Nitric oxide knock out mouse is good disease model for constipation dominant irritable bowel syndrome

장다은,황우성,염수청 한국실험동물학회 2016 한국실험동물학회 학술발표대회 논문집 Vol.2016 No.08

Multiple sgRNAs with overlapping sequences enhance CRISPR/Cas9-mediated knock-in efficiency

장다은,이재용,이재훈,구옥재,배희숙,정민희,배지현,황우성,장유진,이윤호,이한웅,염수청 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-

The CRISPR/Cas9 system is widely applied in genome engineering due to its simplicity and versatility. Although this has revolutionized genome-editing technology, knockin animal generation via homology directed repair (HDR) is not as efficient as nonhomologous end-joining DNA-repair-dependent knockout. Although its double-strand break activity may vary, Cas9 derived from Streptococcus pyogenens allows robust design of single-guide RNAs (sgRNAs) within the target sequence; However, prescreening for different sgRNA activities delays the process of transgenic animal generation. To overcome this limitation, multiple sets of different sgRNAs were examined for their knockin efficiency. We discovered profound advantages associated with single-stranded oligo-donor-mediated HDR processes using overlapping sgRNAs (sharing at least five base pairs of the target sites) as compared with using non-overlapping sgRNAs for knock-in mouse generation. Studies utilizing cell lines revealed shorter sequence deletions near target mutations using overlapping sgRNAs as compared with those observed using non-overlapping sgRNAs, which may favor the HDR process. Using this simple method, we successfully generated several transgenic mouse lines harboring loxP insertions or single-nucleotide substitutions with a highly efficiency of 18–38%. Our results demonstrate a simple and efficient method for generating transgenic animals harboring foreign-sequence knockins or short-nucleotide substitutions by the use of overlapping sgRNAs.

대체 감미료를 활용한 저칼로리 콤부차 제조 및 특성

장다은,홍성진,임현정,안혜수,Doan Thi Thanh Huyen,김명성,정아영,김영민 한국차학회 2023 한국차학회지 Vol.29 No.1

콤부차(Kombucha)는 홍차 또는 녹차 물 추출물에 초산균과 효모 공생체인 symbiotic colony of bacteria and yeast (SCOBY)를 접종하여, 공생 배양 중 당을 먹이원으로 첨가한 대사과정으로 얻어지는 자연 발효음료이다. 콤부차에는 페놀성 화합물(epigallocatechin gallate, epicatechin gallate, gallocatechin gallate, epicatechin, epigallocatechin, gallocatechin, catechin), 비타민류(B1, B2, B6, B12, C)와 유기산(D-gluconic acid, glucuronic acid) 등이 대표적인 활성 성분으로 알려져 있다. 주요 기능성으로는 신진대사 촉진, 항암, 노화 방지, 해독 등 수많은연구를 통해 기능성이 입증된 바 있다. 하지만 이러한 장점과는 별개로 설탕 등의 당이 발효에 있어 중요한 요소로 작용하며, 첨가되는 당 함량에 따라 칼로리가 높아질 수 있다는 문제점이 야기된다. 따라서 본 연구에서는 시판 콤부차의 품질특성을 조사하여 대체 감미료 활용 저칼로리 콤부차 제조연구를 수행하고자 하였다. 비정제원당의 함량을 달리한 대체 감미료(난소화성 말토덱스트린, 효소처리 스테비아 또는 수크랄로스)를 혼합 첨가한 콤부차를 제조하여 콤부차 발효특성, 콤부차 내 생성된 유기산, 카테킨 및 카페인 함량변화를 측정하고, 음료등의 식품소재로서의 특성을 조사하였다. 발효기간이 증감함에 따라 제조된 콤부차의 색도는 전반적으로 커다란 차이를 보이지 않았지만, pH는 5.4-2.81로 감소하였다. 흥미롭게도, 5% sugar cane 첨가 제조 콤부차에서D-gluconic acid가 가장 높게 확인되었다. 카테킨 함량은 발효 기간 중 커다란 변화를 보이지 않았지만, 16일 발효한 감미료 첨가 저칼로리 콤부차에서 카페인은 2.00 ± 0.01 mg/100 mL에서 0.94 ± 0.00 mg/100 mL로 감소하였다. 본 연구를 통해 기존 콤부차에 필수적인 설탕 함량을 낮추고 대체 감미료를 첨가한 저칼로리 콤부차 개발이 가능하였고, 새로운 트렌드의 콤부차로서 가능성을 제시하였다.

재배지역별 감초의 Glycyrrhizic Acid 함량 분석과 항당뇨 효능 평가

장다은(Da Eun Jang),송진(Jin Song),황인국(In Guk Hwang),이상훈(Sang Hoon Lee),최정숙(Jeong-Sook Choe),황경아(Kyung-A Hwang) 한국식품영양과학회 2017 한국식품영양과학회지 Vol.46 No.1

본 연구에서는 생산지역별 감초의 물질 표준화를 위해 주요지표성분 중 glycyrrhizic acid를 지표물질로 선정하고 정량 분석을 위해 분석법 검증을 수행하였다. 감초의 glycyrrhizic acid의 분석법 검증은 특이성, 직선성, 검출 및 정량한계, 정확성, 정밀성을 구하여 확립하였다. 확립된 분석 방법으로 시험한 국내외 생산지역별 감초의 glycyrrhizic acid 정량 분석 결과 국내산 중에서는 제천산이, 수입산 중에서는 우즈베키스탄산이 각각 16.98, 54.22 mg/g으로 높은 함량을 나타내었다. 또한, 현대인의 심각한 대사질환 중 하나인 당뇨병 예방 및 개선에 효과적인 기능성 천연물 소재를 발굴하고 항당뇨 효과 검증을 위해 생산지역별 감초 에탄올 추출물의 α-glucosidase 저해 활성 및 glucose uptake 활성을 평가하였다. α-Glucosidase 저해 활성은 국내외 5종 중 제천, 곡성지역의 감초에서 무처리군 대비 높은 저해 활성을 보였고, 우즈베키스탄산은 유의적인 활성이 나타나지 않았다. Glucose uptake 활성의 경우 인슐린 처리를 한 대조군 대비 제천, 영주, 우즈베키스탄 지역의 감초에서 각각 24.19, 27.18, 26.92% 유의적으로 증가함을 확인하였다. 따라서 본 연구의 지표성분 분석, 분석법 검증 결과 및 생리활성평가는 생산지역별 감초의 물질 표준화 및 품질 평가의 기초자료를 제공하는 데 큰 의미가 있으며, 감초가 대사질환인 당뇨 예방 및 개선에 효과적인 생약식품 소재로의 개발 가능성을 확보하였다. α-Glucosidase 저해 활성 및 glucose uptake 활성 평가뿐만 아니라 감초에 함유된 많은 물질연구를 통해 알려지지 않은 활성에 관한 계속된 추후 연구가 필요할 것으로 보인다. This study investigated the glycyrrhizic acid content and anti-diabetic activities of Glycyrrhiza uralensis (GU) depending on the cultivation region (Jecheon, Youngju, Gokseong, China, and Uzbekistan). Glycyrrhizic acid accuracy recovery and intra- and inter-day precisions (RSD%) of the method were calculated at 99.10∼107.07% and 3.92 and 6.31% for GU samples, respectively, whereas the limits of detection and quantitation were 0.14 and 0.20 μg/mL. Anti-diabetic activity was measured by α-glucosidase and glucose uptake. GU (20 g) was extracted with 70% ethanol at 70°C for 6 h. The Jecheon and Gokseong GU showed good inhibitory activity compared to the control. The Jecheon, Youngju, and Uzbekistan GU ethanol extracts (100 μg/mL) showed glucose uptakes (in C2C12 myotube) of 124.19, 127.18, and 126.92%, respectively, compared to the positive control. In conclusion, these methods were validated for detection of glycyrrhizic acid in GU, and the results indicate that GU might have potential anti-diabetic activities.

재배 지역에 따른 산수유의 Loganin 함량과 분석법 검증

장다은 ( Da Eun Jang ),이상훈 ( Sang Hoon Lee ),황인국 ( In Guk Hwang ),송진 ( Jin Song ),황경아 ( Kyung A Hwang ) 한국산업식품공학회 2016 산업 식품공학 Vol.20 No.2

본 연구는 다양한 활성을 가진 산수유를 건강기능식품기능성 원료를 개발하기 위하여 지표성분으로 loganin을 선정하였다. 분석법 검증 지표로는 특이성, 직선성, 검출 및 정량한계 정확성, 정밀성을 HPLC/DAD분석에 의해 검증하였다. 특이성은 표준용액과 산수유 추출액의 크로마토 그램 비교 결과 peak유지시간이 16.3분으로 일치하였으며 표준용액과 시료추출액의 UV spectrum도 정확하게 일치하는 것을 확인하였다. 직선성은 검량선의 상관계수 (R2)는 1로 우수한 직선성을 보였으며, 검출한계는 0.007 μg/mL, 정량한계는 0.026 μg/mL 수준으로 나타났다. 정확성은 loganin의 회수율로 확인하였으며, RSD (%)는 5% 이내였고 농도별로 50 μg/g에서는 105.45%, 100 μg/g에서는 96.15%, 200 μg/g에서는 95.14%의 회수율(%)을 보였다. 정밀성은 intra- 및 inter-day 분석에서 RSD값이 3% 이내로 일내와 일간의 반복성과 재현성이 우수함을 확인하였다. 검증된 분석방법으로 재배 지역별 산수유의 Loganin 함량을 측정 한 결과 3.41-7.81 mg/g 범위의 함량을 보였다. Loganin 분 석방법의 검증과정을 통한 HPLC/DAD분석법이 산수유 중 존재하는 중요한 약리적 성분의 표준화를 위한 정량에 이 용될 수 있는 충분한 감도, 특이성, 직선성, 정확성 및 정 밀성을 갖고 있음을 알 수 있었다. Loganin 분석법 검증을 통해 loganin 함량이 높은 국내산 산수유의 기능성 원료로 서 산업적 가치가 우수함을 확인 하였다. The aim of this study was to investigate the method validation for the determination of loganin content in Cornus officinalis (CO). This medicinal plant reportedly mainly included loganin. The specificity, linearity, precision, accuracy, limit of detection (LOD), and quantification (LOQ) were measured by HPLC/DAD. Our results showed that high linearity in the coefficient of calibration correlation (R2) for loganin standard was 1. LOD and LOQ for loganin were 0.007 and 0.026 mg/mL, respectively. The recovery rate of loganin was revealed to be in the high range of 95.14-105.45%. The relative standard deviation of intra- and inter-day precision of loganin in CO was 1.8% and 2.3%, respectively. The loganin content of CO from Gurye, Uiseng, Ichoen, and China were 7.81, 3.41, 7.42, and 6.52 mg/g, respectively. In conclusion, these methods were validated for the detection of loganin in CO.

고든 마타-클락의 ‘건물-자르기’에 관한 연구

장다은(Chang Da-eun) 현대미술사학회 2008 현대미술사연구 Vol.23 No.-

Gordon Matta Clark(1943-1978) is a leading artist in the Activism and Antiarchitectural movement during the 1960s and 1970s in the United States. From 1971 to 1978 he made ‘Building-Cuts’, intentionally creating holes in architectures in New York and Europe. This study focuses on why and how he treats obsolete buildings to compel people to see cut and split houses. To answer these questions, I primarily analyze visual qualities of Building-cuts in retrospect to the art and architecture in Matta-Clark’s time period. By looking at his work in this context, I could find that divisions and holes in ruined houses of that time constitute main characteristics of Building-cuts, and I intended to interpret them in the psychological and sociological field. The progressively extended structure, “the physical form includes psychological intention, effects and is finally understood sociologically according to artist's intention”, provides us with the system to synthesize elements of Building-cuts partially mentioned by most critics and art historians. In Chapter I, in comparison to Minimalism, Earth Art, Installation Art in alternative spaces, I make clear that Matta-Clark’s Building-cuts directly reconstruct the whole building differently from others’ works only focusing on architectural scale and materials or just adding minor renovations to the building. This characteristic asks us to examine his works in terms of architecture field. He studied architecture in Cornell University for 5 years, and the architectural conventions he had learned at that time are inscribed in his Building-cuts. However, he un-builds rather than builds, resisting and destroying established architectures. This aspect is connected with the architectural ideas of his father, Roberto Matta who is a surrealism painter and architect. Matta-Clark always wanted people to walk inside and experience the cut buildings. The theme of bodily perception, which is to appreciate artworks while walking in the space where works are installed, originated from minimalism in the late 60’s. In chapter II, comparing Matta-Clark’s Building-cuts to minimalism in the subject of perception, I analyze the unique effects of perception and psychological elements which Matta-Clark intended. In contrast with unitary form in minimalism, Building-cuts makes divisions between the inside and outside of a building, pointing out that people get dispersed perceptions once inside the building. In the course of this appreciation, houses which feel cozy and familiar are experienced in an unfamiliar and fearful way. These kinds of feelings toward houses are linked to the Sigmund Freud’s concept, ‘Uncanny’. Several existing studies mention this perspective in Matta-Clark’s works, but briefly. In this study, I tried to make a close inquiry into this perspective with Freud's original reference and Matta-Clark’s personal history. Matta-Clark transforms ruined houses into uncanny views and says that we should see them. This strategy is connected to his criticism of society. He thought that there were more buildings in New York than were necessary, and old buildings were being removed for the construction of new buildings in pursuit of stimulating capitalism. In chapter Ⅲ, I research Matta-Clark’s attack on metropolitan capitalism with his Building-cuts and how psychological elements are developed into sociological connotations. To accomplish this, I first prepare a theological base through Walter Benjamin’s theory and then verify the development carefully examining Matta-Clark’s works. Matta-Clark wanted to raise the alternative idea or plan for architecture by comparing new and old buildings in the work of group, ‘Anarchitecture’ which he named and led. Making a psychical shock through juxtaposing old and new buildings and using that energy for social revolution was based on Benjamin's ideas. He hoped that people would awaken from the illusion of capitalism by being shocked from seei

Immunoaffinity-HPLC/DAD법을 이용한 과자류와 시리얼류의 비타민 B12 분석 및 검증

장다은,정명근,천지연 경상대학교 농업생명과학연구원 2014 농업생명과학연구 Vol.48 No.6

식품에 존재하는 주요 비타민 B12는 hydroxycobalamin, adenosylcobalamin, methylcobalamin이며 cyanocobalamin은 시리얼과 같은 강화식품에 사용되는 합성형이다. 비타민 B12는 식품 중에 낮은 농도로 존재하기 때문에 추출 및 농축과정이 매우 중요하다. 본 연구에서는 과자와 시리얼 같은 탄수화물 식품 중의 비타민 B12 추출을 위해 α-amylase처리와 산성조건과 cyanide 존재시 열처리, immunoaffinity column(IAC)로 농축과 세척, HPLC/DAD로 분리 검출하였다. 분석법 수행특성지표들을 측정함으로 분석법을 검증하였다. 정확성과 정밀성은 AOAC 가이드라인을 충족시켰고, LOD 0.01 μg/100g과 LOQ 0.03 μg/100 g을 얻었다. 국제정도관리 시험에서 z-score 1.4로 분석수행이 우수함을 확인하였고 QC chart를 이용한 분석품질관리로 모든 분석이 관리하에 있음을 확인하였다. 검증된 분석법으로 과자류 12종과 시리얼 11종의 비타민 B12 함량을 분석하여 각각 0.02-0.26과 0.03-2.67 μg/100 g 범위를 얻 고 이는 신뢰도높은 데이터로 식품성분표에 활용 가능할 것이며 IAC-HPLC/DAD법이 탄수화물 식품의 비타민 B12 데이터베이스 구축에 활용될 수 있을 것으로 보인다. Naturally occurring vitamin B12 include hydroxycobalamin, adenosylcobalamin and methylcobalamin. Cyanocobalamin is a synthetic form observed in fortified food. Because vitamin B12 exists at very low concentration in food, concentration and clean-up are the crucial steps for accurate quantification. In this study, vitamin B12 assay using enzyme and heating extraction under acidic condition with cyanide and immunoaffinity-HPLC/DAD was validated and applied for vitamin B12 determination(μg cyanocobalamin/100 g) in confectionery and cereals. Accuracy and precision were good enough to meet the AOAC guidelines. Limit of detection and quantification were 0.01 and 0.03 μg/100 g, respectively. The z-score of the vitamin B12 assay in a FAPAS Proficiency test was less than 2, assuring reliability of analytical performance. The validated method was applied for determining vitamin B12 from confectionery and cereals: 0.02-0.26 and 0.03-2.67 μg/100 g, respectively. All analysis was under the control based on quality control chart of vitamin B12. This study shows that the IAC-HPLC/DAD is acceptable for the construction of reliable database on vitamin B12 for carbohydrate products.

식품영양성분 DB 구축을 위한 함량 분석 : 비타민 E

김형재,장다은,김미선,문귀임,이준수,이기택,김영국,천지연 한국식품영양과학회 2011 한국식품영양과학회 학술대회발표집 Vol.2011 No.10

Combination of vegetable soup and glucan demonstrates synergistic effects on macrophage-mediated immune responses

이하늘,최주희,박지연,안재훈,장다은,심재군,박종환,김영민 한국식품과학회 2021 Food Science and Biotechnology Vol.30 No.4

Vegetable soup (VS), a plant-based functionalfood, has been used as a traditional folk medicine and isattracting attention for its ability to enhance the immuneresponse. b-Glucan, a well-established and effectiveimmunomodulator, has synergistic effects when used incombination with some bioactive compounds. In the presentstudy, we aimed to evaluate the synergisticimmunomodulatory effects of the combination of VS andb-glucan on macrophage-mediated immune responses. b-Glucan was demonstrated to synergistically enhance theVS-stimulated immune response, including the productionof interleukin-6, tumor necrosis factor-a, and nitric oxide,mainly through the mitogen-activated protein kinasepathway in macrophages. In addition, this combination hasthe potential for further development in functional foodswith immune-enhancing activity.

Generation of inducible cas9 gene knock-in mice for in vivo gene targeting

배지현,어경준,정영희,황우성,장다은,염수청 한국실험동물학회 2016 한국실험동물학회 학술발표대회 논문집 Vol.2016 No.08