication to the designed turn-off system in genome. In this study, we attempted to address the role of DNA methylation by the functional analysis of the methyltransferase molecule, we described the involvement of DMAP1 and DNMTs in cell divistion and the effect of their loss. We also described distinct patterns that DMAP1 and DNMTs are spatially reorganized and displaced from condensing chromosomes as cells progress through mitosis in HeLa cell, COS7, and HIH3T3 cell cycle progressions. DNMT1, DNMT3b, and DMAP1 do not stably contact the genetic material during chromosome compaction and repressive expression. These finding show that the loss of activities of DNMTs and DMAP1 occure stage specifically during the cell cycle, may contribute to the integral balance of global DNA methylation. This is consistent with previous studies resulted in decreased histone acetyltransferases and HDACs, and differs from studies resulted in increased histone methyltransferases. Our results suggest that DNA methylation by DNMTs and DMAP1 during mitosis acts to antagonize hypermethylation by which this mark is epigenetical mitotic-specific methylation.

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Cloning of tlrD, 23S rRNA Monomethyltransferase Gene, Overexpression in Eschepichia coli and Its Activity (235 rRNA Monomethyltransferase인 tlrD의 클로닝, 이의 대장균에서 대량생산과 활성 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.166-172
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    • 2007
  • ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at $37^{\circ}C$ accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as $18^{\circ}C$ did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.

Optimization of Dual Layer Phoswich Detector for Small Animal PET using Monte Carlo Simulation

  • Y.H. Chung;Park, Y.;G. Cho;Y.S. Choe;Lee, K.H.;Kim, S.E.;Kim, B.T.
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2003.09a
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    • pp.44-44
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    • 2003
  • As a basic measurement tool in the areas of animal models of human disease, gene expression and therapy, and drug discovery and development, small animal PET imaging is being used increasingly. An ideal small animal PET should have high sensitivity and high and uniform resolution across the field of view to achieve high image quality. However, the combination of long narrow pixellated crystal array and small ring diameter of small animal PET leads to the degradation of spatial resolution for the source located at off center. This degradation of resolution can be improved by determining the depth of interaction (DOI) in the crystal and by taking into account the information in sorting the coincident events. Among a number of 001 identification schemes, dual layer phsowich detector has been widely investigated by many research groups due to its practicability and effectiveness on extracting DOI information. However, the effects of each crystal length composing dual layer phoswich detector on DOI measurements and image qualities were not fully characterized. In order to minimize the DOI effect, the length of each layer of phoswich detector should be optimized. The aim of this study was to perform simulations using a simulation tool, GATE to design the optimum lengths of crystals composing a dual layer phoswich detector. The simulated small PET system employed LSO front layer LuYAP back layer phoswich detector modules and the module consisted of 8${\times}$8 arrays of dual layer crystals with 2 mm ${\times}$ 2 mm sensitive area coupled to a Hamamatsu R7600 00 M64 PSPMT. Sensitivities and variation of radial resolutions were simulated by varying the length of LSO front layer from 0 to 10 mm while the total length (LSO + LuYAP) was fixed to 20 mm for 10 cm diameter ring scanner. The radial resolution uniformity was markedly improved by using DOI information. There existed the optimal lengths of crystal layers to minimize the variation of radial resolutions. In 10 cm ring scanner configuration, the radial resolution was kept below 3.4 mm over 8 cm FOV while the sensitivity was higher than 7.4% for LSO 5 mm : LuYAP 15 mm phoswich detector. In this study, the optimal length of dual layer phoswich detector was derived to achieve high and uniform radial resolution.

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Change of Dendritic Cell Subsets Involved in Protection Against Listeria monocytogenes Infection in Short-Term-Fasted Mice

  • Young-Jun Ju;Kyung-Min Lee;Girak Kim;Yoon-Chul Kye;Han Wool Kim;Hyuk Chu;Byung-Chul Park;Jae-Ho Cho;Pahn-Shick Chang;Seung Hyun Han;Cheol-Heui Yun
    • IMMUNE NETWORK
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    • v.22 no.2
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    • pp.16.1-16.20
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    • 2022
  • The gastrointestinal tract is the first organ directly affected by fasting. However, little is known about how fasting influences the intestinal immune system. Intestinal dendritic cells (DCs) capture antigens, migrate to secondary lymphoid organs, and provoke adaptive immune responses. We evaluated the changes of intestinal DCs in mice with short-term fasting and their effects on protective immunity against Listeria monocytogenes (LM). Fasting induced an increased number of CD103+CD11b- DCs in both small intestinal lamina propria (SILP) and mesenteric lymph nodes (mLN). The SILP CD103+CD11b- DCs showed proliferation and migration, coincident with increased levels of GM-CSF and C-C chemokine receptor type 7, respectively. At 24 h post-infection with LM, there was a significant reduction in the bacterial burden in the spleen, liver, and mLN of the short-term-fasted mice compared to those fed ad libitum. Also, short-term-fasted mice showed increased survival after LM infection compared with ad libitum-fed mice. It could be that significantly high TGF-β2 and Aldh1a2 expression in CD103+CD11b- DCs in mice infected with LM might affect to increase of Foxp3+ regulatory T cells. Changes of major subset of DCs from CD103+ to CD103- may induce the increase of IFN-γ-producing cells with forming Th1-biased environment. Therefore, the short-term fasting affects protection against LM infection by changing major subset of intestinal DCs from tolerogenic to Th1 immunogenic.

Enhancement of Bioactive Compounds in Mugwort Grown under Hydroponic System by Sucrose Supply in a Nutrient Solution (양액 내 자당 처리에 의한 수경재배 쑥의 생리활성물질 증진)

  • Moon-Sun Yeom;Jun-Soo Lee;Myung-Min Oh
    • Journal of Bio-Environment Control
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    • v.32 no.1
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    • pp.23-33
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    • 2023
  • Sucrose (suc) is a disaccharide that consists of glucose (glu) and fructose (fru). It is a carbohydrate source that acts as a nutrient molecule and a molecular signal that regulates gene expression and alters metabolites. This study aimed to evaluate whether suc-specific signaling induces an increase in bioactive compounds by exogenous suc absorption via roots or whether other factors, such as osmotic stress or biotic stress, are involved. To compare the osmotic stress induced by suc treatment, 4-week-old cultured mugwort plants were subjected to Hoagland nutrient solution with 10 mM, 30 mM, and 50 mM of suc or mannitol (man) for 3 days. Shoot fresh weight in suc and man treatments was not significantly different from the control. Both man and suc treatments increased the content of bioactive compounds in mugwort, but they displayed different enhancement patterns compared to the suc treatments. Mugwort extract treated with suc 50 mM effectively protected HepG2 liver cells damaged by ethanol and t-BHP. To compare the biotic stress induced by suc treatment, 3-week-old mugwort plants were subjected to microorganism and/or suc 30 mM with Hoagland nutrient solution. Microorganisms and/or suc 30 mM treatments showed no difference about the shoot fresh weight. However, sugar content in mugwort treated with suc 30 mM and microorganism with suc 30 mM treatment was significantly higher than that of the control. Suc 30 mM and microorganism with suc 30 mM were effective in enhancing bioactive compounds than microorganism treatment. These results suggest that mugwort plants can absorb exogenous suc via roots and the enhancement of bioactive compounds by suc treatment may result not from osmotic stress or biotic stress because of microorganism, but by suc-specific signaling.

Effect of Experimental Muscle Fatigue on Muscle Pain and Occlusal Pattern (실험적으로 유발되는 근피로가 근통증 및 교합양상에 미치는 영향)

  • Kim, Jae-Chang;Lim, Hyun-Dae;Kang, Jin-Kyu;Lee, You-Mee
    • Journal of Oral Medicine and Pain
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    • v.33 no.3
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    • pp.279-294
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    • 2008
  • This study aimed to make an analysis of the occlusion in the state of muscle fatigue produced by excessive mouth opening and clenching during the dental treatment to control the dental pain and to evaluate the sensory nerve in the muscle pain state. Most of the reasons why patients visit the dental office result in pain-either conceivably the dental origin pain or the non-dental origin pain. The dental offices have many therapeutic actions to produce the masticatory muscle fatigue for the treatment. Dental treatment with long minutes of mouth opening can cause some headaches, masticatory muscle pain and mouth opening difficulties. Patients with mastication problems who visits a dental office to alleviate pain run against another unexpected pain with other aspects. This study uses T-scan II system(Tekscan Co., USA) for the evaluation on the occlusal pattern in the experimental muscle fatigue after clenching, opening the mouth excessively and chewing gum. The occlusal contact pattern is analyzed by the contact timing, namely first, intercuspal, maximum and end point of contact. This inspection was performed at frequencies of 2000Hz, 250 Hz and 5 Hz before and after each experimental muscle pain was produced to 24 subjects who had normal occlusion without the orthodontic treatment or a wide range of the prosthesis by using $neurometer^{\circledR}$ CPT/C(Neurotron, Inc. Baltimore, Maryland, USA). The measuring sites were mandibular nerve experimental muscle fatigue respectively. This study could obtain the following results after the assessment of occlusion and sensory nerve of the experimental muscle fatigue. 1. There were the fastest expression after the excessive mouth opening in muscle fatigue and after tooth clenching in muscle pain. In the visual analog scale that records the subjective level, there was the highest scale after the clenching in the muscle fatigue in jumping off the point of pain. 2. Tooth contact time, contact force, relative contact force on the point of the first contact had no difference, and there were decreases in the contact force after the excessive mouth opening on intercuspal position point, after the excessive mouth opening and the gum chewing on the point of the maximum, and in the contact time after all the experimental muscle fatigue state on the point of the end contact. 3. There was no statistic significance in the current perception threshold before and after the experimental muscle fatigue. 4. There was no significant difference in the contact number, the maximal contact number on the point of the first contact, and the contact number after the mouth opening and gum chewing on the point of the intercuspal position and the contact number after the experimental muscle fatigue on the maximum point, and showed significant decreases. In conclusion, it was found that the occlusal pattern can cause the changes on the case of the clinical muscle weakness by intra-external oral events. It was important that the sedulous attention to details is required during dental treatment in case of excessive mouth opening, mastication and clenching.

Analysis of the Stage and Performance Elements for Bongsudang-jinchan Banquet in Joseon Dynasty (봉수당진찬(奉壽堂進饌)의 무대와 공연 요소 분석)

  • Song, Hye-jin
    • (The) Research of the performance art and culture
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    • no.18
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    • pp.413-444
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    • 2009
  • This paper is an analysis of stage and performance elements for the ceremonial procedures and dance featured in bongsudang-jinchan, a feast celebrating the 60th birthday of Hyegyeong-gung Hongssi (Crown Princess Hong of Hyegyeonggung), the mother of King Jeongjo, which took place in Hwaseong haegung palace in 1795. The primary sources used are data on bongsudangjinchan recorded in Wonhaeng-eulmyojeongriuigwe, Jeongjo-sillok, Hongjae-jeonseo, pictorial sources such as Folding Screens of Hwaseong-neunghaeng and Hwaseong Ilgi, which is a journal in Korean by Yi Hui-pyeong. A court ceremony to offer music, dance, flowers, and food, as well as wine and poetry which express the sentiments of chung (fidelity) and hyo (filial piety) was considered a national ceremony and has constituted a unique musical culture during the 500 years of Joseon dynasty. However, after the fall of Joseon dynasty, ceremonial music and dance, which have been organically linked within the overall symbolic system of ye (courtesy), became scattered to become independent 'pieces.' As a result, all of their philosophy, principles, and the time-space interpretation of court music and dance became greatly reduced, leaving only the artistic expression and formal structure of the music and dance to become emphasized. Since the 1990s, there has been many research and events aiming to re-create the court ceremonial tradition, resulting in the increase of the related performance activities. This is especially true with bongsudang-jinchan, which is now being performed on modern stage in various forms. However there are still many problems to be solved, such as the issue of re-creating and restoring the original, and the question of artistic value found in the traditional pieces. Until now, much focus has been paid to the outer re-construction of uiju document as recorded in Wonhaeng-eulmyo-jeongriuigwe. On the other hand, there lacked an in-depth study which analyzes the stage situation and performance elements. Therefore in this paper, after focusing on the stage structure and performance elements, it is concluded that bongsudang-jinchan, the only court feast to be held in Haeng gung, not only consists of the fundamental aspects of court performance principle as 'governing through ye and ak (music),' but also served as an important occasion to bring together the sovereign and the subject. Bongsudang-jinchan had features of both naeyeon (feast for ladies) and oeyeon (feast for gentlemen). It minimized the use of screens and allowed every guest to enjoy food, music, and dance together, but provided a separate tent for foreign guests, maintaining the ideal balance between equality and distinction among different gender and social status. A screen symbolizing the venue for the feast is placed for all of the government officials. The king then pronounces the beginning of the banquet in which the ideal of gunsin-dongyeon (king and officials dining together) is realized. This indicates that bongsudang-jinchan, compared to other court ceremonies that emphasize the principle of yeak (courtesy and music), focuses more on the spirit of harmony and rapprochement. The king played a more active role in bongsudang-jinchan than in any other royal feasts. Examples as recorded in uiju documents are; Jeongjo's conversation with his retainers after the 7th wine, king's bestowing of food and flowers to the officials, writing his own majesty's poems with regard to the festival, and asking the retainers to write replying poems. All these played an important part in making the occasion more rich, extensive, and meaningful. Moreover, as analysis of the structure of orchestral music and court dance featured in bongsudang-jinchan shows, it was like any other court banquet in that it employed minimal use of extravagance in movements and conversation. However, the colors and tonal texture used in the music and dance were more brilliant in this case. Compared to other banquets that took place before king Jeongjo, the dance style was more diverse, which included some of the latest additions. There were past performances arranged anew. Noteworthy are; the incorporation of "Seonyurak (Boat Dance)" and "Geommu (Sword Dance)," traditionally used for local officials and civilians feast, to suit the court taste; and the use of saenghwang (mouth organ), which was a rising instrument in pungnyubang (literati's private salon), for "Hakmu (Cranes Dance)." This especially indicates the nature of the 'open structure' pursued by the court banquets at the time, which strove to break away from the traditional rules and customs and accept something new.