생물전환을 통한 음나무발효물의 지표성분 설정 및 동시분석법 검증

장원희,이화영,이봉진,김진만,박선주,Jang, Won Hui,Lee, Wha Young,Lee, Bong Jin,Kim, Jean Man,Park, Seon Ju 한국식품영양학회 2019 韓國食品營養學會誌 Vol.32 No.2

The aim of this study was to select compounds for the standardization of fermented Kalopanax pictus Nakai (KP-F), to develop the analysis method using HPLC-PDA and to perform method validation. KP-F is a fermented powder developed to improve the original physiological activities and create a new functionality. Eleutheroside E, Acanthoside B, and Syringaresinol were selected as the standard compounds and developed our own method for simultaneous analysis. The analyte was isolated using C18 column with a gradient elution of 0.05 M phosphoric acid in water and methanol as the mobile phase at a flow rate of 1 mL/min and detected at 210 nm. As a result, all standard compounds showed good linearity with an $R^2$ (coefficient of correlation) of 1.000 and for the limit of detection range of $0.710{\sim}0.831{\mu}g/mL$, and the limit of quantification as $2.150{\sim}2.520{\mu}g/mL$. The precision was RSD (%) of less than 4.80%, while the accuracy was 4.70%>RSD (%) for the range 102.44~110.48%. In conclusion, the developed analysis method is suitable for the detection of Eleutheroside E, Acanthoside B, and Syringaresinol in KP-F.

강화 묘지사지 전면구들-누마루 조합 유형에 관한 고찰

장원희(Jang, Won-Hee),류성룡(Ryoo, Seong-Lyong) 대한건축학회 2023 대한건축학회 학술발표대회 논문집 Vol.43 No.2

This study aims to analyze the entire Ondol-Numaru combination type in Late Goryeo and early Joseon Period architecture and to examine the original type of the entire Ondol-Daecheongmaru combination type that became common in the Joseon Dynasty.There are many differences in the occurrence of Ondol, but the spread of the one-line Gorae during the Three Kingdoms period begins with the early iron period. Around the 13th century of Goryeo, the entire ondol appeared, and a major change occurred due to a new type of floor composition consisting of a combination of the entire ondol-maru. Before the occurrence of the entire Ondol, the composition consisting of an Ondol, a Fireplace, and a Soil floor installed in a part of the ‘Khan’ creates a level difference between the Ondol and the Soil floor in each compartment As a result, this level difference hinders the composition of the wood floor around the Ondol-Khan, so the generation of the entire Ondol can be said to be the basis for the Ondol and the Wood floor to be configured without structural restrictions. Ondol and wood floor are spatial elements whose structure and purpose of use are largely different, but in traditional Korean architecture, these elements are combined and composed of each other in one space. These characteristics are characteristics of traditional Korean architecture that are not well seen in East Asian architecture except Korea, and we would like to consider the original shape of the entire ondol-maru combination through the remains of Late Goryeo and early Joseon Period architecture.

인공피부모델을 이용한 보습파우더의 보습 효능 평가

장원희 ( Won Hee Jang ),정경미 ( Kyoung Mi Jung ),최선경 ( Sun Kyung Choi ),정행선 ( Haeng Sun Jung ),이수현 ( Su Hyon Lee ),최영진 ( Young Jin Choi ),박영호 ( Young Ho Park ),임경민 ( Kyung Min Lim ) 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1

Dry skin is induced by excessive transdermal water loss due to the damages of intracellular lipids. Moisturizer is safe and effective for improving dry skin, however, there is few efficient alternative method to evaluate moisturizing activity of new cosmetic ingredients. Here, we investigated the moisturizing activity of a face powder coated with mannosylerithritol lipids (MEL) that are similar to ceramides in structure and intermediates in the biosynthesis of intracellular lipids, without using animals or human but Keraskin™, a three-dimensional reconstructed human epidermis. After Keraskin™ was incubated, MEL-coated sericite or uncoated sericite was applied and then sodium lauryl sulphate (SLS) was treated with a nylon mesh covering to spread it uniformly. After incubation for 15 min, Keraskin™ was washed and it was post-incubated for 24 hr. At the end of post-incubation, Keraskin™ was incubated with MTT for 3 hr to measure viability. Formed purple formazan was extracted with isopropanol for 2 hr. Absorbance of extract(A590) was measured using spectrophotometer. Cell viability was calculated by dividing with the absorbance of negative control. The viability of reconstructed epidermis treated 10% SLS was determined to be 12.3 ± 4.87%. When SLS was co-treated with sericite or MEL-coated sericite, cell viability was increased to 23.7 ± 2.92% or 71.5 ± 20% (p<0.05), respectively. MEL-coated sericite inhibited SLS-induced cytotoxicity substantially in comparison with sericite alone. Based on these results, reconstructed human epidermis can be used for the evaluation of the efficacy of cosmetic ingredients or products.

MRI 영상 및 Deep Learning을 이용한 병변 인식 알고리즘 개발

장원희(WonHee Jang),이승규(Seungkyu Lee) 한국정보과학회 2018 한국정보과학회 학술발표논문집 Vol.2018 No.6

세포내형 동결보존액을 이용한 사람 간세포의 동결보존 개선

장원희 ( Won Hee Jang ),김형인 ( Hyeong In Kim ),이우정 ( Woo Jeong Lee ),서지연 ( Ji Yeon Seo ),최창수 ( Chang Soo Choi ),최영길 ( Young Kil Choi ),허찬영 ( Chan Young Hur ),김기훈 ( Ki Hoon Kim ),박정익 ( Jeong Ik Park ),김광희 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4

Primary human hepatocytes are used for cell transplantation and bioartificial liver system for supporting patients with intractable liver diseases. Efficient cryopreservation of human hepatocytes is of great interest. To improve the cryopreservation outcome of human hepatocytes, effects of intracellular-type cryopreservation solutions and apoptosis inhibitor were investigated. Hepatocytes were isolated from the resected livers from 10 patients with intrahepatic stones or tumors. Isolated hepatocytes were cryopreserved usingeither culture media with 10% DMSO, CryoStor CS5, or University of Wisconsin(UW) solution with 10% DMSO. CryoStor CS5 significantly improved the post-thaw cell viability assessed by MTT test, the recovery of viable cells, and the plating efficiency compared to the culture media-based solution. UW solution also showed some beneficial effects on the cryopreservation outcome, but the effects were not statistically significant except the improved 24 hours post-thaw viability. On the other hand, inhibition of apoptosis by a pan-caspase inhibitor Z-VAD-fmk showed no effects on the cryopreservation outcome. These results demonstrate the usefulness of the intracellular-type cryopreservation solutions and CryoStor CS5 is superior to UW solution for the cryopreservation of human hepatocytes.

연구논문 : 인공피부모델을 이용한 보습파우더의 보습 효능 평가

장원희 ( Won Hee Jang ),정경미 ( Kyoung Mi Jung ),최선경 ( Sun Kyung Choi ),정행선 ( Haeng Sun Jung ),이수현 ( Su Hyon Lee ),최영진 ( Young Jin Choi ),박영호 ( Young Ho Park ),임경민 ( Kyung Min Lim ) 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1

Dry skin is induced by excessive transdermal water loss due to the damages of intracellular lipids. Moisturizer is safe and effective for improving dry skin, however, there is few efficient alternative method to evaluate moisturizing activity of new cosmetic ingredients. Here, we investigated the moisturizing activity of a face powder coated with mannosylerithritol lipids (MEL) that are similar to ceramides in structure and intermediates in the biosynthesis of intracellular lipids, without using animals or human but Keraskin™, a three-dimensional reconstructed human epidermis. After Keraskin™ was incubated, MEL-coated sericite or uncoated sericite was applied and then sodium lauryl sulphate (SLS) was treated with a nylon mesh covering to spread it uniformly. After incubation for 15 min, Keraskin™ was washed and it was post-incubated for 24 hr. At the end of post-incubation, Keraskin™ was incubated with MTT for 3 hr to measure viability. Formed purple formazan was extracted with isopropanol for 2 hr. Absorbance of extract(A590) was measured using spectrophotometer. Cell viability was calculated by dividing with the absorbance of negative control. The viability of reconstructed epidermis treated 10% SLS was determined to be 12.3 ± 4.87%. When SLS was co-treated with sericite or MEL-coated sericite, cell viability was increased to 23.7 ± 2.92% or 71.5 ± 20% (p<0.05), respectively. MEL-coated sericite inhibited SLS-induced cytotoxicity substantially in comparison with sericite alone. Based on these results, reconstructed human epidermis can be used for the evaluation of the efficacy of cosmetic ingredients or products.

한옥 교보재에서 결구 방식 표현 한계에 관한 사례 연구

장원희(Jang, Won-Hee),류성룡(Ryoo, Seong-Lyong) 대한건축학회 2023 대한건축학회 학술발표대회 논문집 Vol.43 No.1

Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs)

Won Hee Jang(장원희),Young Joo Jeong(정영주),Won Hee Lee(이원희),Mooseong Kim(김무성),Sang-Jin Kim(김상진),Sang-Hwa Urm(엄상화),Il Soo Moon(문일수),Dae-Hyun Seog(석대현) 한국생명과학회 2016 생명과학회지 Vol.26 No.6

세포내소기관과 단백질 복합체의 운반은 kinesin superfamily proteins (KIFs)에 의해 매개된다. 처음으로 밝혀진 kinesin인 kinesin 1은 motor단백질로서 세포 내에서 미세소관을 따라 이동하며, 다양한 세포내 소기관이나 단백질복합체를 운반한다. Kinesin 1은 장쇄(KHC, 또한 KIF5s로도 통칭) 2분자와 단쇄(KLCs) 2분자로 구성된 4합체(tetramer) 구조를 가진다. KIF5s의 운반체 결합영역을 포함하는 말단영역은 다수의 운반체와 결합하지만, 결합운반체에 관하여 아직 충분히 밝혀지지 않았다. 본 연구에서 KIF5A의 결합 단백질을 동정하기 위하여 효모 two-hybrid screening을 수행하였고 철 저장 및 해독 기능을 하는 단백질인 ferritin heavy chain (Frt-h)을 찾아내었다. Frt-h은 KIF5A의 아미노산 800번과 940번 사이의 부위와 결합하며, 다른 KIF5s와도 결합함을 효모 two-hybrid assay로 확인하였다. 또한 Frt-h의 coiled-coil 도메인이 KIF5A와의 결합에 필수영역임을 밝혔다. 한편, ferritin light chain (Frt-l) 또한 KIF5s와 결합함을 효모 two-hybrid assay로 확인하였다. 이러한 단백질간의 결합을 glutathione S-transferase (GST) pull-down assay를 통하여 검증하였다. 추가적으로 생쥐의 뇌 파쇄액을 항 KHC 항체로 면역침강을 행한 결과, KLC1뿐만 아니라 Frt-h와 Frt-l도 같이 침강하였다. 이러한 결과들은 세포 내에서 kinesin 1이 ferritin 복합체를 운반함을 시사한다. The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

PtdIns(3,5)P2 5-phosphatase Fig4 Interacts with Kinesin Superfamily 5A (KIF5A)

Won Hee Jang(장원희),Dae-Hyun Seog(석대현) 한국생명과학회 2014 생명과학회지 Vol.24 No.1

Kinesin-1은 2개의 장쇄(KHCs, 또는 KIF5s)와 2개의 단쇄(KLCs)가 결합한 복합체로 되어 있다. 본 연구에서 효모 two-hybrid system을 이용하여 중추신경계의 신경세포에서 주로 발현되는 KIF5A와 결합하는 단백질을 탐색한 결과 phosphatidylinositol-3,5-bisphosphate (PI(3,5)P₂)의 5번 위치 인산을 제거하는 탈인산화효소 Fig4(Sac3)를 분리하였다. KIF5A는 Fig4의 C-말단과 결합함을 효모 two-hybrid assay로 확인하였다. Fig4는 KIF5A의 C-말단과 결합하지만, 두 개의 다른 장쇄인 KIF5B와 KIF5C 그리고 KLC1와는 결합하지 않았다. 단백질 간 결합을 glutathione S-transferase pull-down assay와 공동면역침강으로 추가 검증하였다. 생쥐의 뇌 파쇄액을 KIF5A 항체로 면역 침강한 결과 Fig4가 같이 침강하였다. 이러한 결과들은 kinesin-1이 Fig4와 결합한 단백질 복합체 혹은 운반체를 세포 내에서 운반함을 시사한다. Kinesin-1 consists of two heavy chains (KHCs), also called KIF5s, and two light chains (KLCs) that form a heterotetrameric complex. Here, we demonstrate the binding of a neuronal KHC, KIF5A, to the carboxyl (C)-terminal tail region of Fig4 (also known as Sac3), a phosphatase that removes the 5-phosphate from phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)P₂). Fig4 bound to the C-terminal region of KIF5A but not to other KHCs (KIF5B and KIF5C) and KLC1 in yeast two-hybrid assays. The interaction was further confirmed in a glutathione S-transferase pull-down assay and by co-immunoprecipitation. Anti-KIF5A antibody co-immunoprecipitated Fig4 with KIF5A from mouse brain extracts. These results suggest that kinesin-1 could transport the Fig4-associated protein complex or cargo in cells.

Cadms/SynCAMs/Necls/TSLCs Interact with Multi-PDZ Domain Protein MUPP1

Won Hee Jang(장원희),Young Joo Jeong(정영주),Sun Hee Choi(최선희),Sang-Jin Kim(김상진),Sang-Hwa Urm(엄상화),Il Soo Moon(문일수),Dae-Hyun Seog(석대현) 한국생명과학회 2014 생명과학회지 Vol.24 No.12

조직의 구조 안정성을 유지하는 세포 사이 연접복합체는 multi-PDZ domain protein 1 (MUPP1)을 포함하여 50종류 이상의 단백질로 이루어져 있다. MUPP1은 13개의 PDZ 도메인을 가지는 단백질로서 막경유(transmembrane) 단백질과 세포골격단백질이나 신호단백질 사이에서 scaffold로 작용한다고 알려져 있지만, MUPP1이 어떻게 세포막인접 단백질들을 연결하고 구조 안정화에 기여하는지에 대해 아직 명확히 밝혀지지 않았다. 본 연구에서 MUPP1의 PDZ 도메인과 상호 작용하는 단백질을 규명하기 위하여 효모 two-hybrid 방법을 이용, cell adhesion molecule 1 (Cadm1; SynCAM1, Necl-2 또는 TSLC1로도 알려짐)이 MUPP1과 결합하는 것을 확인하였다. Cadm1은 MUPP1의 2번째 PDZ 도메인과 결합하며, Cadm1의 C-말단에 존재하는 II 형 PDZ-결합모티프(-Y-F-I)가 MUPP1과의 결합에 필수적임을 확인하였다. 또한 MUPP1은 다른 Cadm family 단백질들인 Cadm2, Cadm3, 그리고 Cadm4와도 결합하며, 이러한 단백질간 결합은 glutathione S-transferase (GST) pull-down assay와 공동면역침강으로도 추가 확인하였다. 생쥐의 뇌 파쇄액을 MUPP1 항체로 면역침강하였을 때 Cadm1과 Cadm4가 같이 침강하였다. 이러한 결과들은 MUPP1이 세포 사이 연접에서 Cadms와 세포골격 단백질 사이를 연결한다는 것을 시사한다. Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by coimmunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.